scholarly journals Problems associated with the assay of arylsulphatases A and B of rat tissues

1973 ◽  
Vol 134 (1) ◽  
pp. 183-190 ◽  
Author(s):  
Mark Worwood ◽  
Kenneth S. Dodgson ◽  
Gary E. R. Hook ◽  
Frederick A. Rose
Keyword(s):  
Rat Liver ◽  
Liver Enzymes ◽  
Gel Filtration ◽  
Precise Determination ◽  
Approximate Determination ◽  
Rat Tissues ◽  
Human Tissues ◽  
Separation And Purification ◽  
The Individual

A method for the separation and purification of rat liver arylsulphatases A and B by gel filtration on Sephadex G-200 is described. The properties of the A enzyme and its molecular weight are similar to those of the corresponding ox liver enzyme. The B enzymes were found to be dissimilar. The method already developed for the assay of the corresponding enzymes from human tissues was shown to be unsuitable for the assay of the enzymes of rat tissues. A method of assay was developed which permits an approximate determination of the individual rat liver enzymes in a mixture of the two, but precise determination requires prior separation of the enzymes by gel-filtration chromatography.

scholarly journals Properties of subunits of the multicatalytic proteinase complex revealed by the use of subunit-specific antibodies

1991 ◽  
Vol 278 (1) ◽  
pp. 171-177 ◽  
Author(s):  
A J Rivett ◽  
S T Sweeney
Keyword(s):  
Molecular Mass ◽  
Rat Liver ◽  
Gel Filtration ◽  
Immunoblot Analysis ◽  
Rat Tissues ◽  
Specific Antibodies ◽  
Multicatalytic Proteinase ◽  
Cell Extracts ◽  
Liver And Kidney ◽  
Lysosomal Proteinase

The multicatalytic proteinase (MCP) is a high-molecular-mass non-lysosomal proteinase that gives rise to a characteristic pattern of bands of molecular mass 22-34 kDa on SDS/PAGE gels. Isoelectric-focusing gels of the enzyme purified from rat liver show 16 bands with isoelectric points in the range of pH 5-8.5. Two-dimensional PAGE gels reveal that there are more than the previously reported 13 polypeptides associated with the MCP from rat liver and show a pattern of 15-20 major spots and several minor ones, similar to that of MCP isolated from some other sources. Possible relationships between the different polypeptides were investigated by immunoblot analysis of electrophoretically purified proteinase subunits with affinity-purified subunit-specific antibodies as well as antibodies raised against individual denatured subunits of the complex. The results demonstrate that many of the major polypeptide components of the MCP complex are antigenically distinct. Moreover comparison of immunoreactive material in crude cell extracts with that in purified MCP preparations has shown that the polypeptides are not derived from a smaller number of higher-molecular-mass subunits. Also, individual subunits have the same apparent molecular mass in a variety of rat tissues, suggesting close similarity between MCPs of different tissues. The highest concentrations of MCP subunits occur in liver and kidney. Gel-filtration analysis of crude extracts has demonstrated that MCP polypeptides are also associated with a higher-molecular-mass complex, which may be the 26 S proteinase that has been implicated in the degradation of ubiquitin-protein conjugates.

Fuzzy Quality Measures for Use in a Hierarchical Fuzzy Controller

2000 ◽  
Author(s):  
Alan Carlson ◽  
Dean B. Edwards ◽  
Michael J. Anderson
Keyword(s):  
Fuzzy Controller ◽  
Structure Optimization ◽  
Quality Measures ◽  
Quality Measure ◽  
Approximate Determination ◽  
Rule Base ◽  
System Parameters ◽  
The Individual ◽  
Monolithic Structure

Abstract This paper presents a control strategy that uses a hierarchical structure to arbitrate between recommendations from lower level modules. The lower level modules represent lower level tasks or behaviors. Each lower level module provides its own control recommendation based on its limited perception of the environment. We introduce the concept of a fuzzy quality measure that may be used by the hierarchical controller to determine how best to fuse the individual recommendations. The Quality Measure provides an approximate determination of each control recommendations potential value. The hierarchical partitioning reduces the cardinality of the rule base and decreases the number of system parameters, as compared to a monolithic structure. Optimization of the reduced parameter set is simpler and requires less time.

scholarly journals Biological Relevance of a Stable Biochemical Interaction between the Tombusvirus-Encoded P19 and Short Interfering RNAs

2006 ◽  
Vol 80 (6) ◽  
pp. 3000-3008 ◽  
Author(s):  
Rustem Omarov ◽  
Kim Sparks ◽  
Lindsay Smith ◽  
Jelena Zindovic ◽  
Herman B. Scholthof
Keyword(s):  
Gel Filtration ◽  
Rna Degradation ◽  
Precise Determination ◽  
Viral Rna ◽  
Model Organisms ◽  
Wild Type ◽  
Viral Spread ◽  
Rnai Suppressor ◽  
Biochemical Interaction

