The main dose-limiting side effect of cisplatin is nephrotoxicity. The utilization of cisplatin is an issue of balancing tumour toxicity versus platinum-induced nephrotoxicity. In this study, we focused on intraorgan distribution of common essential trace elements zinc, copper, and iron in healthy mouse kidneys and distribution of platinum after cisplatin treatment. Renal distribution in 12 nontreated Nu-Nu mice (males) was assessed by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS). Furthermore, 9 Nu-Nu mice were treated with cisplatin. The order of elements concentration in kidneys was as follows: Fe > Zn > Cu. All three metals showed the higher concentrations at the cortex and medulla (28.60, 3.35, and 93.83 μg/g for Zn, Cu, and Fe, respectively) and lower concentration at the pelvis and the urinary tract (20.20, 1.93, and 62.48 μg/g for Zn, Cu, and Fe, respectively). No statistically significant difference between cortex and medulla was observed for these elements. After platinum treatment, the concentration of platinum in kidneys was enhanced more than 60-times, p < 0.001. Platinum significantly showed the highest accumulation in cortex (2.11 μg/g) with a gradient distribution. Platinum was less accumulated in medulla and pelvis than in cortex, and the lowest accumulation occurred in the urinary tract (1.13 μg/g). Image processing has been successfully utilized to colocalize metal distribution using LA-ICP-MS and histological samples images.
- MeSH
- buňky PC-3 MeSH
- cisplatina škodlivé účinky farmakologie toxicita MeSH
- hmotnostní spektrometrie metody MeSH
- ledviny účinky léků metabolismus patologie MeSH
- lidé MeSH
- měď analýza MeSH
- myši nahé MeSH
- myši MeSH
- platina analýza MeSH
- spektrální analýza metody MeSH
- železo analýza MeSH
- zinek analýza MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Antracyklinová cytostatika lze pozorovat na úrovni organel, buněk i celých organizmů díky jejich fluorescenčním vlastnostem. Zobrazovací techniky na bázi detekce fluorescence mohou být použity nejen pro pozorování interakce léčiva s nádorovými buňkami, ale také mohou sloužit pro cílení léčby nádorů nanočásticemi s obsahem antracyklinového cytostatika. V klinické praxi je využíván doxorubicin a daunorubicin uzavřený do lipozomů jako zástupce nanočástic vhodných pro cílenou léčbu. Hlavní výhodou lipozomálních preparátů je omezení nežádoucích účinků v důsledku rozdílné farmakokinetiky a distribuce léku v organizmu. Vzhledem k tomu, že stále nejsou prozkoumány všechny biologické mechanizmy účinku antracyklinových léčiv, nabízejí moderní zobrazovací techniky nástroj pro in vivo studium těchto mechanismů.
Anthracycline cytostatics can be observed at the level of organelles, cells and whole organisms due to their fluorescent properties. Imaging techniques based on detection of fluorescence can be used not only for observation of drug interaction with tumor cells, but also for targeting therapy of tumors with nanoparticles containing anthracycline cytostatics. Doxorubicin and daunorubicin, enclosed in liposomes, as representatives of nanoparticles suitable for targeted therapy, are used in clinical practice. The main advantage of liposomal drugs is to reduce the side effects due to differences in pharmacokinetics and distribution of the drug in the body. Due to the fact that all biological mechanisms of action of anthracycline drugs are not still fully understood, modern imaging techniques offer tool for in vivo studies of these mechanisms.
