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Průměrný učitel vypráví. Dobrý učitel vysvětluje. Výborný učitel ukazuje. Nejlepší učitel inspiruje.

Charles Farrar Browne

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Zprávy z laboratoře / Laboratory information

liposon liposon liposon

Picture of the week

Pravidelný seminář (29 T) Movember (combination of words moustache and November) is event organized every November since 1999. Thanks to encouragement by men (referred as Mo Bros), Movember aims to increase the early detection, diagnosis and treatment effectivity of prostate cancer. Anybody can donate and money is distributed to organizations supporting prostate carcinoma research, such as Prostate Cancer Foundation, (over $18 million). During its activity Movember earned more than $180 million and in 2012 was classified in world's top 100 non-government organization. The goal of Movember is to "change the face of men's health."

BBC News interview in our laboratory.



Interaction of Heavy Metal Ions with Carbon and Iron Based Particles
Pravidelný seminář (29 T)Metal ions are still a threat, polluting environment and having great bioaccumulation potential. Heavy metals can enter human body through digestive or respiratory tract and their effects may be very serious. Heavy metals can cause a disruption of function of the kidneys, bones, central nervous system, and hematopoietic system and have adverse biochemical, histological, neuropsychological and reproductive effects. Designing of new technological methods for effective removal of heavy metals from the environment is the leading point in a considerable number of research institutions [5-10]. In recent years, nanotechnologies have recorded a great development with a potential to be used also in this case. These highly advanced and revolutionary technologies are focused on the study and application of materials based on individual particles having a size in the order of nano and/or micrometers.

Již před 10 lety se Česká republika spolu s dalšími devíti evropskými zeměmi staly členy Evropské unie. Ačkoliv si Česká republika oficiálně připomene svůj vstup do Evropské unie 1. května, Úřad vlády ČR ve spolupráci se Zastoupením Evropské komise v ČR a dalšími partnery přichystaly řadu akcí, které budou probíhat po celé jaro. Europe Day is observed annually on May 05, 2014. There are two separate designations of Europe Day: The Council of Europe's day was established in 1949, while the European Union's (EU) day celebrates the day the EU's predecessor was proposed in 1950. For the EU, the day is also known as Schuman Day, comme morating the historical declaration by French foreign minister Robert Schuman.

Journal of Metallomics and Nanotechnologies

Pravidelný seminář (29 T) Třetí číslo právě vyšlo
Časopis Journal of Metallomics and Nanotechnologies vychází pouze elektronicky, čtvrtletně. Jeho obsahové zaměření je v oblastí nano-biochemie, nanotechonologie, biomedicína a nanomedicína. Časopis vychází bez regionálních mutací v českém, slovenském a anglickém jazyce. Vydavatel: Laboratoř metalomiky a nanotechnologií Mendelova univerzita v Brně, Zemědělská 1, 613 00, Brno, Česká republika http://web2.mendelu.cz/af_239_nanotech/J_Met_Nano/index.html

Pravidelný seminář (29 T)

