Příručka se zaměřuje na analýzu biologických dat pomocí softwaru R. Určeno odborné veřejnosti.; Kniha je zaměřena na regresní modely, konkrétně jednorozměrné zobecněné lineární modely (GLM).

• MeSH
• Data Analysis MeSH
• Biology statistics & numerical data   MeSH
• Numerical Analysis, Computer-Assisted MeSH
• Models, Statistical MeSH
• Publication type
• Handbook MeSH

• Conspectus
• Biologické vědy
• NML Fields
• biologie
• statistika, zdravotnická statistika

Scanning electron microscopes are useful biological tools that can be used to image the surface of whole organisms, tissues, cells, cellular components, and macromolecules. Processes and structures that exist at surfaces can be imaged in pseudo, or real 3D at magnifications ranging from about 10× to 1,000,000×. Therefore a whole multicellular organism, such as a fly, or a single protein embedded in one of its cell membranes can be visualized. In order to identify that protein at high resolution, or to see and quantify its distribution at lower magnifications, samples can be labeled with antibodies. Any surface that can be exposed can potentially be studied in this way. Presented here is a generic method for immunogold labeling for scanning electron microscopy, using two examples of specimens: isolated nuclear envelopes and the cytoskeleton of mammalian culture cells. Various parameters for sample preparation, fixation, immunogold labeling, drying, metal coating, and imaging are discussed so that the best immunogold scanning electron microscopy results can be obtained from different types of specimens.

• MeSH
• Antigens genetics   metabolism   MeSH
• Staining and Labeling methods   MeSH
• Cell Membrane metabolism   ultrastructure   MeSH
• Cytoskeleton metabolism   ultrastructure   MeSH
• Epoxy Resins chemistry   MeSH
• Gene Expression MeSH
• Tissue Fixation methods   MeSH
• Fixatives chemistry   MeSH
• Formaldehyde chemistry   MeSH
• Immunohistochemistry methods   MeSH
• Nuclear Envelope metabolism   ultrastructure   MeSH
• Gold Colloid chemistry   MeSH
• Nuclear Pore Complex Proteins genetics   metabolism   MeSH
• Microscopy, Electron, Scanning methods   MeSH
• Microtomy MeSH
• Oocytes metabolism   ultrastructure   MeSH
• Polymers chemistry   MeSH
• Antibodies chemistry   MeSH
• Xenopus laevis MeSH
• Tissue Embedding methods   MeSH
• Animals MeSH
• Check Tag
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

The small bacterial laccase from the actinobacterium Streptomyces coelicolor which lacks the second of the three domains of the laccases structurally characterized to date was crystallized. This multi-copper phenol oxidase crystallizes in a primitive tetragonal lattice, with unit-cell parameters a = b = 179.8, c = 175.3 A. The crystals belong to either space group P4(1)2(1)2 or P4(3)2(1)2. The self-rotation function shows the presence of a noncrystallographic threefold axis in the structure. Phases will be determined from the anomalous signal of the natively present copper ions.

• MeSH
• X-Ray Diffraction MeSH
• Financing, Organized MeSH
• Crystallization MeSH
• Laccase genetics   chemistry   metabolism   MeSH
• Molecular Weight MeSH
• Streptomyces coelicolor enzymology   genetics   MeSH

Cieľom práce je poskytnúť stručný prehľad o aktuálnej situácii v oblasti skríningu cytotoxického účinku a mechanizmu pôsobenia prírodných a syntetických látok využiteľných v chemoterapii nádorov. Medzi v súčasnosti najčastejšie sa vyskytujúce a najzávažnejšie ochorenia patria onkologické ochorenia, ktoré predstavujú pre človeka život ohrozujúce riziko. Vážnou komplikáciou protinádorovej terapie je neadekvátna terapeutická odpoveď, ktorá je zapríčinená rezistenciou ľudského organizmu na používané liečivá, chemoterapeutiká. A práve rezistencia je jednou z hnacích síl, ktoré nás neustále nútia vyhľadávať nové protinádorovo účinné liečivá, či už prírodného alebo syntetického pôvodu. Dnes sa bežne a rutinne používa na vyhľadávanie nových potenciálnych cytostatík primárny skríning in vitro, pri ktorom sa rôznymi metódami sleduje citlivosť nádorových buniek rastúcich in vitro na cytotoxické látky. Vlastnosti potenciálnej protinádorovej zlúčeniny sú charakterizované rôznymi parametrami, medzi ktoré patrí antiproliferačná aktivita, sledovanie štrukturálnych a funkčných zmien cytoplazmovej membrány, zmien v obsahu bunkových proteínov, resp. nukleových kyselín, zmien metabolizmu buniek, zmien bunkového cyklu, indukcia apoptózy, enzýmová aktivita (dihydrofolátreduktázy, proteináz, proteínkináz, topoizomeráz, tymidylátsyntetázy), vplyv na mitochondrie, cytoskelet bunky, telomerázová aktivita atď. Sledovaním týchto parametrov súčasne sledujeme aj mechanizmus účinku cytotoxicky účinnej látky.

