The endothelium of different organs displays a remarkable heterogeneity, although it presents many common functional and morphological features. However, despite our knowledge of heterogeneity among endothelial cells from different sites, the differences between brain microvascular endothelial cells (BMEC) and coronary microvascular endothelial cells (CMEC) are poorly defined. The aim of this study was to investigate whether BMEC are distinct from CMEC at the protein level. Using the proteomic approach, we comparatively analyzed the proteome of cultured BMEC and CMEC. We reproducibly separated over 2000 polypeptides by using two-dimensional electrophoresis (2-DE) at pH range of 3-10. Using PDQuest software to process the 2-DE gel images, forty-seven protein spots were differentially expressed in the two-endothelial cells. Of these, thirty-five proteins are highly expressed in BMEC, whereas twelve proteins are highly expressed in CMEC. Fifteen proteins in BMEC and seven proteins in CMEC were identified with high confidence by matrix-associated laser desorption ionization time-of-flight mass spectrometer (MALDI-TOF-MS). Our data suggested that BMEC and CMEC were different in several aspects including cytokine and growth-related molecules, stress-related proteins, metabolic enzymes, signal transduction proteins and others. The identification of a set of proteins preferentially expressed in BMEC and CMEC provided new data on the heterogeneity of the endothelium., L. Lu, P.-Y. Yang, Y.-Ch. Rui, H. Kang, J. Zhang, J.-P. Zhang, W.-H. Feng., and Obsahuje bibliografii a bibliografické odkazy
Cytokiny jsou proteiny, které regulují růst, diferenciaci a aktivaci buněk. Jejich účinek se široce využívá pro léčbu zánětlivých a nádorových onemocnění. Anti-cytokinová léčba je velmi slibná, ale přinese v blízké budoucnosti velké finanční zatížení zdravotnictví., Cytokines are proteins that regulate the growth, differentiation and activation of cells. Their functions are used in therapy of inflammatory diseases and cancer. Antibody blockade of cytokines seems to be very promising, but it will result in considerable financial burden for healthcare in future., and Ilja Trebichavský.
Ischemia can contribute to the inner ear pathology and hearing loss. To determine the susceptibility of inner and outer hair cells (IHCs/OHCs) to ischemic and post-ischemic period, we used organotypic cultures of the organ of Corti isolated from P3 rats as an in vitro model of inner ear ischemia (oxygen-glucose deprivation, OGD). We identified the hair cells (HCs) by phalloidin staining. The cells with damaged cellular membrane integrity were identified by propidium iodide (PI)-exclusion assay. The cells with fragmented chromosomal DNA were detected by TUNEL assay. Organotypic cultures were subjected to a mild (3 h duration) or severe (4 h duration ) OGD, followed by a recovery period of 21 h and 20 h, respectively. Mild OGD induced a loss of 10-20 % HCs, whereas severe OGD induced loss of 35 % HCs. We confirmed that OHCs are less vulnerable to OGD than IHCs. Of all missing OHCs, 80-90 % was lost during the OGD period and 10-20 % during the recovery period. In contrast, the loss of IHCs was equal during both experimental periods. The OGD period was mainly associated with PI-positive nuclei. TUNEL-positive nuclei were a minor frac tion during the OGD period and increased during the recovery period, indicating the progression of DNA fragmentation. Our results implicate a differential susceptibility of IHCs and OHCs during and after ischemia-like insult, which may be of therapeutic consequence., N. Amarjargal ... [et al.]., and Obsahuje seznam literatury
An important mechanism underlying cochlear hair cell (HC) susceptibility to hypoxia/ischemia is the influx of Ca2+. Two main ATP-dependent mechanisms contribute to maintaining low Ca2+ levels: uptake of Ca2+ into intracellular stores via smooth endoplasmic reticulum calcium ATPase (SERCA) and extrusion of Ca2+ via plasma membrane calcium ATPase (PMCA). The effects of the SERCA inhibitors thapsigargin (10 nM-10 μM) and cyclopiazonic acid (CPA; 10-50 μM) and of the PMCA blockers eosin (1.5-10 μM) and o-vanadate (1-5 mM) on inner and outer hair cells (IHCs/OHCs) were examined in normoxia and ischemia using an in vitro model of the newborn rat cochlea. Exposure of the cultures to ischemia resulted in a significant loss of HCs. Thapsigargin and CPA had no effect. Eosin decreased the numbers of IHCs and OHCs by up to 25 % in normoxia and significantly aggravated the ischemia-induced damage to IHCs at 5 and 10 μM and to OHCs at 10 μM. o-Vanadate had no effect on IHC and OHC counts in normoxia, but aggravated the ischemia-induced HC loss in a dose-dependent manner. The effects of eosin and o-vanadate indicate that PMCA has an important role to play in protecting the HCs from ischemic cell death., N. Amarjargal, B. Mazurek, H. Haupt, N. Andeeva, J. Fuchs, J. Gross., and Obsahuje bibliografii a bibliografické odkazy
Huntingtonova choroba (Huntington´s disease, HD) je smrtelné dědičné neurodegenerativní onemocnění s nástupem projevů až v dospělosti. Onemocnění je způsobeno expanzí cytozin-adenin-guanin (CAG) repetic v genu pro protein huntingtin (Htt), který je exprimován ve většině tkání. HD je charakteristická především rozsáhlou degenerací buněk centrální nervové soustavy, ale mutace má velký dopad i na další orgány a tkáně. Mechanismy těchto změn nejsou stále dostatečně popsány. Jednou z nezbytných součástí výzkumu HD jsou zvířecí modely., Huntington's disease (HD) is a fatal inherited neurodegenerative disease with onset of symptoms in adulthood. The disease is caused by the expansion of CAG repeats in the gene for the huntingtin protein, which is expressed in most tissues. HD is characterized by extensive degeneration of the cells of the central nervous system, but the mutation has a large impact on other organs and tissues too. The mechanisms of these changes have not yet been adequately described. Animal models are one of the fundamental approaches in HD research., and Daniela Pallová ... [et al.].