Zprávy z výzkumu/ Scientific Reports

Orion Recovery
W3 Star-forming Region (Dec. 5, 2014) --- NASA's Orion spacecraft awaits the U.S. Navy's USS Anchorage for a ride home. Orion launched into space on a two-orbit, 4.5-test flight at 7:05 am EST on Dec. 5, and safely splashed down in the Pacific Ocean, where a combined team from NASA, the Navy and Orion prime contractor Lockheed Martin retrieved it for return to shore on board the Anchorage. It is expected to be off loaded at Naval Base San Diego on Monday. Photo credit: U.S. Navy
Paramagnetic Beads and Magnetically Mediated Strain Enhance Cardiomyogenesis in Mouse Embryoid Bodies
W3 Star-forming RegionMechanical forces play an important role in proper embryologic development, and similarly such forces can directly impact pluripotency and differentiation of mouse embryonic stem cells (mESC) in vitro. In addition, manipulation of the embryoid body (EB) microenvironment, such as by incorporation of microspheres or microparticles, can similarly influence fate determination. In this study, we developed a mechanical stimulation regimen using permanent neodymium magnets to magnetically attract cells within an EB. Arginine-Glycine-Aspartic Acid (RGD)-conjugated paramagnetic beads were incorporated into the interior of the EBs during aggregation, allowing us to exert force on individual cells using short-term magnetization. EBs were stimulated for one hour at different magnetic field strengths, subsequently exerting a range of force intensity on the cells at different stages of early EB development. Our results demonstrated that following exposure to a 0.2 Tesla magnetic field, ESCs respond to magnetically mediated strain by activating Protein Kinase A (PKA) and increasing phosphorylated extracellular signal-regulated kinase 1/2 (pERK1/2) expression. The timing of stimulation can also be tailored to guide ESC differentiation: the combination of bone morphogenetic protein 4 (BMP4) supplementation with one hour of magnetic attraction on Day 3 enhances cardiomyogenesis by increasing contractile activity and the percentage of sarcomeric ?-actin-expressing cells compared to control samples with BMP4 alone. Interestingly, we also observed that the beads alone had some impact on differentiation by increasingly slightly, albeit not significantly, the percentage of cardiomyocytes. Together these results suggest that magnetically mediated strain can be used to enhance the percentage of mouse ESC-derived cardiomyocytes over current differentiation protocols.
Metal Ions, Not Metal-Catalyzed Oxidative Stress, Cause Clay Leachate Antibacterial Activity
Aqueous leachates prepared from natural antibacterial clays, arbitrarily designated CB-L, release metal ions into suspension, have a low pH (3.4–5), generate reactive oxygen species (ROS) and H2O2, and have a high oxidation-reduction potential. To isolate the role of pH in the antibacterial activity of CB clay mixtures, we exposed three different strains of Escherichia coli O157:H7 to 10% clay suspensions. The clay suspension completely killed acid-sensitive and acid-tolerant E. coli O157:H7 strains, whereas incubation in a low-pH buffer resulted in a minimal decrease in viability, demonstrating that low pH alone does not mediate antibacterial activity. The prevailing hypothesis is that metal ions participate in redox cycling and produce ROS, leading to oxidative damage to macromolecules and resulting in cellular death. However, E. coli cells showed no increase in DNA or protein oxidative lesions and a slight increase in lipid peroxidation following exposure to the antibacterial leachate. Further, supplementation with numerous ROS scavengers eliminated lipid peroxidation, but did not rescue the cells from CB-L-mediated killing. In contrast, supplementing CB-L with EDTA, a broad-spectrum metal chelator, reduced killing. Finally, CB-L was equally lethal to cells in an anoxic environment as compared to the aerobic environment. Thus, ROS were not required for lethal activity and did not contribute to toxicity of CB-L. We conclude that clay-mediated killing was not due to oxidative damage, but rather, was due to toxicity associated directly with released metal ions.
Single Arginine Mutation in Two Yeast Isocitrate Dehydrogenases: Biochemical Characterization and Functional Implication
W3 Star-forming Region Isocitrate dehydrogenase (IDH), a housekeeping gene, has drawn the attention of cancer experts. Mutation of the catalytic Arg132 residue of human IDH1 (HcIDH) eliminates the enzyme's wild-type isocitrate oxidation activity, but confer the mutant an ability of reducing ?-ketoglutarate (?-KG) to 2-hydroxyglutarate (2-HG). To examine whether an analogous mutation in IDHs of other eukaryotes could cause similar effects, two yeast mitochondrial IDHs, Saccharomyces cerevisiae NADP+-IDH1 (ScIDH1) and Yarrowia lipolytica NADP+-IDH (YlIDH), were studied. The analogous Arg residues (Arg148 of ScIDH1 and Arg141 of YlIDH) were mutated to His. The Km values of ScIDH1 R148H and YlIDH R141H for isocitrate were determined to be 2.4-fold and 2.2-fold higher, respectively, than those of the corresponding wild-type enzymes. The catalytic efficiencies (kcat/Km) of ScIDH1 R148H and YlIDH R141H for isocitrate oxidation were drastically reduced by 227-fold and 460-fold, respectively, of those of the wild-type enzymes. As expected, both ScIDH1 R148H and YlIDH R141H acquired the neomorphic activity of catalyzing ?-KG to 2-HG, and the generation of 2-HG was confirmed using gas chromatography/time of flight-mass spectrometry (GC/TOF-MS). Kinetic analysis showed that ScIDH1 R148H and YlIDH R141H displayed 5.2-fold and 3.3-fold higher affinities, respectively, for ?-KG than the HcIDH R132H mutant. The catalytic efficiencies of ScIDH1 R148H and YlIDH R141H for ?-KG were 5.5-fold and 4.5-fold, respectively, of that of the HcIDH R132H mutant. Since the HcIDH Arg132 mutation is associated with the tumorigenesis, this study provides fundamental information for further research on the physiological role of this IDH mutation in vivo using yeast

