scholarly journals A METHOD FOR THE HISTOCHEMICAL DEMONSTRATION OF TRIPHOSPHOPYRIDINE NUCLEOTIDE-LINKED l-HEXONATE DEHYDROGENASE ACTIVITY IN KIDNEYS OF SEVERAL SPECIES

1965 ◽  
Vol 13 (7) ◽  
pp. 533-540 ◽  
Author(s):  
KÁROLY BALOGH
Keyword(s):  
Enzyme Activity ◽  
Dehydrogenase Activity ◽  
Guinea Pigs ◽  
Renal Cortex ◽  
Histochemical Demonstration ◽  
Tetrazolium Salt ◽  
Histochemical Reaction ◽  
Mammalian Tissues ◽  
Histochemical Evidence ◽  
Enzyme Distribution

The described technique utilizes a tetrazolium salt to localize TPN2-linked l-hexonate dehydrogenase activity in unfixed frozen sections of mammalian kidneys. Enzyme diffusion was kept at a minimum by adding PVP to the incubation medium in a final concentration of 20%. Nevertheless, the sites of enzyme activity could not be localized precisely at the cell level. Histochemical evidence for l-hexonate specificity of the enzyme has been presented. A survey of various mammalian tissues revealed strong enzyme activity in the renal cortex of hamsters, rabbits, dogs and humans. In guinea pigs, the histochemical reaction was considerably weaker in heterozygous albino males than in pigmented ones. No enzyme activity was demonstrable in any of the female guinea pigs. Otherwise, the intensity of the histochemical reaction was consistent for a particular species. Although striking species differences were seen in the enzyme distribution pattern, activity was always limited to the epithelial cells of the renal cortex. Other tissues, known to contain the enzyme, failed to reveal an unequivocal histochemical reaction.

scholarly journals EFFECT OF INFECTION WITH M. TUBERCULOSIS AND OF TUBERCULIN SHOCK ON THE SUCCINIC DEHYDROGENASE ACTIVITY OF GUINEA PIG TISSUES

1953 ◽  
Vol 98 (2) ◽  
pp. 99-105 ◽  
Author(s):  
S. N. Chaudhuri ◽  
Samuel P. Martin
Keyword(s):  
Enzyme Activity ◽  
Guinea Pig ◽  
Dehydrogenase Activity ◽  
Guinea Pigs ◽  
Normal Tissue ◽  
Succinic Dehydrogenase ◽  
Tissue Extract ◽  
Succinic Dehydrogenase Activity ◽  
H37rv Strain ◽  
Sublethal Doses

The kidney of guinea pigs infected with the H37Rv and BCG strains of M. tuberculosis showed a diminution in succinic dehydrogenase activity when measured by the tetrazolium technique. This effect was also seen in the liver and spleen of animals infected with the BCG strain. Sensitized animals showed similar results when given tuberculin in sublethal doses. The succinic oxidase was also low in the kidneys of animals infected with the H37Rv strain. The depressed enzyme activity of the tissues of infected animals could be restored to normal by addition of normal tissue extract or dialysate. This suggests that the alteration in tissue metabolism observed in tuberculosis may depend upon the loss of some as yet unidentified factor important for succinic dehydrogenase activity.

HISTOCHEMICAL DEMONSTRATION OF 11β-HYDROXYSTEROID DEHYDROGENASE

1965 ◽  
Vol 33 (1) ◽  
pp. 119-125 ◽  
Author(s):  
A. H. BAILLIE ◽  
M. M. FERGUSON ◽  
K. C. CALMAN ◽  
D. McK. HART
Keyword(s):  
Dehydrogenase Activity ◽  
Leydig Cells ◽  
Hydroxysteroid Dehydrogenase ◽  
Histochemical Demonstration ◽  
Postnatal Life ◽  
Interstitial Tissue ◽  
Corpora Lutea ◽  
Histochemical Reaction ◽  
Mouse Ovary ◽  
Theca Interna

