Hot spot mapping of protein surfaces with TEMPOL: Bovine pancreatic RNase A as a model system

2017 ◽  
Vol 1865 (2) ◽  
pp. 201-207 ◽  
Author(s):  
Neri Niccolai ◽  
Edoardo Morandi ◽  
Simone Gardini ◽  
Valentino Costabile ◽  
Roberta Spadaccini ◽  
...  
Keyword(s):  
Hot Spot ◽  
Model System ◽  
Rnase A ◽  
Protein Surfaces ◽  
Pancreatic Rnase ◽  
Spot Mapping

Crowdsourcing photograph locations for debris flow hot spot mapping

2017 ◽  
Vol 90 (3) ◽  
pp. 1259-1276 ◽  
Author(s):  
Hone-Jay Chu ◽  
Yi-Chin Chen
Keyword(s):  
Debris Flow ◽  
Hot Spot ◽  
Spot Mapping

scholarly journals Isolation of the Ascobolus immersus spore color gene b2 and study in single cells of gene silencing by methylation induced premeiotically.

Genetics ◽  
1995 ◽  
Vol 141 (4) ◽  
pp. 1299-1314
Author(s):  
V Colot ◽  
J L Rossignol
Keyword(s):  
Gene Silencing ◽  
Physical Mapping ◽  
Meiotic Recombination ◽  
Hot Spot ◽  
Single Cells ◽  
Model System ◽  
Ascobolus Immersus ◽  
First Time ◽  
Mutant Rescue ◽  
Color Gene

Abstract The ascomycete Ascobolus immersus has been extensively used as a model system for the genetic study of meiotic recombination. More recently, an epigenetic process, known as methylation induced premeiotically (MIP), that acts on duplicated sequences has been discovered in A. immersus and has raised a new interest in this fungus. To try and extend these studies, we have now cloned the A. immersus spore color gene b2, a well characterized recombination hot-spot. Isolation of the whole gene was verified by physical mapping of four large b2 alterations, followed by transformation and mutant rescue of a null b2 allele. Transformation was also used to duplicate b2 and subject it to MIP. As a result, we were able for the first time to observe gene silencing as early as just after meiosis and in single cells. Furthermore, we have found evidence for a modulating effect of MIP on b2 expression, depending on the region of the gene that is duplicated and hence subjected to MIP.

Turbulence permitting air pollution simulation for the Stuttgart metropolitan area - A winter case study

2021 ◽  
Author(s):  
Thomas Schwitalla ◽  
Kirsten Warrach-Sagi ◽  
Hans-Stefan Bauer ◽  
Volker Wulfmeyer
Keyword(s):  
Air Pollution ◽  
Urban Space ◽  
Metropolitan Area ◽  
Complex Dynamics ◽  
Hot Spot ◽  
Vertical Mixing ◽  
Urban Canopy ◽  
Model System ◽  
Emission Data

<p>Currently a strong discussion is ongoing in Germany and Europe whether to ban vehicles from downtown areas in order to lower particle concentrations of e.g. PM<sub>10</sub> and NO<sub>2</sub>. As often only few measurements exist inside city centers, little to nothing is known about the horizontal and vertical distributions of air pollutants. Within the EU demonstration project Open Forecast (https://open-forecast.eu/), we applied the WRF-Chem model system version 4.0.3 in order to close this knowledge gap. We zoom in the urban area of Stuttgart, a hot spot of air pollution in Germany. The outermost domain with convection-permitting resolution of 1.25 km encompasses parts of Central Europe in order to provide boundary conditions for the inner two domains.</p><p>The model system was improved in many ways, e.g., with respect to the representation of land cover, urban canopy, and soil properties, which turned out to be key for an acceptable performance. Furthermore, we developed a sophisticated infrastructure to ingest the required high-resolution emission data, which turned out to be very challenging.</p><p>We show that this model approach is likely the best means to understand and to predict air pollution, as the distribution of their constituents depends strongly and simultaneously on the vertical mixing by turbulence, the mesoscale circulation in the complex urban environment, and orographic environment.</p><p>The model system was operated and investigated for a case study of January 21, 2019 during which an alert with respect to the exceedance of PM<sub>10</sub> was issued. We present the simulations of meteorological variables as well as PM<sub>10</sub> and NO<sub>2</sub> and show the complexity of their distribution in the nighttime stable and daytime shallow boundary layer in dependence of the temporal variability of the traffic in the Stuttgart metropolitan area.</p><p>To the best of our knowledge, the results reveal for the first time the complex dynamics of air pollution in complex urban space of Stuttgart at a very high spatial and temporal resolution that cannot currently be achieved with measurements.</p>

Immunological studies of ribonuclease C from human urine.

