Bactrocera invadens Drew, Tsuruta & White, Bactrocera papayae Drew & Hancock, and Bactrocera philippinensis Drew & Hancock, key pest species within the Bactrocera dorsalis species complex, have been recently synonymized under the name Bactrocera dorsalis (Hendel). The closely related Bactrocera carambolae Drew & Hancock remains as a discrete taxonomic entity. Although the synonymizations have been accepted by most researchers, debate about the species limits remains. Because of the economic importance of this group of taxa, any new information available to support or deny the synonymizations is valuable. We investigated the chemical epicuticle composition of males and females of B. dorsalis, B. invadens, B. papayae, B. philippinensis, and B. carambolae by means of one- and two-dimensional gas chromatography-mass spectrometry, followed by multiple factor analyses and principal component analysis. Clear segregation of complex cuticule profiles of both B. carambolae sexes from B. dorsalis (Hendel) was observed. In addition to cuticular hydrocarbons, abundant complex mixtures of sex-specific oxygenated lipids (three fatty acids and 22 fatty acid esters) with so far unknown function were identified in epicuticle extracts from females of all species. The data obtained supports both taxonomic synonymization of B. invadens, B. papayae, and B. philippinensis with B. dorsalis, as well as the exclusion of B. carambolae from B. dorsalis.
Melatonin is a hormone with strong antioxidant properties. In this experiment, Freund's complete adjuvant was used as a stressogenic substance given to laboratory outbred mice, whereas melatonin was investigated as a protectant against the stressogenic effect. Levels of low molecular weight antioxidants, thiobarbituric acid reactive substances, and tumor necrosis factor α and activity of glutathione reductase were determined in blood from the animals. Surprisingly, melatonin was not involved in direct regulation of antioxidants, thiobarbituric acid reactive substances and tumor necrosis factor α. On the other hand, melatonin regulated glutathione reductase activity. We can conclude on regulation of metabolism caused by melatonin in the model. The effect was more important than the expected regulation of immunity and basal oxidative homeostasis.
- MeSH
- adjuvancia imunologická farmakologie MeSH
- antioxidancia farmakologie fyziologie MeSH
- Freundovo adjuvans farmakologie MeSH
- melatonin farmakologie fyziologie MeSH
- myši MeSH
- oxidační stres účinky léků fyziologie MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
DNA nanotechnology is a rapidly growing research area, where DNA may be used for wide range of applications such as construction of nanodevices serving for large scale of diverse purposes. Likewise a panel of various purified fluorescent proteins is investigated for their ability to emit their typical fluorescence spectra under influence of particular excitation. Hence these proteins may form ideal donor molecules for assembly of fluorescence resonance emission transfer (FRET) constructions. To extend the application possibilities of fluorescent proteins, while using DNA nanotechnology, we developed nanoconstruction comprising green fluorescent protein (GFP) bound onto surface of surface active nanomaghemite and functionalized with gold nanoparticles. We took advantage of natural affinity between gold and thiol moieties, which were modified to bind DNA fragment. Finally we enclosed doxorubicin into fullerene cages. Doxorubicin intercalated in DNA fragment bound on the particles and thus we were able to connect these parts together. Because GFP behaved as a donor and doxorubicin as an acceptor using excitation wavelength for GFP (395 nm) in emission wavelength of doxorubicin (590 nm) FRET was observed. This nanoconstruction may serve as a double-labeled transporter of doxorubicin guided by force of external magnetic force owing to the presence of nanomaghemite. Further nanomaghemite offers the possibility of using this technology for thermotherapy.
- MeSH
- DNA chemie MeSH
- doxorubicin chemie MeSH
- fullereny MeSH
- luminescentní proteiny chemie MeSH
- magnetické nanočástice chemie MeSH
- nanotechnologie metody MeSH
- rezonanční přenos fluorescenční energie metody MeSH
- zelené fluorescenční proteiny chemie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
It has been already three decades, since the fluorescent nanocrystals called quantum dots (QDs) appeared and attracted attention of a broad scientific community. Their excellent not only optical but also electronic properties predetermined QDs for utilization in a variety of areas. Besides lasers, solar cells, and/or computers, QDs have established themselves in the field of (bio)chemical labeling as well as medical imaging. However, due to the numerous application possibilities of QDs, there are high demands on their properties that need to be precisely controlled and characterized. CE with its versatile modes and possibilities of detection was found to be an effective tool not only for characterization of QDs size and/or surface properties but also for monitoring of their interactions with other molecules of interest. In this minireview, we are giving short insight in analysis of QDs by CE, and summarizing the advantages of this method for QDs characterization.