The paper reviews the current approaches to cytotoxic effect screening and mode of action of natural and synthetic compounds usable in tumour chemotherapy. Oncological diseases belong to the most frequently occurring and the most serious diseases that threaten millions of human lives. A serious complication of anticancer therapy is an inadequate therapeutic answer which is caused by the resistance of the human organism to the employed drugs, chemotherapeutic agents. Therefore the resistance is one of the driving forces which constantly force us to search for new anticancer effective drugs of natural or synthetic origin. Primary screening in vitro, in which by different methods the sensitivity of cancer cells growing in vitro to cytotoxic compounds is monitored, is commonly and routinely used for searching for new potential cytostatics today. The properties of a potential anticancer compound are characterized by different parameters, which include antiproliferative activity, monitoring of structural and functional changes in the cytoplasmic membrane, changes in cell proteins and nucleic acids content, cell metabolism changes, cell cycle changes, induction of apoptosis, enzymatic activity (dihydrofolate reductase, proteinases, proteinkinases, topoisomerases, tymidylatsynthetase), effect on mitochondria, cell cytoskeleton, telomerase activity, etc. By monitoring these parameters, the mode of action of a cytotoxically effective compound can be followed up.

• MeSH
• Apoptosis genetics   MeSH
• Cell Death physiology   drug effects   MeSH
• Cell Cycle physiology   drug effects   MeSH
• Cytoskeleton physiology   drug effects   MeSH
• Research Support as Topic MeSH
• Antineoplastic Agents, Phytogenic administration & dosage   adverse effects   MeSH
• Humans MeSH
• Cell Transformation, Neoplastic MeSH
• Drug-Related Side Effects and Adverse Reactions MeSH
• Mass Screening methods   MeSH
• Review Literature as Topic MeSH
• Antineoplastic Agents administration & dosage   chemical synthesis   adverse effects   MeSH
• Check Tag
• Humans MeSH

Insulin is stored in vivo inside the pancreatic β-cell insulin secretory granules. In vitro studies have led to an assumption that high insulin and Zn2+ concentrations inside the pancreatic β-cell insulin secretory granules should promote insulin crystalline state in the form of Zn2+-stabilized hexamers. Electron microscopic images of thin sections of the pancreatic β-cells often show a dense, regular pattern core, suggesting the presence of insulin crystals. However, the structural features of the storage forms of insulin in native preparations of secretory granules are unknown, because of their small size, fragile character and difficult handling. We isolated and investigated the secretory granules from MIN6 cells under near-native conditions, using cryo-electron microscopic (Cryo-EM) techniques. The analysis of these data from multiple intra-granular crystals revealed two different rhomboidal crystal lattices. The minor lattice has unit cell parameters (a ≃ b ≃ 84.0 Å, c ≃ 35.2 Å), similar to in vitro crystallized human 4Zn2+-insulin hexamer, whereas the largely prevalent unit cell has more than double c-axis (a ≃ b ≃ c ≃ 96.5 Å) that probably corresponds to two or three insulin hexamers in the asymmetric unit. Our experimental data show that insulin can be present in pancreatic MIN6 cell granules in a microcrystalline form, probably consisting of 4Zn2+-hexamers of this hormone.

