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991.
992.
Molecular cloning and characterization of a novel angiopoietin family protein, angiopoietin-3 总被引:9,自引:0,他引:9
Using homology-based PCR, we have isolated cDNA encoding a novel member (491 amino acids) of the angiopoietin (Ang) family from human adult heart cDNA and have designated it angiopoietin-3 (Ang3). The NH2-terminal and COOH-terminal portions of Ang-3 contain the characteristic coiled-coil domain and fibrinogen-like domain that are conserved in other known Angs. Ang3 has a highly hydrophobic region at the N-terminus (approximately 21 amino acids) that is typical of a signal sequence for protein secretion. Ang3 mRNA is most abundant in adrenal gland, placenta, thyroid gland, heart and small intestine in human adult tissues. Additionally, Ang3 is a secretory protein, but is not a mitogen in endothelial cells. 相似文献
993.
In order to develop a serum-free (SF) medium for the production of erythropoietin (EPO) by suspension culture of recombinant Chinese hamster ovary (rCHO) cells, a statistical optimization approach based on a Plackett-Burman design was adopted. A basal medium was prepared by supplementing Iscove's modified Dulbecco's medium (IMDM) with Fe(NO3)3.9H2O, CuCl2 and ZnSO4.7H2O which are generally contained in SF medium formulations. Insulin, transferrin and ethanolamine were also supplemented to the basal medium to determine their optimal concentrations. From this statistical analysis, glutamate, serine, methionine, phosphatidylcholine, hydrocortisone and pluronic F68 were identified as positive determinants for cell growth. The SF medium was formulated by supplementing the basal medium with components showing positive effects on cell growth in suspension culture. An EPO titer in this optimized SF medium was 79% of that in IMDM supplemented with 5% dialyzed fetal bovine serum (dFBS). Furthermore, the in vitro and in vivo biological activities of EPO produced in the SF medium were comparable to those produced in the serum-supplemented medium. Taken together, the results obtained here show that a Plackett-Burman design facilitates the development of SF media for the production of EPO by suspension culture of rCHO cells. 相似文献
994.
L-arabinose isomerase (EC 5.3.1.4) mediates the isomerization of D-galactose into D-tagatose as well as the conversion of L-arabinose into L-ribulose. To investigate the properties of L-arabinose isomerase as a biocatalyst for the conversion of galactose to tagatose, the L-arabinose isomerase of Escherichia coli was characterized. The substrate specificity for L-arabinose was 166-fold higher than that for D-galactose. The optimal pH and temperature for the galactose isomerization reaction were 8.0 and 30 °C, respectively. The enzyme activity was stable for 1 h at temperatures below 35 °C and within a pH range of 8–10. The Michaelis constant, K
m, for galactose was 1480 mM, which is 25-fold higher than that for arabinose. The addition of Fe2+ and Mn2+ ions enhanced the conversion of galactose to tagatose, whereas the addition of Cu2+, Zn2+, Hg2+, and Fe3+ ions inhibited the reaction completely. In the presence of 1 mM Fe2+ ions, the K
m for galactose was found to be 300 mM. 相似文献
995.
996.
Maxime Willems Mieke Boone Marjolein Couvreur Katrien De Mulder Jelka Van Ranst Tom Artois Gaëtan Borgonie 《Development genes and evolution》2009,219(5):273-279
A method for studying whole mount flatworm embryos based on freeze-cracking of the eggs is described. This method allows successful
immunohistological and immunocytological studies of whole mount embryos. It does not require the use of sharpened needles
or a microinjection system to puncture the eggshell. Moreover, this method is more practical and less time-consuming than
classical puncturing and much cheaper than the use of a microinjection system. The advantages of this method are illustrated
by results of several immunolocalisation experiments in the macrostomid flatworm Macrostomum lignano. The optimal procedure and crucial steps for this method are discussed.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献
997.
