6 research outputs found
The changes in expression of selected drug-metabolizing enzymes in parasites after exposition to sublethal doses of drugs
Univerzita Karlova Farmaceutická fakulta v Hradci KrálovĂ© Katedra biochemickĂ˝ch vÄ›d Kandidát: Bc. Michael Pasák Ĺ kolitel: prof. RNDr. Lenka Skálová, Ph.D. Název diplomovĂ© práce: ZmÄ›ny v expresi vybranĂ˝ch enzymĹŻ metabolizujĂcĂch lĂ©ÄŤiva u parazita po kontaktu se subletálnĂmi dávkami lĂ©ÄŤiv Parazitem, kterĂ˝ zpĹŻsobuje vĂ˝znamnĂ© ztráty v chovu hospodářskĂ˝ch zvĂĹ™at skrze onemocnÄ›nĂ zvanĂ© hemonchĂłza, je vlasovka slĂ©zová (Haemonchus contortus). JednĂm z anthelmintik pouĹľĂvanĂ˝m ve veterinárnĂ medicĂnÄ› v boji proti hemochĂłze je i ivermektin (IVM). PĹ™ĂÄŤinou problĂ©mĹŻ s tĂmto parazitem je ÄŤastĂ˝ vĂ˝skyt rezistence na IVM i dalšà podávaná anthelmintika. PĹ™edpokládá se, Ĺľe urÄŤitou roli v rezistenci u vlasovky mohou hrát enzymy z rodiny cytochromĹŻ P450 (CYP) a efluxnĂ transportĂ©ry P-glykoproteiny (P-gp). Tato diplomová práce se vÄ›nuje vlivu sub-letálnĂch koncentracĂ IVM na zmÄ›nu exprese genĹŻ pro jednotlivĂ© CYP a P-gp. PĹ™ed vlastnĂm experimentem se dospÄ›lci kmene ISE H. contortus rozdÄ›lili na samce a samice. Po inkubaci dospÄ›lcĹŻ se 3 rĹŻznĂ˝mi koncentracemi IVM, byla izolována RNA a zmÄ›na exprese vybranĂ˝ch genĹŻ se stanovila metodou qPCR. Byly zjištÄ›ny vĂ˝raznĂ© rozdĂly v expresi testovanĂ˝ch genĹŻ u samcĹŻ a samic. Kontaktem vlasovek s IVM došlo k signifikantnĂm zmÄ›nám v expresi testovanĂ˝ch genĹŻ jak pro CYP tak i P-gp....Charles University Faculty of Pharmacy in Hradec KrálovĂ© Department of Biochemical Sciences Candidate: Bc. Michael Pasák Supervisor: prof. RNDr. Lenka Skálová, Ph.D. Title of diploma thesis: The changes in expression of selected drug-metabolizing enzymes in parasites after exposition to sublethal doses of drugs The parasite, which causes significant losses of small ruminants through a disease called haemonchosis, is the abomasal nematode Haemonchus contortus. IVM is one of the anthelmintics used in veterinary medicine to eliminate haemonchosis. The main problem with this parasite is the presence of drug resistance to almost all administered anthelmintics including IVM. It is believed that xenobiotic-metabolizing enzymes such as cytochromes P450 (CYPs) and membrane efflux transporters P- glycoproteins (P-gps), play the role in resistance in H. contortus. This study focuses on the effect of sublethal doses of IVM on expression of selected CYPs and P-gps. Females and males of susceptible ISE strain H. contortus were separated before our experiment. After incubation of nematodes with three different concentrations of IVM, RNA was isolated and the changes in expression of selected genes were analyzed using qPCR. Significant sex-differences were observed in inducibility of tested genes. Significant...Katedra biochemickĂ˝ch vÄ›dDepartment of Biochemical SciencesFaculty of Pharmacy in Hradec KrálovĂ©Farmaceutická fakulta v Hradci Králov
The changes in expression of selected drug-metabolizing enzymes in parasites after exposition to sublethal doses of drugs
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Michael Pasák Supervisor: prof. RNDr. Lenka Skálová, Ph.D. Title of diploma thesis: The changes in expression of selected drug-metabolizing enzymes in parasites after exposition to sublethal doses of drugs The parasite, which causes significant losses of small ruminants through a disease called haemonchosis, is the abomasal nematode Haemonchus contortus. IVM is one of the anthelmintics used in veterinary medicine to eliminate haemonchosis. The main problem with this parasite is the presence of drug resistance to almost all administered anthelmintics including IVM. It is believed that xenobiotic-metabolizing enzymes such as cytochromes P450 (CYPs) and membrane efflux transporters P- glycoproteins (P-gps), play the role in resistance in H. contortus. This study focuses on the effect of sublethal doses of IVM on expression of selected CYPs and P-gps. Females and males of susceptible ISE strain H. contortus were separated before our experiment. After incubation of nematodes with three different concentrations of IVM, RNA was isolated and the changes in expression of selected genes were analyzed using qPCR. Significant sex-differences were observed in inducibility of tested genes. Significant..