- MeSH
- cytostatické látky MeSH
- doxorubicin * farmakologie MeSH
- farmaceutická technologie MeSH
- fullereny MeSH
- lidé MeSH
- liposomy MeSH
- nanočástice * využití MeSH
- nanotechnologie * MeSH
- nosiče léků * MeSH
- systémy cílené aplikace léků * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy MeSH
- Publikační typ
- abstrakt z konference MeSH
Metallothionein (MT) as a potential cancer marker is at the center of interest and its properties, functions and behavior under various conditions is intensively studied. In the present study, two major mammalian MT isoforms (MT-1 and MT-2) were separated using capillary electrophoresis (CE) coupled with UV detector in order to describe their basic behavior. Under the optimized conditions, the separation of both isoforms was enabled as well as estimation of detection limits as subunits and units of ng per μL for MT-2 and MT-1, respectively. Further, the effects of thermal treatment and the presence of denaturing agent such as urea on MT-1 and MT-2 isoforms were studied by CE-UV. Thermal treatment caused an increase in the signals of both isoforms. A new parameter called precipitation rate has been defined based on this finding. This parameter can be expressed as a slope of the linear regression of the time dependency curve recalculated on the MT concentration. The thermal precipitation rate for MT-1 and MT-2 was determined as 1.1 and 0.9 ng of MT/min, respectively. The chemical precipitation rate calculated from the linear regression for both isoforms provided the same value of 0.25 ng of MT/min. The results were confirmed by manual spectrometric measurements and by differential pulse voltammetry Brdicka reaction. Based on these results, a model of MT behavior under the conditions studied was suggested.
- MeSH
- biochemické jevy MeSH
- chemická precipitace MeSH
- chemické modely MeSH
- denaturace proteinů MeSH
- elektroforéza kapilární metody MeSH
- králíci MeSH
- lineární modely MeSH
- metalothionein chemie metabolismus MeSH
- močovina chemie MeSH
- molekulární sekvence - údaje MeSH
- nádorové biomarkery analýza chemie MeSH
- protein - isoformy MeSH
- sekvence aminokyselin MeSH
- sekvenční seřazení MeSH
- senzitivita a specificita MeSH
- spektrofotometrie ultrafialová MeSH
- vysoká teplota MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Currently, the influenza virus infects millions of individuals every year. Since the influenza virus represents one of the greatest threats, it is necessary to develop a diagnostic technique that can quickly, inexpensively, and accurately detect the virus to effectively treat and control seasonal and pandemic strains. This study presents an alternative to current detection methods. The flow-injection analysis-based biosensor, which can rapidly and economically analyze a wide panel of influenza virus strains by using paramagnetic particles modified with glycan, can selectively bind to specific viral A/H5N1/Vietnam/1203/2004 protein-labeled quantum dots. Optimized detection of cadmium sulfide quantum dots (CdS QDs)-protein complexes connected to paramagnetic microbeads was performed using differential pulse voltammetry on the surface of a hanging mercury drop electrode (HMDE) and/or glassy carbon electrode (GCE). Detection limit (3 S/N) estimations based on cadmium(II) ions quantification were 0.1 μg/mL or 10 μg/mL viral protein at HMDE or GCE, respectively. Viral protein detection was directly determined using differential pulse voltammetry Brdicka reaction. The limit detection (3 S/N) of viral protein was estimated as 0.1 μg/mL. Streptavidin-modified paramagnetic particles were mixed with biotinylated selective glycan to modify their surfaces. Under optimized conditions (250 μg/mL of glycan, 30-min long interaction with viral protein, 25°C and 400 rpm), the viral protein labeled with quantum dots was selectively isolated and its cadmium(II) content was determined. Cadmium was present in detectable amounts of 10 ng per mg of protein. Using this method, submicrogram concentrations of viral proteins can be identified.
- MeSH
- biosenzitivní techniky přístrojové vybavení metody MeSH
- biotin chemie metabolismus MeSH
- elektrochemické techniky přístrojové vybavení metody MeSH
- elektrody MeSH
- hemaglutininové glykoproteiny viru chřipky analýza metabolismus MeSH
- kadmium analýza MeSH
- kvantové tečky MeSH
- limita detekce MeSH
- lineární modely MeSH
- magnetické nanočástice chemie MeSH
- průtoková injekční analýza přístrojové vybavení metody MeSH
- rtuť chemie MeSH
- sloučeniny kadmia chemie MeSH
- streptavidin chemie metabolismus MeSH
- sulfidy chemie MeSH
- uhlík chemie MeSH
- virus chřipky A, podtyp H5N1 chemie izolace a purifikace MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Zinc(II) as the only transition metal lacking redox activity is an essential part of approximately 10% proteins as a cofactor of these proteins. Considering the fact that there are numerous zinc(II) containing proteins, proteomics and metallomics studies aimed on them require accurate methods for preparation of real biological samples prior to their subsequent analysis using 2DE and MS. For this purpose, we suggested a new method based on chicken anti-zinc antibodies and magnetizable particles. Antibodies were covalently immobilized to the surface of paramagnetic beads activated with tosyl group. Binding of the antibody to the beads was confirmed by secondary anti-chicken antibody conjugated with horseradish peroxidase. The immunoextraction conditions, such as concentration of the beads (6-18 μg/mL of the sample), time of immunoextraction (6-34 min), pH and composition of the elution buffer, and time of extraction (48-300 s) were optimized. Subsequently, zinc proteins were extracted from human plasma and total concentration of zinc was monitored by electrochemical detection in the extracts. Under optimal conditions it was possible to monitor the proteins and zinc removal from the sample by chip CE, SDS-PAGE, and indirectly using electrochemistry.
