全文获取类型
收费全文 | 884篇 |
免费 | 46篇 |
国内免费 | 4篇 |
出版年
2023年 | 1篇 |
2022年 | 7篇 |
2021年 | 7篇 |
2020年 | 3篇 |
2019年 | 10篇 |
2018年 | 10篇 |
2017年 | 17篇 |
2016年 | 17篇 |
2015年 | 18篇 |
2014年 | 26篇 |
2013年 | 60篇 |
2012年 | 44篇 |
2011年 | 74篇 |
2010年 | 37篇 |
2009年 | 36篇 |
2008年 | 64篇 |
2007年 | 66篇 |
2006年 | 54篇 |
2005年 | 64篇 |
2004年 | 49篇 |
2003年 | 53篇 |
2002年 | 46篇 |
2001年 | 6篇 |
2000年 | 5篇 |
1999年 | 8篇 |
1998年 | 14篇 |
1997年 | 14篇 |
1996年 | 12篇 |
1995年 | 11篇 |
1994年 | 15篇 |
1993年 | 10篇 |
1992年 | 6篇 |
1991年 | 3篇 |
1990年 | 5篇 |
1989年 | 8篇 |
1988年 | 1篇 |
1987年 | 4篇 |
1986年 | 3篇 |
1985年 | 3篇 |
1984年 | 3篇 |
1983年 | 12篇 |
1982年 | 5篇 |
1981年 | 6篇 |
1980年 | 6篇 |
1979年 | 5篇 |
1978年 | 1篇 |
1977年 | 1篇 |
1975年 | 1篇 |
1974年 | 2篇 |
1971年 | 1篇 |
排序方式: 共有934条查询结果,搜索用时 15 毫秒
851.
Construction of a Library of Human Glycosyltransferases Immobilized in the Cell Wall of Saccharomyces cerevisiae
下载免费PDF全文
![点击此处可从《Applied microbiology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Yoh-ichi Shimma Fumie Saito Fumi Oosawa Yoshifumi Jigami 《Applied microbiology》2006,72(11):7003-7012
Fifty-one human glycosyltransferases were expressed in Saccharomyces cerevisiae as immobilized enzymes and were assayed for enzymatic activities. The stem and catalytic regions of sialyl-, fucosyl-, galactosyl-, N-acetylgalactosaminyl-, and N-acetylglucosaminyltransferases were fused with yeast cell wall Pir proteins, which anchor glycosyltransferases at the yeast cell wall glucan. More than 75% of expressed recombinant glycosyltransferases retained their enzymatic activities in the yeast cell wall fraction and will be used as a human glycosyltransferase library. In increasing the enzymatic activities of immobilized glycosyltransferases, several approaches were found to be effective. Additional expression of yeast protein disulfide isomerase increased the expression levels and activities of polypeptide N-acetylgalactosaminyltransferases and other glycosyltransferases. PIR3 and/or PIR4 was more effective than PIR1 as a cell wall anchor when the Pir-glycosyltransferase fusions were expressed under the control of the constitutive glyceraldehyde-3-phosphate dehydrogenase promoter. Oligosaccharides such as Lewis x, Lewis y, and H antigen were successfully synthesized using this immobilized glycosyltransferase library, indicating that the Pir-fused glycosyltransferases are useful for the production of various human oligosaccharides. 相似文献
852.
Hisaya Matsunami Makoto Kobayashi Shin-ichi Tsuruta Yoshifumi Terajima Hiroko Sato Masumi Ebina Shotaro Ando 《Bioenergy Research》2018,11(3):467-479
Erianthus arundinaceus, a warm-season perennial species in the Gramineae family, is currently being considered as a bioenergy crop candidate due to its capacity for high yields. Several experiments to explore this possibility have been conducted in Nasushiobara, Japan, where the mean minimum air temperature in January over the last three decades has been ??4.4 °C. Some accessions and breeding lines have demonstrated overwintering abilities, and annual dry matter yields of up to 52 t ha?1 were recorded in the 7th year after planting. Cutting to a 0.05-m height in February contributed more to subsequent regrowth and yield than did similar cutting in November. However, when performed in November, cutting to a 0.3-m height was more beneficial to subsequent regrowth and yield than cutting to a 0.05-m height. Allowing the foliage to run dry during winter led to a dry (approximately 70% dry matter ratio) biomass harvest in late winter. During winter, nutrient remobilization within the plants decreases nutrient removal from the soil. Although nitrogen fertilizer use efficiency was quite high (60%), only 20% of the nitrogen in an individual plant in the 2nd year after planting originated from fertilizer. This was likely due to a large amount of nitrogen obtained from non-fertilizer sources, i.e., soil and stored in the stubble from the previous harvest. Future experiments should focus on designing a fertilizer application program that could lead to sustainable and long-term high-yield E. arundinaceus biomass production in temperate zones. 相似文献
853.
