The importance of sharing experimental data in neuroscience grows with the amount and complexity of data acquired and various techniques used to obtain and process these data. However, the majority of experimental data, especially from individual studies of regular-sized laboratories never reach wider research community. A graphical user interface (GUI) engine called Neurovascular Network Explorer 2.0 (NNE 2.0) has been created as a tool for simple and low-cost sharing and exploring of vascular imaging data. NNE 2.0 interacts with a database containing optogenetically-evoked dilation/constriction time-courses of individual vessels measured in mice somatosensory cortex in vivo by 2-photon microscopy. NNE 2.0 enables selection and display of the time-courses based on different criteria (subject, branching order, cortical depth, vessel diameter, arteriolar tree) as well as simple mathematical manipulation (e.g. averaging, peak-normalization) and data export. It supports visualization of the vascular network in 3D and enables localization of the individual functional vessel diameter measurements within vascular trees. NNE 2.0, its source code, and the corresponding database are freely downloadable from UCSD Neurovascular Imaging Laboratory website1. The source code can be utilized by the users to explore the associated database or as a template for databasing and sharing their own experimental results provided the appropriate format.
- MeSH
- databáze faktografické MeSH
- mozková kůra metabolismus MeSH
- myši MeSH
- neuronové sítě (počítačové) MeSH
- somatosenzorické korové centrum metabolismus MeSH
- vazomotorický systém patologie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- audiovizuální média MeSH
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
Isolation of DNA using magnetic particles is a field of high importance in biotechnology and molecular biology research. This protocol describes the evaluation of DNA-magnetic particles binding via dynamic light scattering (DLS) and electrophoretic light scattering (ELS). Analysis by DLS provides valuable information on the physicochemical properties of particles including particle size, polydispersity, and zeta potential. The latter describes the surface charge of the particle which plays major role in electrostatic binding of materials such as DNA. Here, a comparative analysis exploits three chemical modifications of nanoparticles and microparticles and their effects on DNA binding and elution. Chemical modifications by branched polyethylenimine, tetraethyl orthosilicate and (3-aminopropyl)triethoxysilane are investigated. Since DNA exhibits a negative charge, it is expected that zeta potential of particle surface will decrease upon binding of DNA. Forming of clusters should also affect particle size. In order to investigate the efficiency of these particles in isolation and elution of DNA, the particles are mixed with DNA in low pH (~6), high ionic strength and dehydration environment. Particles are washed on magnet and then DNA is eluted by Tris-HCl buffer (pH = 8). DNA copy number is estimated using quantitative polymerase chain reaction (PCR). Zeta potential, particle size, polydispersity and quantitative PCR data are evaluated and compared. DLS is an insightful and supporting method of analysis that adds a new perspective to the process of screening of particles for DNA isolation.
The proper way of breathing is important for everyone. Healthy people often do not follow respiration until breathing problems start-during stress or during sport activity in physiological cases. More serious cases are stroke, injury, or surgery of the chest and others. So, learning to breathe correctly and/or breathing diagnosis is considerable for many reasons. Two novel methods of breath analysis suitable for diagnostics and rehabilitation are presented. The first technique utilizes pressure belts fastened to the patient's belly and chest, and the second method relies on a SwissRanger SR-4000 time-of-flight camera. The measurement principles are described together with the advantages and disadvantages of the applied techniques. The SwissRanger camera depth calibration is proposed to facilitate better results during the breath analysis. The methods are tested on a group of students to provide a comparison of their individual performances. As it was demonstrated, presented methods proved to work reliably. The method based on time-of-flight camera seems to be more suitable for diagnosis, while the method based on pressure belts is more suitable for rehabilitation and biofeedback applications.
- MeSH
- audiovizuální záznam přístrojové vybavení metody MeSH
- dechové testy přístrojové vybavení metody MeSH
- design vybavení MeSH
- dospělí MeSH
- dýchání MeSH
- kalibrace MeSH
- lidé MeSH
- mladý dospělý MeSH
- tlak MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
This article is focused on reviewing the benefit and limitations of utilization of doxorubicin, which is one of the most widely used anticancer drug even though it causes severe side effects. The mechanism of action of common cytostatic drugs is described and special attention is paid to doxorubicin. Its pathway through the body is highlighted and its toxicity is emphasized. Finally, strategy of elimination of the negative effects induced by doxorubicin therapy is mentioned and the most widely approach – the use of liposomes – is summarized.
