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991.
DNA polymerase from Thermus aquaticus has become a common reagent in molecular biology because of its utility in DNA amplification and DNA sequencing protocols. A simplified method is described here for isolating the recombinant Taq enzyme after overproduction in Escherichia coli. Purification requires 8 to 10 h and entails heat treating and clearing the E. coli lysate, followed by precipitation of the enzyme with polyethyleneimine and elution from Bio Rex 70 ion exchange resin in a single salt step. The resulting enzyme preparation contains a single, nearly homogeneous protein consistent with the previously established size of the Taq DNA polymerase in a yield of 40-50 mg of protein per liter of cell culture. 相似文献
992.
†M. J. Blanco-Príeto C. Durieux V. Daugé †E. Fattal †P. Couvreur B. P. Roques 《Journal of neurochemistry》1996,67(6):2417-2424
Abstract: Neuropeptides have been shown to play a critical role in adaptational processes, probably by long-term modulation of neuronal pathways. It could therefore be interesting to study behavioral changes induced by chronic local stimulation of neuropeptide receptors. With this aim poly(lactide-co-glycolide) microspheres loaded with a highly potent, peptidase-resistant, cholecystokinin (CCK)-B-selective CCK peptidomimetic agonist (pBC 264) were prepared by a water in oil in water emulsion solvent evaporation method and stereotaxically implanted into the anterior part of the rat nucleus accumbens. Two different kinds of loaded polymeric microspheres differing only by the stabilizing agent [ovalbumin (OVA) or Pluronic F 68] added to the inner emulsion were used. The histological and behavioral studies done 24 h and 8 days after implantation of nonloaded microspheres in the nucleus accumbens indicated that the microspheres were well tolerated. The in vivo release of the selective CCK-B agonist pBC 264 (associated with a tracer dose of [3 H]pBC 264) from microspheres prepared with OVA was very fast (92% after 6 h), whereas only 26% (88 pmol) of pBC 264 was released from the formulation with Pluronic F 68 after 24 h. Eight days after implantation 36% of pBC 264 had diffused from the microspheres, and 8% (∼30 pmol) was still present in the brain concentrated around the site of administration. In all cases the released material was found to correspond to intact pBC 264, thus demonstrating the possibility of obtaining a slow controlled release of peptide in vivo. This method opens up interesting perspectives to study the long-term effects of neuropeptides. 相似文献
993.
E. R. R. Rochedo L. F. C. Conti H. G. Paretzke 《Radiation and environmental biophysics》1996,35(4):243-261
The structure and mathematical model of PARATI, a detailed computer programme developed for the assessment of the radiological
consequences of an accidental contamination of urban areas, is described with respect to the scenarios used for the estimation
of exposure fields in a village or town, the models for the initial and secondary contamination with the radionuclide 137Cs, the concepts for calculating the resulting radiation exposures and the changes with time of the contamination and radiation
fields. Kerma rates at various locations in tropical urban areas are given, and the contribution of different contaminated
surfaces to these rates after dry or wet deposition are discussed.
Received: 12 April 1996 / Accepted in revised form: 30 August 1996 相似文献
994.
The involvement of gibberellins in the control of flowering of sunflower was studied by direct application of GA3 to the apex of the plants, analysis of the endogenous levels of gibberellin-like substances at different plant ages, and indirectly by the application of paclobutrazol, an inhibitor of gibberellin synthesis. GA3 speeded-up flower initiation and floral apex development. The time of GA3 application was more critical than the amount of GA3 applied. The endogenous levels of gibberellin-like compounds increased significantly by day 15 after sowing. The application of paclobutrazol markedly delayed floral initiation and this effect was also depedent on plant age. Both GA3 and paclobutrazol had their greatest effects between 10 and 20 days after sowing suggesting that an increase in gibberellins in that time period plays a role in floral initiation. 相似文献
995.
996.
997.
998.
