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1.
2.
Summary An acetyl esterase was purified from Trichoderma reesei by cation and anion exchange chromatography. The enzyme had a molecular weight of 45 000 as determined by SDS-electrophoresis, or 67 000 as determined by gel filtration. In chromatofocusing the enzyme was shown to consist of two isoenzymes with isoelectric points of 6.8 and 6.0. The enzyme showed activity towards naphthyl acetate, triacetin and glucose-and xylose acetates. However, it liberated acetic acid from acetylated xylo-oligomers only to a small extent. The liberation of acetic acid from the oligomeric substrate was enhanced by addition of endoxylanase and -xylosidase. 相似文献
3.
Identification of Pneumocystis carinii in immunodeficient mice 总被引:3,自引:0,他引:3
Various procedures were utilized to determine the most sensitive, cost and labor effective techniques for detection of Pneumocystis carinii in immunologically compromised mice. Immunoperoxidase staining techniques that utilized polyclonal antibodies directed against purified rat or mouse P. carinii were more sensitive and specific than staining with Gomori's methenamine silver. Indirect immunofluorescence microscopy on frozen sections was comparable to immunoperoxidase staining, but lacked fine cytologic detail. Impression smears were of limited value when stained with Diff-Quik Stain, Harleco's Hemacolor, Wright-Giemsa or Wright-Leishman stains. However, cysts could be detected consistently in imprints stained with Gomori's methanamine silver. Transmission electron microscopy showed ultrastructural detail of P. carinii, but this technique was too costly and time consuming for routine use. Thus, because of its sensitivity and specificity, immunohistochemistry on paraffin sections was the most satisfactory method for screening and identifying P. carinii in lungs of immunocompromised mice. 相似文献
4.
Total body burdens of Zn, Pb, Al and Cu were measured in Swedish marine nemerteans sampled from an area close to a major source
of pollution. Accumulation patterns varied both seasonally and spatially. The results suggest that nemerteans may be useful
bioindicator species in environmental monitoring programmes. 相似文献
5.
The essential mitotic target of calmodulin is the 110-kilodalton component of the spindle pole body in Saccharomyces cerevisiae. 总被引:17,自引:0,他引:17 下载免费PDF全文
J R Geiser H A Sundberg B H Chang E G Muller T N Davis 《Molecular and cellular biology》1993,13(12):7913-7924
Two independent methods identified the spindle pole body component Nuf1p/Spc110p as the essential mitotic target of calmodulin. Extragenic suppressors of cmd1-1 were isolated and found to define three loci, XCM1, XCM2, and XCM3 (extragenic suppressor of cmd1-1). The gene encoding a dominant suppressor allele of XCM1 was cloned. On the basis of DNA sequence analysis, genetic cosegregation, and mutational analysis, XCM1 was identified as NUF1/SPC110. Independently, a C-terminal portion of Nuf1p/Spc110p, amino acid residues 828 to 944, was isolated as a calmodulin-binding protein by the two-hybrid system. As assayed by the two-hybrid system, Nuf1p/Spc110p interacts with wild-type calmodulin and triple-mutant calmodulins defective in binding Ca2+ but not with two mutant calmodulins that confer a temperature-sensitive phenotype. Deletion analysis by the two-hybrid system mapped the calmodulin-binding site of Nuf1p/Spc110p to amino acid residues 900 to 927. Direct binding between calmodulin and Nuf1p/Spc110p was demonstrated by a modified gel overlay assay. Furthermore, indirect immunofluorescence with fixation procedures known to aid visualization of spindle pole body components localized calmodulin to the spindle pole body. Sequence analysis of five suppressor alleles of NUF1/SPC110 indicated that suppression of cmd1-1 occurs by C-terminal truncation of Nuf1p/Spc110p at amino acid residues 856, 863, or 881, thereby removing the calmodulin-binding site. 相似文献
6.
