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81.
Abstract The antifungal antibiotic papulacandin β inhibited B(1,3)glucan-synthase activity, in vitro, from Schizosaccharomyces pombe . Levels of β(1,3)glucan-synthase from antibiotic-treated cultures were lower than the control cultures whereas mannan-synthase and β(1,3)glucanase activities were almost unaffected. The presence of an osmotic stabilizer reduced the inhibition of growth caused by the antibiotic. Addition of papulacandin β to a culture of S. pombe specifically inhibited incorporation of glucose into the β-glucan cell wall fraction. The fatty acids as well as the hydroxyl groups on the phenol residue of the papulacandin β molecule were essential for the inhibitory activity. 相似文献
82.
A birefringence technique is used to determine the average magnetic moments <μ> of magnetotactic bacteria in culture. Differences in <μ> are noted between live and dead bacteria, as well as between normal density and high density samples of live bacteria. 相似文献
83.
Characterization of immune type interferon (IFN gamma)-induced cytoplasmic protein kinase activity in mouse L cells 总被引:1,自引:0,他引:1
K Ohtsuki B A Torres H M Johnson 《Biochemical and biophysical research communications》1982,104(2):422-429
Mouse immune interferon (IFNγ) induced double-stranded RNA-independent protein kinase activity in the cytoplasmic fraction of mouse L cells as measured against a histone substrate. Chromatographic purification separated the activity into three distinct kinases of molecular weights of approximately 100K (kinase I), 70K (kinase II), and 70K (kinase III). Partially purified IFNγ was as effective as crude in inducing protein kinase activity. Induction was blocked by treatment of IFNγ with specific anti-IFNγ antibody or by treatment of the L cells with actinomycin D. Kinase II activity was different from that of kinases I and III in that it showed Ca-dependence in the absence of Mg2+, was inhibited in activity by the SH binding agent N-ethylmaleimide, and could use the cellular enzymes RNase A and hexokinase as substrates. The described protein kinases could play an important role in mediation of IFNγ effects, particularly where phosphorylated enzyme substrates were shown to have altered activity. 相似文献
84.
85.
Detection of salmonellas by DNA hybridization with a fluorescent alkaline phosphatase substrate. 总被引:2,自引:0,他引:2
R J Cano M J Torres R E Klem J C Palomares J Casadesus 《The Journal of applied bacteriology》1992,72(5):393-399
This study evaluates a DNA hybridization assay for salmonella with AttoPhos (JBL Scientific, San Luis Obispo, CA), a fluorescent substrate for alkaline phosphatase. The probe used (50 ng/ml) was a biotinylated 600 bp fragment consisting of a tandem repeat of an insertion sequence (IS200) found in most Salmonella spp. evaluated. The hybridization was carried out at 65 degrees C for 2 h without prior prehybridization and hybrids were detected by the addition of a streptavidin-alkaline phosphatase conjugate. Circles (5 mm) were cut from the membrane and placed in a cuvette containing 1 ml of 1 mmol/l AttoPhos. The reaction was evaluated after 30 min at 37 degrees C with a fluorometer with an excitation wavelength of 440 nm and an emission wavelength of 550 nm. The sensitivity of the probe was estimated to be 10,000 copies of target DNA or 5 x 10(-20) mol of DNA. All 74 salmonella strains tested reacted with the probe but none of the 98 heterologous species tested gave positive results. The results of this study indicate that our assay method, which employs a biotinylated tandem repeat of IS200 and AttoPhos, is a specific and highly sensitive quantitative method for the detection of salmonellas. 相似文献
86.
Calvet S Clapés P Vigo JP Xaus N Jorba X Mas RM Torres JL Valencia G Serralheiro ML Cabral JM Empis JM 《Biotechnology and bioengineering》1992,39(5):539-549
The influence of five different N-terminal protecting groups (For, Ac, Boc, Z, and Fmoc) and reaction conditions (temperature and dimethylformamide content) on the alpha-chymotrypsin-catalyzed synthesis of the dipeptide derivative X-Phe-Leu-NH(2) was studied. Groups such as For, Ac, Boc, and Z always rendered good peptide yields (82% to 85%) at low reaction temperatures and DMF concentrations, which depended on the N-alpha protection choice. Boc and Z were the most reactive N-alpha groups and, in addition, the most suitable for peptide synthesis. On the other hand, the use of empirical design methodologies allowed, with minimal experimentation and by multiple regression, to deduce an equation, which correlates the logarithm of the first order kinetic constant (log k') with reaction temperature, DMF concentration, and hydrophobicity (log P values) of the different protecting groups. The predictive value of the equation was tested by comparing the performance of another protective group, such as Aloc, with the performance predicted by said equation. Experimental and calculated k' values were found to be in good agreement. 相似文献
87.
