HYDROLYSIS OF CONJUGATED ESTROGEN FRACTIONS IN HUMAN PREGNANCY URINE

1961 ◽  
Vol 39 (10) ◽  
pp. 1501-1514 ◽  
Author(s):  
Sidsel Bugge ◽  
Mona Nilsen ◽  
Ann Metcalfe-Gibson ◽  
R. Hobkirk
Keyword(s):  
Helix Pomatia ◽  
Enzyme Hydrolysis ◽  
Digestive Juice ◽  
Human Pregnancy ◽  
Mammalian Liver ◽  
Pregnancy Urine ◽  
Snail Enzyme ◽  
17Β Estradiol ◽  
Snail Helix Pomatia ◽  
Hydrolysis Of

The release of six estrogen fractions from conjugation in human pregnancy urines has been studied using various hydrolytic methods. The estrogens concerned were estrone, estradiol-17β (estradiol), 2-methoxyestrone, 16-epiestriol, and a ring D ketolic fraction (mainly 16α-hydroxyestrone). Considerable amounts of urinary estrone and ring D ketolic estrogens may be conjugated in a non-glucuronide form. In these cases an enzyme preparation containing β-glucuronidase and sulphatase, prepared from the digestive juice of the snail Helix pomatia, proved to be superior to β-glucuronidase enzymes of bacterial or mammalian liver origin. Conventional hot acid hydrolysis yielded levels of estrone, estradiol, estriol, and 16-epiestriol which agreed fairly well with those obtained following snail enzyme hydrolysis. In some urines, hot acid treatment was not suitable for hydrolysis of conjugated 2-methoxyestrone. Optimum hydrolytic conditions for both normal and diabetic pregnancy urines were realized by incubating for 24 hours with 500 units of the snail β-glucuronidase and 250 units of sulphatase/ml of urine at pH 5.2 and 37–38 °C.

The Histochemical Localization of β-Glucuronidase in the Digestive Gland of the Roman Snail (Helix pomatia)

1955 ◽  
Vol s3-96 (33) ◽  
pp. 35-48
Author(s):  
F. BILLETT ◽  
S. M. McGEE-RUSSELL
Keyword(s):  
Prussian Blue ◽  
Digestive Gland ◽  
Secretory Granules ◽  
Helix Pomatia ◽  
Histochemical Localization ◽  
Enzymic Hydrolysis ◽  
Histochemical Technique ◽  
Original Technique ◽  
Snail Helix Pomatia ◽  
Hydrolysis Of

A modification of the histochemical technique for the localization of β-glucuronidase originally suggested by Friedenwald and Becker (1948) has been applied to the digestive gland of the gastropod Helix pomatia. In the original technique the ferric 8-hydroxyquinoline formed by the enzymic hydrolysis of quinolyl-8-glucuronide, in a saturated solutionof ferric 8-hydroxyquinoline, was converted to Prussian blue. The Prussian blue conversion is omitted in the technique described in this paper as it appears to introduce errors in localization. The ferric 8-hydroxyquinoline crystals are sufficiently characteristic to be used as the end-point of the technique. The results obtained suggest that β-glucurcnidase is confined to the digestive cells in the digestive gland of the snail, and is associated with secretory granules in them.

STUDIES ON THE HYDROLYSIS OF GEL-FILTERED URINARY OESTROGENS

1968 ◽  
Vol 57 (1) ◽  
pp. 49-68 ◽  
Author(s):  
H. Adlercreutz
Keyword(s):  
Sodium Acetate ◽  
Enzyme Inhibitors ◽  
Helix Pomatia ◽  
Gel Filtration ◽  
Enzyme Concentration ◽  
Enzyme Hydrolysis ◽  
Sodium Acetate Buffer ◽  
Quantitative Results ◽  
Hydrolysis Of ◽  
Great Advance

