scholarly journals Use of procainamide gels in the purification of human and horse serum cholinesterases

1983 ◽  
Vol 211 (1) ◽  
pp. 243-250 ◽  
Author(s):  
J S Ralston ◽  
A R Main ◽  
B F Kilpatrick ◽  
A L Chasson
Keyword(s):  
Human Serum ◽  
Large Scale ◽  
Small Volume ◽  
Horse Serum ◽  
Retention Capacity ◽  
Relative Binding ◽  
Human Enzyme ◽  
Sepharose 4B ◽  
Human Cholinesterase

Two large-scale methods based primarily on the use of procainamide-Sepharose gels were developed for the purification of horse and human serum non-specific cholinesterases. With method I, the procainamide-Sepharose 4B gel was used in the first step to handle large volumes of serum. With method II, the procainamide-Sepharose 4B gel was used in the final step to obtain pure enzyme. Although both methods gave electrophoretically pure cholinesterase preparations in good yields, they were significantly more efficient at purifying the horse enzyme than the human enzyme. To study this problem, the relative binding of human and horse cholinesterases to procainamide-, methylacridinium (MAC)-, m-trimethylammoniophenyl (m-PTA)- and p-trimethylammoniophenyl (p-PTA)-Sepharose 4B gels were measured, by using two approaches. In one, binding was measured by a procedure involving equilibration of pure cholinesterase in a small volume of diluted gel slurry (4%, v/v). A partially purified preparation of Electrophorus acetylcholinesterase was included. Pure human cholinesterase bound consistently more tightly to each of the gels than did horse cholinesterase, and the acetylcholinesterase appeared to bind the gels 10-100 times more tightly than did the non-specific cholinesterases. The order of binding for the cholinesterases, beginning with the tightest, was: procainamide-Sepharose 4B, MAC-Sepharose 4B, p-PTA-Sepharose 4B and m-PTA-Sepharose 4B. For the acetylcholinesterase the order was: MAC-Sepharose 4B, procainamide-Sepharose 4B, p-PTA-Sepharose 4B and m-PTA-Sepharose 4B. The second approach involved passing native sera or partially purified sera fractions through 1 ml test columns of each of the four affinity gels to determine their retention capacity for the cholinesterases. With these impure samples, the MAC-Sepharose 4B gels proved superior to the procainamide-Sepharose 4B gels at retaining human cholinesterase, but the opposite was true for the horse cholinesterase.

Effective pollutant retention capacity of permeable pavements for infiltrated road runoffs determined by laboratory tests

2005 ◽  
Vol 51 (2) ◽  
pp. 37-45 ◽  
Author(s):  
S. Fach ◽  
W.F. Geiger
Keyword(s):  
Large Scale ◽  
Urban Runoff ◽  
Assessment Procedure ◽  
Retention Capacity ◽  
Test Installation ◽  
Permeable Pavements ◽  
Batch Tests ◽  
Different Types ◽  
Stormwater Infiltration

The infiltration of urban runoff always implies an entrance of pollutants into the soil and ground water. Due to legal regulations in many communes there is no longer any permission needed for stormwater infiltration, if administrative regulations and the requirements of standards are observed. The results of a research project carried out under the heading “Development of an assessment procedure for permeable pavements” show, that the pollutant retention capacity of permeable pavements varies considerably, depending on the material and the specific reactive surface. The objective of the study was to work out recommendations of suitable permeable pavements for different types of urban runoff. Selected data about the quality of urban runoff was compiled into a runoff matrix, which was used for defining characteristic dilutions. In batch tests, the material of the infiltration devices is penetrated with the dilutions. A test installation in large scale is used to calibrate the sorption coefficients derived from the batch experiment.

scholarly journals Esterases in Serum-Containing Growth Media Counteract Chloramphenicol Acetyltransferase Activity In Vitro

1999 ◽  
Vol 43 (3) ◽  
pp. 655-660 ◽  
Author(s):  
Charles D. Sohaskey ◽  
Alan G. Barbour
Keyword(s):  
Human Serum ◽  
Horse Serum ◽  
Chloramphenicol Acetyltransferase ◽  
Rabbit Serum ◽  
Growth Media ◽  
Chloramphenicol Resistance ◽  
E Coli ◽  
Cell Extracts