ABSTRACT The Tomato bushy stunt virus (TBSV)-encoded p19 protein (P19) is widely used as a robust tool to suppress RNA interference (RNAi) in various model organisms. P19 dimers appropriate 21-nucleotide (nt) duplex short interfering RNAs (siRNAs) generated by Dicer presumably to prevent programming of the RNA-induced silencing complex (RISC). In the context of virus infection, this model predicts that P19 mutants compromised for siRNA binding cannot prevent RISC-mediated degradation of TBSV RNA and thus reduce viral pathogenicity. To test this, we used P19/43 (R→W), which is less pathogenic than wild-type P19 (wtP19), and P19/75-78 (RR→GG), with pathogenicity properties (i.e., viral spread and symptom induction) comparable to those of a P19-null mutant. We demonstrate that P19/43 still suppresses RNAi-mediated viral RNA degradation in infected Nicotiana benthamiana, while P19/75-78 is unable to prevent this clearance of viral RNA, leading to an irreversible recovery phenotype. Gel filtration and immunoprecipitation assays show that at the onset of the infection, wtP19, P19/43, and P19/75-78 readily accumulate, and they form dimers. The wtP19 is stably associated with duplex ∼21-nt TBSV siRNAs, while P19/75-78 does not bind these molecules, and the electrostatic interaction of P19/43 with siRNAs is perturbed for ∼21-nt duplexes but not for longer siRNAs. This is the first clear demonstration of a direct correlation between a novel structurally orchestrated siRNA binding of an RNAi suppressor and its roles in viral pathogenesis. The findings should be particularly valuable for the RNAi field in general because the P19 mutants enable precise determination of siRNA appropriation effects.

Assessing Alzheimer Severity With a Global Clinical Scale

1992 ◽  
Vol 4 (1) ◽  
pp. 55-74 ◽  
Author(s):  
J. Wesson Ashford ◽  
Vinod Kumar ◽  
Mary Barringer ◽  
Marion Becker ◽  
Jami Bice ◽  
...  
Keyword(s):  
Rating Scale ◽  
Disease Patient ◽  
Nursing Home Residents ◽  
Precise Determination ◽  
Clinical Dementia Rating ◽  
Dementia Severity ◽  
Accurate Indication ◽  
Diagnosis Of Dementia ◽  
The Individual

Diagnosis of dementia needs to be complemented by precise determination of disease severity across the broad spectrum of disease progression. The Mini-Mental State Exam (MMS), the Activities-of-Daily-Living assessment (ADL) and the Clinical Dementia Rating scale (CDR) were modified for direct comparability and administred to 112 outpatients and 45 nursing home residents with a range of dementia severity from mild to profound. The scales showed the highest correlations for the probable Alzheimer's disease patient group (62) (Global Assessment of Dementia; GAD vs. ADL: r = 0.91; Extended Mini-Mental Assessment; EMA vs. GAD: r = 0.91; ADL vs. EMA: r = 0.86). For these patients, scores on the individual scales tended to be similar. Disparity among the three scores for individual cases was associated with the presence of comorbidities. The high correlations and correspondence among these scales demonstrate their reliability, validity, and utility in the assessment of dementia severity. The use of an average of these measures, with their increased precision, may give a more accurate indication of dementia severity over a broader range of impairment.

scholarly journals Simple spectrophotometric methods for the determination of amlodipine and atorvastatin in bulk and tablets

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Imad Osman Abu Reid ◽  
Hind Mohamed Farid ◽  
Sara Osman Eltayeb
Keyword(s):  
Individual Component ◽  
Analytical Procedure ◽  
Precise Determination ◽  
Analytical Challenge ◽  
Statistical Comparison ◽  
Spectrophotometric Methods ◽  
Relative Standard ◽  
Significant Difference ◽  
The Individual

Abstract Background Simultaneous spectrophotometric determination of samples containing more than one analyte presents analytical challenge; the choice of an analytical procedure is strictly related to the extent of overlapping between the individual absorption peaks of these components; if the absorption peaks are satisfactorily resolved, the determination is not problematic, but if the individual component signals are partly or totally overlapped, then powerful techniques are needed. Combined amlodipine and atorvastatin are typical example where special techniques are needed to resolve bands overlapping. Results Application of multiwavelength regression and absorbance factor methods to the analysis of atorvastatin and amlodipine combination proved to be satisfactorily capable of accurate and precise determination of the two analytes. The two methods recoveries were very close to the expected analytes concentrations, and the precision of the methods was < 2% relative standard deviation. Statistical comparison indicated that there is no significant difference between the assay results obtained by the two method as the calculated t values 0.91 and 1.13 for amlodipine and atorvastatin, respectively, were less than the tabulated t value 2.23 at 95% confidence level. Conclusion The proposed methods are accurate, precise, simple and inexpensive. They can be applied successfully to the analysis of the two drugs in combined dosage form.