Hanan H. Abdelbaky Ragab M. Fereig Yoshifumi Nishikawa 《Parasitology international》2018,67(6):675-678
Dense granule protein 7 (NcGRA7) is a potent diagnostic antigen of Neospora caninum. Some studies have reported on the difficult expression, low yield, and variable degree of solubility of recombinant NcGRA7. We aimed to unravel the possible causes for these issues and tested NcGRA7 antigenicity in enzyme linked immunosorbent assays (ELISAs). The NcGRA7 coding sequence (217 amino acids) was split into five amino acid regions: NcGRA7m (27–217), NcGRA7m3 (27–160), NcGRA7m4 (27–135), NcGRA7m5 (161–190), and NcGRA7m6 (188–217). Three fragments, NcGRA7m, NcGRA7m3 and NcGRA7m4, exhibited high antigenic properties when tested against experimentally-infected mice and dog sera by ELISA. High levels of IgG2 antibodies against NcGRA7m3 were observed in field dog sera. In experimentally and naturally-infected cattle, the N. caninum-specific sera only reacted with NcGRA7m, indicating that this antigenic region differs among the three animal species. This study presents valuable information about the antigenic properties and topology of NcGRA7, and highlights its suitability for diagnostic purposes. 相似文献
854.
855.
Yoshifumi Iwamaru Takato Takenouchi Morikazu Imamura Yoshihisa Shimizu Kohtaro Miyazawa Shirou Mohri Takashi Yokoyama Hiroshi Kitani 《Journal of virology》2013,87(15):8745-8755
The molecular mechanisms of prion-induced cytotoxicity remain largely obscure. Currently, only a few cell culture models have exhibited the cytopathic changes associated with prion infection. In this study, we introduced a cell culture model based on differentiated neurosphere cultures isolated from the brains of neonatal prion protein (PrP)-null mice and transgenic mice expressing murine PrP (dNP0 and dNP20 cultures). Upon exposure to mouse Chandler prions, dNP20 cultures supported the de novo formation of abnormal PrP and the resulting infectivity, as assessed by bioassays. Furthermore, this culture was susceptible to various prion strains, including mouse-adapted scrapie, bovine spongiform encephalopathy, and Gerstmann-Sträussler-Scheinker syndrome prions. Importantly, a subset of the cells in the infected culture that was mainly composed of astrocyte lineage cells consistently displayed late-occurring, progressive signs of cytotoxicity as evidenced by morphological alterations, decreased cell viability, and increased lactate dehydrogenase release. These signs of cytotoxicity were not observed in infected dNP0 cultures, suggesting the requirement of endogenous PrP expression for prion-induced cytotoxicity. Degenerated cells positive for glial fibrillary acidic protein accumulated abnormal PrP and exhibited features of apoptotic death as assessed by active caspase-3 and terminal deoxynucleotidyltransferase nick-end staining. Furthermore, caspase inhibition provided partial protection from prion-mediated cell death. These results suggest that differentiated neurosphere cultures can provide an in vitro bioassay for mouse prions and permit the study of the molecular basis for prion-induced cytotoxicity at the cellular level. 相似文献
856.
Yoshifumi Tanaka 《Ocean Development & International Law》2013,44(2):205-220
In the M/V “Louisa” case of 28 May 2013, the International Tribunal for the Law of the Sea held that it had no jurisdiction, even though it had established prima facie jurisdiction at the provisional measures stage. The M/V “Louisa” case thus gives rise to questions regarding the relationship between prima facie jurisdiction and jurisdiction on the merits. Moreover, the M/V “Louisa” Judgment also sheds some light on the applicability of the doctrine of abuse of rights provided in Article 300 of the UN Convention on the Law of the Sea. 相似文献
857.
