Prolamellar bodies (PLBs) isolated from dark-grown, 6.5-d-old leaves of wheat (Triticum aesíivum L. cv. Kosack) were treated with the carboxylic acid cross-linker l-ethyl-3-[3-(diniethylaniino)propyl]carbodiimide (EDC) or with the lysině specific cross-linker 2-iniinothiolane. SDS-PAGE showed that the most prominenent cross- linked product was a dimer of the NADPH-protochlorophyllide oxidoreductase (PCR), but also larger aggregates of the polypeptide were identified by inununological detection on electro-blots. A two-dimensional diagonál gel showed that much of the cross-linking was between the PCR polypeptides. The cross-linkers induced a shift of the fluorescence peak to shorter wavelengths, a bandwidth increase of the fluorescence peak, and an increase of the fluorescence yield. In the presence of NADPH the blue shift was reduced, but the increase in the fluorescence yield still occmred. A cross-linker treatment of PLBs prior to solubilization with 1-0-n-octyl-P -D-glucopyranoside (octylglucoside) delayed, but did not prevent the spectral shifts from 657 to 646 nm and from 646 to 635 nm observed in non-cross-linked detergent- treated PLBs. The cross-linking did not prevent a spectral shift, corresponding to the Shibata shift, of Chlide. Thus the spectral shifts are not strictly coupled to disaggregation of the PCR polypeptides.
The aim of our work was to compare the effect of D-galactosamine (GalN) on primary cultures of lean and steatotic rat hepatocytes isolated from intact and fatty liver, respectively. GalN caused more severe injury to steatotic hepatocytes than to lean cells as documented by lactate dehydrogenase leakage. Necrotic mode of cell death strongly prevails over apoptosis since we did not observe any significant increase in activities of caspase 3, 8 and 9 in any group of hepatocytes treated with GalN. Reactive oxygen species (ROS) formation and lipid peroxidation were elevated in a dose-dependent manner by GalN and were significantly more pronounced in fatty hepatocytes. A decrease in the percentage of hepatocytes with energized mitochondria was observed from 30 mM and 10 mM GalN in lean and steatotic hepatocytes, respectively. Our results undoubtedly indicate that steatotic hepatocytes exert higher sensitivity to the toxic effect of GalN. This sensitivity may be caused by more intensive GalN-induced ROS production and lipid peroxidation and by higher susceptibility of mitochondria to loss of mitochondrial membrane potential in steatotic hepatocytes. In our experimental arrangement, apoptosis does not seem to participate considerably on hepatotoxic action of GalN in either group of hepatocytes., O. Kučera, H. Lotková, O. Sobotka Z. Červinková., and Obsahuje bibliografii
DPC played an important role in regulating the production, translocation and partítioning of i‘*C-assimilates in cotton {Gossypium hirsutum L.) plants. Seed soaking with DPC increased the partítioning of cotton assimilates into roots aitd main stem, and decreased the partítioning into seedling tip which was beneficial for the seedling. After the appearance of a square, spraying with DPC decreased the partítioning of assimilates into the main stem, branches and their growing points, and increased the partítioning into reproductíve organs and roots. This helped to avoid or reduce spindling, ensured a steady growth, coordination of the relatíon between vegetatíve and reproductíve organs, and improved the development of floral buds. From bloom to boll-setting,. sprayings with DPC greatly increased the partítioning of assimilates into reproductíve organs and decreased the partítioning into vegetatíve organs, which was usefiil for the growth and development of squares and bolls.
