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51.
Veronique Blanchard Rita Raisman-Vozari Marc Savasta Etienne Hirsch France Javoy-Agid Claude Feuerstein Yves Agid 《Journal of neurochemistry》1993,61(2):617-626
Abstract: We developed a rapid and sensitive radioimmunohistochemical method for the quantification of tyrosine hydroxylase (TH) at both the anatomical and cellular level. Coronal tissue sections from fresh-frozen rat brains were incubated in the presence of a TH monoclonal antibody. The reaction was revealed with a 35 S-labeled secondary antibody. TH content was quantified in catecholaminergic brain areas by measuring optical density on autoradiographic films or silver grain density on autoradiographic emulsion-coated sections. Regional TH concentrations determined in the locus ceruleus (LC), substantia nigra pars compacta (SNC), and ventral tegmental area (VTA) were significantly increased by 45% after reserpine treatment in the LC but unchanged in the SNC and VTA. Microscopic examination of TH radioimmunolabeling showed a heavy accumulation of silver grains over catecholaminergic cell bodies. In the LC, grain density per cell was heterogeneous and higher in the ventral than in the dorsal part of the structure. After reserpine treatment, TH levels were significantly increased (57%) in the neurons of the LC but not in those of the SNC or VTA. The data support the validity of this radioimmunohistochemical method as a tool for quantifying TH protein at the cellular level and they confirm that TH protein content is differentially regulated in noradrenergic and dopaminergic neurons in response to reserpine. 相似文献
52.
Chesapeake Bay was investigated as a source of actinomycetes to screen for production of novel bioactive compounds. The presence of relatively large populations of actinoplanetes (chemotype II/D actinomycetes) in Chesapeake Bay sediment samples indicates that it is an eminently suitable ecosystem from which to isolate actinomycetes for screening programs. Actinomycetes were isolated from sediment samples collected in Chesapeake Bay with an isolation medium containing nalidixic acid, which proved to be more effective than heat pretreatment of samples. Actinomycete counts ranged from a high of 1.4 × 105 to a low of 1.8 × 102 CFU/ml of sediment. Actinomycetes constituted 0.15 to 8.63% of the culturable microbial community. The majority of isolates from the eight stations studied were actinoplanetes (i.e., chemotype II/D), and 249 of these isolates were obtained in a total of 298 actinomycete isolates. Antimicrobial activity profiles indicated that diverse populations of actinoplanetes were present at each station. DNA hybridization studies showed considerable diversity among isolates between stations, but indicated that actinoplanete strains making up populations at nearby stations were more similar to each other than to populations sampled at distant stations. The diversity of actinoplanetes and the ease with which these organisms were isolated from Chesapeake Bay sediments make this a useful source of these actinomycetes. 相似文献
53.
Neuza Domingues André R. A. Marques Rita Diogo Almeida Calado Inês S. Ferreira Cristiano Ramos José Ramalho Maria I. L. Soares Telmo Pereira Luís Oliveira José R. Vicente Louise H. Wong Inês C. M. Simões Teresa M. V. D. Pinho e Melo Andrew Peden Cláudia Guimas Almeida Clare E. Futter Rosa Puertollano Winchil L. C. Vaz Otília V. Vieira 《Traffic (Copenhagen, Denmark)》2023,24(7):284-307
54.
Summary Previous studies have suggested that gap junctions may have a role in various uterine functions, including parturition. Because
nickel has been demonstrated to increase uterine contractility in vitro, the effect of nickel (II) chloride on gap junctional
communication was assessed in a tumorigenic uterine cell line, SK-UT-1 (ATCC HTB 114). Cells were exposed in vitro to 25 and
50 μM NiCl2 for 24 h or 100 μM NiCl2 for 3, 12, and 24 h, then functional gap junctional communication was measured as the transfer of Lucifer yellow dye from
microinjected donor cells to their primary neighbor cells. Dye transfer was significantly increased only in cell cultures
exposed to 100 μM NiCl2 for 24h, compared to untreated controls, lower doses, and shorter exposure periods. This response was inhibited by the simultaneous
co-treatment of SK-UT-1 cells with magnesium by adding 100 μM MgSO4 to the dosing medium. Possible mechanisms and implications for these findings are discussed. 相似文献
55.