ABSTRACT The effect of pH, enzyme concentration, administration of corticotrophin and cortisol on the enzyme hydrolysis of gel-filtered urinary oestrogens was studied. Special attention was directed to the hydrolysis of oestrogen conjugates in the first fraction (peak I) following gel filtration. In addition, the effect on enzyme hydrolysis of D-glucosaccharic acid added to urine before gel filtration was studied and some comparisons between acid and enzyme hydrolysis were also carried out. It was found that for peak I oestrogen conjugates, higher enzyme concentrations of β-glucuronidase (600 Fishman units per ml) are needed to give quantitative results than for peak II conjugates (second fraction obtained by gel filtration) (200 Fishman units per ml), but otherwise the same conditions for hydrolysis can be used for both fractions. These conditions are 0.15 m sodium acetate buffer, pH 4.1–4.2, 37° C, and overnight incubation with a Helix pomatia extract. It was also found that if the Kober reaction is used as the final step for oestriol estimation by the method of Beling, impurities occurring in the final extract decrease the recoveries to some extent by interfering both with the development of the Kober reaction and with the Allen correction of the estimated absorbancies. This phenomenon occurs predominantly if peak I oestrogens are estimated. In addition, some inhibition of the β-glucuronidase occurs in peak I, varying in degree from one urine to another, while the administration of ACTH and cortisol increases both the amount of impurities interfering with the Kober reaction and the amount of enzyme inhibitors present in urine and occurring mostly before, but also partially in the peak I fraction. It is concluded that peak I and peak II oestrogens following gel filtration should be hydrolysed separately and with a higher enzyme concentration for peak I. In addition, it is concluded that both glucose, D-glucosaccharic acid and a large number of impurities are eliminated from the oestrogen fractions by gel filtration, and that this procedure represents a great advance in oestrogen methodology.

scholarly journals Direct deamination of AMP, ADP, ATP and NADH by non-specific adenylate deaminase in the foot muscle of the snail Helix pomatia

1983 ◽  
Vol 215 (1) ◽  
pp. 39-44 ◽  
Author(s):  
A J Stankiewicz
Keyword(s):  
Helix Pomatia ◽  
Analytical Tool ◽  
Fresh Tissue ◽  
High Affinity ◽  
Adenylate Deaminase ◽  
Snail Enzyme ◽  
Snail Helix Pomatia ◽  
Species Specific ◽  
Very High ◽  
Millimolar Concentration

Homogeneous adenylate deaminase from snail foot muscle deaminated 5′-AMP, 5′-ADP, 5′-ATP and NADH with similar velocity and affinity to all substrates. At millimolar concentration NAD+ was also deaminated to a comparable extent, but NADP+, NADPH and FAD were not substrates for the snail enzyme. The amount of deaminase activity per g of fresh tissue is 5-10 times greater than in the muscle of any other species studied. The activity of the snail deaminase is regulated by pH, KCl and buffer concentrations, and Pi; however, regulation seems to be very poor in comparison with that of muscle deaminases from other species, specific to 5′-AMP. Snail enzyme appears as the first animal deaminase so far described that has such characteristics. It offers also some opportunities as an analytical tool as a consequence of its very high affinity toward adenylates.

ON THE ORIGIN OF THE CELLULASE AND CHITINASE OF HELIX POMATIA

1963 ◽  
Vol 41 (1) ◽  
pp. 1621-1626 ◽  
Author(s):  
G. A. Strasdine ◽  
D. R. Whitaker
Keyword(s):  
Soluble Protein ◽  
Specific Activity ◽  
Helix Pomatia ◽  
Cellulase Activity ◽  
Chitinase Activity ◽  
Intestinal Wall ◽  
Total Soluble Protein ◽  
Digestive Juice ◽  
Viable Bacteria ◽  
Snail Helix Pomatia

Specimens of the snail Helix pomatia were fed for 2 days at room temperature after emergence from hibernation at 4° and total soluble protein, total units of cellulase activity towards carboxymethyl cellulose, and total units of chitinase activity towards glycol chitin were determined in the digestive juice, hepatopancreas, and intestinal wall. Comparisons of these values with those for hibernating snails showed that emergence from hibernation and resumption of feeding was accompanied by a substantial decrease in soluble protein in the hepatopancreas, and a substantial increase in soluble protein cellulase and chitinase in the digestive juice. Cellulase and chitinase in the digestive juice showed little change in specific activity and were present in approximately the same ratio as in the hepatopancreas and intestinal wall. The volume of digestive juice in snails which had been deprived of food after a brief posthibernation feeding increased significantly with starvation times up to at least 5 days but the concentration of protein, cellulase, chitinase, and helicorubin showed no significant change. Plate counts for viable bacteria in the hepatopancreas indicated that the hepatopancreas was virtually sterile. The difficulty of reconciling these findings with the hypothesis of a bacterial origin for the cellulase and chitinase in snail digestive juice is discussed.