The aim of the present study was to define the stress-induced pattern of cytosolic glucocorticoid receptor (GR) and Hsp70 protein in the liver of male Wistar rats exposed to different stress models: acute (2 h/day) immobilization or cold (4 °C); chronic (21 days) isolation, crowding, swimming or isolation plus swimming and combined (chronic plus acute stress). Changes in plasma levels of corticosterone were studied by radioimmunoassay (RIA). The results obtained by Western immunoblotting showed that both acute stressors led to a significant decrease in cytosolic GR and Hsp70 levels. Compared to acute stress effects, only a weak decrease in the levels of GR and Hsp70 was demonstrated in chronic stress models. Chronically stressed rats, which were subsequently exposed to novel acute stressors (immobilization or cold), showed a lower extent of GR down-regulation when compared to acute stress. The exception was swimming, which partially restores this down-regulation. The observed changes in the levels of these major stress-related cellular proteins in liver cytosol lead to the conclusion that chronic stressors compromise intracellular GR down-regulation in the liver., D. Filipović, L. Gavrilović, S. Dronjak, M. Demajo, M. B. Radojčić., and Obsahuje bibliografii a bibliografické odkazy
Mor je zoonóza, jejíž epidemie sužují lidstvo od starověku. Od objevu bakteriálního původce moru A. Yersinem a S. Kitasatem uplynulo 120 let, během kterých byla tato choroba velmi dobře popsána jak z epidemiologického, tak z molekulárně mikrobiologického a evolučního hlediska. Studiem DNA izolované z ostatků obětí moru byl původce moru přímo prokázán u epidemií starých až 650 let. Vysoká mortalita při moru je dána neefektivním přenosem mezi hostiteli pomocí blechy jako vektoru., Plague is a zoonotic disease, the epidemics of which have troubled mankind since ancient times. During the last 120 years that have passed since the discovery of the plague bacillus Y. pestis by A. Yersin and S. Kitasato this infectious disease was described in detail, including its epidemiology, molecular microbiology and evolution. Ancient DNA isolated from the remains of plague victims have enabled us to establish Y. pestis as the causative agent in epidemics more than 650 years old. The high mortality of the plague is caused by an ineffective transfer by its flea vector., and Ivo Konopásek.
Komunikace mezi buňkami mnohobuněčného organismu je nezbytná k zajištění přežití organismu, správné funkce tkání a orgánů, tvorby energie, růstu a vývoje. Bílkoviny sekretované z buněk jsou hlavními molekulami, které zprostředkovávají mezibuněčnou komunikaci na malé i velké vzdálenosti. Většina sekretovaných bílkovin je z buněk uvolňována cestou přes endoplasmatické retikulum a Golgiho aparát. Vývoj nových laboratorních technik pro studium sekretovaných bílkovin umožnil v posledním desetiletí studovat a popsat sekreci mnoha typů buněk., Communication among cells in a multicellular organism is fundamental for the correct functioning of organs and tissues, energy production, growth and development, to assure survival and reproduction of the organism. Proteins secreted by cells are principal molecules for intercellular communication at both short and long distances. Most of the secreted proteins are released through the endoplasmic reticulum – the Golgi pathway. The significant development of analytical techniques for detection of secreted proteins in the last 10 years has enabled us to explore the secretion of various cell types., and Helena Kupcová Skalníková.
Tuto otázku si kladou lidé již od počátku lidstva. Pro členy Laboratoře biochemie a molekulární biologie zárodečných buněk v Ústavu živočišné fyziologie a genetiky AV ČR v Liběchově je odpověď jasná, jelikož na počátek vzniku nového jedince nahlíží prostřednictvím molekulární biologie. První bylo vajíčko. and Denisa Jansová.