SUMMARY 11β-Hydroxysteroid dehydrogenase can be demonstrated histochemically by incubating tissues with nitro blue tetrazolium (2,2′-di-p-nitrophenyl-5,5′-diphenyl-3,3′-(3,3′-dimethoxy-4,4′-diphenylene) ditetrazolium chloride), NAD or NADP and an appropriate steroid. Suitable steroid substrates are: (1) 11β-hydroxyandrost-4-ene-3,17-dione (11β-hydroxyandrostenedione), (2) 3,11β-dihydroxyoestra-1,3,5(10)-trien-17-one (11β-hydroxyoestrone), (3) 3α,11β-dihydroxy-5α-androstan-17-one, (4) 3α,11β-dihydroxy-5β-androstan-17-one and (5) 11β-hydroxypregn-4-ene-3,20-dione(11β-hydroxyprogesterone). 11β-Hydroxysteroid dehydrogenase activity was found in the Leydig cells of six human testes from subjects ranging in age from 12 to 57 yr. with all five substrates. The Leydig cells of the mouse testis contain demonstrable 11β-hydroxysteroid dehydrogenase activity and the volume of reactive tissue increases regularly between birth and the end of the 10th week of postnatal life; this growth curve is sigmoid in form. An extremely weak histochemical reaction with human placenta obtained at term was observed, 11β-hydroxyandrostenedione being the only substrate utilized to any extent. A specimen of hydatid mole, however, showed intense 11β-hydroxysteroid dehydrogenase activity with all substrates surveyed. 11β-Hydroxysteroid dehydrogenase was also found in the ova, granulosa, theca interna, interstitial tissue and corpora lutea of the mouse ovary.

3β-HYDROXYSTEROID DEHYDROGENASE IN THE ADRENAL GLAND AND PLACENTA

1965 ◽  
Vol 31 (3) ◽  
pp. 227-NP ◽  
Author(s):  
A. H. BAILLIE ◽  
E. H. D. CAMERON ◽  
K. GRIFFITHS ◽  
D. McK. HART
Keyword(s):  
Adrenal Gland ◽  
Dehydrogenase Activity ◽  
Adrenal Glands ◽  
Hydroxysteroid Dehydrogenase ◽  
Histochemical Demonstration ◽  
Zona Fasciculata ◽  
Histochemical Reaction ◽  
Tissue Sections ◽  
Adrenal Zona Fasciculata ◽  
Human Placentae

SUMMARY 3β-Hydroxysteroid dehydrogenase activity was studied histochemically in human, monkey, and rat adrenal glands and in human placentae. Tissue sections were incubated separately with each of the following substrates: (1) 3β-hydroxypregn-5-en-20-one (pregnenolone); (2) sodium 3β-sulphoxypregn-5-en-20-one (pregnenolonesulphate); (3) 3β-acetoxypregn-5-en-20 one (pregnenoloneacetate); (4) 3β,16α-dihydroxypregn-5-en-20-one (16α-hydroxypregnenolone); (5) 3β,17α-dihydroxypregn-5-en-20-one (17α-hydroxypregnenolone); (6) ammonium 3β-sulphoxy-17α-hydroxypregn-5-en-20-one (17α-hydroxypregnenolone ammonium sulphate); (7) 3β-hydroxyandrost-5-en-17-one (DHA); (8) 3β-sulphoxyandrost-5-en-17-one (DHA sulphate); (9) 3β-acetoxyandrost-5-en-17-one (DHA acetate); (10) androst-5-ene-3β, 17β-diol (androstenediol). The histochemical results obtained with pregnenolone and DHA as substrates resemble those described by other workers. Using pregnenolone sulphate and 17α-hydroxypregnenolone sulphate, a strong histochemical reaction with diformazan deposition was found in the zona fasciculata of the adrenals of all species and in the placental syntrophoblast. With DHA sulphate an extremely weak histochemical reaction was obtained with the adrenal zona fasciculata, monoformazan only being deposited. The syntrophoblast, however, showed intense 3β-hydroxysteroid dehydrogenase activity when incubated with DHA sulphate. These results accord with recent findings regarding the secretion and metabolism of 3β-sulphoxysteroids. A strong histochemical reaction was also obtained in both adrenal and placental tissues using 17α-hydroxypregnenolone, 16α-hydroxypregnenolone, androstenediol, pregnenolone acetate, and DHA acetate. These steroids have not previously been described as substrates for the histochemical demonstration of 3β-hydroxysteroid dehydrogenase in the adrenal or placenta.