The biological effects of photodynamic therapy of cancer
Department of Biochemical SciencesKatedra biochemických vědFarmaceutická fakulta v Hradci KrálovéFaculty of Pharmacy in Hradec Králov
The changes in expression of selected drug-metabolizing enzymes in parasites after exposition to sublethal doses of drugs
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Michael Pasák Supervisor: prof. RNDr. Lenka Skálová, Ph.D. Title of diploma thesis: The changes in expression of selected drug-metabolizing enzymes in parasites after exposition to sublethal doses of drugs The parasite, which causes significant losses of small ruminants through a disease called haemonchosis, is the abomasal nematode Haemonchus contortus. IVM is one of the anthelmintics used in veterinary medicine to eliminate haemonchosis. The main problem with this parasite is the presence of drug resistance to almost all administered anthelmintics including IVM. It is believed that xenobiotic-metabolizing enzymes such as cytochromes P450 (CYPs) and membrane efflux transporters P- glycoproteins (P-gps), play the role in resistance in H. contortus. This study focuses on the effect of sublethal doses of IVM on expression of selected CYPs and P-gps. Females and males of susceptible ISE strain H. contortus were separated before our experiment. After incubation of nematodes with three different concentrations of IVM, RNA was isolated and the changes in expression of selected genes were analyzed using qPCR. Significant sex-differences were observed in inducibility of tested genes. Significant..
The changes in expression of selected drug-metabolizing enzymes in parasites after exposition to sublethal doses of drugs
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Michael Pasák Supervisor: prof. RNDr. Lenka Skálová, Ph.D. Title of diploma thesis: The changes in expression of selected drug-metabolizing enzymes in parasites after exposition to sublethal doses of drugs The parasite, which causes significant losses of small ruminants through a disease called haemonchosis, is the abomasal nematode Haemonchus contortus. IVM is one of the anthelmintics used in veterinary medicine to eliminate haemonchosis. The main problem with this parasite is the presence of drug resistance to almost all administered anthelmintics including IVM. It is believed that xenobiotic-metabolizing enzymes such as cytochromes P450 (CYPs) and membrane efflux transporters P- glycoproteins (P-gps), play the role in resistance in H. contortus. This study focuses on the effect of sublethal doses of IVM on expression of selected CYPs and P-gps. Females and males of susceptible ISE strain H. contortus were separated before our experiment. After incubation of nematodes with three different concentrations of IVM, RNA was isolated and the changes in expression of selected genes were analyzed using qPCR. Significant sex-differences were observed in inducibility of tested genes. Significant..
Mitochondrial Dysfunction in a High Intraocular Pressure-Induced Retinal Ischemia Minipig Model
Purpose: Retinal ischemia (RI) and progressive neuronal death are sight-threatening conditions. Mitochondrial (mt) dysfunction and fusion/fission processes have been suggested to play a role in the pathophysiology of RI. This study focuses on changes in the mt parameters of the neuroretina, retinal pigment epithelium (RPE) and choroid in a porcine high intraocular pressure (IOP)-induced RI minipig model. Methods: In one eye, an acute IOP elevation was induced in minipigs and compared to the other control eye. Activity and amount of respiratory chain complexes (RCC) were analyzed by spectrophotometry and Western blot, respectively. The coenzyme Q10 (CoQ10) content was measured using HPLC, and the ultrastructure of the mt was studied via transmission electron microscopy. The expression of selected mt-pathway genes was determined by RT-PCR. Results: At a functional level, increased RCC I activity and decreased total CoQ10 content were found in RPE cells. At a protein level, CORE2, a subunit of RCC III, and DRP1, was significantly decreased in the neuroretina. Drp1 and Opa1, protein-encoding genes responsible for mt quality control, were decreased in most of the samples from the RPE and neuroretina. Conclusions: The eyes of the minipig can be considered a potential RI model to study mt dysfunction in this disease. Strategies targeting mt protection may provide a promising way to delay the acute damage and onset of RI