Q41871498
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Urinary metabolomic profiles have recently drawn a lot of attention owing to a debate regarding their possible role as potential clinical markers for prostate cancer. As was shown, amino acid metabolism in cancer patients differs from that in healthy people, and it can be thus utilized in early diagnostics. In this study, we monitored the behavior of potential noninvasive biomarker for prostate carcinoma, sarcosine, involved in the folate metabolism and DNA methylation processes, linked to the progression of prostate carcinoma. To obtain the maximum amount of information, the biochemical parameters (total protein, creatinine, ions, conductivity) were determined using spectrophotometry and electrochemistry. All results were subjected to statistical processing for revealing different correlations between the studied parameters. These metabolites were observed in the urine obtained from healthy subjects and influence of storage conditions (freezing and thawing) on the concentration of addition of sarcosine was monitored.
- MeSH
- časové faktory MeSH
- chromatografie iontoměničová * statistika a číselné údaje MeSH
- elektrochemické techniky statistika a číselné údaje MeSH
- lidé MeSH
- metabolomika MeSH
- nádorové biomarkery metabolismus moč MeSH
- nádory prostaty metabolismus moč MeSH
- odběr biologického vzorku * metody MeSH
- sarkosin * metabolismus moč MeSH
- teplota MeSH
- termodynamika MeSH
- zmrazování MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- hodnotící studie MeSH
- práce podpořená grantem MeSH
- MeSH
- kongresy jako téma MeSH
- nanomedicína MeSH
- Publikační typ
- práce podpořená grantem MeSH
- zprávy MeSH
A novel microfluidic label-free bead-based metallothionein immunosensors was designed. To the surface of superparamagnetic agarose beads coated with protein A, polyclonal chicken IgY specifically recognizing metallothionein (MT) were immobilized via rabbit IgG. The Brdicka reaction was used for metallothionein detection in a microfluidic printed 3D chip. The assembled chip consisted of a single copper wire coated with a thin layer of amalgam as working electrode. Optimization of MT detection using designed microfluidic chip was performed in stationary system as well as in the flow arrangement at various flow rates (0-1800 μL/min). In stationary arrangement it is possible to detect MT concentrations up to 30 ng/mL level, flow arrangement allows reliable detection of even lower concentration (12.5 ng/mL). The assembled miniature flow chip was subsequently tested for the detection of MT elevated levels (at approx. level 100 μg/mL) in samples of patients with cancer. The stability of constructed device for metallothionein detection in flow arrangement was found to be several days without any maintenance needed.
- MeSH
- design vybavení MeSH
- elektrochemické techniky přístrojové vybavení metody MeSH
- elektrody MeSH
- imobilizační protilátky chemie metabolismus MeSH
- imunoglobulin G chemie metabolismus MeSH
- imunoglobuliny chemie metabolismus MeSH
- imunomagnetická separace přístrojové vybavení metody MeSH
- králíci MeSH
- kur domácí MeSH
- lidé středního věku MeSH
- lidé MeSH
- metalothionein krev MeSH
- nádory hlavy a krku krev MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Metallothioneins (MT) are small (molecular weight in range 500 – 1400 Da) intracellular proteins rich in cysteine content. In humans four main isoforms of MTs have been discovered so far – MT-1, MT-2, MT-3 and MT-4. All of MTs functions – heavy metals binding, antioxidative, regulation and mmunomodulation are involved in MTs roles in cancer. MTs role is doubleedged in carcinogenesis; in healthy cells with high MT content it has protective effects, while in healthy cells with low MT content, carcinogenesis via environmental factors occurs, on the other hand, at cancer-transformed cell the high MT content increases the tumour malignancy and chemotherapeutics resistance. Expression of MT can be used as marker of tumour diseases. Increased level of MT in serum or full blood was found in many cancer diseases and can be used as early stage biomarker. To decrease tumour resistance, or on the other hand, to improve non-target cells tolerance to chemotherapeutic multiple approaches are tested, including specific increasing or decreasing of MT expression based on specific compounds or targeted gene therapy.