- Klíčová slova
- stealth, enkapsulace,
- MeSH
- cytostatické látky farmakokinetika farmakologie škodlivé účinky MeSH
- doxorubicin * farmakokinetika farmakologie škodlivé účinky MeSH
- lidé MeSH
- liposomy metabolismus MeSH
- nádory farmakoterapie MeSH
- nemoci jater etiologie MeSH
- nemoci srdce chemicky indukované MeSH
- nežádoucí účinky léčiv * prevence a kontrola MeSH
- polyethylenglykoly MeSH
- systémy cílené aplikace léků * metody MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy MeSH
Quantum dots (QDs) are small semiconductor nanoparticles with great optical properties. Their behaviour enables the usage of QDs in in vitro and in vivo experiments and they are promising tools in disease treatment and targeted therapy. The limitation of their usage is the toxicity. Quantum dots consist of different metals, which have various effects on the health. To decrease their toxicity, different surface coatings are used. The effect of QDs on the organism can be tested on chicken embryos. Chicken embryos represent great model for QDs toxicity studies, because there is no need of any permission for the work with embryos and the experiments are low cost and fast.
Human papillomavirus (HPV) selectively infects the epithelium of the skin and mucous membranes. Specific HPV types are associated with squamous cell carcinoma, adenocarcinoma, and dysplasias of the cervix, penis, anus, vagina and vulva1. The term head and neck cancer includes malignancy in an area that comprises the skin, oral cavity, salivary glands, lip, pharynx, larynx, nasal cavity, paranasal sinuses and soft tissues of the neck and ear2. The first association of HPV with head and neck cancer was published in 1985 3. HPV was also shown to play a role in the pathogenesis of a subset of head and neck squamous cell carcinomas (HNSCCs)4. Almost 650,000 patients worldwide are diagnosed with head or neck cancer each year and 350,000 patients die of this disease as this cancer is the sixth most prevalent type of cancer worldwide. The ratio of males to females is approximately 2:12, 3. From the point of view of the infection, HPVs have developed several molecular mechanisms to enable infected cells to suppress apoptosis5, 6. Based on their potential for oncogenesis, HPV types can be classified both as high-risk or low-risk7. Precancerous lesions of the oral mucosa are epithelial changes that are able to undergo malignant transformation more likely than normal tissue at other mucosal sites8. A total of 150 HPV genotypes have been identified9. The HPV 16 and 18 strains, which are known to cause nearly all cases of cervical cancer, also raise the risk of developing oropharyngeal cancer10. The evident similarities between both cervical and head and neck tumors prompted the utilization of the same HPV diagnostic procedures. There is now compelling evidence that specially designed methodologies must be employed for prognosis11, 12.
