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J M Wallach

Publications and source records attributed to J M Wallach.

23 records · Page 2Linked to original sources

Elastolysis of normal and partially cross-linked elastin.

Rate of elastolysis by pancreatic and leukocyte elastases of normal and copper deficient porcine aortic elastins were measured using a conductimetric method. Kinetics obey to Michaelis-Menten model for both substrates and enzymes. KM and Vmax values derived from Lineweaver-Burk plots indicate that, if a near uniformity exists in KM, differences were observed in catalytic rates, kcat increasing approximately 40% for copper deficient elastin elastolysis by leukocyte elastase. This higher susceptibility to proteolysis may have implications for understanding turnover of elastin in tissues.

Animals↗

Ca2+ and Mg2+ protection against thermal denaturation of pancreatic elastase.

Thermal denaturation of porcine pancreatic elastase was studied by difference spectrophotometry. At 293 nm, and pH 8.0, the thermal transition of elastase occurs with a midpoint temperature (Tm) of (58.0 +/- 0.5) degrees C. Mg2+ and Ca2+ stabilize the native form in increasing the midpoint temperature of the transition, Ca2+ being more effective than Mg2+ in the 0-0.02 M concentration range. Furthermore, Ca2+ protects pancreatic elastase against the destabilizing effect of Cu2+. Whatever be the temperature between 40 degrees C and 55 degrees C, Ca2+ protects pancreatic elastase against loss of enzymatic activity.

Animals↗

Microconductimetric assay of butyrylcholinesterase activity.

A conductimetric method for the assay of butyrylcholinesterase is described. A new conductimetric cell has been used which allows increased sensitivity. The physicochemical parameters (pH, buffer concentration) have been optimized. With 1 mM butyrylcholine, butyrylcholinesterase activities down to 2 x 10(-4) U ml-1 may be measured. Serum volumes needed for this assay are in the microliter range (1-5 microliters).

Animals↗

Lipase assay in duodenal juice using a conductimetric method.

Lipase activity in duodenal juice is known to undergo important variations in pathologic states, especially in cases of chronic pancreatitis. Almost all of the current assay methods are based on the measurement of hydrolysis of olive oil or triolein, mainly by potentiometry. As we have developed a conductimetric method for enzyme activity measurements, we have applied it to lipase assay. A higher experimental conductimetric sensitivity is obtained when liberated acids have a short chain (higher limiting equivalent conductivity). We have therefore used triacetin as a substrate and compared out method with potentiometry (pH-stat) and spectrophotometry. The correlation coefficients of both methods with conductimetry were 0.94 and 0.97, respectively, indicating that the conductimetric method may be used for lipase assay in duodenal juice, using triacetin as a substrate.

Body Fluids↗

Age-dependent variations of human PMNL elastase activity as a function of pH, ionic strength and calcium concentration.

Human leukocyte elastase (HLE) has been implicated in the pathomechanism of various diseases, such as emphysema and atherosclerosis. The incidence of these diseases is increasing with aging. Therefore, it can be supposed that the HLE activity is changing with aging according to the well known age-related physiological alterations. Thus, the effects of pH, NaCl and calcium concentrations on HLE activity, separated from polymorphonculear leucocytes (PMNLs) of healthy, young (<35 years) and elderly (>75 years) subjects, were studied by measuring the activity with synthetic substrate and with bovine and human atherosclerotic and non-atherosclerotic aortic elastin. From our results, it may be concluded, that the elastolytic activity of HLE separated from PMNLs of elderly subjects is more sensitive to ionic strength, to pH and to the calcium concentration of the medium, than the HLE activity of young subjects. The elastolytic activity of HLE, of both young and old subjects, increases dramatically on atherosclerotic aortic elastin in the presence of calcium. These findings might explain, at least partly, the increased incidence of atherosclerosis with aging.

Journal Article↗