Minimal requirements for the human immunodeficiency virus type 1 V3 domain to support the syncytium-inducing phenotype: analysis by single amino acid substitution. 总被引:11,自引:29,他引:11 下载免费PDF全文
The third variable domain (V3) of the human immunodeficiency virus type 1 external envelope contains determinants of cell tropism, cytopathicity, and infectivity and elicits antibodies able to block infectivity in vitro and in vivo. Our study encompassed point-mutational analysis of HXB-2 viruses containing patient-derived V3 regions and expressing a non-syncytium-inducing, low-replicating phenotype in T-cell line SupT1. The mutation within V3 of a serine at position 306 into an also naturally occurring arginine (S to R) required an additional, naturally occurring mutation at position 320 (aspartate to glutamine, D to Q) or 324 (aspartate to asparagine, D to N) for full expression of the syncytium-inducing, high-replicating (SI) phenotype. The naturally occurring mutation of an aspartate into an arginine at position 320 (D to R) was sufficient for production of the SI phenotype. This study proves that introduction of a positively charged amino acid at position 306 or 320, previously shown to be strongly associated with the SI phenotype in field isolates (R.A.M. Fouchier, M. Groenink, N.A. Kootstra, M. Tersmette, H.G. Huisman, F. Miedema, and H. Schuitemaker, J. Virol. 66:3183-3187, 1992), is minimally required for production of SI viruses. In addition, naturally occurring mutations at residue 324 also modulate the virus phenotype. 相似文献
998.
Differential expression analysis for sequence count data 总被引:22,自引:0,他引:22
High-throughput sequencing assays such as RNA-Seq, ChIP-Seq or barcode counting provide quantitative readouts in the form
of count data. To infer differential signal in such data correctly and with good statistical power, estimation of data variability
throughout the dynamic range and a suitable error model are required. We propose a method based on the negative binomial distribution,
with variance and mean linked by local regression and present an implementation, DESeq, as an R/Bioconductor package. 相似文献
999.
The toxicities of the herbicide glufosinate-ammonium to three predatory insect and two predatory mite species of Tetranychus urticae Koch were determined in the laboratory by the direct contact application. At a concentration of 540 ppm (a field application rate for weed control in apple orchards), glufosinate-ammonium was almost nontoxic to eggs of Amblyseius womersleyi Schicha, Phytoseiulus persimilis Athias-Henriot, and T. urticae but highly toxic to nymphs and adults of these three mite species, indicating that a common mode of action between predatory and phytophagous mites might be involved. In tests with predatory insects using 540 ppm, glufosinate-ammonium revealed little or no harm to larvae and pupae of Chrysopa pallens Rambur but was slightly harmful to eggs (71.2% mortality), nymphs (65.0% mortality), and adults (57.7% mortality) of Orius strigicollis Poppius. The herbicide showed no direct effect on eggs and adults of Harmonia axyridis (Pallas) but was harmful, slightly harmful, and harmless to first instars (100% mortality), fourth instars (51.1% mortality), and pupae (24.5% mortality), respectively. The larvae and nymphs of predators died within 12 h after treatment, suggesting that the larvicidal and nymphicidal action may be attributable to a direct effect rather than an inhibitory action of chitin synthesis. On the basis of our data, glufosinate-ammonium caused smaller effects on test predators than on T. urticae with the exception of P. persimilis, although the mechanism or cause of selectivity remains unknown. Glufosinate-ammonium merits further study as a key component of integrated pest management. 相似文献
1000.
Corti A 《Cellular and molecular neurobiology》2010,30(8):1163-1170
Chromogranin A (CgA) is an acidic glycoprotein belonging to a family of regulated secretory proteins stored in the dense core
granules of the adrenal medulla and of many other neuroendocrine cells and neurons. This protein is frequently used as a diagnostic
and prognostic serum marker for a range of neuroendocrine tumors. Circulating CgA is also increased in patients with other
diseases, including subpopulations of patients with non-neuroendocrine tumors, with important prognostic implications. A growing
body of evidence suggests that CgA is more than a diagnostic/prognostic marker for cancer patients. Indeed, results of in
vitro experiments and in vivo studies in animal models suggest that this protein and its fragments can affect several elements
of the tumor microenvironment, including fibroblasts and endothelial cells. In this article, recent findings implicating CgA
as a modulator of the tumor microenvironment and suggesting that abnormal secretion of CgA could play important roles in tumor
progression and response to therapy in cancer patients are reviewed and discussed. 相似文献