ABSTRACT The spirochete Borrelia burgdorferi was unexpectedly found to be as susceptible to diacetyl chloramphenicol, the product of the enzyme chloramphenicol acetyltransferase, as it was to chloramphenicol itself. The susceptibilities of Escherichia coli and Bacillus subtilis, as well as that ofB. burgdorferi, to diacetyl chloramphenicol were then assayed in different media. All three species were susceptible to diacetyl chloramphenicol when growth media were supplemented with rabbit serum or, to a lesser extent, human serum. Susceptibility ofE. coli and B. subtilis to diacetyl chloramphenicol was not observed in the absence of serum, when horse serum was used, or when the rabbit or human serum was heated first. In the presence of 10% rabbit serum, a strain of E. colibearing the chloramphenicol acetyltransferase (cat) gene had a fourfold-lower resistance to chloramphenicol than in the absence of serum. A plate bioassay for chloramphenicol activity showed the conversion by rabbit, mouse, and human sera but not bacterial cell extracts or heated serum of diacetyl chloramphenicol to an inhibitory compound. Deacetylation of acetyl chloramphenicol by serum components was demonstrated by using fluorescent substrates and thin-layer chromatography. These studies indicate that esterases of serum can convert diacetyl chloramphenicol back to an active antibiotic, and thus, in vitro findings may not accurately reflect the level of chloramphenicol resistance by cat-bearing bacteria in vivo.

Comparison of effects of human serum and horse serum onin vitrosusceptibility testing of echinocandins

2013 ◽  
Vol 26 (1) ◽  
pp. 62-63 ◽  
Author(s):  
Anna Prigitano ◽  
Maria Carmela Esposto ◽  
Anna Maria Tortorano
Keyword(s):  
Human Serum ◽  
Horse Serum

Gold–copper–bismuth mineralization in hedenbergitic skarn, Tombstone Mountains, Yukon

1987 ◽  
Vol 24 (12) ◽  
pp. 2362-2372 ◽  
Author(s):  
Isobel J. Brown ◽  
Bruce E. Nesbitt
Keyword(s):  
Large Scale ◽  
Small Volume ◽  
Gold Mineralization ◽  
Rock Interaction ◽  
The North ◽  
Mineral Assemblages ◽  
Intrusive Rocks ◽  
Porosity And Permeability ◽  
Relationship Of ◽  
The Relationship

Gold mineralization on the Marn property, Yukon, occurs in two pyroxene skarn bodies, which are adjacent to the Mount Brenner Stock in the Ogilvie Mountains. The skarns are separated by a 600 m wide monzonite intrusion and show contrasting mineralogical and geochemical characteristics in addition to quite different metal values. Significant but uneconomic Au, Ag, W, and Cu mineralization is found in skarn on the north side of the intrusion, while very low Au grades (0.052 g/t) occur at the southern contact. The mineral assemblages of both skarns are dominated by iron-rich pyroxenes. The iron content of the pyroxenes varies between Hd40 and Hd80 in the northern location and Hd80 and Hd100 in the southern skarn. A well-developed sequence of retrograde alteration affected only the northern skarn. This was probably the result of porosity and permeability differences in the early, high-temperature pyroxene skarn, which permitted greater fluid–rock interaction in the northern skarn during cooling. A small volume of diopsidic, aluminous, wollastonite-bearing skarn occurs in both the northern and southern localities. The relationship of this type of skarn to the hedenbergitic skarn is ambiguous, since there is no large-scale mineralogical zoning. The Marn is similar to hedenbergitic, auriferous skarns of Japan, where the oxidation state of the intrusive rocks is believed to be the controlling factor in the development of skarn mineralogy.

scholarly journals OBSERVATIONS ON THE PRESENCE OF A TOXIC SUBSTANCE IN THE BLOOD AND URINE OF PATIENTS WITH SCARLET FEVER

1924 ◽  
Vol 40 (3) ◽  
pp. 381-395 ◽  
Author(s):  
James D. Trask ◽  
Francis G. Blake
Keyword(s):  
Human Serum ◽  
Scarlet Fever ◽  
Horse Serum ◽  
Toxic Substance ◽  
Local Infection ◽  
Hemolytic Streptococcus ◽  
Normal Horse Serum