Sven Göbel Karim E. Jaén Rita P. Fernandes Manfred Reiter Jennifer Altomonte Udo Reichl Yvonne Genzel 《Biotechnology and bioengineering》2023,120(11):3335-3346
The development of efficient processes for the production of oncolytic viruses (OV) plays a crucial role regarding the clinical success of virotherapy. Although many different OV platforms are currently under investigation, manufacturing of such viruses still mainly relies on static adherent cell cultures, which bear many challenges, particularly for fusogenic OVs. Availability of GMP-compliant continuous cell lines is limited, further complicating the development of commercially viable products. BHK21, AGE1. CR and HEK293 cells were previously identified as possible cell substrates for the recombinant vesicular stomatitis virus (rVSV)-based fusogenic OV, rVSV-NDV. Now, another promising cell substrate was identified, the CCX.E10 cell line, developed by Nuvonis Technologies. This suspension cell line is considered non-GMO as no foreign genes or viral sequences were used for its development. The CCX.E10 cells were thus thoroughly investigated as a potential candidate for OV production. Cell growth in the chemically defined medium in suspension resulted in concentrations up to 8.9 × 106 cells/mL with a doubling time of 26.6 h in batch mode. Cultivation and production of rVSV-NDV, was demonstrated successfully for various cultivation systems (ambr15, shake flask, stirred tank reactor, and orbitally shaken bioreactor) at vessel scales ranging from 15 mL to 10 L. High infectious virus titers of up to 4.2 × 108 TCID50/mL were reached in orbitally shaken bioreactors and stirred tank reactors in batch mode, respectively. Our results suggest that CCX.E10 cells are a very promising option for industrial production of OVs, particularly for fusogenic VSV-based constructs. 相似文献
56.
Joana Pais De Faria Vitor H. Paiva Sara N. Veríssimo Catarina S. Lopes Rita Soares João Oliveira Ivo dos Santos Ana C. Norte Jaime A. Ramos 《Ibis》2023,165(1):312-321
Gulls, as largely flexible opportunistic individuals, have been increasingly breeding in many cities around the world, but it is still unclear whether urban habitats are of equal or higher quality than traditional natural habitats or represent an ecological trap with immediate reproductive benefits but longer-term detrimental consequences to health. Here we present a study of breeding parameters (nest density, egg dimensions, clutch size, hatching success and adult body condition) and physiological parameters (erythrocyte sedimentation rate, heterophil/lymphocyte ratio, haemoglobin concentration and measurements of oxidative stress) as indicators of the general health condition of Yellow-legged Gull Larus michahellis adults and chicks from natural and urban colonies. Yellow-legged Gulls in the largest urban area (Porto) laid smaller eggs and clutches, showed a significantly lower occurrence of inflammatory processes in chicks, and showed a slower early chick growth than in the natural colony of Deserta. This suggests that urban gulls might be facing important trade-offs between the advantages of breeding in lower density urban colonies, with fewer intraspecific interactions and a lower disease transmission probability, and the disadvantages of having an anthropogenic diet usually lower in nutritional value. 相似文献
57.
Pedro Lamosa David L Turner Rita Ventura Christopher Maycock Helena Santos 《European journal of biochemistry》2003,270(23):4606-4614
Heteronuclear NMR relaxation measurements and hydrogen exchange data have been used to characterize protein dynamics in the presence or absence of stabilizing solutes from hyperthermophiles. Rubredoxin from Desulfovibrio gigas was selected as a model protein and the effect of diglycerol phosphate on its dynamic behaviour was studied. The presence of 100 mM diglycerol phosphate induces a fourfold increase in the half-life for thermal denaturation of D. gigas rubredoxin. A model-free analysis of the protein backbone relaxation parameters shows an average increase of generalized order parameters of 0.015 reflecting a small overall reduction in mobility of fast-scale motions. Hydrogen exchange data acquired over a temperature span of 20 degrees C yielded thermodynamic parameters for the structural opening reactions that allow for the exchange. This shows that the closed form of the protein is stabilized by an additional 1.6 kJ x mol(-1) in the presence of the solute. The results seem to indicate that the stabilizing effect is due mainly to a reduction in mobility of the slower, larger-scale motions within the protein structure with an associated increase in the enthalpy of interactions. 相似文献
58.