1981 ◽  
Vol 27 (8) ◽  
pp. 1362-1367 ◽  
Author(s):  
J W Cranston ◽  
H C Hoover ◽  
E R Crisp
Keyword(s):  
Pancreatic Carcinoma ◽  
Human Urine ◽  
Rnase A ◽  
Serum Samples ◽  
Positive Correlation ◽  
Alternative Method ◽  
Normal Individuals ◽  
Human Sera ◽  
Pancreatic Rnase

Abstract Antiserum to human urinary RNase C [ribonuclease (pancreatic), EC 3.1.27.5], developed in rabbits, was used to characterize this enzyme through studies of inhibition of RNase C-catalyzed poly(C) hydrolysis and of competition in a RIA. By either assay, the antiserum failed to cross react with human urinary RNase U (EC 3.1.27.-) or bovine pancreatic RNase A (EC 3.1.27.5). RNase C is immunologically identical to the poly(C)-active RNase in various human sera, including samples obtained from normal individuals, patients with pancreatic carcinoma, pancreatitis, or other malignant and nonmalignant diseases. This conclusion is based on the finding of superimposable antibody dose-inhibition curves for poly(C) hydrolysis and parallel competition RIA curves for RNase C and the various sera. There was a positive correlation (r = 0.73; p less than 0.001) between concentrations of RNase C as determined by poly(C) hydrolysis and competition RIA in serum samples from 102 patients. Therefore, the latter technique provides al alternative method for measuring RNase C in sera.

The Utilization of Crime Hot Spot Mapping: Košice — a Case Study

2015 ◽  
Vol 7 (2) ◽  
pp. 43-52
Author(s):  
Monika Blišťanová
Keyword(s):  
Hot Spot ◽  
Spot Mapping

Partial purification and characterization of the components of the neutral ribonucleage II – inhibitor system of normal and dystrophic mouse skeletal muscle

1981 ◽  
Vol 59 (3) ◽  
pp. 220-231 ◽  
Author(s):  
Brian W. Little ◽  
William L. Meyer
Keyword(s):  
Molecular Weight ◽  
Rnase A ◽  
Free Form ◽  
Partial Purification ◽  
Mouse Muscle ◽  
Molecular Weights ◽  
Dystrophic Mouse ◽  
Inhibitor System ◽  
Pancreatic Rnase ◽  
Rnase Ii

The complexes between a proteinaceous inhibitor and neutral ribonuclease II (EC 3.1.27.5) purified from low ionic strength extracts of normal and dystrophic mouse muscle are essentially indistinguishable in (a) purification behavior, (b) apparent molecular weights of approximately 50 000, (c) thermal denaturation (50% loss of activity in 5 min at 73.5 °C), (d) isoelectric points (pH 4.8), and (e) procedures for reversible resolution into free inhibitor and free RNase II.The free RNase II species are also similar whether obtained by resolution of the purified complexes or by direct isolation of free enzyme from dystrophic muscle. All have apparent molecular weights of 11 500 compared with 13 700 for bovine pancreatic RNase A; all retain 80% of activity after 5 min at 95 °C. The active RNases II prepared directly from muscle, by resolution of inhibitor complexes or by organic mercurial treatment of the inhibitor complexes, all have identical pH–activity profiles in 200 mM KCl with an optimum near pH 7.0. In comparison RNase A has an optimum pH near 7.5 and its activity decreases more rapidly as KCl concentration is increased above 50 mM KCl.RNase II inhibitor obtained by resolution of the purified complexes or by direct isolation in the free form from normal muscle extracts has an apparent molecular weight of 42 000 and is very sensitive to heat; it loses all activity at 40 °C in 5 min.These studies (a) provide methods for obtaining useful amounts of the components of the neutral RNase II – inhibitor system from muscle, (b) provide the first method reported for the reversible resolution of RNase II – inhibitor complexes, (c) fail to show any distinct difference between corresponding components of the system from normal and dystrophic mice, (d) establish interesting differences between the apparently homologous enzymes, murine muscle neutral RNase II, and bovine pancreatic RNase A, and (e) provide a substantially lower molecular weight estimate for RNase II inhibitor from muscle than has been reported for the inhibitor from liver, kidney, and placenta.

Enzymatic and Physical Studies on the Triplex dTn∙dAn∙rUn

1973 ◽  
Vol 51 (4) ◽  
pp. 436-449 ◽  
Author(s):  
Nancy L. Murray ◽  
A. Richard Morgan
Keyword(s):  
Buoyant Density ◽  
Minor Groove ◽  
Model System ◽  
Rational Approach ◽  
Major Groove ◽  
Dna And Rna ◽  
Pancreatic Rnase ◽  
Mixing Curves ◽  
Transcription And Replication

The triplex dTn∙dAn∙rUn was studied as a model system both enzymatically and physically with a view that a rational approach for attempting to isolate triplexes from in vivo situations might emerge. The triplex was characterized by mixing curves and by its equilibrium buoyant density. In 5 mM Na phosphate, pH 7.3, for KCl concentrations below 0.4 M the triplex dissociated into dTn∙dAn + rUn, dissociation being complete at about 0.3 M KCl. If MgCl2 replaced KCl a strongly cooperative dissociation occurred at 1 mM MgCl2. Whereas with alkaline titration of dTn∙dAn∙rUn, rUn was dissociated first followed by dTn∙dAn, acidic titration resulted in the whole triplex dissociating together. Both transcription and replication of dTn∙dAn were strongly inhibited by formation of a triplex. The DNA and RNA moieties in the triplex are somewhat protected against DNase I and pancreatic RNase degradation, when compared with duplex dTn∙dAn or rUn. Spermine binds equally well to dTn∙dAn and dTn∙dAn∙rUn which is consistent with spermine binding in the minor groove and RNA in the major groove of DNA.

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