• MeSH
• Insulin-Secreting Cells * MeSH
• Microscopy, Electron MeSH
• Insulin MeSH
• Islets of Langerhans * MeSH
• Humans MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

The development of nanomedicines for the treatment of neurodegenerative disorders demands innovative nanoarchitectures for combined loading of multiple neuroprotective compounds. We report dual-drug loaded monoolein-based liquid crystalline architectures designed for the encapsulation of a therapeutic protein and a small molecule antioxidant. Catalase (CAT) is chosen as a metalloprotein, which provides enzymatic defense against oxidative stress caused by reactive oxygen species (ROS) such as hydrogen peroxide (H2O2). Curcumin (CU), solubilized in fish oil, is co-encapsulated as a chosen drug with multiple therapeutic activities, which may favor neuro-regeneration. The prepared self-assembled biomolecular nanoarchitectures are characterized by biological synchrotron small-angle X-ray scattering (BioSAXS) at multiple compositions of the lipid/co-lipid/water phase diagram. Constant fractions of curcumin (an antioxidant) and a PEGylated agent (TPEG1000) are included with regard to the lipid fraction. Stable cubosome architectures are obtained for several ratios of the lipid ingredients monoolein (MO) and fish oil (FO). The impact of catalase on the structural organization of the cubosome nanocarriers is revealed by the variations of the cubic lattice parameters deduced by BioSAXS. The outcome of the cellular uptake of the dual drug-loaded nanocarriers is assessed by performing a bioassay of catalase peroxidatic activity in lysates of nanoparticle-treated differentiated SH-SY5Y human cells. The obtained results reveal the neuroprotective potential of the in vitro studied cubosomes in terms of enhanced peroxidatic activity of the catalase enzyme, which enables the inhibition of H2O2 accumulation in degenerating neuronal cells.

• MeSH
• Liquid Crystals chemistry   MeSH
• Catalase chemistry   MeSH
• Curcumin chemistry   MeSH
• Humans MeSH
• Scattering, Small Angle MeSH
• Nanostructures chemistry   MeSH
• Hydrogen Peroxide chemistry   MeSH
• Polyethylene Glycols chemistry   MeSH
• Reactive Oxygen Species MeSH
• Synchrotrons MeSH
• Imaging, Three-Dimensional MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

Analysis of C cross-polarization magic angle spinning (CP/MAS) nuclear magnetic resonance (NMR), differential scanning calorimetry (DSC), Fourier transform infrared (FTIR), and X-ray powder diffraction data of trospium chloride (TCl) products crystallized from different mixtures of water-ethanol [φ(EtOH) = 0.5-1.0] at various temperatures (0°C, 20°C) and initial concentrations (saturated solution, 30%-50% excess of solvent) revealed extensive structural variability of TCl. Although (13) C CP/MAS NMR spectra indicated broad variety of structural phases arising from molecular disorder, temperature-modulated DSC identified presence of two distinct components in the products. FTIR spectra revealed alterations in the hydrogen bonding network (ionic hydrogen bond formation), whereas the X-ray diffraction reflected unchanged unit cell parameters. These results were explained by a two-component character of TCl products in which a dominant polymorphic form is accompanied by partly separated nanocrystalline domains of a secondary phase that does not provide clear Bragg reflections. These phases slightly differ in the degree of molecular disorder, in the quality of crystal lattice and hydrogen bonding network. It is also demonstrated that, for the quality control of such complex products, (13) C CP/MAS NMR spectroscopy combined with factor analysis (FA) can satisfactorily be used for categorizing the individual samples: FA of (13) C CP/MAS NMR spectra found clear relationships between the extent of molecular disorder and crystallization conditions. © 2013 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 102:1235-1248, 2013.

• MeSH
• Benzilates chemistry   MeSH
• Calorimetry, Differential Scanning MeSH
• X-Ray Diffraction MeSH
• Crystallization MeSH
• Magnetic Resonance Spectroscopy MeSH
• Nortropanes chemistry   MeSH
• Powder Diffraction MeSH
• Spectroscopy, Fourier Transform Infrared MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

• MeSH
• Physical Phenomena MeSH
• Magnetic Resonance Imaging MeSH

• Conspectus
• Patologie. Klinická medicína
• NML Fields
• radiologie, nukleární medicína a zobrazovací metody
• NML Publication type
• kolektivní monografie