J P Deans H M Serra J Shaw Y J Shen R M Torres L M Pilarski 《Journal of immunology (Baltimore, Md. : 1950)》1992,148(6):1898-1905
High molecular mass isoforms of CD45 protein tyrosine phosphatase, predominantly CD45RA, are expressed on naive T cells with increasing density during the first 2 days of cellular activation, and subsequently down-regulated concurrent with increasing expression of the low Mr isoform, CD45RO. We show this to be de novo synthesis of CD45RA dependent upon both RNA and protein synthesis. Using a probe shown to detect mRNA encoding the alternatively spliced abc, ab, bc, and b exon isoforms of CD45, the expression of CD45 was analyzed. Kinetic studies of the transition from high to low Mr CD45 mRNA indicate an immediate decrease in the level of high Mr CD45 mRNA after mitogen stimulation of T cells, quickly followed at 4 h by an increase to above steady state levels. This is consistent with the transitory increase in cell-surface density of CD45RA observed at 1 to 2 days poststimulation. Within 24 h of stimulation the level of high Mr CD45 mRNA declines precipitously such that little or no mRNA encoding any of the alternatively spliced exons is detectable. High levels of CD45 mRNA are detectable at later points but this does not hybridize with the Sfa N1 probe recognizing the abc exons suggesting that only the CD45RO mRNA splicing pattern is operative. We also show that CD45 mRNA have a relatively short t1/2. In mitogen-stimulated cells the t1/2 of high and low m.w. CD45 mRNA was estimated at 2.25 h and 3.5 h, respectively. In unstimulated T cells the t1/2 of high Mr CD45 mRNA was estimated at 2.8 h. CD45 mRNA is super-induced in the presence of the protein synthesis inhibitor, cycloheximide, indicating that the degradation and/or splicing of CD45 mRNA is controlled by a labile pathway, and suggesting that mechanisms may exist in vivo to prolong synthesis of CD45RA. The kinetics of accumulation for high Mr CD45 mRNA are very similar to those of the TCR beta-chain and pp56lck suggesting that these functionally linked signaling molecules are regulated in tandem. This implicates stringent molecular control mechanisms on the production of mRNA encoding either high or low m.w. isoforms, consistent with the fundamental role of CD45 in signal transduction, and the apparent need to selectively and sequentially express functionally distinct external CD45 domains. 相似文献
88.
In vivo and in vitro effects of boron on the plasma membrane proton pump of sunflower roots 总被引:3,自引:0,他引:3
Marta Roldán rés Belver Pilar Rodríguez-Rosales Nuria Ferrol Juan Pedro Donaire 《Physiologia plantarum》1992,84(1):49-54
The effect of boron excess and deficiency on H+ efflux from excised roots from sunflower ( Heliarahus annuus L. cv. Enano) seedlings and on plasma membrane H+ -ATPase (EC 3.6.1.35) in isolated KI-washed microsomes has been investigated. When seedlings were grown in media with toxic levels of H3 BO3 (5 m M ) or without added boron and exposed to light conditions, an inhibition of the capacity for external acidification by excised roots was observed as compared to roots from seedlings grown with optimal H3 BO3 concentration (0.25 m M ). Toxic and deficient boron conditions also inhibited the vanadate-sensitive H+ -ATPase of microsomes isolated from the roots. The mechanism of boron toxicity was investigated in vitro with microsorne vesicles. A strong effect of boron on the vanadate-sensitive, ATP-dependent H+ transport was found, but the vanadate-sensitive phospho-bydrolase activity was not affected. These results suggest that boron could exert an effect on the plasma membrane properties, directly or indirectly regulating, proton transport. 相似文献
89.
Soledad Gaitán Elvira Cuenllas Pilar Sancho Juan A. Bueren Concepción Tejero 《Bioscience reports》1992,12(4):281-292
This paper analyzes the long-term (6 and 12 months) function of mouse granulocytes after total body irradiation with a single dose (5 Gy) of X-rays. Superoxide anion production has been investigated in granulocytes from peripheral blood, and also in those harvested from long term bone marrow cultures, with the aim of correlating the environmental damage induced by radiation with the functional properties of granulocytes. Anin vivo andin vitro enhancement of superoxide anion production and protein levels in granulocytes from irradiated mice is described. The presence of some colony stimulating factor in the supernatant of cultures from irradiated mice could play an important role in the priming of granulocytes. 相似文献
90.
Pilar Galan Helene Thibault Paul Preziosi Serge Hercberg 《Biological trace element research》1992,32(1-3):421-426
The relationship between iron status and capacity for IL-2 production by lymphocytes was assessed in 81 children from 6 mo
to 3 yr of age selected at random from a population with low socioeconomic status, undergoing free systematic examination
in four children's health centers in the Paris area. Iron deficiency was defined by the existence of at least two abnormal
values among the three indicators of iron status: serum ferritin level ≤12 μg/L, transferrin saturation <12%, and erythrocyte
protoporphyrin concentration >3 μg/g hemoglobin. According to this definition, 53 children were classified as iron deficient
and 28 as iron sufficient. No differences were observed between the iron-deficient and iron-sufficient groups in terms of
the IL-2 concentration without stimulation by PHA. IL-2 production by lymphocytes stimulated with PHA, as well as the stimulation
index (ratio of IL-2 concentration following stimulation by PHA to that of IL-2 concentration without stimulation by PHA)
were significantly lower in iron-deficient children. The reduction in IL-2 production by activated lymphocytes observed in
our study of iron-deficient children may be responsible for impairments in immunity found by other authors, particularly in
cell-mediated immunity. 相似文献