Ilana Agmon Tamar Auerbach David Baram Heike Bartels Anat Bashan Rita Berisio Paola Fucini Harly A S Hansen Joerg Harms Maggie Kessler Moshe Peretz Frank Schluenzen Ada Yonath Raz Zarivach 《European journal of biochemistry》2003,270(12):2543-2556
High-resolution crystal structures of large ribosomal subunits from Deinococcus radiodurans complexed with tRNA-mimics indicate that precise substrate positioning, mandatory for efficient protein biosynthesis with no further conformational rearrangements, is governed by remote interactions of the tRNA helical features. Based on the peptidyl transferase center (PTC) architecture, on the placement of tRNA mimics, and on the existence of a two-fold related region consisting of about 180 nucleotides of the 23S RNA, we proposed a unified mechanism integrating peptide bond formation, A-to-P site translocation, and the entrance of the nascent protein into its exit tunnel. This mechanism implies sovereign, albeit correlated, motions of the tRNA termini and includes a spiral rotation of the A-site tRNA-3' end around a local two-fold rotation axis, identified within the PTC. PTC features, ensuring the precise orientation required for the A-site nucleophilic attack on the P-site carbonyl-carbon, guide these motions. Solvent mediated hydrogen transfer appears to facilitate peptide bond formation in conjunction with the spiral rotation. The detection of similar two-fold symmetry-related regions in all known structures of the large ribosomal subunit, indicate the universality of this mechanism, and emphasizes the significance of the ribosomal template for the precise alignment of the substrates as well as for accurate and efficient translocation. The symmetry-related region may also be involved in regulatory tasks, such as signal transmission between the ribosomal features facilitating the entrance and the release of the tRNA molecules. The protein exit tunnel is an additional feature that has a role in cellular regulation. We showed by crystallographic methods that this tunnel is capable of undergoing conformational oscillations and correlated the tunnel mobility with sequence discrimination, gating and intracellular regulation. 相似文献
59.
The membrane-bound F1 sector of the H+–ATPase complex (F-type ATPase) in dark-adapted photosynthetic chromatophores is endowed with MgATP- and CaATP-dependent ATPase
activities, both sensitive to inhibitors such as oligomycin and venturicidin. Because of contatamination of free Mg2
+ and Ca2+ ions in chromatophore preparations, kinetic characterization of the two hydrolitic reactions can be performed only in the
presence of both substrates, using a model for two alternative substrates. The two activities are characterized by similar
maximal rates and affinity constants [VMgATP and VCaATP: 13±1 and 10±1 nmol s–1 ATP hydrolyzed (μmol BChl)–1; KMgATP and KCaATP: 0.22±0.06 and 0.20±0.05 mm]. However, only the MgATP-dependent ATPase is coupled to Δ*H
+ generation. In this process CaATP acts as an alternative substrate and a competitive inhibitor of the proton pump, with a
KI coincident with KCaATP for the hydrolytic activity. This finding highlights the central role that the coordination chemistry of the ion-nucleotide
complex plays in determining the proton gating mechanism at the catalytic site(s) of the enzyme complex. These results are
discussed on the basis of the coordination properties of the ions and of the available information on the protein structure.
Received: 5 December 1995 / Accepted: 7 March 1996 相似文献
60.
Nery N. Lima Carem G.V. Rechia Joana Lea M.S. Ganter Fany Reicher Maria Rita Sierakowski 《International journal of biological macromolecules》1995,17(6):413-415
On aqueous extraction, Hymenaea courbaril var. stilbocarpa, known in Brazil as jatobá, furnishes a high yield of viscous xyloglucan (45%) from its seeds. The crude polysaccharide (B1) was hydrolysed and the products, analysed as alditol acetates, were glucose, xylose, galactose and arabinose in the ratio 50:35:13:2. After further fractionation on a DEAE-cellulose column (chloride form), the main fraction (70% yield, B2) was obtained. The basic structure of the xyloglucan was determined as a cellulose-type (1 → 4)-linked β-d-glucan backbone partially substituted with side chains at 06 of -d-xylopyranose, some of which were themselves substituted at 02 by the units of β-d-galactopyranose. Treatment of the xyloglucan (B2) with commercial cellulase from Trichoderma sp. yielded six oligosaccharides. These oligosaccharides were isolated by preparative paper chromatography, and their structures were determined by gas-liquid chromatography-mass spectroscopy of the derived partially O-methylated alditol acetates. These results confirm the structure proposed for jatobá seed xyloglucan. 相似文献