- MeSH
- chemorezistence MeSH
- karcinogeneze MeSH
- lidé MeSH
- metalothionein * fyziologie genetika metabolismus MeSH
- nádorové biomarkery * krev MeSH
- nádorové proteiny fyziologie genetika metabolismus MeSH
- nádory * diagnóza krev terapie MeSH
- protein - isoformy MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy MeSH
The anti-DNA antibodies are produced in patients with autoimmune disease called systemic lupus erythematodes. They can be reactive against double or single stranded DNA or DNA modified with some other molecules. Using the variety of antibodies it is possible to determine the structure of studied DNA. In this work, we used 4 anti-DNA antibodies produced in egg yolk after immunization of hens with DNA-mBSA antigen – anti-dsDNA, anti-ssDNA, afi-dsDNA and afi-ssDNA. The reactivity of these antibodies was evaluated using the dot blot method with different lengths and concentrations of DNA antigen. The most reactive antibodies (anti-ssDNA) were modified with carbon quantum dots synthesized from multiwall carbon nanotubes and this modification was verified by ELISA-like method with fluorescent detection and fluorescence resonance energy transfer between DNA and quantum dots was observed, increasing the sensitivity of the DNA detection.
There is still a lot of unknown related to our perceptiveness to civilization and other illnesses including tumour ones often connected with environmental changes. There is also still an enormous field for cutting-edge research necessary to establish a role the unique tiny particles playing in the whole concert leading to our fitness or illness or, telling in other words, to normal or pathological functioning of our body cells. Studying metallome as the whole picture composed from metals, peptides, proteins and cell parts belongs to the most challenging issues of present biomedicine. Here, we summarize the omics advances in this field with special focus on in vivo imaging systems
- Klíčová slova
- metalomika, metaloproteomika,
- MeSH
- kovy * metabolismus MeSH
- lidé MeSH
- metaloproteiny MeSH
- metalothionein MeSH
- molekulární zobrazování MeSH
- nádory * genetika MeSH
- nanočástice * MeSH
- proteomika MeSH
- stopové prvky MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy MeSH
Currently, metallothioneins (MTs) are extensively investigated as the molecular biomarkers and the significant positive association of the MT amount was observed in tumorous versus healthy tissue of various types of malignant tumors, including head and neck cancer. Thus, we proposed a biosensor with fluorescence detection, comprising paramagnetic nanoparticles (nanomaghemite core with gold nanoparticles containing shell) for the magnetic separation of MT, based on affinity of its sulfhydryl groups toward gold. Biosensor was crafted from PDMS combined with technology of 3D printing and contained reservoir with volume of 50 μL linked to input (sample/detection components and washing/immunobuffer) and output (waste). For the immunolabeling of immobilized MT anti-MT antibodies conjugated to CdTe quantum dots through synthetic heptapeptide were employed. After optimization of fundamental conditions of the immunolabeling (120 min, 20°C, and 1250 rpm) we performed it on a surface of paramagnetic nanoparticles in the biosensor reservoir, with evaluation of fluorescence of quantum dots (λexc 400 nm, and λem 555 nm). The developed biosensor was applied for quantification of MT in cell lines derived from spinocellular carcinoma (cell line 122P-N) and fibroblasts (122P-F) and levels of the biomarker were found to be about 90 nM in tumor cells and 37 nM in fibroblasts. The proposed system is able to work with low volumes (< 100 μL), with low acquisition costs and high portability.
- MeSH
- 3D tisk * MeSH
- biosenzitivní techniky MeSH
- dimethylpolysiloxany chemie MeSH
- fluorescence MeSH
- kovové nanočástice MeSH
- kvantové tečky MeSH
- lidé MeSH
- magnetismus MeSH
- metalothionein analýza MeSH
- nádorové buněčné linie MeSH
- nádory patologie MeSH
- sloučeniny kadmia chemie MeSH
- telur chemie MeSH
- zlato chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Dot-blot is one of biological methods that are normally used in research laboratories and especially in the diagnostics. It is the most commonly used method for identification and immunodetection of particular proteins which may be markers of various diseases. The main aim of the experiment was to develop a simple, inexpensive and rapid method for specific detection of nucleic acids, especially DNA, and then this procedure apply to the detection of DNA modified by platinum cytostatic drugs. Despite platinum cytostatic drugs‘ common use in chemotherapy of various cancer types, their biochemical effect is still not completely clear. The generally accepted opinion is that the drug binds to cellular DNA. It was observed that cisplatin is bound to the DNA the most compared to oxaliplatin and carboplatin.
- Klíčová slova
- oxaliplatina,
- MeSH
- cisplatina MeSH
- DNA * imunologie účinky léků MeSH
- imunoblotting * metody statistika a číselné údaje MeSH
- karboplatina MeSH
- organoplatinové sloučeniny MeSH
- polymerázová řetězová reakce MeSH
- protilátky MeSH
- protinádorové látky * MeSH
- Publikační typ
- hodnotící studie MeSH
- práce podpořená grantem MeSH