- MeSH
- dospělí MeSH
- imunoglobuliny chemie metabolismus MeSH
- imunomagnetická separace metody MeSH
- koncentrace vodíkových iontů MeSH
- kur domácí MeSH
- lidé MeSH
- metaloproteiny krev chemie izolace a purifikace metabolismus MeSH
- molekulová hmotnost MeSH
- protilátky chemie metabolismus MeSH
- sérový albumin hovězí chemie metabolismus MeSH
- vaječný žloutek chemie imunologie MeSH
- western blotting MeSH
- zinek krev chemie MeSH
- zvířata MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Prostate cancer with altered zinc(II) cell metabolism is the second most frequently diagnosed cancer in developed countries. The alterations of zinc(II) metabolism can influence metabolism of other metal ions and can also be associated with the expression and translation of metal-binding proteins including metallothioneins. The aim of this article was to optimize immunoseparation protocol based on paramagnetic beads conjugated with protein G for the isolation of metallothionein. Isolated metallothionein was determined by differential pulse voltammetry Brdicka reaction and SDS-PAGE. Optimal conditions: antigen-binding time - 60 min, temperature - 70°C, and buffer composition and pH - acetate buffer, pH 4.3, were determined. Under the optimized conditions, lysates from 22Rv1 prostate cancer cells treated with various concentrations of cadmium(II) and copper(II) ions were analyzed. We observed strong correlation in all experimental groups and all lysate types (r>0.83 at p<0.041) between metallothionein concentration related to viability and concentration of copper(II) ions and cadmium(II) ions in medium. Moreover, the results were compared with standard sample preparation as heat treatment and SDS-PAGE analysis.
- MeSH
- buněčné extrakty chemie MeSH
- časové faktory MeSH
- elektrická impedance MeSH
- elektroforéza v polyakrylamidovém gelu metody MeSH
- fluorescenční mikroskopie MeSH
- imunoglobulin G chemie metabolismus MeSH
- imunoprecipitace metody MeSH
- koncentrace vodíkových iontů MeSH
- lidé MeSH
- lineární modely MeSH
- magnety MeSH
- metalothionein izolace a purifikace metabolismus MeSH
- mikrosféry MeSH
- monoklonální protilátky chemie MeSH
- nádorové buněčné linie MeSH
- nádory prostaty chemie metabolismus patologie MeSH
- teplota MeSH
- těžké kovy MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Klíčová slova
- diagnostika, biomarker,
- MeSH
- elektroforéza kapilární statistika a číselné údaje využití MeSH
- financování organizované MeSH
- lidé MeSH
- moč MeSH
- nádory prostaty diagnóza klasifikace moč MeSH
- sarkosin analýza diagnostické užití moč MeSH
- stupeň závažnosti nemoci MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Klíčová slova
- diagnostika, fingerprint, agregáty,
- MeSH
- adenokarcinom diagnóza genetika MeSH
- elektroforéza metody statistika a číselné údaje využití MeSH
- financování organizované MeSH
- lidé MeSH
- metalothionein analýza diagnostické užití MeSH
- nádory prostaty diagnóza genetika MeSH
- peptidové mapování metody statistika a číselné údaje využití MeSH
- sérum MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