A series of observations on the blood of patients acutely ill with scarlet fever has shown that a toxic substance can be demonstrated in the serum by means of intracutaneous injections of the serum in persons who have not had scarlet fever and whose serums fail to blanch the rash in scarlet fever. The reaction caused by this substance consists of a bright red local erythema, varying from 20 to 70 mm. in diameter, of 1 to 4 days duration. The severer reactions are moderately indurated and tender, and are followed bypigmentation and desquamation. Control injections in persons whose serums blanch the rash in scarlet fever cause no reaction. The toxic substance is not neutralized by mixture with a human serum which gives a negative blanching test but is readily neutralized by a human serum which gives a positive blanching test. It is not neutralized by normal horse serum, but is completely neutralized by Dochez's scarlatinal antistreptococcic serum. In a limited number of observations on the urine of patients with scarlet fever a similar toxic substance has been found in two out of five cases studied. Since the toxic substance described appears to resemble the toxic substance found in the filtrates of scarlatinal hemolytic streptococcus cultures by Dick and Dick and since it is neutralized not only by a blanching human serum but also by Dochez's scarlatinal antistreptococcic horse serum, the experiments reported support the conception that scarlet fever is a local infection of the throat by a particular type of Streptococcus hæmolyticus capable of producing a toxin which is absorbed and is the cause of the general manifestations of the disease.

scholarly journals Demonstration of alpha 1-antitrypsin by immunofluorescence on paraffin-embedded hepatic and pancreatic tissue.

1979 ◽  
Vol 27 (3) ◽  
pp. 794-796 ◽  
Author(s):  
M J McElrath ◽  
R M Galbraith ◽  
R C Allen
Keyword(s):  
Human Serum ◽  
Solid Phase ◽  
Islet Cells ◽  
Pancreatic Tissue ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Normal Human ◽  
Liver Biopsies ◽  
Alpha 1 Antitrypsin ◽  
Sepharose 4B

Studies were performed in an attempt to improve current immunohistological techniques for the demonstration of alpha 1-antitrypsin (A1AT) in formalin-fixed paraffin-embedded tissues. The unwanted fluorescence (UF) commonly occurring in such procedures was found to be effectively eliminated by immunoadsorption of A1AT antisera with human serum lacking A1AT (Pi-null phenotype) coupled in solid phase to glutaraldehyde-activated aminohexyl-Sepharose 4B. Specificity of the antisera for A1AT was established by subsequent solid phase immunoadsorption against normal human serum bound to AH-Sepharose 4B. Using these techniques, immunoreactive A1AT was demonstrated in the cytoplasm of hepatocytes in liver biopsies obtained from patients with Z and MZ serum phenotypes, and in the cytoplasm of normal pancreatic islet cells.

The Development of a Radioimmunoassay Procedure for the Estimation of Tamm—Horsfall Glycoprotein in Human Serum

1977 ◽  
Vol 52 (2) ◽  
pp. 183-191 ◽  
Author(s):  
P. J. G. Avis
Keyword(s):  
Human Serum ◽  
Serum Proteins ◽  
Solid Phase ◽  
Normal Serum ◽  
Small Sample ◽  
Agarose Beads ◽  
Assay Procedure ◽  
Series Of Experiments ◽  
Sepharose 4B ◽  
Cyanogen Bromide Activation

1. A quantitative radioimmunoassay was developed for the measurement of nanogram amounts of Tamm—Horsfall (TH) glycoprotein in the presence of serum proteins at concentrations above 30 mg/ml. 2. Specific anti-(TH-glycoprotein) antibodies were labelled with 125I and these were usable for a period of 8 weeks. 3. Agarose beads (Sepharose 4B), to which TH-glycoprotein had been coupled via cyanogen bromide activation of the Sepharose, were used as the solid phase in the assay. This proved stable upon storage at 4°C for periods in excess of 4 months. 4. The dissociation of the glycoprotein in the presence of serum proteins that was necessary for quantification was achieved by subjecting the sample to ultrasonication. 5. Assays conducted on a small sample of normal serum produced evidence that normal serum contained amounts (50–180 ng/ml) of a substance that reacted similarly to TH-glycoprotein in the assay procedure and in a series of experiments conducted to confirm the presence of this substance in human serum.

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