- MeSH
- elektroforéza kapilární metody MeSH
- kvantové tečky chemie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Microfluidic techniques have been developed intensively in recent years due to lower reagent consumption, faster analysis, and possibility of the integration of several analytical detectors into one chip. Electrochemical detectors are preferred in microfluidic systems, whereas liposomes can be used for amplification of the electrochemical signals. The aim of this study was to design a nanodevice for targeted anchoring of liposome as transport device. In this study, liposome with encapsulated Zn(II) was prepared. Further, gold nanoparticles were anchored onto the liposome surface allowing binding of thiol moiety-modified molecules (DNA). For targeted capturing of the transport device, DNA loops were used. DNA loops were represented by paramagnetic microparticles with oligo(DT)25 chain, on which a connecting DNA was bound. Capturing of transport device was subsequently done by hybridization to the loop. The individual steps were analyzed by electrochemistry and UV/Vis spectrometry. For detection of Zn(II) encapsulated in liposome, a microfluidic system was used. The study succeeded in demonstrating that liposome is suitable for the transport of Zn(II) and nucleic acids. Such transporter may be used for targeted binding using DNA anchor system.
- MeSH
- kovové nanočástice chemie MeSH
- liposomy chemie metabolismus ultrastruktura MeSH
- mikrofluidní analytické techniky MeSH
- nanomedicína přístrojové vybavení MeSH
- nanostruktury chemie ultrastruktura MeSH
- nukleové kyseliny chemie metabolismus MeSH
- zinek chemie MeSH
- zlato chemie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Sarcosine has been identified as a potential prostate cancer marker. To provide determination of this compound, a number of methods are developing. In this study, we optimized a method for its separation by hydrophilic interaction LC with electrochemical detection (ED). Due to the fact that mobile phases commonly used for this type of separation altered the LODs measured by electrochemical detectors, we applied postcolumn dosing of buffer suitable for ED. The optimized conditions were mobile phase A acetonitrile, mobile phase B water in the ratio A/B 70:30, with postcolumn addition of mobile phase C (200 mM phosphate buffer pH 9). The optimal mixing ratio was A + B/C 1:1 with a flow rate of 0.80 mL/min (0.40 + 0.40 mL/min) and detection potential of 1000 mV. Due to the optimization of the parameters for effective separation, which had to meet the optimal parameters of ED, we reached a good resolution for separation also with a good LOD (100 nM). In addition, we successfully carried out sarcosine analysis bound on our modified paramagnetic microparticles with the ability to preconcentrate sarcosine isolated from artificial urine.
Cisplatin, carboplatin, and oxaliplatin represent three generations of platinum based drugs applied successfully for cancer treatment. As a consequence of the employment of platinum based cytostatics in the cancer treatment, it became necessary to study the mechanism of their action. Current accepted opinion is the formation of Pt-DNA adducts, but the mechanism of their formation is still unclear. Nanomaterials, as a progressively developing branch, can offer a tool for studying the interactions of these drugs with DNA. In this study, fluorescent CdTe quantum dots (QDs, λem = 525 nm) were employed to investigate the interactions of platinum cytostatics (cisplatin, carboplatin, and oxaliplatin) with DNA fragment (500 bp, c = 25 μg/mL). Primarily, the fluorescent behavior of QDs in the presence of platinum cytostatics was monitored and major differences in the interaction of QDs with tested drugs were observed. It was found that the presence of carboplatin (c = 0.25 mg/mL) had no significant influence on QDs fluorescence; however cisplatin and oxaliplatin quenched the fluorescence significantly (average decrease of 20%) at the same concentration. Subsequently, the amount of platinum incorporated in DNA was determined by QDs fluorescence quenching. Best results were reached using oxaliplatin (9.4% quenching). Linear trend (R(2) = 0.9811) was observed for DNA platinated by three different concentrations of oxaliplatin (0.250, 0.125, and 0.063 mg/mL). Correlation with differential pulse voltammetric measurements provided linear trend (R(2) = 0.9511). As a conclusion, especially in the case of oxaliplatin-DNA adducts, the quenching was the most significant compared to cisplatin and nonquenching carboplatin.
