全文获取类型
收费全文 | 285篇 |
免费 | 27篇 |
出版年
2023年 | 3篇 |
2022年 | 4篇 |
2021年 | 8篇 |
2020年 | 4篇 |
2019年 | 3篇 |
2018年 | 6篇 |
2017年 | 13篇 |
2016年 | 18篇 |
2015年 | 16篇 |
2014年 | 16篇 |
2013年 | 19篇 |
2012年 | 15篇 |
2011年 | 20篇 |
2010年 | 19篇 |
2009年 | 10篇 |
2008年 | 19篇 |
2007年 | 14篇 |
2006年 | 13篇 |
2005年 | 15篇 |
2004年 | 13篇 |
2003年 | 9篇 |
2002年 | 8篇 |
2001年 | 5篇 |
2000年 | 3篇 |
1999年 | 5篇 |
1998年 | 4篇 |
1997年 | 4篇 |
1996年 | 2篇 |
1995年 | 4篇 |
1994年 | 2篇 |
1993年 | 2篇 |
1990年 | 2篇 |
1988年 | 1篇 |
1987年 | 2篇 |
1983年 | 1篇 |
1982年 | 2篇 |
1978年 | 1篇 |
1977年 | 3篇 |
1974年 | 1篇 |
1968年 | 1篇 |
1961年 | 1篇 |
1958年 | 1篇 |
排序方式: 共有312条查询结果,搜索用时 46 毫秒
1.
High level of divergence of male-reproductive-tract proteins, between Drosophila melanogaster and its sibling species, D. simulans 总被引:1,自引:0,他引:1
We compared male-reproductive-tract polypeptides of Drosophila melanogaster
and D. simulans by using two-dimensional gel electrophoresis. Approximately
64% of male-reproductive-tract polypeptides were identical between two
randomly chosen isofemale lines from these two species, compared with 83%
identity for third-instar imaginal wing-disc polypeptides. Qualitatively
similar differences were found between reproductive tracts and imaginal
discs when D. sechellia was compared with D. melanogaster and with D.
simulans. When genic polymorphism was taken into account, approximately 10%
of male- reproductive-tract polypeptides were apparently fixed for
different alleles between D. melanogaster and D. simulans; this proportion
is the same as that found for soluble enzymes by one-dimensional gel
electrophoresis. Strikingly, approximately 20% of male-reproductive- tract
polypeptides of either D. melanogaster or D. simulans had no detectable
homologue in the other species. We propose that proteins of the Drosophila
male reproductive tract may have diverged more extensively between species
than have other types of proteins and that much of this divergence may
involve large changes in levels of polypeptide expression.
相似文献
2.
RS Fisher 《The Journal of general physiology》1977,69(5):571-604
When the outer surface of short-circuited frog skin was penetrated with microelectrodes, stable negative potentials that averaged near -100 mV were recorded consistently, confirming the results of Nagel (W. Nagel. 1975. Abstracts of the 5th International Biophysics Congress, Copenhagen. P-147.). The appearance of these stable potentials, V(O), concurrent with the observations that (a) a high resistance outer barrier R(O) accounting for approximately 75 percent or more of the transcellular resistance of control skins had been penetrated and that (b) 10(-5) M amiloride and reduced [Na] outside caused the values of V(O) to increase towards means value near -130 mV while the values of percent R(O) increased to more than 90 percent. It was of relationships were the same as the values of E(1) observed in studies of the current-voltage relationships were the same as the values of E’(1) defined as the values of voltage at the inner barrier when the V(O) of the outer barrier was reduced to zero by voltage clamping of the skins. Accordingly, these data are interpreted to mean that the values of E(1), approximately 130 mV, represent the E(Na) of the sodium pump at the inner barrier. 2,4-DNP was observed to decrease the values of transepithelial voltage less than E(1) the V(O) was negative. These data can be interpreted with a simple electrical equivalent circuit of the active sodium transport pathway of the frog skin that includes the idea that the outer membrane behaves as an electrical rectifier for ion transport. 相似文献
3.
4.