Berit Backlund Hk Anette M. Johansson Stephan Hjorth Staffan Sundell Uli Hacksell 《Chirality》1993,5(3):112-119
Racemic 5-methoxy-2-methyl-2-dipropylaminotetralin ( 3 ) has been prepared by a short synthetic route, in which the N,N-dipropyliminium perchlorate of 5-methoxy-2-tetralone ( 4 ) is a key intermediate. Racemic 3 was resolved by crystallization of the corresponding diastereomeric di-p-toluoyltartrates. The enantiomeric excess (%ee) of the phenolic derivatives of (+)-(R)- and (?)-(S)-3 [(+)-(R)- and (?)-(S)-2] was determined by 1HNMR spectroscopic analysis of the corresponding diastereomeric (?)-(R)-1,1′-binaphthyl-2,2′-diylphosphoric acid salts utilizing 13C satellites. X-ray crystallography established the absolute configuration of (?)-(S)-2 · HCl. The enantiomers of 2 were tested for hippocampal output of 5-hydroxytryptamine, 5-hydroxyindoleacetic acid, and dihydroxyphenylacetic acid in rats by use of in vivo microdialysis. The (?)-(S)-enantiomer appeared to affect 5-HT-turnover, whereas (+)-(R)- 2 was inactive. Results obtained provide support for the previously reported hypothesis that the inactivity of (?)-(S)- 2 at central DA receptors is caused by the steric bulk of the C(2)-methyl group. This makes it possible to define a “DA D2 receptor essential volume.” © 1993 Wiley-Liss, Inc. 相似文献
7.
Berit M. Mortensen Hanne W. Lund Greg Jablonski Ruth H. Paulssen Jan O. Gordeladze 《Bioscience reports》1995,15(3):135-150
In normal rats treated with 1,25(OH)2D3 or 24,25(OH)2D3, serum Ca2+, ALP, PRL and GH are significantly altered. In order to study the primary effect of vitamin D3 analogues on target organ function, rat UMR 106 osteosarcoma and GH3 pituitary adenoma cells in monolayer culture were exposed accordingly.Surprisingly, prolonged exposure of these cell lines to physiological levels of either 1,25(OH)2D3 or 24,25(OH)2D3 did not significantly affect the secretory parameters (ALP, PRL or GH) tested. However, 1,25(OH)2D3 exposure significantly reduced PTH- and Gpp(NH)p-elicited AC as well as Gpp(NH)p-stimulated PLC activities in the UMR 106 cells. These changes were accompanied by an increase and decrease in the membrane contents of the G-protein subunits G36 and Gq/11, respectively. In contrast, 24,25(OH)2D3 remained without significant biological effect on these signalling systems despite concomitantly augmented levels of G36. TRH- and Gpp(NH)p-elicited PLC activities in the GH3 cells were significantly reduced by 1,25(OH)2D3 with a concurrent reduction in cellular amounts of Gq/11, however, 24,25(OH)2D3 did not significantly alter any signalling systems nor G-proteins analyzed.It is concluded that the osteoblastic and pituitary cell secretion of ALP, PRL and GH remain unaffected by the presence of 1,25(OH)2D3 and 24,25(OH)2D3, despite distinct alterations in components of G-protein mediated signalling pathways. Hence, other factors like ambient Ca2+ may be responsible for the perturbed secretory patterns of ALP and PRL seen in vitamin D3 treated rats.Abbreviations AC
adenylate cyclase
- ALP
alkaline phosphatase
- BGP
osteocalcin
- BSA
bovine serum albumin
- DA
dopamine
- DAG
diacylglycerol
- GH
growth hormone
- GHRH
growth hormone releasing hormone
- Gpp(NH)p
guanosine 5-[-imido]triphosphate
- G-protein
guanine nucleotide-binding regulatory protein
- Gs etc.