Kosaku Hirota Yukio Kitade Tetsuo Tomishi Yoshifumi Maki 《Nucleosides, nucleotides & nucleic acids》2013,32(5):681-694
Abstract Reaction of 2′,5′-dichloro-2′,5′-dideoxyuridine (1) with ammonia and benzylamine afforded the corresponding 2-N-substituted 1-(5-chloro-5-deoxy-β-D-arabinofuranosyl)-isocytosine derivatives (2 and 10). Reaction of 1 with ammonia, methylamine, cyclohexylamine, and benzylamine followed by treatment with methanolic sodium methoxide gave the corresponding 2-N-substituted 1-(2,5-anhydro-β-D-arabino-furanosyl)isocytosine derivatives (6, 11, and 12). 相似文献
858.
Akihito Hagihara Manabu Hasegawa Takeru Abe Yoshifumi Wakata Takashi Nagata Yoshihiro Nabeshima 《PLoS medicine》2013,10(2)
Background
No studies have evaluated whether administering intravenous lactated Ringer''s (LR) solution to patients with out-of-hospital cardiac arrest (OHCA) improves their outcomes, to our knowledge. Therefore, we examined the association between prehospital use of LR solution and patients'' return of spontaneous circulation (ROSC), 1-month survival, and neurological or physical outcomes at 1 month after the event.Methods and Findings
We conducted a prospective, non-randomized, observational study using national data of all patients with OHCA from 2005 through 2009 in Japan. We performed a propensity analysis and examined the association between prehospital use of LR solution and short- and long-term survival. The study patients were ≥18 years of age, had an OHCA before arrival of EMS personnel, were treated by EMS personnel, and were then transported to hospitals. A total of 531,854 patients with OHCA met the inclusion criteria. Among propensity-matched patients, compared with those who did not receive pre-hospital intravenous fluids, prehospital use of LR solution was associated with an increased likelihood of ROSC before hospital arrival (odds ratio [OR] adjusted for all covariates [95% CI] = 1.239 [1.146–1.339] [p<0.001], but with a reduced likelihood of 1-month survival with minimal neurological or physical impairment (cerebral performance category 1 or 2, OR adjusted for all covariates [95% CI] = 0.764 [0.589–0.992] [p = 0.04]; and overall performance category 1 or 2, OR adjusted for all covariates [95% CI] = 0.746 [0.573–0.971] [p = 0.03]). There was no association between prehospital use of LR solution and 1-month survival (OR adjusted for all covariates [95% CI] = 0.960 [0.854–1.078]).Conclusion
In Japanese patients experiencing OHCA, the prehospital use of LR solution was independently associated with a decreased likelihood of a good functional outcome 1 month after the event, but with an increased likelihood of ROSC before hospital arrival. Prehospital use of LR solution was not associated with 1-month survival. Further study is necessary to verify these findings. Please see later in the article for the Editors'' Summary 相似文献859.
Yuko Ushiki‐Kaku Yoshifumi Iwamaru Kentaro Masujin Morikazu Imamura Shigeyoshi Itohara Kiyoko Ogawa‐Goto Shunji Hattori Takashi Yokoyama 《Microbiology and immunology》2013,57(11):792-796
Limited information is available about conformational differences between the abnormal isoform of prion protein (PrPSc) and cellular prion protein (PrPC) under native conditions. To clarify conformational differences between these two isoforms, PrP‐deficient mice were immunized with brain homogenates of normal and scrapie‐infected animals. All mice generated anti‐PrP antibodies. Peptide array analysis of these serum samples revealed a distinctive epitope of PrPSc consisting of QGSPGGN (PrP41–47) at the N‐terminus. This study demonstrated a conformational dissimilarity at the N‐terminus between PrPSc and PrPC, a finding that may provide novel information about conformational features of PrPSc. 相似文献
860.
Yoshifumi Jigami Toshio Omori Yasuji Minoda Koichi Yamada 《Bioscience, biotechnology, and biochemistry》2013,77(2):467-469
Equimolar aqueous solutions of d-glucose and n-butylamine were heated at 95°C for various times at pH 4, 6.5 and 11.48. The resulting brown solutions were extracted with ether. The volatile components in the ether extracts were analyzed by gas chromatography and gas chromatography-mass spectrometry, with a fused silica capillary column. The major components formed were identified as three alcohols, four N-butylpyrroles, N-butylacetamide, N-butylformamide, N-butylsuccinimide, one pyranone and 5-(hydroxymethyl)-2-furfural. In addition, eleven minor components were also identified. The relative amount of each component changed markedly with pH. At pH 4.0, higher-boiling heterocyclic compounds without C-C fission of glucose were largely formed, and at pH 11.48, lower-boiling fission compounds were mainly formed. Both were observed in the reaction at pH 6.5. 相似文献