- Klíčová slova
- imunohistochemické značení,
- MeSH
- Alphapapillomavirus * izolace a purifikace MeSH
- hybridizace in situ MeSH
- imunohistochemie MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- lidé MeSH
- nádorové proteiny MeSH
- nádory hlavy a krku * diagnóza MeSH
- polymerázová řetězová reakce MeSH
- Southernův blotting MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy MeSH
Determination of biomarkers in body fluids provides information about the overall state of the organism 1. Among commonly determined markers in clinical laboratories belong AAs. The AAs (Fig. 1), basic structural units of peptides, as well as their derivatives occupy many metabolic and biochemical roles in human body 2,3. The AAs belong to zwitterions containing acidic and basic functional groups (carboxyl groups and amino, respectively). Zwitterions have neutral character at a certain pH known as the isoelectric point due the dissociation of acidic and basic groups. Therefore, the charge of AAs depend on the number of functional groups and pH environment 4. Determination of AAs levels in the body fluids such as blood and/or plasma 5,6, cerebrospinal fluid 6,7, urine 6,8, saliva 6,7 or amniotic fluid 9 represents the significant clinical indicator not only for control of the nutritional state of the organism, but for a number of metabolic disorders 10. Examples can be inherited metabolic disorders such as phenylketonuria (PKU) (an enzyme deficiency in phenylalanine hydroxylase (PAH) (Fig. 2)) 11 or maple syrup urine disease (an enzyme deficiency in branched-chain α-ketoacid dehydrogenase (BCKD) (Fig. 3)) 12. There are many methods for AAs determination employing various techniques such as high pressure liquid chromatography (HPLC) 13 or gas chromatography (GC) 14. During the 80's of the 20th century CE became a routine laboratory technique 15. Recently, this method has ranked among very useful tools for separation and determination of AAs 2,9,16,17, biogenic amines 18 or peptides 19 and is often used as an alternative to HPLC or GC due to its electrophoretic separation mechanism 16,20. In addition, a main advantage is very low sample and solvent consumption, speed of analysis and excellent separation efficiency 2,16,21-23. Besides, due to the high resolving power is CE a promising method for the analysis of biological fluids (complex mixtures of many metabolites) 2,22. The essential part of the CE instrumentation is a detector. In photometric and fluorescence detection is problem with a limited presence of chromophores and fluorofores. Because of that, the derivatization procedures prior CE are usually used, which is the one of the limiting factors 24. Therefore, there are attempts to analyze AAs without derivatization employing various strategies such as indirect detection 25, electrochemical detection (conductometric detection 6,8, amperometric detection 26) or CE in combination with mass spectrometry (MS) 5,11.
- MeSH
- aminokyseliny * krev metabolismus moč MeSH
- biologické markery MeSH
- elektroforéza kapilární * MeSH
- lidé MeSH
- tělesné tekutiny * chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy MeSH
Increasing emissions of heavy metals such as cadmium, mercury, and arsenic into the environment pose an acute problem for all organisms. As a mass of protection, many of them, develop mechanisms of full resistance or at least exhibit partially resisting toward these effects. In this way, based on the chemical similarity of the involved metallic species, they are able, to replace them with viable metals necessary for the effective functioning of the cell. These heavy metals may be bound to the functional groups of proteins and modify their structure and through this also affect their physiological function 1, 2. Higher plants, algae, certain yeasts and animals are able to respond to heavy metals by synthesizing phytochelatins (PCs) and related cysteine-rich polypeptides. Phytochelatin synthases are γ-glutamylcysteine (γ-Glu-Cys) dipeptidyl transpeptidases that catalyze the synthesis of heavy metal-binding PCs 3, 4. PCs, cysteine-rich peptides, are produced from glutamine, cysteine and glycine. Unlike commonmetal-binding structures, MT and GSH, PCs are not gene-encoded, but enzymatically synthesized peptides 5. PCs have been identified in a wide variety of plant species, microorganisms and some invertebrates 6-10. They are structurally related to glutathione (GSH) and were presumed to be the products of a biosynthetic pathway. Numerous physiological, biochemical and genetic studies have confirmed GSH as the substrate for PCs biosynthesis 11, 12. The general structure of PCs is (c-Glu-Cys)n-Gly, with increasing repetitions of the dipeptide Glu-Cys linked through a c-carboxylamide bond (Fig 1), where n varies from 2 to 11, but typically reaching not further than five 13. Except glycine, also other amino acid residues can be found on C-terminal end of (γ-Glu-Cys)n peptides. Examples of which, like Ser, Glu, Gln and Ala are often found at this position in some plant species, and they are assumed to be functionally analogous and synthesised via essentially similar biochemical pathways 14, 15. In in vitro studies of PC synthase expressed in E. coli or in S. cerevisiae, the enzyme was activated to varying extents by Cd, Cu, Ag, Hg, Zn and Pb ions 16-18. PC synthase genes were also isolated in A.thaliana 16 and T.aestivum 18. Genes homologous to those from A.thaliana and T.aestivum were also found in S.pombe and C.elegans, suggesting the existence of PC synthase genes in more species 19.
- MeSH
- chemické techniky analytické metody MeSH
- fytochelatiny * biosyntéza chemie metabolismus MeSH
- rostliny metabolismus MeSH
- těžké kovy * metabolismus MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy MeSH
