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991.
Background
Serological surveys for disease investigation of wild animal populations require obtaining blood samples for analysis, which has logistic, ethic and economic difficulties. Applying serological test to fluids collected from dead animals is an alternative. The aim of this study was to assess if antibodies could be detected in two types of fluids collected from 56 carcasses of red foxes (Vulpes vulpes): pleural fluid and lung extract.Findings
In 22 (39%) foxes antibodies against Sarcoptes scabiei were detected in both fluid types by ELISA and Western blot. In 46 (82%) foxes, antibodies against Toxoplasma gondii were detected in pleural fluid and in 41 (73%) in lung extract applying a Toxo-screen test (DAT). Antibodies were still detectable in the same fluids kept at room temperature for 28 days, although in fewer foxes (16 and 14 foxes tested for T. gondii in lung extract and pleural fluid respectively; and 1 and 4 tested for S. scabiei in lung extract and pleural fluid respectively.Conclusions
These results indicate the potential utility of using fluids from carcasses for antibody screening of wild animals at the population level. 相似文献992.
Assessing petroleum biodegradation rates is an important part of predicting natural attenuation in subsurface sediments. Monitoring carbon dioxide (CO2) and methane (CH4) produced in situ, and their radiocarbon 14C), stable carbon (13C) and deuterium (D). signature provide a novel method to assess anaerobic microbial processes. Our objectives were to: (1) estimate the rate of anaerobic petroleum hydrocarbon (PH) mineralization by monitoring the production of soil gas CH4 and CO2 in the vadose zone of low-permeability sediment, (2) evaluate the dominant microbial processes using δ13C and δD, and (3) determine the proportion of CH4 and CO2 attributable to anaerobic mineralization of PH using 14C analysis. Argon was sparged into the subsurface to dilute existing CO2 and CH4 concentrations. Vadose zone CO2, CH4, oxygen, total combustible hydrocarbons, and argon concentrations were measured for 75 days. CO2 and CH4 samples were collected on day 86 and analyzed for 14C, δ13C, and δD. Based on CH4 soil gas production, the anaerobic biodegradation rate was estimated between 0.017 to 0.055 mg/kg soil-d. CH4 14C (2.6 pMC), δ13C (-45.64‰), and δD (-316‰) values indicated that fermentation of PH was the sale source of CH4 in the vadose zone. CO2 14C (62 pMC) indicated that approximately 47% of the total CO2 was from PH mineralization and 53% from plant root respiration. Although low-permeability sediment increases the difficulty of completely replacing in situ soil gas and assuring anaerobic conditions, this novel respiration method distinguished between anaerobic processes responsible for PH degradation. 相似文献
993.
We used the natural abundance of 15N in soils in forests, pastures and cultivated lands in the Menagesha and Wendo-Genet areas of Ethiopia to make inferences
about the N cycles in these ecosystems. Since we have described the history of these sites based on variations in 13C natural abundance, patterns of δ15N and δ13C values were compared to determine if shifts of 15N correlate with shifts of vegetation. At Menagesha, a > 500-yr-old planted forest, we found δ15N values from −8.8 to +3.5‰ in litter, from −3.5 to +4.5‰ in 0–10 cm soil layer, and from −1.5 to +6.8‰ at >20 cm soil depth.
The low δ15N in litter and surface mineral soils suggests that a closed N cycle has operated for a long time. At this site, the low δ13C of the surface horizon and the high δ13C of the lower soil horizons is clear evidence of a long phase of C4 grass dominance or cultivation of C4 crops before the establishment of the forest >500 years ago. In contrast, at Wendo-Genet, high δ13C of soils reveals that most of the land has been uncovered by forests until recently. Soil δ15N was high throughout (3.4–9.8‰), and there were no major differences between forested, cultivated and pasture soils in δ15N values of surface mineral soils. The high δ15N values suggest that open N cycles operate in the Wendo-Genet area. From the points of view of soil fertility management,
it is interesting that tall forest ecosystems with relatively closed N cycling could be established on the fairly steep slopes
at Menagesha after a long period of grass vegetation cover or cultivation.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
994.
