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Studies of esterase 6 in Drosophila melanogaster. XIII. Purification and characterization of the two major isozymes 总被引:4,自引:0,他引:4
Esterase-6 (EST 6; carboxylic-ester hydrolase; EC 3.1.1.1) from Drosophila melanogaster was purified to homogenity. Purified enzyme occurs as two closely moving isozymes, slow (EST 6S) and fast (EST 6F), on native polyacrylamide gel electrophoresis. Except for slight differences in their mobility, the two isozymes share similar molecular and catalytic properties. Both isozymes are glycoproteins and have an apparent molecular weight of 62,000 to 65,000 as judged by analytical gel filtration and sodium dodecyl sulfate (SDS) electrophoresis. They have identical mobility on SDS-polyacrylamide gels and an isoelectric point of 4.5. Each isozyme has a single active catalytic site as confirmed by titration with 0,0-diethyl-p-nitrophenyl phosphate (Paraoxon). We conclude that EST 6 is a monomeric enzyme. The amino acid composition of the two isozymes is very similar and both variants lack half-cystine residues. The low pI of the enzyme is due in part to a relatively high proportion of glutamic and aspartic amino acid residues. Characterization of the kinetic parameters of the isozymes using beta-naphthyl and p-nitrophenyl esters revealed no statistically significant differences in catalytic efficiency. There is, however, a suggestion that the two isozymes may differ in their substrate specificity. 相似文献
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Urinary proteome alterations in HER2 enriched breast cancer revealed by multipronged quantitative proteomics
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Khushman Taunk Garikapati Vannuruswamy Sourav RoyChoudhury Ragini Adhav Shubhendu Seal Anupama Mane Santhakumari Bayatigeri Manas K. Santra Koel Chaudhury Srikanth Rapole 《Proteomics》2016,16(17):2403-2418
Globally, breast cancer is the second most common cancer among women. Although biomarker discoveries through various proteomic approaches of tissue and serum samples have been studied in breast cancer, urinary proteome alterations in breast cancer are least studied. Urine being a noninvasive biofluid and a significant source of proteins, it has the potential in early diagnosis of breast cancer. This study used complementary quantitative gel‐based and gel‐free proteomic approaches to find a panel of urinary protein markers that could discriminate HER2 enriched (HE) subtype breast cancer from the healthy controls. A total of 183 differentially expressed proteins were identified using three complementary approaches, namely 2D‐DIGE, iTRAQ, and sequential window acquisition of all theoretical mass spectra. The differentially expressed proteins were subjected to various bioinformatics analyses for deciphering the biological context of these proteins using protein analysis through evolutionary relationships, database for annotation, visualization and integrated discovery, and STRING. Multivariate statistical analysis was undertaken to identify the set of most significant proteins, which could discriminate HE breast cancer from healthy controls. Immunoblotting and MRM‐based validation in a separate cohort testified a panel of 21 proteins such as zinc‐alpha2‐glycoprotein, A2GL, retinol‐binding protein 4, annexin A1, SAP3, SRC8, gelsolin, kininogen 1, CO9, clusterin, ceruloplasmin, and α1‐antitrypsin could be a panel of candidate markers that could discriminate HE breast cancer from healthy controls. 相似文献
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Na Li Lakshman Subrahmanyan Emily Smith Xiaoqing Yu Samir Zaidi Murim Choi Shrikant Mane Carol Nelson-Williams Mohaddeseh Behjati Mohammad Kazemi Mohammad Hashemi Mohsen Fathzadeh Anand Narayanan Likun Tian Farhad Montazeri Mitra Mani Michael L. Begleiter Brian G. Coon Henry T. Lynch Eric N. Olson Hongyu Zhao Jürgen Ruland Richard P. Lifton Arya Mani 《American journal of human genetics》2016,98(6):1082-1091
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Bingbing Nie Matthew Brian Panzer Adwait Mane Alexander Ritz Mait John-Paul Donlon Jason Lee Forman 《Computer methods in biomechanics and biomedical engineering》2016,19(12):1254-1265
Ligament sprains account for a majority of injuries to the foot and ankle complex, but ligament properties have not been understood well due to the difficulties in replicating the complex geometry, in situ stress state, and non-uniformity of the strain. For a full investigation of the injury mechanism, it is essential to build up a foot and ankle model validated at the level of bony kinematics and ligament properties. This study developed a framework to parameterize the ligament response for determining the in situ stress state and heterogeneous force–elongation characteristics using a finite element ankle model. Nine major ankle ligaments and the interosseous membrane were modeled as discrete elements corresponding functionally to the ligamentous microstructure of collagen fibers and having parameterized toe region and stiffness at the fiber level. The range of the design variables in the ligament model was determined from existing experimental data. Sensitivity of the bony kinematics to each variable was investigated by design of experiment. The results highlighted the critical role of the length of the toe region of the ligamentous fibers on the bony kinematics with the cumulative influence of more than 95%, while the fiber stiffness was statistically insignificant with an influence of less than 1% under the given variable range and loading conditions. With the flexibility of variable adjustment and high computational efficiency, the presented ankle model was generic in nature so as to maximize its applicability to capture the individual ligament behaviors in future studies. 相似文献
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Peccoud J Blauvelt MF Cai Y Cooper KL Crasta O DeLalla EC Evans C Folkerts O Lyons BM Mane SP Shelton R Sweede MA Waldon SA 《PloS one》2008,3(7):e2671
Background
The design and construction of novel biological systems by combining basic building blocks represents a dominant paradigm in synthetic biology. Creating and maintaining a database of these building blocks is a way to streamline the fabrication of complex constructs. The Registry of Standard Biological Parts (Registry) is the most advanced implementation of this idea.Methods/Principal Findings
By analyzing inclusion relationships between the sequences of the Registry entries, we build a network that can be related to the Registry abstraction hierarchy. The distribution of entry reuse and complexity was extracted from this network. The collection of clones associated with the database entries was also analyzed. The plasmid inserts were amplified and sequenced. The sequences of 162 inserts could be confirmed experimentally but unexpected discrepancies have also been identified.Conclusions/Significance
Organizational guidelines are proposed to help design and manage this new type of scientific resources. In particular, it appears necessary to compare the cost of ensuring the integrity of database entries and associated biological samples with their value to the users. The initial strategy that permits including any combination of parts irrespective of its potential value leads to an exponential and economically unsustainable growth that may be detrimental to the quality and long-term value of the resource to its users. 相似文献119.
Esophageal cancer involves multiple genetic alternations. A systematic codon usage bias analysis was completed to investigate the bias among the esophageal cancer responsive genes. GC-rich genes were low (average effective number of codon value was 49.28). CAG and GTA are over-represented and under-represented codons, respectively. Correspondence analysis, neutrality plot, and parity rule 2 plot analysis confirmed the dominance over mutation pressure in modulating the codon usage pattern of genes linked with esophageal cancer. 相似文献
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