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51.
Candiano G Santucci L Bruschi M Petretto A D'Ambrosio C Scaloni A Righetti PG Ghiggeri GM 《Journal of Proteomics》2012,75(3):796-805
A new method is here reported for facile elution of the human urinary proteome after being captured with combinatorial peptide ligand libraries (CPLL, ProteoMiner). It consists in challenging the beads with 100 mM Tris, pH 7.4, or with 100 mM Lys, pH 7.4 or even better with a mixture of Lys, Arg, Asp and Glu (150 mM final concentration). These elutions permit recovery of species in a native form, for monitoring any biological activity of the eluted species, while avoiding the noxious presence of sodium dodecyl sulphate (SDS), reported as the best eluant so far from CPLL beads. SDS, albeit permitting quantitative recovery from the beads, has to be removed from the sample prior to mass spectrometry analysis. This unorthodox elution, which most likely will work only for urine samples, seems to be due to the fact that bile salts and urinary pigments are massively adsorbed by the beads, thus masking the hydrophobic binding sites of aromatic and non-aromatic amino acids. The binding thus occurs mostly via ionic and hydrogen bond interactions via the “Grand Catchers” Arg, Lys, His, which can then be easily challenged by positively charged species, such a Tris, free Lys and free Arg in the eluant as well as by negatively charged compounds, such as Glu and Asp. When eluting with the four-amino acid mix, at least 3300 spots can be visualized in a 2D map. 相似文献
52.
Background
Listeria monocytogenesis a well-characterized food-borne pathogen that infects pregnant women and immunocompromised individuals. Listeriolysin O (LLO) is the major virulence factor of the pathogen and is often used as a diagnostic marker for detection of L. monocytogenes. In addition, LLO represents a potent antigen driving T cell-mediated immunity during infection. In the present work, Lactococcus lactisNZ9000 was used as an expression host to hyper-produce LLO under inducible conditions using the NICE (NIsin Controlled Expression) system. We created a modified pNZ8048 vector encoding a six-His-tagged LLO downstream of the strong inducible PnisA promoter. 相似文献53.
Perissé P y GM Tourn 《Phyton》2015,84(1):184-189
The morphological characteristics of the seeds of Vicia dasycarpa Ten. cv. Tolse FCA were studied in relation to the area of imbibition water entry and were considered the presence of areoles. Seeds were analyzed using a stereo, optical and scanning electron microscope (SEM). The determination of the initial water entry area was carried out by immersing the seeds in a solution of tetrazolium (1%). This study showed that this species has seeds with a halo framing the hilum, an inconspicuous dry aril and a deltoid micropyle. The seedcoat pattern is papillose. The tracheid bar is surrounded by a ring of parenchymatous cells, and the tracheids show slight warty and non-vestured pits. It was confirmed the presence of an endospermic radicle pocket that surrounds and protect the radical tip. Two pairs of cotyledonar areoles were identified. It was established that the entry of water during imbibition starts in the area of the lens -having cracks- and moves in the sagittal plane. Both citological characteristic of tracheid bar and areoles presence show an apomorfous state between the Papiplionoids. 相似文献
54.
We developed a new and simple method to collect sections of a whole brown rice kernel for investigation of histological properties. A single kernel of rice was dehydrated through a graded ethanol series, transferred to xylene, and embedded in paraffin. During sectioning of the blocks using a rotary microtome, we used a special adhesive tape to collect and place the sections on slides so they remained flat. The use of the adhesive tape technique combined with autofluorescence characteristics allowed us to visualize cell walls throughout an entire longitudinal or transverse section of a whole rice kernel. We obtained scanning electron microscopy images of the sections to determine section quality. 相似文献
55.
Eliane R Popa Coen A Stegeman Cees GM Kallenberg Jan Willem Cohen Tervaert 《Arthritis research & therapy》2001,4(2):77-3
Wegener's granulomatosis (WG) is a form of systemic vasculitis. It is characterized by granulomatous inflammation in the upper and lower airways, vasculitis and necrotizing glomerulonephritis, and is strongly associated with antineutrophil cytoplasmic antibodies against proteinase 3. Since the etiology of the disease is not clear, treatment, consisting of corticosteroids and immunosuppressives, is nonspecific and associated with severe side effects. Pinpointing the trigger(s) of the disease would highly improve treatment. Clinical evidence shows that an infectious agent, the bacterium Staphylococcus aureus, is a risk factor for disease relapse, suggesting its involvement in the pathogenesis of WG. Here we review both clinical and experimental data that either indicate or support a role for S. aureus in WG. 相似文献
56.