Claire J. Sarell Theodoros K. Karamanos Simon J. White David H. J. Bunka Arnout P. Kalverda Gary S. Thompson Amy M. Barker Peter G. Stockley Sheena E. Radford 《The Journal of biological chemistry》2014,289(39):26859-26871
Although amyloid fibrils assembled in vitro commonly involve a single protein, fibrils formed in vivo can contain multiple protein sequences. The amyloidogenic protein human β2-microglobulin (hβ2m) can co-polymerize with its N-terminally truncated variant (ΔN6) in vitro to form hetero-polymeric fibrils that differ from their homo-polymeric counterparts. Discrimination between the different assembly precursors, for example by binding of a biomolecule to one species in a mixture of conformers, offers an opportunity to alter the course of co-assembly and the properties of the fibrils formed. Here, using hβ2m and its amyloidogenic counterpart, ΔΝ6, we describe selection of a 2′F-modified RNA aptamer able to distinguish between these very similar proteins. SELEX with a N30 RNA pool yielded an aptamer (B6) that binds hβ2m with an EC50 of ∼200 nm. NMR spectroscopy was used to assign the 1H-15N HSQC spectrum of the B6-hβ2m complex, revealing that the aptamer binds to the face of hβ2m containing the A, B, E, and D β-strands. In contrast, binding of B6 to ΔN6 is weak and less specific. Kinetic analysis of the effect of B6 on co-polymerization of hβ2m and ΔN6 revealed that the aptamer alters the kinetics of co-polymerization of the two proteins. The results reveal the potential of RNA aptamers as tools for elucidating the mechanisms of co-assembly in amyloid formation and as reagents able to discriminate between very similar protein conformers with different amyloid propensity. 相似文献
995.
Sangita Choudhury Soochan Bae Qingen Ke Ji Yoo Lee Sylvia S. Singh René St-Arnaud Federica del Monte Peter M. Kang 《PloS one》2014,9(9)
Aim
Altered vitamin D signaling is associated with cardiac dysfunction, but the pathogenic mechanism is not clearly understood. We examine the mechanism and the role of vitamin D signaling in the development of cardiac dysfunction.Methods and Results
We analyzed 1α-hydroxylase (1α-OHase) knockout (1α-OHase−/−) mice, which lack 1α-OH enzymes that convert the inactive form to hormonally active form of vitamin D. 1α-OHase−/− mice showed modest cardiac hypertrophy at baseline. Induction of pressure overload by transverse aortic constriction (TAC) demonstrated exaggerated cardiac dysfunction in 1α-OHase−/− mice compared to their WT littermates with a significant increase in fibrosis and expression of inflammatory cytokines. Analysis of calcium (Ca2+) transient demonstrated profound Ca2+ handling abnormalities in 1α-OHase−/− mouse cardiomyocytes (CMs), and treatment with paricalcitol (PC), an activated vitamin D3 analog, significantly attenuated defective Ca2+ handling in 1α-OHase−/− CMs. We further delineated the effect of vitamin D deficiency condition to TAC by first correcting the vitamin D deficiency in 1α-OHase−/− mice, followed then by either a daily maintenance dose of vitamin D or vehicle (to achieve vitamin D deficiency) at the time of sham or TAC. In mice treated with vitamin D, there was a significant attenuation of TAC-induced cardiac hypertrophy, interstitial fibrosis, inflammatory markers, Ca2+ handling abnormalities and cardiac function compared to the vehicle treated animals.Conclusions
Our results provide insight into the mechanism of cardiac dysfunction, which is associated with severely defective Ca2+ handling and defective vitamin D signaling in 1α-OHase−/− mice. 相似文献996.
Raquel Ordó?ez Gabriel Gallo-Oller Soledad Martínez-Soto Sheila Legarra Noémie Pata-Merci Justine Guegan Giselle Danglot Alain Bernheim Bárbara Meléndez Juan A. Rey Javier S. Castresana 《PloS one》2014,9(11)
Neuroblastoma has a very diverse clinical behaviour: from spontaneous regression to a very aggressive malignant progression and resistance to chemotherapy. This heterogeneous clinical behaviour might be due to the existence of Cancer Stem Cells (CSC), a subpopulation within the tumor with stem-like cell properties: a significant proliferation capacity, a unique self-renewal capacity, and therefore, a higher ability to form new tumors. We enriched the CSC-like cell population content of two commercial neuroblastoma cell lines by the use of conditioned cell culture media for neurospheres, and compared genomic gains and losses and genome expression by array-CGH and microarray analysis, respectively (in CSC-like versus standard tumor cells culture). Despite the array-CGH did not show significant differences between standard and CSC-like in both analyzed cell lines, the microarray expression analysis highlighted some of the most relevant biological processes and molecular functions that might be responsible for the CSC-like phenotype. Some signalling pathways detected seem to be involved in self-renewal of normal tissues (Wnt, Notch, Hh and TGF-β) and contribute to CSC phenotype. We focused on the aberrant activation of TGF-β and Hh signalling pathways, confirming the inhibition of repressors of TGF-β pathway, as SMAD6 and SMAD7 by RT-qPCR. The analysis of the Sonic Hedgehog pathway showed overexpression of PTCH1, GLI1 and SMO. We found overexpression of CD133 and CD15 in SIMA neurospheres, confirming that this cell line was particularly enriched in stem-like cells. This work shows a cross-talk among different pathways in neuroblastoma and its importance in CSC-like cells. 相似文献
997.