a1_Renal transplantation is associated with a large number of risk factors that can have an influe nce on early renal graft function (ERGF). One of these factors could be the increasing number of obese kidney donors. The mechanisms of reduced ERGF in obese kidney donors are still poorly understood. To that end, w e compared ERGF in recipients with body mass index (BMI), perivascular fat and plasma inflammation markers of live kidney donors. We hypothesi zed that the BMI of donors would negatively correlate with an average increase of glomerular filtration rate (GFR ) and that it would also be associated wi th increased perivascular and plasma inflammation markers in the first seven days after transplantation. Between January 2013 and December 2014, some 58 living kidney transplantation pairs were included in the study. Donor and recipient demographic data, preoperative BMI, blood C -reactive protein (CRP) and adiponectin levels, perivascular adipose tissue (PAT) samples and recipient blood creatinine levels were analy zed. The median CRP of donors was 0.68 mg/l (max: 8.66 mg/l, min: 0.33 mg/l), the median of M1 macrophages (CD14+CD16+) in one gram of PAT was 5940 (max: 41 100, min: 248) and the median of adiponectin was 411 930 pg/ml (max: 14 217 000, min: 167 300) in plasma. We did not find any association between early renal graft function and the percentage o f M1 macrophages in donor perirenal adipose tissue (p=0.83, r=0.03, n=58), adiponectin (p=0.65, r=0.06, n=58) or CRP (p=0.16, r=0.2, n=58) in plasma. The obesity level of donors, expressed as BMI, did not correlate with early renal graft function in the first seven days after transplantation. The associations between ERGF and plasma and perivascular fat inflammation markers were not significant., a2_We confirmed a negative correlation between the BMI of recipients and an average increase of GFR in the first sev en days after transplantation (p<0.02, r= -0.325, N=58). We confirmed a negative correlation of adiponectin plasma concentration to the BMI of donors., F. Thieme, L. Janousek, J. Fronek, A. Kralova, S. Cejkova, I. Kralova Lesna, R. Poledne., and Obsahuje bibliografii
The effect of enhanced air CO2 concentrations (C520 and 0^50 = 520 and 650 cm^ m"^) on the growth of Lamium galeobdolon and Stellaria holostea and on the competition between the two species was examined. After five months growth imder CO2 enrichment the dry masses of both species increased when the plants were grown in monoculture, but the increase in biomass was much more pronounced in Stellaria. When the plants were grown together in competition, the measured shoot masses of Stellaria were again higher under C520 and 0^50 than at ambient CO2 concentration (C390 = 390 cm^ m'^), while the shoot masses of Lamium strongly decreased at Cgso- The effect of CO2 enrichment on the two plant species in monoculture differed significantly from that observed in mixed cultures. In terms of plant relative yield, Stellaria benefitted slightly but insignificantly from competition, while Lamium was significantly suppressed imder c^sq. Total community production of the mixed culture was optimum at C520, while that of the monocultures was highest at c^sq. At C390 and C520, growth of Stellaria depended strongly on irradiance in all types of culture. At C650 no such dependence could be demonstrated.
The effect of exposure to miracidia of Schistosoma mansoni Sambon on mortality rates of five different age-groups of Biomphalarìa pfeifferi (Krauss) was investigated under carefully controlled conditions in the laboratory. The per capita mortality rates were determined for each age-group and the mean life expectancy and mean snail mortalities per week calculated. It was demonstrated that the cohorts of snails exposed to three miracidia per snail at an age of one week suffered the highest mortalities during the prepatent period and had the shortest life-span. No significant differences could be indicated in these respects between cohorts of snails exposed at an age of three, four and five weeks to the same number of miracidia. The results of this investigation could make a significant contribution towards establishing and maintaining laboratory cultures of S. mansoni.
In morphological and biochemical studies we demonstrated that the development of eight-day old dark-grown beán seedlings was drastically influenced by prolonged irradiation with far red (FR) irradiation. The synthetic processes associated with leaf expansion were fiilly active, but the accumulation of chJorophyll (Chl) was very slow and the development of the photosynthetic apparatus was much prolonged. The capacity for oxygen evolution and photophosphorylation began at about the 2''‘* day. Low temperature spectroscopy showed the accumulation of three Chl a forms with absorption maxima at 670, 677 and 683 nm from the veiy beginning of the greening process and the appearance of two longer wavelength forms (with maxima at 690 and 698 nm) at the onset of photosynthetic activity. Electron microscopy showed unfiised stacks of primary thylakoids without grana formation. When the FR treated seedlings were subsequently placed in "white light", grana differentiated from these primary thylakoids.