Rick Brouwer Wilma TM Vree Egberts Gerald JD Hengstman Reinout Raijmakers Baziel GM van Engelen Hans Peter Seelig Manfred Renz Rudolf Mierau Ekkehard Genth Ger JM Pruijn Walther J van Venrooij 《Arthritis research & therapy》2001,4(2):1-5
The autoantigenic polymyositis/scleroderma (PM/Scl) complex was recently shown to be the human homologue of the yeast exosome, which is an RNA-processing complex. Our aim was to assess whether, in addition to targeting the known autoantigens PM/Scl-100 and PM/Scl-75, autoantibodies also target recently identified components of the PM/Scl complex. The prevalence of autoantibodies directed to six novel human exosome components (hRrp4p, hRrp40p, hRrp41p, hRrp42p, hRrp46p, hCsl4p) was determined in sera from patients with idiopathic inflammatory myopathy (n = 48), scleroderma (n = 11), or the PM/Scl overlap syndrome (n = 10). The sera were analyzed by enzyme-linked immunosorbent assays and western blotting using the affinity-purified recombinant proteins. Our results show that each human exosome component is recognized by autoantibodies. The hRrp4p and hRrp42p components were most frequently targeted. The presence of autoantibodies directed to the novel components of the human exosome was correlated with the presence of the anti-PM/Scl-100 autoantibody in the sera of patients with idiopathic inflammatory myopathy (IIM), as was previously found for the anti-PM/Scl-75 autoantibody. Other clear associations between autoantibody activities were not found. These results further support the conception that the autoimmune response may initially be directed to PM/Scl-100, whereas intermolecular epitope spreading may have caused the autoantibody response directed to the associated components. 相似文献
57.
Giovanni Candiano Rosanna Gusmano Paola Altieri Roberta Bertelli Fabrizio Ginevri Domenico A. Coviello Adalberto Sessa Gianluca Caridi Gian Marco Ghiggeri 《Virchows Archiv. B, Cell pathology including molecular pathology》1993,63(1):1-9
Cells from the cysts of patients with autosomal dominant polycystic kidney disease (PKD) were grown in vitro under standard
conditions without the aid of collagen-pretreated surfaces, and both the synthesis and composition of the extracellular matrix
were investigated. At confluence, PKD cells presented the typical features of epithelial cells, but showed a different collagen
composition from fibroblasts. Compared with normal tubular epithelia (NTE), PKD monolayers produced an excess of extracellular
matrix, which accounted for 30% of the total incorporation of [3H] proline, although this value was considerably lower (by a factor of 10) in the case of NTE. Immunohistochemical and electrophoretic
techniques revealed a complex collagen composition in the extracellular matrix which included [α(III)]3 and collagen IV. However, part of the collagen components remained unidentified in spite of the fact that they exhibited
a typical Mr of α1(I) and α2(I) in the presence of urea. Immunoprecipitation with monospecific antibodies and Northern blotting with specific probes failed
to recognize α1(I) and α2(I), but demonstrated their presence in fibroblasts. Purification and cyanogen bromide digestion demonstrated a strong interhomology
in fingerprint peptide composition among the uncharacterized collagens synthesized by PKD cells, thus suggesting a common
identity. These observations document a markedly augmented production of extracellular matrix by PKD cultured cells in vitro,
and show the presence of collagens which do not share homologies with the major collagen molecules. A better characterization
of extracellular matrix composition is central to any comprehension of the cystogenetic mechanisms in vivo. 相似文献
58.
GianMarco Ghiggeri Gianluca Caridi Paola Altieri Annalisa Pezzolo Giorgio Gimelli Orsetta Zuffardi 《Human genetics》1993,91(2):175-177
The COL5A1 gene, which encodes the pro 1(V) chain, was recently mapped to 9q34.3 in the same region as the nail-patella locus. This was taken as an indication that the nail-patella syndrome may be an inherited connective tissue disorder. We demonstrate COL5A1 heterozygous deletion and fibroblast under-expression of 1(V) chains in a girl with an unbalanced translocation resulting in 9q32qter monosomy. The patient presents dysplastic nails, a sign typical of nail-patella syndrome, but normal patella. Moreover, she has skin and bone disorders similar to those found in the Goltz syndrome. We suggest that monosomy for the COL5A1 gene is responsible for these connective tissue disorders. Accordingly, the nail-patella syndrome could be attributable to mutations inside the COL5A1 gene rather than to a deletion of it. 相似文献
59.
G. Caridi Annalisa Pezzolo Roberta Bertelli G. Gimelli A. DiDonato G. Candiano G. M. Ghiggeri 《Human genetics》1992,90(1-2):174-176
Summary A 353-bp region encoding for the NH2 terminus of the noncollagenic part of the l(V) chain was amplified by the polymerase chain reaction (PCR), subcloned and sequenced. The subcloned PCR product (pGC1) presented the same nucleotide sequence as the original fragment from the published sequence of COL5A1. In situ hybridization, using pGC1 as a probe, mapped the COL5A1 gene to chromosome 9q34.3. This assignement shows that COL5A1 is not synthenic with COL5A2, which is localized together with other collagen genes on chromosome 2. 相似文献
60.
Polymorphism and divergence at the prune locus in Drosophila melanogaster and D. simulans 总被引:3,自引:0,他引:3
The prune locus of Drosophila melanogaster lies at the tip of the X
chromosome, in a region of reduced recombination in which nearby loci show
reduced variation relative to evolutionary divergence from D. simulans. DNA
sequencing of prune alleles from D. melanogaster and D. simulans reveals
extremely low variation in D. melanogaster but greater variation in D.
simulans. Divergence between the two species is not reduced. This pattern
may be explained by either positive selection leading to hitchhiking of
neutral variation or background selection against deleterious mutations.
The pattern of silent versus replacement polymorphism and divergence at
prune is consistent with either a model of weakly deleterious selection
against amino acid substitutions or balancing selection.
相似文献