W3 Star-forming Region Increasing biomedical workforce diversity remains a persistent challenge. Recent reports have shown that biomedical sciences (BMS) graduate students become less interested in faculty careers as training progresses; however, it is unclear whether or how the career preferences of women and underrepresented minority (URM) scientists change in manners distinct from their better-represented peers. We report results from a survey of 1500 recent American BMS Ph.D. graduates (including 276 URMs) that examined career preferences over the course of their graduate training experiences. On average, scientists from all social backgrounds showed significantly decreased interest in faculty careers at research universities, and significantly increased interest in non-research careers at Ph.D. completion relative to entry. However, group differences emerged in overall levels of interest (at Ph.D. entry and completion), and the magnitude of change in interest in these careers. Multiple logistic regression showed that when controlling for career pathway interest at Ph.D. entry, first-author publication rate, faculty support, research self-efficacy, and graduate training experiences, differences in career pathway interest between social identity groups persisted. All groups were less likely than men from well-represented (WR) racial/ethnic backgrounds to report high interest in faculty careers at research-intensive universities (URM men: OR 0.60, 95% CI: 0.36–0.98, p = 0.04; WR women: OR: 0.64, 95% CI: 0.47–0.89, p = 0.008; URM women: OR: 0.46, 95% CI: 0.30–0.71, p0.001), and URM women were more likely than all other groups to report high interest in non-research careers (OR: 1.93, 95% CI: 1.28–2.90, p = 0.002). The persistence of disparities in the career interests of Ph.D. recipients suggests that a supply-side (or “pipeline”) framing of biomedical workforce diversity challenges may limit the effectiveness of efforts to attract and retain the best and most diverse workforce. We propose incorporation of an ecological perspective of career development when considering strategies to enhance the biomedical workforce and professoriate through diversity.

W3 Star-forming Region DNA topoisomerases are enzymes responsible for the relaxation of DNA torsional strain, as well as for the untangling of DNA duplexes after replication, and are important cancer drug targets. One class of topoisomerase inhibitors, “poisons”, binds to the transient enzyme-DNA complex which occurs during the mechanism of action, and inhibits the religation of DNA. This ultimately leads to the accumulation of DNA double strand breaks and cell death. Different types of topoisomerases occur in human cells and several poisons of topoisomerase I and II are widely used clinically. However, their use is compromised by a variety of side effects. Recent studies confirm that the inhibition of the ?-isoform of topoisomerase II is responsible for the cytotoxic effect, whereas the inhibition of the ß-isoform leads to development of adverse drug reactions. Thus, the discovery of agents selective for topoisomerase II? is an important strategy for the development of topoisomerase II poisons with improved clinical profiles. Here, we present a computer-aided drug design study leading to the identification of structurally novel topoisomerase II? poisons. The study combines ligand- and structure-based drug design methods including pharmacophore models, homology modelling, docking, and virtual screening of the National Cancer Institute compound database. From the 8 compounds identified from the computational work, 6 were tested for their capacity to poison topoisomerase II in vitro: 4 showed selective inhibitory activity for the ?- over the ß-isoform and 3 of these exhibited cytotoxic activity. Thus, our study confirms the applicability of computer-aided methods for the discovery of novel topoisomerase II poisons, and presents compounds which could be investigated further as selective topoisomerase II? inhibitors.

