PURIFICATION OF REACTIVE SULFHYDRYL ENZYMES BY BIOSELECTIVE ADSORPTION ON MONOMERIC AVIDIN: PURIFICATION OF SULFHYDRYL OXIDASE1

ABSTRACT A biotinylation-bioselective adsorption procedure was developed for single-step purification of bovine sulfhydryl oxidase from solubilized skim milk membrane vesicles. Sulfhydryl oxidase was specifically biotinylated by reaction of its chemically reactive sulfhydryl group with a disulfide-containing biotinylation reagent giving a mixed disulfide between the enzyme and the biotinyl moiety. The biotinylated enzyme was then selectively adsorbed on a monomeric avidin matrix. After washing all nonspecifically bound proteins from the matrix, the enzyme was released by reduction of the disulfide with dithiothreitol. Both the biotinylation and the isolation are performed at pH 7.0 under extremely mild conditions. The resulting enzyme preparation appeared to be homogeneous by gel electrophoresis and the specific activity increased over 3000 fold in comparison to whey. The monomeric avidin matrix can be regenerated and used indefinitely. This purification procedure should be generally applicable to proteins with a chemically reactive sulfhydryl group.     

AN ALTERNATIVE PERSPECTIVE ON FUZZY SET-BASED APPROXIMATE REASONING

Abstract

On the subject of fuzzy set-based approximate reasoning, this paper attempts to present a perspective that is both rigorously stated and different from the standard approach as stated by Mamdani and many others. Functionally speaking, this alternative perspective produces results essentially the same as the standard one, and in fact we intend to prove that subject to certain assumptions the results will be exactly die same. We feel however that the alternative perspective offers advantages in that it makes a more appropriate and more understandable use of such concepts as fuzzy relations, the extension principle, and fuzzy composition. If indeed it can lead to a clearer and more widespread understanding of the approximate reasoning formalism then this could conceivably spark broader interest in fuzzy control and other applications.     

Estimation of storage space requirements using multiple regression

Two equations hove been obtained using regression analysis techniques which can be applied to determine storage space requirements with acceptable accuracy and minimum effort. By defining all items by means of a common parameter—packing factor—it was possible to ignore the individual weights of each item, yet still load the transport and storage unit to some practical load limits. The method used in this paper is general and can be applied to similar problems with different boundary conditions.     

Correlation of Ethanol Concentration with Sensory Classification of Decomposition in Canned Salmon

Canned salmon samples of four species (sockeye, pink, coho, and chum) were analyzed for ethanol content. A highly significant correlation (r = 0.71, p < 0.001, Pearson;r = 0.80, p = 0.001, Spearman) was found between the ethanol content and the sensory classification of decomposition in canned salmon. This relationship has potential use as a means of objectively confirming the initial sensory classification of canned salmon.     

The Sterols and Triterpenes of Banana Pulp

SUMMARY: Using thin-layer chromatography and gas-liquid chromatography the following substances were identified in banana pulp (Musa saplentum L.): Cycloartenol, cycloeucalenol, 24-methylene cycloartanol, campesterol, β-sitosterol and stigmasterol. The major triterpene was 24-methylene cycloartanol while β-sitosterol accounted for greater than 72% of the sterol fraction.     

PURIFICATION AND PARTIAL CHARACTERIZATION OF TWO α-AMYLASE INHIBITORS FROM BLACK BEAN (Phaseolus vulgaris)1

Two α-amylase inhibitors from black bean (Phaseolus vulgaris) were purified to homogencity using ammonium sulfate fractionation, DEAE-Sephadex chromatography, phenyl-Sepharose hydrophobic interaction chromatography and gel filtration with Sephadex G-100. The inhibitors were designated I–1 and I–2 based on their order of elution from the phenyl-Sepharose column. Both inhibitors are mannose containing glycoproteins, composed of subunits; active against porcine pancreatic, human salivary, and insect α-amylases and inactive against bacterial, mold, and plant α-amylases. The inhibitors I-1 and I–2 have molecular weights of 49,000 and 47,000 and isoelectric points 4.93 and 4.86, respectively. Both inhibitors have similar amino acid compositions and are rich in aspartic acid, serine, glutamic acid, valine, and threonine and are low in sulfur containing amino acids. I–2 is more resistant to heat denaturation than I-1.     

BEHAVIOR MODIFICATION AND THE BRAT SYNDROME.

DESCRIBES A PROGRAM OF BEHAVIOR MODIFICATION TO TRAIN A MOTHER TO CONTROL HER 81/2-YR-OLD EMOTIONALLY DISTURBED BOY WHO WAS A SEVERE DISCIPLINARY PROBLEM. TRAINING WAS BASED ON LEARNING PRINCIPLES, AND BEHAVIORAL FEEDBACK TO THE MOTHER WAS PROVIDED VIA CLOSED-CIRCUIT TV. MOTHER-SON INTERACTIONS WERE VIDEOTAPED AS THE MOTHER PRACTICED STEP-BY-STEP INSTRUCTIONS, AND THE INTERACTION TAPES WERE VIEWED BY THE MOTHER FOR APPRAISAL OF SUCCESS. WITHIN A FEW WEEKS, THE BOY'S ABUSIVE BEHAVIOR WAS REDUCED, AND TREATMENT EFFECTS WERE STILL EVIDENT 23 WK. AFTER INITIATION OF THE PROGRAM. (PsycINFO Database Record (c) 2010 APA, all rights reserved)