BINDING OF CORTICOSTERONE IN RAT LIVER

1970 ◽  
Vol 47 (2) ◽  
pp. 149-158 ◽  
Author(s):  
R. S. SNART ◽  
N. N. SANYAL ◽  
M. K. AGARWAL
Keyword(s):  
Rat Liver ◽  
Binding Sites ◽  
Microsomal Fraction ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Gel Filtration ◽  
Nuclear Fraction ◽  
Binding Characteristics ◽  
Mole Percentage

SUMMARY The binding characteristics of corticosterone by rat liver were studied by a displaceable binding technique. The binding of corticosterone to protein fractionated by gel filtration and density gradient centrifugation has been carried out as a preliminary determination of the nature of the binding sites. The results were analysed and showed three types of binding sites for corticosterone with the characteristic association constants at 0° of K1 = 1·2 × 1010, K2 = 1 × 108 and K3 = 1 × 104 1./mole. Percentage displacement of corticosterone from the nuclear fraction did not differ significantly from that from tissue or the mitochondrial-microsomal fraction. The K1 and K2 sites persisted in separated buffer-soluble fractions but were destroyed on mild heating leaving only the K3 sites.

scholarly journals An approximate determination of the boiling points of metals

1909 ◽  
Vol 82 (556) ◽  
pp. 396-408 ◽  
Keyword(s):  
High Temperatures ◽  
Electric Heating ◽  
Approximate Determination ◽  
Published Data ◽  
Arc Furnace ◽  
Boiling Points ◽  
Loss Of Weight ◽  
General Investigation ◽  
The Individual

Despite the facility with which high temperatures can be reached and maintained constant by means of electric heating, no general investigation of the boiling points of the metals has yet been carried out, and such information as is available has in many cases been obtained by considerable extrapolation. Moreover, the published data are remarkably discordant, as will be seen from the individual results quoted below. In the course of an extended experimental investigation, H. Moissan has made observations on the vaporisation of metals at high temperatures by observing the loss of weight of a considerable mass of metal heated for definite periods of time in his arc furnace. O. P. Watts has attempted to deduce from these experiments approximate values for the boiling points of the metals. In addition to the uncertainty due to the fact that many metals possess a high vapour tension at temperatures much below their actual boiling points, considerable errors are caused by the fact that Moissan does not appear to have measured the expenditure of energy in the furnace, which varies widely according to the conductivity of the vapours surrounding the arc. Also, in many of his experiments the temperature of ebullition must have been altogether modified by carburisation.

scholarly journals Die Trennung der Hämolymphe-Proteine bei Vorpuppen und Puppen der Galleria mellonella L. mittels Gel-Filtration auf Sephadex G-200, Bestimmung ihrer isoelektrischen Punkte und ihrer Molekulargewichte / Separation of Haemolymph Proteins of Prepuae and Pupae of Galleria mellonella L. by means of Gel Filtration on Sephadex 200, Determination of their Isoelectric Points and their Molecular Weights

1969 ◽  
Vol 24 (6) ◽  
pp. 732-740 ◽  
Author(s):  
Milan Marek
Keyword(s):  
Gel Electrophoresis ◽  
Galleria Mellonella ◽  
Gel Filtration ◽  
Starch Gel Electrophoresis ◽  
Molecular Weights ◽  
Isoelectric Points ◽  
Starch Gel ◽  
The Individual ◽  
Naphthyl Acetate

Gel filtration on Sephadex G-200 was carried out on haemolymph proteins of prepupae. ligatured prepupae, male and female pupae and cooled pupae of Galleria mellonella L.The proteins were separated into two main fractions. The esterase activity of the eluated haemolymph was determined by means of beta-naphthyl acetate after filtration.After elution the samples were condensed and additionally separated on horizontal starch-gel electrophoresis.The “cooling protein” of pupae and the “ligature protein” of ligatured larvae of Galleria mellonella were shown by means of starch-gel electrophoresis to be new proteins, so far not described.The isoelectric point and molecular weight were determined for the individual protein fractions.They were then stained with amido black 10 B for the proof of proteins, and with alpha-naphthyl butyrate with Fast-Blue BB salt for the identification of esterases.

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