W3 Star-forming Region Osteosarcoma (OS) is the most common primary malignant bone tumor and prevalently occurs in the second decade of life. Etoposide, a chemotherapeutic agent used in combined treatments of recurrent human OS, belongs to the topoisomerase inhibitor family and causes DNA breakage. In this study we evaluated the cascade of events determined by etoposide-induced DNA damage in OS cell lines with different p53 status focusing on methylation status and expression of miR-34a that modulate tumor cell growth and cell cycle progression. Wild-type p53 U2-OS cells and U2-OS cells expressing dominant-negative form of p53 (U2- OS175) were more sensitive to etoposide than p53-deficient MG63 and Saos-2 cells, showing increased levels of unmethylated miR-34a, reduced expression of CDK4 and cell cycle arrest in G1 phase. In contrast, MG63 and Saos-2 cell lines presented aberrant methylation of miR-34a promoter gene with no miR-34a induction after etoposide treatment, underlining the close connection between p53 expression and miR-34a methylation status. Consistently, in p53siRNA transfected U2-OS cells we observed loss of miR-34a induction after etoposide exposure associated with a partial gain of gene methylation and cell cycle progress towards G2/M phase. Our results suggest that the open and unmethylated conformation of the miR-34a gene may be regulated by p53 able to bind the gene promoter. In conclusion, cell response to etoposide-induced DNA damage was not compromised in cells with dominant-negative p53 expression.

W3 Star-forming Region The molecular function of the cellular prion protein (PrPC) and the mechanism by which it may contribute to neurotoxicity in prion diseases and Alzheimer's disease are only partially understood. Mouse neuroblastoma Neuro2a cells and, more recently, C2C12 myocytes and myotubes have emerged as popular models for investigating the cellular biology of PrP. Mouse epithelial NMuMG cells might become attractive models for studying the possible involvement of PrP in a morphogenetic program underlying epithelial-to-mesenchymal transitions. Here we describe the generation of PrP knockout clones from these cell lines using CRISPR-Cas9 knockout technology. More specifically, knockout clones were generated with two separate guide RNAs targeting recognition sites on opposite strands within the first hundred nucleotides of the Prnp coding sequence. Several PrP knockout clones were isolated and genomic insertions and deletions near the CRISPR-target sites were characterized. Subsequently, deep quantitative global proteome analyses that recorded the relative abundance of>3000 proteins (data deposited to ProteomeXchange Consortium) were undertaken to begin to characterize the molecular consequences of PrP deficiency. The levels of ~120 proteins were shown to reproducibly correlate with the presence or absence of PrP, with most of these proteins belonging to extracellular components, cell junctions or the cytoskeleton

W3 Star-forming Region Serous ovarian cancer (SEOC) is the deadliest gynecologic malignancy. MicroRNAs (miRNAs) are a class of small noncoding RNAs which regulate gene expression and protein translation. MiRNAs are also encoded by viruses with the intent of regulating their own genes and those of the infected cells. This is the first study assessing viral miRNAs in SEOC. MiRNAs sequencing data from 487 SEOC patients were downloaded from the TCGA website and analyzed through in-house sequencing pipeline. To cross-validate TCGA analysis, we measured the expression of miR-H25 by quantitative immunofluorescence in an additional cohort of 161 SEOC patients. Gene, miRNA expression, and cytotoxicity assay were performed on multiple ovarian cancer cell lines transfected with miR-H25 and miR-BART7. Outcome analysis was performed using multivariate Cox and Kaplan-Meier method. Viral miRNAs are more expressed in SEOC than in normal tissues. Moreover, Herpetic viral miRNAs (miR-BART7 from EBV and miR-H25 from HSV-2) are significant and predictive biomarkers of outcome in multivariate Cox analysis. MiR-BART7 correlates with resistance to first line chemotherapy and early death, whereas miR-H25 appears to impart a protective effect and long term survival. Integrated analysis of gene and viral miRNAs expression suggests that miR-BART7 induces directly cisplatin-resistance, while miR-H25 alters RNA processing and affects the expression of noxious human miRNAs such as miR-143. This is the first investigation linking viral miRNA expression to ovarian cancer outcome. Viral miRNAs can be useful to develop biomarkers for early diagnosis and as a potential therapeutic tool to reduce SEOC lethality.