Gs protein -subunit
- IP3
inositol 1,4,5 trisphosphate
- OAF
osteoclast activating factor
- PGE2
prostaglandin E2
- PKA & PKC
protein kinase A & C
- PLC
phospholipase C
- PRL
prolactin
- PTH
parathyroid hormone
- SRIF
somatostatin
- TRH
thyrotropin releasing hormone
- VIP
vasoactive intestinal peptide
- 25(OH)D3
25 hydroxy vitamin D3
- 1,25(OH)2D3
1·25 dihydroxy vitamin D3
- 24,25(OH)2D3
24,25 dihydroxy vitamin D3 相似文献
8.
Central neural peptides and catecholamines in spontaneous and DOCA/salt hypertension 总被引:1,自引:0,他引:1
Hypothalamic and neurophypophyseal levels of catecholamines and peptides were measured in spontaneous and deoxycorticosterone (DOCA)/salt hypertension. Catecholamines, norepinephrine, epinephrine and dopamine were measured by electrochemical detection while the peptides, vasopressin, oxytocin, luteinizing hormone-releasing hormone (LHRH), the enkephalins and somatostatin (SRIF) were measured by radioimmunoassay. Blood pressure was significantly elevated in both groups as compared to their controls. Marked changes in central neural peptides were observed in the SHR, while no differences were seen in DOCA/salt hypertension. Hypothalamic vasopressin, oxytocin, LHRH and SRIF were significantly decreased. In the posterior pituitary, enkephalins were increased twofold in the SHR. With regard to catecholamines, there was no change in hypothalamic content. However, a dramatic decrease in neurohypophyseal dopamine was observed in SHR. Plasma levels of vasopressin were significantly elevated in both types of hypertension while oxytocin was increased only in the DOCA/salt model. These result show that (1) a wide spectrum of neuroendocrine changes are associated with genetic hypertension, (2) there are CNS differences between DOCA/salt and spontaneous hypertension, and (3) central aminergic changes may be involved in th neuroendocrine alterations seen in the SHR. 相似文献
9.
10.
Jeemeng Lao Ai Oikawa Jennifer R. Bromley Peter McInerney Anongpat Suttangkakul Andreia M. Smith‐Moritz Hector Plahar Tsan‐Yu Chiu Susana M. González Fernández‐Niño Berit Ebert Fan Yang Katy M. Christiansen Sara F. Hansen Solomon Stonebloom Paul D. Adams Pamela C. Ronald Nathan J. Hillson Masood Z. Hadi Miguel E. Vega‐Sánchez Dominique Loqué Henrik V. Scheller Joshua L. Heazlewood 《The Plant journal : for cell and molecular biology》2014,79(3):517-529
The glycosyltransferases (GTs) are an important and functionally diverse family of enzymes involved in glycan and glycoside biosynthesis. Plants have evolved large families of GTs which undertake the array of glycosylation reactions that occur during plant development and growth. Based on the Carbohydrate‐Active enZymes (CAZy) database, the genome of the reference plant Arabidopsis thaliana codes for over 450 GTs, while the rice genome (Oryza sativa) contains over 600 members. Collectively, GTs from these reference plants can be classified into over 40 distinct GT families. Although these enzymes are involved in many important plant specific processes such as cell‐wall and secondary metabolite biosynthesis, few have been functionally characterized. We have sought to develop a plant GTs clone resource that will enable functional genomic approaches to be undertaken by the plant research community. In total, 403 (88%) of CAZy defined Arabidopsis GTs have been cloned, while 96 (15%) of the GTs coded by rice have been cloned. The collection resulted in the update of a number of Arabidopsis GT gene models. The clones represent full‐length coding sequences without termination codons and are Gateway® compatible. To demonstrate the utility of this JBEI GT Collection, a set of efficient particle bombardment plasmids (pBullet) was also constructed with markers for the endomembrane. The utility of the pBullet collection was demonstrated by localizing all members of the Arabidopsis GT14 family to the Golgi apparatus or the endoplasmic reticulum (ER). Updates to these resources are available at the JBEI GT Collection website http://www.addgene.org/ . 相似文献