AIMS: Authors studied potential side effects of fetal calf serum (FCS) in cultivation media on human dental pulp stem cells (DPSC) during long-term cultivation. METHODS: Two lines of DPSC obtained healthy donors (male 22 years, female 23 years) were used. Both lines were cultivated under standard cultivation conditions in four different media containing 10% or 2% FCS and substituted with growth factors. During long-term cultivation proliferation ability, karyotype and phenotype of DPSC were measured. RESULTS: Both lines of DPSC cultivated in a media containing 2% FCS and ITS supplement showed the highest number of population doublings. On the other hand the proliferation rate of DPSC cultivated in a media with 2% FCS without ITS supplement was slowest. Proliferation rate of DPSC cultivated in 10% FCS media with or without FGF-2 was comparable. DPSC cultivated in a media with 10% FCS showed a significantly higher amount of chromosomal aberrations. These chromosomal aberrations do not seem to be clonal but surprisingly we found large amounts of tetraploid cells in the 9th passage in both media containing 10% FCS. CONCLUSIONS: Our study proved that cultivation of DPSC in media containing higher concentration of FCS has critical side effects on cell chromosomal stability. and J. Suchánek, TS. Kleplová, M. Kapitán, T. Soukup
Changes of chlorophyll (Chl) a fluorescence and photosynthetic pigment contents were analysed in galled leaves (visibly damaged and undamaged parts) and intact leaves. The values of minimal fluorescence of the dark-adapted state, maximal quantum yield of PSII photochemistry, effective quantum yield of PSII photochemical conversion, and photochemical quenching coefficient decreased in Ulmus pumila L. leaves galled by Tetraneura ulmi (L.) and in U. glabra Huds. galled by Eriosoma ulmi (L.). Colopha compressa (Koch.) feeding affected these parameters only in damaged parts of U. laevis Pall. galled leaves. The increasing number of T. ulmi galls progressively decreased photosynthetic performance. In gall tissues of all analysed aphid species, the lowest photosynthetic pigment content was found, indicating that the photosynthetic capacity must have been low in galls. Significant reduction of Chl and carotenoid contents were observed in damaged and undamaged portions of galled leaves only in the case of T. ulmi feeding., K. Kmieć, K. Rubinowska, W. Michałek, H. Sytykiewicz., and Obsahuje bibliografii
Increased and prolonged postprandial lipemia has been identified as a risk factor of cardiovascular disease. However, there is no consensus on how to test postprandial lipemia, especia lly with respect to the composition of an experimental meal. To address this question of how glucose, when added to a fat load, affects the selected parameters of postprandial lipemia, we carried out a study in 30 healthy male volunteers. Men consumed an experimental meal containing either 75 g of fat + 25 g of glucose (F+G meal) or 75 g of fat (F meal) in a control experiment. Blood was taken before the meal and at selected time points within the following 8 h. Glucose, when added to a fat load, induced an increase of glycemia and insulinemia and, surprisingly, a 20 % reduction in the response of both total and active glucagon -like peptide -1 (GLP -1) concentration. The addition of glucose did not affect the magnitude of postprandial triglyceridemia and TRL -C and TRL -TG concentrations but stimulated a faster response of chylomicrons to the test meal, evaluated by changes in apolipoprotein B -48 concentrations. The addition of glucose induced the physiological response of insulin and the lower response of GLP -1 to the test meal during the early postprandial phase, but had no effect on changes of TRL -cholesterol and TRL -TG within 8 h after the meal., K. Zemánková, J. Mrázková, J. Piťha, J. Kovář., and Obsahuje bibliografii