W3 Star-forming Region The ATP-gated P2X7 has been shown to play an important role in invasiveness and metastasis of some tumors. However, the possible links and underlying mechanisms between P2X7 and prostate cancer have not been elucidated. Here, we demonstrated that P2X7 was highly expressed in some prostate cancer cells. Down-regulation of P2X7 by siRNA significantly attenuated ATP- or BzATP-driven migration and invasion of prostate cancer cells in vitro, and inhibited tumor invasiveness and metastases in nude mice. In addition, silencing of P2X7 remarkably attenuated ATP- or BzATP- driven expression changes of EMT/invasion-related genes Snail, E-cadherin, Claudin-1, IL-8 and MMP-3, and weakened the phosphorylation of PI3K/AKT and ERK1/2 in vitro. Similar effects were observed in nude mice. These data indicate that P2X7 stimulates cell invasion and metastasis in prostate cancer cells via some EMT/invasion-related genes, as well as PI3K/AKT and ERK1/2 signaling pathways. P2X7 could be a promising therapeutic target for prostate cancer.

W3 Star-forming Region Resident bacterial communities (microbiota) and host antimicrobial peptides (AMPs) are both essential components of normal host innate immune responses that limit infection and pathogen induced inflammation. However, their interdependence has not been investigated in the context of urinary tract infection (UTI) susceptibility. Here, we explored the interrelationship between the urinary microbiota and host AMP responses as mechanisms for UTI risk. Using prospectively collected day of surgery (DOS) urine specimens from female pelvic floor surgery participants, we report that the relative abundance and/or frequency of specific urinary microbiota distinguished between participants who did or did not develop a post-operative UTI. Furthermore, UTI risk significantly correlated with both specific urinary microbiota and ß-defensin AMP levels. Finally, urinary AMP hydrophobicity and protease activity were greater in participants who developed UTI, and correlated positively with both UTI risk and pelvic floor symptoms. These data demonstrate an interdependency between the urinary microbiota, AMP responses and symptoms, and identify a potential mechanism for UTI risk. Assessment of bacterial microbiota and host innate immune AMP responses in parallel may identify increased risk of UTI in certain populations..


12.10.2014: Sondu brněnských a slovenských vědců vynese do stratosféry balón. Bude zkoumat, jak kosmické záření poškozuje DNA a jak ovlivňuje život ve vesmíru

Journal of Metallomics and Nanotechnologie
laboratorní časopis
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- Special Issue Information

Upozornění pro cestující týkající se výskytu horečky způsobené virem EBOLA - pdf

Kontrola a úprava informačního systému, uživatelské zpracování – XIV
Petr Čapek,
16. 12. 2014, od 14:00 – 18:00 h

Nanotransportéry pro přenos doxorubicinu
Iva Blažková
19. 12. 2014, od 12:00

An Effect of Heavy Metals on Escherichia coli with the Cloned Gene for Metallothioneins Revealed by Electrochemistry
Ana María Jiménez Jiménez
19. 12. 2014, od 12:00

Účinky kationických antimikrobiálních peptidů
Markéta Komínková
pátek 19. 12. 2014, od 12:00

Regulace genové exprese pomocí oligonukleotidů, zvláště aptamerů
Monika Kremplová
19. 12. 2014, od 12:00

Nanopóry a DNA sekvenování
Jiří Kudr
19. 12. 2014, od 12:00

Studium interakce receptoru pro hemaglutinin – přehledový seminář
Petr Michálek
19. 12. 2014, od 12:00

Dálkově řízené robotické systémy a jejich aplikace
Lukáš Zima
19. 12. 2014, od 12:00

Sumarizace výstupů projektu excelentní mladí vědci na VUT v Brně v roce 2014
Markéta Vaculovičová
19. 12. 2014, od 12:00


Současná kosmonautika a kosmické technologie-fotografie

Den AIDS - 1/12/2014

Journal of Metallomics and Nanotechnologies


Mechanisms of Uptake and Interaction of Platinum Based Drugs in Eukaryotic Cells

17 estradiol-containing liposomes as a novel delivery system for the antisense therapy of ER-positive breast cancer: An in vitro study on the MCF-7 cell line

Modulation of Induced Cytotoxicity of Doxorubicin by Using Apoferritin and Liposomal Cages

Formation of DNA Adducts by Ellipticine and Its Micellar Form in Rats — A Comparative Study

Use of green fluorescent proteins for in vitro biosensing

The effect of metal ions on Staphylococcus aureus revealed bybiochemical and mass spectrometric analyses

Synthesis of carbon quantum dots for DNA labeling and its electrochemical, fluorescent and electrophoretic characterization

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