Labelling of DNA in replicating cells using 5-bromo-2'- deoxyuridine (BrdU) is widely used, however the rapid clearance and metabolisation of BrdU in the living organism is a critical issue. Although the pharmacokinetic of BrdU in experimental animals is empirically approximated, the exact time-curve remains unknown. Here we present novel method for estimation of the BrdU content in the blood serum. The application is based on the in vitro cocultivation of tumour cells with the examined serum and the subsequent quantification of the incorporated BrdU in the DNA using flow cytometry analysis. Our results demonstrate that this approach can quantify the BrdU concentration in serum at 1 μmol.dm-3 and might represent an attractive alternative to conventional chromatographic analysis. The employment of tumour cells as "detectors" of the BrdU content in serum provides an advantage over high pressure liquid chromatography (HPLC), as this approach allows us to approximate not only the concentration of BrdU, but also to determine, whether BrdU is present in the blood serum in effective concentration to reliable label all cells undergoing the S-phase of the cell cycle. The presented application might be a helpful tool for studies on pharmacokinetics of BrdU or other thymidine analogues when testing various administration routes or protocols., J. Mikeš, J. Ševc, J. Košuth, A. Matiašová, Z. Daxnerová, P. Fedoročko., and Obsahuje bibliografii
The document deals with the research of flow in the cross-over channel of the high-pressure stage with partial admission of first stage. In the opening part there is a description of the experimental model and the method of measuring flow fields with high velocity gradient using the stereo PIV principle. The second part describes the design of the model inlet part. The third chapter deals with measuring in the cross-over channel outlet slot with no influence of the stator blade cascade of the flollowing stage. The stator blade cascade feedback is described in the fourth part. During all the measurements we observe the influence of the shaft revoluton on the flow field at the cross-over channel outlet. and Obsahuje seznam literatury
The cavitations in turbo-machines decrease their efficiency and have a bad impact of their work-life and therefore constructors are always trying to avoid or minimize the cavitation risks in hydraulic machines. In this paper we have done the LDV measurement of flow in the model pump suction sump with baffle. In precedent works we did the PIV (Particles Image Velocimetry) measurement with and without baffle. Around the pump bell there are very complex vortices, which can affect seriously the flow characteristics of the pump. They can also cause vibrations, cavitations and decrease the efficiency of the system. In this experiment the pump suction sump and the pump bell with particular structure are used to search the flow situation in the surrounding of the bell. Both LDV and PIV experiments were done with steady flow using appropriate materials. and Kavitace v turbogenerátorech a čerpacích systémech snižuje jejich účinnost a negativně ovlivňuje jejich životnost. Snahou konstruktérů je proto vyloučit nebo minimalizovat riziko kavitace v hydraulických strojích. V článku jsou prezentována měření rychlostních polí pomocí LDV v modelové sací jímce s tokovým usměrňovačem a výsledky jsou porovnány s dřívějšími měřeními pomocí PIV. V blízkosti sacího zvonu vznikají komplikované vírové struktury, které mohou následně ovlivňovat charakteristiku čerpadla a které mohou být příčinou vzniku vibrací, kavitace a snížení účinnosti. Pozornost proto byla zaměřena na měření rychlostních polí v okolí sacího zvonu.
The flow behaviour of coarse-grained slurry depends on particle size, shape, density and concentration, and on the density and rheological properties of the carrier liquid. The present paper describes the results of an experimental investigation and flow visualisation of model coarse-grained particle-water mixtures in a closed pipe loop with smooth stainless steel pipes of inner diameter 36 mm. Glass balls and washed graded pebble gravel of mean diameter d50 = 6 mm were used as model coarse-grained material. The effect of slurry velocity and particle concentration on the slurry flow behaviour and pressure drop in the turbulent regime was evaluated. Particle distribution in the pipe cross-section and motion of particles along the pipe invert, particle saltation and particle conveying in the carrier liquid were investigated in a transparent pipe viewing section and motion of individual particles was described. Velocity profiles of the carrier liquid and conveyed particles were determined. and Tokové chování hrubozrnných suspenzí závisí na velikosti, tvaru a hustotě částic, koncentraci pevné fáze a hustotě a reologických vlastnostech nosné kapaliny. Článek popisuje výsledky experimentálního výzkumu a vizualizace proudění modelové hrubozrnné suspenze v experimentální potrubní lince s hladkým nerezovým potrubím s vnitřním průměrem 36 mm. Skleněné kuličky a praný oblý štěrk (kačírek) se středním zrnem d50 = 6 mm byly použity jako modelový materiál. Byl vyhodnocen vliv rychlosti proudění suspenze a koncentrace pevné fáze na chování a tlakové ztráty suspenze. Rozdělení částic v příčném průřezu potrubí a pohyb částic podél dna potrubí, jejich saltace a unášení v nosné kapalině byly zkoumány v transparentní části potrubí a byl popsán pohyb jednotlivých částic a pro vybrané případy byly stanoveny rychlostní profily nosné kapaliny a unášených částic.