Brian Effer Guilherme Meira Lima Sindy Cabarca Adalberto Pessoa Jorge G. Farías 《Preparative biochemistry & biotechnology》2013,43(7):679-685
AbstractL-Asparaginase (L-ASNase) is an important enzyme used to treat acute lymphoblastic leukemia, recombinantly produced in a prokaryotic expression system. Exploration of alternatives production systems like as extracellular expression in microorganisms generally recognized as safe (such as Pichia pastoris Glycoswitch®) could be advantageous, in particular, if this system is able to produce homogeneous glycosylation. Here, we evaluated extracellular expression into Glycoswitch® using two different strains constructions containing the asnB gene coding for Erwinia chrysanthemi L-ASNase (with and without His-tag), in order to find the best system for producing the extracellular and biologically active protein. When the His-tag was absent, both cell expression and protein secretion processes were considerably improved. Three-dimensional modeling of the protein suggests that additional structures (His-tag) could adversely affect native conformation and folding from L-ASNase and therefore the expression and cell secretion of this enzyme. 相似文献
5.
Luciana P. Malpiedi César A. Díaz Bibiana B. Nerli Adalberto Pessoa 《Process Biochemistry》2013,48(8):1242-1251
At present, single-chain variable fragments (scFv) of antibodies are considered one of the most important tools in human therapies. Wide applications of antibodies are being exploited in different medical, pharmaceutical and research areas. These molecules maintain the same binding functionality that full length antibodies but possess several advantageous features as quickness to penetrate the tissues, easy manipulation, fast elimination of their immunocomplex and the possibility of being produced in simple expression systems like bacteria and yeast. The increasing demand in antibody based methodologies is driving advances in the production and purification of genetically engineered antibodies and antibody fragments. While advances in expression systems allow the production of high titers of antibodies, there exist some limits imposed by the downstream methodologies which are not efficient enough to ensure their industrialization.The main aim of this review is to highlight the principal characteristics of single-chain variable fragments of antibodies addressing advances and perspectives on scFv purification. 相似文献
6.
Rayana Carla Silva de Morais Suênia da Cunha Gonçalves Pietra Lemos Costa Kamila Gaudêncio da Silva Fernando José da Silva Rômulo Pessoa e Silva Maria Edileuza Felinto de Brito Sinval Pinto Brandão-Filho Filipe Dantas-Torres Milena de Paiva-Cavalcanti 《Experimental & applied acarology》2013,59(4):473-481
Visceral leishmaniosis (VL) is a parasitic disease caused by Leishmania infantum, which is primarily transmitted by phlebotomine sandflies. However, there has been much speculation on the role of other arthropods in the transmission of VL. Thus, the aim of this study was to assess the presence of L. infantum in cats, dogs and their ectoparasites in a VL-endemic area in northeastern Brazil. DNA was extracted from blood samples and ectoparasites, tested by conventional PCR (cPCR) and quantitative real time PCR (qPCR) targeting the L. infantum kinetoplast DNA. A total of 280 blood samples (from five cats and 275 dogs) and 117 ectoparasites from dogs were collected. Animals were apparently healthy and not previously tested by serological or molecular diagnostic methods. Overall, 213 (76.1 %) animals and 51 (43.6 %) ectoparasites were positive to L. infantum, with mean parasite loads of 795.2, 31.9 and 9.1 fg in dogs, cats and ectoparasites, respectively. Concerning the positivity between dogs and their ectoparasites, 32 (15.3 %) positive dogs were parasitized by positive ectoparasites. The overall concordance between the PCR protocols used was 59.2 %, with qPCR being more efficient than cPCR; 34.1 % of all positive samples were exclusively positive by qPCR. The high number of positive animals and ectoparasites also indicates that they could serve as sentinels or indicators of the circulation of L. infantum in risk areas. 相似文献
7.