Joan Gannon Julia Fernandez-Rodriguez Hussam Alamri Shi Bo Feng Fariba Kalantari Sarita Negi Amy H. Y. Wong Alexander Mazur Lennart Asp Ali Fazel Ayat Salman Anthoula Lazaris Peter Metrakos John J. M. Bergeron Tommy Nilsson 《PloS one》2014,9(11)
The ARF GTPase Activating Protein 1 (ARFGAP1) associates mainly with the cytosolic side of Golgi cisternal membranes where it participates in the formation of both COPI and clathrin-coated vesicles. In this study, we show that ARFGAP1 associates transiently with lipid droplets upon addition of oleate in cultured cells. Also, that addition of cyclic AMP shifts ARFGAP1 from lipid droplets to the Golgi apparatus and that overexpression and knockdown of ARFGAP1 affect lipid droplet formation. Examination of human liver tissue reveals that ARFGAP1 is found associated with lipid droplets at steady state in some but not all hepatocytes. 相似文献
998.
Targeted inactivation of the plastid ndhB gene in tobacco results in an enhanced sensitivity of photosynthesis to moderate stomatal closure 总被引:12,自引:0,他引:12
Horváth EM Peter SO Joët T Rumeau D Cournac L Horváth GV Kavanagh TA Schäfer C Peltier G Medgyesy P 《Plant physiology》2000,123(4):1337-1350
The ndh genes encoding for the subunits of NAD(P)H dehydrogenase complex represent the largest family of plastid genes without a clearly defined function. Tobacco (Nicotiana tabacum) plastid transformants were produced in which the ndhB gene was inactivated by replacing it with a mutant version possessing translational stops in the coding region. Western-blot analysis indicated that no functional NAD(P)H dehydrogenase complex can be assembled in the plastid transformants. Chlorophyll fluorescence measurements showed that dark reduction of the plastoquinone pool by stromal reductants was impaired in ndhB-inactivated plants. Both the phenotype and photosynthetic performance of the plastid transformants was completely normal under favorable conditions. However, an enhanced growth retardation of ndhB-inactivated plants was revealed under humidity stress conditions causing a moderate decline in photosynthesis via stomatal closure. This distinctive phenotype was mimicked under normal humidity by spraying plants with abscisic acid. Measurements of CO(2) fixation demonstrated an enhanced decline in photosynthesis in the mutant plants under humidity stress, which could be restored to wild-type levels by elevating the external CO(2) concentration. These results suggest that the plastid NAD(P)H:plastoquinone oxidoreductase in tobacco performs a significant physiological role by facilitating photosynthesis at moderate CO(2) limitation. 相似文献
999.
Hu B Finsinger D Peter K Guttenberg Z Bärmann M Kessler H Escherich A Moroder L Böhm J Baumeister W Sui SF Sackmann E 《Biochemistry》2000,39(40):12284-12294
We report the synthesis of a new integrin alpha(IIb)beta(3)-specific cyclic hexapeptide that contains an Arg-Gly-Asp (RGD) sequence and is coupled to a dimyristoylthioglyceryl anchor. We demonstrate that this ligand is useful to study specific integrin binding to membrane surfaces. With the help of biotinylated analogues of the peptide, a spacer of optimal length between the peptide and lipid moieties was searched for by evaluating the binding strength with an enzyme-coupled immunosorbent assay (ELISA) and by surface plasmon resonance (SPR). It was found to be strongly dependent on the length of the spacer introduced between the biotin and peptide moieties of the ligands, which consisted either of epsilon-aminohexanoic acid (epsilonAhx) or of epsilonAhx with two additional glycine units. Best results were obtained with c[Arg-Gly-Asp-D-Phe-Lys(Biot-Ahx-Gly-Gly)-Gly-] with dissociation constants of K(D) = 0.158 microM from ELISA and K(D) = 1.1 microM from SPR measurements. The analogous lipopeptide, c[Arg-Gly-Asp-D-Phe-Lys([dimyristoyl-3-thioglyceryl-succinimido -propanoyl]Ahx-Gly-Gly)-Gly], was used as a membrane-anchored integrin ligand. It is shown by fluorescence microscopy and cryo electron microscopy that integrin reconstituted into phospholipid vesicles binds to vesicles decorated with the lipopeptide, forming regularly spaced bridges between the two kinds of vesicles. The novel integrin-specific ligand allows establishment of new model systems for systematic studies of the self-organization of integrin clusters and focal adhesion complexes. 相似文献
1000.