Fluid inclusions in carbonate-dominated veinlets from selected Czech Upper Paleozoic basins display large variations in salinity (0-25 wt. % eq. NaCl/CaCl2) and smaller variations in the homogenization temperatures (41-112 °C). We suggest that the trapped fluids represent a mixture dominated by heated formation fluids., Jiří Zachariáš and Jiří Pešek., and Obsahuje bibliografii
An influence of soil drought (7 or 14 d) and 7 d recovery on changes of leaf fluorescence excitation spectra at wavelengths of 450, 520, 690, and 740 nm (F450, F520, F690, F740) for drought resistant and sensitive genotypes of triticale and maize was compared. In non-stressed plants the differences between maize and triticale were observed for F450 and F520, but not for F690 and F740. Drought caused the increase in F450, F520, and F690 and this increase was more distinct for drought sensitive genotypes. After re-hydration, chlorophyll fluorescence mostly recovered to values of control plants. Drought caused significant increase in F690/F740 but not in F450/F690 and F450/F520. For triticale, highest increase in F690/F740 was observed in the 4th and 7th leaves of resistant genotype and contrarily in maize for the sensitive one. After recovery, the F450/F520, F450/F690, and F690/F740 ratios mostly returned to values of control plants. and M. T. Grzesiak ... [et al.].
We report the observation of two types of changes in fluorescence spectra of LHCII at 4.2 K following intense illumination of the sample with a spectrally narrow laser beam at wavelengths between 678 and 686 nm. Nonspecific changes (burning-wavelength independent) are characterized by two relatively broad bands: a positive one at - 678.7 nm and a negative one at - 680.8 nm. These changes reveal a -1.3-nm blue shift of the distribution of final emitters in LHCII, from 680.3 nm to - 679.0 nm independent of the excitation wavelength. Specific fluorescence changes (burning-wavelength dependent) are characterized by a sharp hole exactly at the burning wavelength, and positive changes directly to the shorter-and longer-wavelength side of the narrow hole. The negative changes are interpreted as zero-phonon holes, while the positive features are assigned to non-photochemical products. In the low-burning intensity experiment, in addition to the zero-phonon holes, we observed also the holes to the longer wavelength of the zero-phonon hole, which were assigned to a sum of phonon and pseudo-phonon side bands. The shapes of these extra holes are identical to the shapes of the holes revealed in the fluorescence line narrowing experiment. On the basis of the low-burning intensity experiment we estimated the upper limit of the electron-phonon coupling strength for LHCII, characterized by a Huang-Rhys factor of 1.5. and K. Gibasiewicz, M. Rutkowski, R. van Grondelle.
In the pursuit of knowledge on the biological behavior of Brazilian Atlantic Forest tree species, this study evaluated the susceptibility of the light-demanding species, Schinus terebinthifolia Raddi., Pseudobombax grandiflorum (Cav.) A. Robyns and Joannesia princeps Vell., and of the shade-tolerant species, Hymenaea courbaril L. var. stilbocarpa and Lecythis pisonis Camb, to photoinhibition and acclimation capacity. These species were first cultivated under two irradiance conditions, I20 (20% direct sunlight radiation) and I100 (all-sky or direct sunlight) and then transferred from I20 to I100. The effects of the sudden increase in light radiation intensity on photosynthetic activity were then evaluated through chlorophyll (Chl) fluorescence imaging, HPLC xanthophylls analysis, and cell membrane lipid peroxidation measurements. Light-demanding species were found to present a higher photochemical efficiency and higher acclimation capacity under high light irradiance than shade-tolerant species. The higher photoinhibition tolerance observed in light-demanding species was associated to their higher capacity for photochemical dissipation and dissipation of excess excitation energy via the xanthophyll cycle, leading to a lower ROS generation. The obtained results suggested that a knowledge of acclimation capacity, by means of Chl fluorescence imaging yields, is a useful indicator of species successional grouping., L. Dos Anjos, M. A. Oliva, and K. N. Kuki., and Obsahuje bibliografii
The kinetics of bacteriochlorophyll fluorescence in intact cells of the purple nonsulfur bacterium Rhodobacter sphaeroides were measured under continuous and pulsed actinic laser diode (808 nm wavelength and maximum 2 W light power) illumination on the micro- and millisecond timescale. The fluorescence induction curve was interpreted in terms of a combination of photochemical and triplet fluorescence quenchers and was demonstrated to be a reflection of redox changes and electron carrier dynamics. By adjustment of the conditions of single and multiple turnovers of the reaction center, we obtained 11 ms-1 and 120 μs-1 for the rate constants of cytochrome c23+ detachment and cyclic electron flow, respectively. The effects of cytochrome c2 deletion and chemical treatments of the bacteria and the advantages of the fluorescence induction study on the operation of the electron transport chain in vivo were discussed., G. Sipka, M. Kis, J. L. Smart, P. Maróti., and Obsahuje bibliografické odkazy
We describe an instrument that allows the rapid measurement of fluorescence lifetime-resolved images of leaves as well as sub-cellular structures of intact plants or single cells of algae. Lifetime and intensity fluorescence images can be acquired and displayed in real time (up to 55 lifetime-resolved images per s). Our imaging technique therefore allows rapid measurements that are necessary to determine the fluorescence lifetimes at the maximum (P level) fluorescence following initial illumination during the chlorophyll (Chl) a fluorescence transient (induction) in photosynthetic organisms. We demonstrate the application of this new instrument and methodology to measurements of: (1) Arabidopsis thaliana leaves showing the effect of dehydration on the fluorescence lifetime images; (2) Zea mays leaves showing differences in the fluorescence lifetimes due to differences in the bundle sheath cells (having a higher amount of low yield photosystem 1) and the mesophyll cells (having a higher amount of high yield photosystem 2); and (3) single cells of wild type Chlamydomonas reinhardtii and its non-photochemical quenching mutant NPQ2 (where the conversion of zeaxanthin to violaxanthin is blocked), with NPQ2 showing lowered lifetime of Chl a fluorescence. In addition to the lifetime differences referred to in (1) and (2), structural dependent heterogeneities in the fluorescence lifetimes were generally observed when imaging mesophyll cells in leaves. and O. Holub ... [et al.].