- MeSH
- adukty DNA analýza chemie MeSH
- cisplatina analýza metabolismus MeSH
- cytostatické látky analýza metabolismus MeSH
- elektrochemie metody MeSH
- elektroforéza kapilární metody MeSH
- elektroforéza v agarovém gelu metody MeSH
- fluorescence MeSH
- glutathion chemie MeSH
- karboplatina analýza metabolismus MeSH
- kvantové tečky * MeSH
- organoplatinové sloučeniny analýza chemie metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Pathogenic bacteria have become a serious socio-economic concern. Immunomagnetic separation-based methods create new possibilities for rapidly recognizing many of these pathogens. The aim of this study was to use superparamagnetic particles-based fully automated instrumentation to isolate pathogen Staphylococcus aureus and its Zn(II) containing proteins (Zn-proteins). The isolated bacteria were immediately purified and disintegrated prior to immunoextraction of Zn-proteins by superparamagnetic beads modified with chicken anti-Zn(II) antibody. S. aureus culture was treated with ZnCl(2). Optimal pathogen isolation and subsequent disintegration assay steps were carried out with minimal handling. (i) Optimization of bacteria capturing: Superparamagnetic microparticles composed of human IgG were used as the binding surface for acquiring live S. aureus. The effect of antibodies concentration, ionic strength, and incubation time was concurrently investigated. (ii) Optimization of zinc proteins isolation: pure and intact bacteria isolated by the optimized method were sonicated. The extracts obtained were subsequently analyzed using superparamagnetic particles modified with chicken antibody against zinc(II) ions. (iii) Moreover, various types of bacterial zinc(II) proteins precipitations from particle-surface interactions were tested and associated protein profiles were identified using SDS-PAGE. Use of a robotic pipetting system sped up sample preparation to less than 4 h. Cell lysis and Zn-protein extractions were obtained from a minimum of 100 cells with sufficient yield for SDS-PAGE (tens ng of proteins). Zn(II) content and cell count in the extracts increased exponentially. Furthermore, Zn(II) and proteins balances were determined in cell lysate, extract, and retentate.
- MeSH
- bakteriální proteiny chemie izolace a purifikace metabolismus MeSH
- imunoglobulin G metabolismus MeSH
- imunomagnetická separace přístrojové vybavení metody MeSH
- kur domácí MeSH
- lidé MeSH
- limita detekce MeSH
- metaloproteiny chemie izolace a purifikace metabolismus MeSH
- protilátky bakteriální metabolismus MeSH
- robotika přístrojové vybavení metody MeSH
- Staphylococcus aureus chemie izolace a purifikace metabolismus MeSH
- zinek chemie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
In the food industry, in the process of creating new agricultural plant products, and in the testing of anti-cancer drugs there is often a need to assay multiple samples of low molecular weight antioxidants, plant samples and foods rich in antioxidants, with minimal additional costs and low degrees of uncertainty. With these demands in mind, we decided to study the fully automated assay of antioxidants using not only automated sample measurements but also automated processing of samples and application of reagents. The automated pipetting system epMotion 5075 and the automated spectrophotometer BS 400 were chosen for the assay purposes. Five methods were introduced for the automation: 2-diphenyl-1-picrylhydrazyl (DPPH) test, ferric reducing antioxidant power (FRAP) method, 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) based test, N,N-dimethyl-1,4-diaminobenzene (DMPD) based test and the free radicals method. Samples containing one of the four antioxidants (standard rutin, quercitrin, ferulic and gallic acid) in a range 1–1000 μg/ml were used throughout. All of the tested methods were found suitable for implementation in an automated assay. However, some of them, such as the ABTS test failed to assay all tested antioxidants. The coefficients of determination were also unequal. From the analytical point of view, FRAP methods provided the most reliable results in the automated assay; because of the capacity of the method, approximately 240 samples per hour (one sample per 15 seconds) can be assayed using the automated protocol. We were encouraged by the data received and we expect further interest in the practical performance of such automation. As a mean of testing the robustness of our method, in the next step of our study, oxidative status was assessed in model cell lines derived from prostate cancer (PC-3, PNT1A and 22RV1) that were cultured on ellipticine (0, 0.5, 1, 1.5, 2, 2.5, 5, 7.5, 10, 15 μmol/l) supplemented agar. Antioxidant activity was assessed (DPPH, ABTS, FRAP, DMPD, FR) and calculated on the phenolic antioxidant level (rutin, quercitrin, ferulic and gallic acid), and thus an estimation was formulated of the oxidative stress as a result of the impact of anti-cancer drugs. It can be demonstrated that the new method has wide applicability.
- MeSH
- antioxidancia analýza MeSH
- antitumorózní látky chemie metabolismus toxicita MeSH
- chemické techniky analytické metody přístrojové vybavení statistika a číselné údaje MeSH
- elipticiny analýza chemie metabolismus MeSH
- financování organizované MeSH
- FRAP MeSH
- kalibrace MeSH
- kyselina gallová analýza chemie MeSH
- kyseliny kumarové analýza chemie MeSH
- laboratorní automatizace metody přístrojové vybavení MeSH
- léky antitumorózní - screeningové testy metody statistika a číselné údaje MeSH
- luminiscenční měření MeSH
- nádorové buněčné linie MeSH
- oxidační stres účinky léků MeSH
- quercetin analogy a deriváty analýza chemie MeSH
- reprodukovatelnost výsledků MeSH
- rutin analýza chemie MeSH
- spektrofotometrie metody přístrojové vybavení statistika a číselné údaje MeSH
- viabilita buněk účinky léků MeSH
- volné radikály analýza MeSH
- vztahy mezi strukturou a aktivitou MeSH
- Publikační typ
- hodnotící studie MeSH
- statistiky MeSH
- tabulky 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