THE EXCRETION OF NEUTRAL STEROIDS IN THE URINE OF NEWBORN INFANTS

1963 ◽  
Vol 27 (1) ◽  
pp. 53-75 ◽  
Author(s):  
D. M. CATHRO ◽  
KATHLEEN BIRCHALL ◽  
F. L. MITCHELL ◽  
CONSTANCE C. FORSYTH
Keyword(s):  
Premature Infants ◽  
Newborn Infants ◽  
Enzyme Hydrolysis ◽  
Adult Women ◽  
Blue Tetrazolium ◽  
Female Urine ◽  
Pregnancy Urine ◽  
Steroid Excretion ◽  
Hydrolysis Of ◽  
In Pregnancy

SUMMARY Following enzyme hydrolysis of the urine of infants aged 1 to 6 days, the neutral steroids have been extracted and their excretion patterns determined using a method based on paper chromatography. Three groups of compounds have been estimated—those reacting with Zimmermann reagent, those reducing blue tetrazolium and those giving sodium fluorescence. The first group includes the 17-oxosteroids and the latter two the corticoids. The steroid excretion patterns in the urine of these infants have been compared with those demonstrated by the same technique in adult female urine and in the urine of mothers in the 24 hr. following delivery. For all groups of compounds, the excretion patterns in the urine of infants are different from those found in the urine of their mothers or in that of adult women. The outstanding feature in infant urine is the dominance of compounds which behave like steroids, and which either do not occur in adult urine, or are present in insignificant amounts. Less obvious changes in the excretion patterns of 17-oxosteroids and of corticoids in pregnancy urine are also described. Many of the Zimmermann chromogens which are dominant in the urine in the first days of life have greatly diminished, or have disappeared, by the 6th day. This applies both to full-term and premature infants. The pattern of excretion of blue tetrazolium-reducing material in the urine of newborn infants is extremely complicated and, apparently, unique. Tetrahydrocortisol and tetrahydrocortisone, which are the major corticoid metabolites in adult urine, are much less prominent in the urine in the first 3 days of life. A blue tetrazolium-reducing steroid isolated from the urine of infants has been provisionally identified as 3β, 21-dihydroxypregn-5-ene-20-one. The altered steroid excretion patterns found in the urine of newborn infants are thought to indicate that there are fundamental differences in the metabolism of adrenal and placental steroids which are peculiar to this period and comparable with those established within recent years for oestrogens and progesterone.

ON THE ORIGIN OF THE CELLULASE AND CHITINASE OF HELIX POMATIA

1963 ◽  
Vol 41 (7) ◽  
pp. 1621-1626 ◽  
Author(s):  
G. A. Strasdine ◽  
D. R. Whitaker
Keyword(s):  
Soluble Protein ◽  
Specific Activity ◽  
Helix Pomatia ◽  
Cellulase Activity ◽  
Chitinase Activity ◽  
Intestinal Wall ◽  
Total Soluble Protein ◽  
Digestive Juice ◽  
Viable Bacteria ◽  
Snail Helix Pomatia

Specimens of the snail Helix pomatia were fed for 2 days at room temperature after emergence from hibernation at 4° and total soluble protein, total units of cellulase activity towards carboxymethyl cellulose, and total units of chitinase activity towards glycol chitin were determined in the digestive juice, hepatopancreas, and intestinal wall. Comparisons of these values with those for hibernating snails showed that emergence from hibernation and resumption of feeding was accompanied by a substantial decrease in soluble protein in the hepatopancreas, and a substantial increase in soluble protein cellulase and chitinase in the digestive juice. Cellulase and chitinase in the digestive juice showed little change in specific activity and were present in approximately the same ratio as in the hepatopancreas and intestinal wall. The volume of digestive juice in snails which had been deprived of food after a brief posthibernation feeding increased significantly with starvation times up to at least 5 days but the concentration of protein, cellulase, chitinase, and helicorubin showed no significant change. Plate counts for viable bacteria in the hepatopancreas indicated that the hepatopancreas was virtually sterile. The difficulty of reconciling these findings with the hypothesis of a bacterial origin for the cellulase and chitinase in snail digestive juice is discussed.

scholarly journals The hydrolysis of the combined forms of oestrone and oestriol present in human pregnancy urine

1935 ◽  
Vol 29 (7) ◽  
pp. 1577-1585 ◽  
Author(s):  
Saul Louis Cohen ◽  
Guy Frederic Marrian
Keyword(s):  
Human Pregnancy ◽  
Pregnancy Urine ◽  
Hydrolysis Of

COMPARATIVE STUDIES ON HYDROLYSIS OF CONJUGATED ESTROGENS IN HUMAN PREGNANCY URINE

Endocrinology ◽  
1955 ◽  
Vol 57 (1) ◽  
pp. 87-95 ◽  
Author(s):  
RICHARD F. STRAW ◽  
PHILIP A. KATZMAN ◽  
EDWARD A. DOISY
Keyword(s):  
Comparative Studies ◽  
Conjugated Estrogens ◽  
Human Pregnancy ◽  
Pregnancy Urine ◽  
Hydrolysis Of
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