Andréa Helena Vidal André Mariano Batista Ellen Cordeiro Bento da Silva Wilton Arruda Gomes Marina Arruda Pelinca Sildivane Valcácia Silva Maria Madalena Pessoa Guerra 《Small Ruminant Research》2013,109(1):47-51
The aim of this study was to evaluate the effect of different concentrations of soybean lecithin (SL) in extenders for sperm goat cryopreservation. Sexually mature male Saanen goats (n = 4) were used, and the ejaculates were obtained using an artificial vagina method. The semen samples were pooled and diluted in a skim milk-based extender (control group; CG) or Tris extender supplemented with SL at different concentrations (G1 = 0.04%, SL G2 = 0.08% SL and G3 = 0.16%) for a final concentration of 240 × 106 spermatozoa/mL. The semen samples were packed in straws (0.25 mL), frozen using an automated system and stored in liquid nitrogen (?196 °C). After thawing (37 °C/30 s), the samples were evaluated for sperm quality parameters, including sperm motility, membrane integrity, acrosome integrity and mitochondrial activity. No significant difference was observed among the experimental and control groups for all of the parameters (P > 0.05). However, even though the control group presented a significantly lower mitochondrial membrane potential compared to fresh semen (P < 0.05), the same did not occur for the extender supplemented with soybean lecithin, that is, it did not differ from fresh sperm (P > 0.05). The extender containing soybean lecithin at different concentrations preserved the sperm quality parameters in a manner similar to the conventional skim milk-based extender. Thus, it is concluded that an extender containing soybean lecithin as the lipoprotein source can be used for freezing goat semen. 相似文献
8.
Paula M. Nogueira Rafael R. Assis Ana C. Torrecilhas Elvira M. Saraiva Natália L. Pessoa Marco A. Campos Eric F. Marialva Cláudia M. Ríos-Velasquez Felipe A. Pessoa Nágila F. Secundino Jer?nimo N. Rugani Elsa Nieves Salvatore J. Turco Maria N. Melo Rodrigo P. Soares 《PLoS neglected tropical diseases》2016,10(8)
The immunomodulatory properties of lipophosphoglycans (LPG) from New World species of Leishmania have been assessed in Leishmania infantum and Leishmania braziliensis, the causative agents of visceral and cutaneous leishmaniasis, respectively. This glycoconjugate is highly polymorphic among species with variation in sugars that branch off the conserved Gal(β1,4)Man(α1)-PO4 backbone of repeat units. Here, the immunomodulatory activity of LPGs from Leishmania amazonensis, the causative agent of diffuse cutaneous leishmaniasis, was evaluated in two strains from Brazil. One strain (PH8) was originally isolated from the sand fly and the other (Josefa) was isolated from a human case. The ability of purified LPGs from both strains was investigated during in vitro interaction with peritoneal murine macrophages and CHO cells and in vivo infection with Lutzomyia migonei. In peritoneal murine macrophages, the LPGs from both strains activated TLR4. Both LPGs equally activate MAPKs and the NF-κB inhibitor p-IκBα, but were not able to translocate NF-κB. In vivo experiments with sand flies showed that both stains were able to sustain infection in L. migonei. A preliminary biochemical analysis indicates intraspecies variation in the LPG sugar moieties. However, they did not result in different activation profiles of the innate immune system. Also those polymorphisms did not affect infectivity to the sand fly. 相似文献
9.
Lilian S. Catenacci Michaele S. Pessoa Sérgio L. G. Nogueira-Filho Kristel M. De Vleeschouwer 《International journal of primatology》2016,37(2):136-157
Habitat disturbance alters plant diversity and food resource availability, affecting the ecology, and ultimately the survival and reproduction, of species depending on those plants. Studies in degraded areas serve to improve our understanding of the consequences of habitat modification for endangered species and to guide conservation actions. We studied diet composition, monthly variation in feeding behavior and fruit feeding time, and dietary diversity in two golden-headed lion tamarin (Leontopithecus chrysomelas) groups ranging in a degraded area of the Atlantic Forest in South-Bahia, Brazil, over a 12-mo period. We recorded feeding behavior and food items consumed through continuous observations and focal animal sampling, and performed dietary analyses on samples from consumed fruits. Substrate manipulation to search for animal prey was the feeding activity most frequently observed, followed by fruit consumption. We observed nectar drinking and exudate feeding at low frequencies from April through July. Bromeliads were the principal foraging substrate. We observed the use of 114 plant species, confirming the large dietary diversity reported for the species. Individual trees from the family Melastomataceae, common in degraded forests, accounted for the highest visiting frequencies (40%). Fruit availability was the main factor explaining variation in monthly fruit feeding time, despite the absence of climatic seasonality. Nutritional or energetic characteristics did not affect fruit choice. Differences in floristic composition appear to be a major determinant of the species’ diet in different study areas. Regional forest restoration programs should consider including advanced forest species, to improve both forest quality and animal mobility between fragments. 相似文献
10.