M. G. Rosenblum Lawrence Cheung S. K. Kim Kalpana Mujoo Nicholas J. Donato James L. Murray 《Cancer immunology, immunotherapy : CII》1996,42(2):115-121
The development of cellular resistance to immunotoxins has been demonstrated in a variety of models and can involve a number
of mechanisms. For the present study, an immunotoxin was utilized composed of an antimelanoma antibody ZME-018 recognizing
a 240-kDa surface glycoprotein (gp 240) and the plant toxin gelonin. Human melanoma cells (A375-M) were grown in the presence
of increasing amounts of ZME-gelonin and a clonal variant (A-375-ZR) was developed that was 100-fold resistant to ZME-gelonin
compared to parental cells. Scatchard analysis showed that the A375-M parental cells had 260×103 ZME-gelonin-binding sites/cell with relatively low affinity (5 nM). In contrast, resistant A375-ZR cells demonstrated a reduced
number of low-affinity sites (160×103/cell), but showed a small number (47×103) of higher-affinity sites (0.8 nM). Internalization rates and degradation rates of 125I-labeled ZME-gelonin were identical in both the parental and resistant cells. A375-ZR cells were found to be more resistant
to vincristine and doxorubicin than were parental cells. Both cell lines were almost equally sensitive to native gelonin,
5-fluorouracil (5-FU), cisplatin, melphalan, carmustine, interferon γ (IFNγ) and IFNα. In addition, both cell lines were equally
sensitive to another gelonin-antibody conjugate that binds to cell-surface, GD2 (antibody 14G2A). However, resistant cells were twice as sensitive to the cytotoxic effects of etoposide than were parental cells. Finally,
a variety of agents were tested in combination with ZME-gelonin against A375-ZR cells in an attempt to identify agents to
augment immunotoxin cytotoxic effects against resistant cells. The agents 5-FU, cisplatin, IFNγ, IFNα, and etoposide were
the most effective in augmenting the cytotoxicity of ZME-gelonin against resistant cells. These studies suggest that development
of resistance to one immunotoxin does not cause development of cross-resistance to other gelonin immunotoxins. Further, specific
biological response modifiers and chemotherapeutic agents may be effective in augmenting the effectiveness of immunotoxins
and specifically targeting or reducing the emergence of immunotoxin-resistant cells.
Received: 15 March 1995 / Accepted: 28 November 1995 相似文献
999.
1000.
T. Hannedouche P. Landais B. Goldfarb N. el Esper A. Fournier M. Godin D. Durand J. Chanard F. Mignon J. M. Suo et al. 《BMJ (Clinical research ed.)》1994,309(6958):833-837
OBJECTIVE--To compare the ability of angiotensin converting enzyme inhibitors and beta blockers to slow the development of end stage renal failure in non-diabetic patients with chronic renal failure. DESIGN--Open randomised multicentre trial with three year follow up. SETTING--Outpatient departments of six French hospitals. PATIENTS--100 hypertensive patients with chronic renal failure (initial serum creatinine 200-400 mumol/l. 52 randomised to enalapril and 48 to beta blockers (conventional treatment). INTERVENTIONS--Enalapril or beta blocker was combined with frusemide and, if necessary, a calcium blocker or centrally acting drug in patients whose diastolic pressure remained above 90 mm Hg. RESULTS--17 patients receiving conventional treatment and 10 receiving enalapril developed end stage renal failure. The cumulative renal survival rate was significantly better in the enalapril group than in the conventional group (P < 0.05). The slope of the reciprocal serum creatinine concentration was steeper in the conventionally treated patients (-6.89 x 10(-5)l/mumol/month) than in the enalapril group (-4.17 x 10(-5)l/mumol/month; P < 0.05). No difference in blood pressure was found between groups. CONCLUSION--In hypertensive patients with chronic renal failure enalapril slows progression towards end stage renal failure compared with beta blockers. This effect was probably not mediated through controlling blood pressure. 相似文献