FURTHER OBSERVATIONS ON 3β-HYDROXYSTEROID DEHYDROGENASE ACTIVITY IN THE MOUSE LEYDIG CELL

1965 ◽  
Vol 31 (3) ◽  
pp. 207-NP ◽  
Author(s):  
A. H. BAILLIE ◽  
K. GRIFFITHS
Keyword(s):  
Dehydrogenase Activity ◽  
Growth Curve ◽  
Leydig Cells ◽  
Age Groups ◽  
Growth Curves ◽  
Hydroxysteroid Dehydrogenase ◽  
Histochemical Demonstration ◽  
Seminiferous Tubules ◽  
Postnatal Life ◽  
Histochemical Reaction

SUMMARY One hundred and ten male Swiss white mice were killed in batches of ten weekly between birth and the end of the 10th week of postnatal life. To demonstrate 3β-hydroxysteroid dehydrogenase activity histochemically, sections of testis from every animal were incubated with the following steroid substrates: (1) sodium 3β-sulphoxypregn-5-en-20-one (pregnenolone sulphate), (2) sodium 3β-sulphoxy-17α-hydroxypregn-5-en-20-one (17α-hydroxypregnenolone sulphate), (3) sodium 3β-sulphoxyandrost-5-en-17-one (DHA sulphate), (4) 3β,16α-dihydroxypregn-5-en-20-one (16α-hydroxypregnenolone), (5) pregn-5-ene-3β,20β-diol (pregnenediol), (6) androst-5-ene-3β, 17β-diol (androstenediol). Pregnenolone sulphate was rapidly used by the entire interstitium at all ages. 17α-Hydroxypregnenolone sulphate was metabolized by some Leydig cells of all age groups. DHA sulphate was not utilized histochemically by the Leydig cells of the various age groups, but formazan deposition occurred in the mature seminiferous epithelium. This is the only steroid so far investigated to give an histochemical reaction with the germinal epithelium, and 3β-hydroxysteroid dehydrogenase activity has not previously been described in the seminiferous tubules. The utilization of steroid sulphates differently from the free steroids in the histochemical demonstration of 3β-hydroxysteroid dehydrogenase activity suggests that the presence of a sulphate group may affect enzyme-substrate binding. With 16α-hydroxypregnenolone and pregnenediol as substrates, 3β-hydroxysteroid dehydrogenase activity was demonstrable at birth, increased progressively until the 6th week of postnatal life, and subsequently decreased during the ensuing 4 weeks. This growth curve closely resembles the growth curve obtained with pregnenolone. Androstenediol gave a histochemical reaction with the Leydig cells of all age groups studied, and the sigmoid growth curve resembles that obtained with 3β-hydroxyandrost-5-en-17-one (DHA). These differing growth curves are regarded as further evidence of substrate-specific 3β-hydroxysteroid dehydrogenases.

THE INFLUENCE OF HAEMOGLOBIN-S AND G6PD DEFICIENCY ON THE ACTIVITY OF THE 17β-HYDROXYSTEROID DEHYDROGENASE OF INTACT HUMAN ERYTHROCYTES

1974 ◽  
Vol 75 (4) ◽  
pp. 793-800
Author(s):  
A. O. Sogbesan ◽  
O. A. Dada ◽  
B. Kwaku Adadevoh
Keyword(s):  
Enzyme Activity ◽  
Dehydrogenase Activity ◽  
Sex Steroids ◽  
G6pd Deficiency ◽  
Incubation Medium ◽  
Hydroxysteroid Dehydrogenase ◽  
G6pd Activity ◽  
Reverse Transformation ◽  
Oxidative Transformation ◽  
Haemoglobin S

ABSTRACT The 17β-hydroxysteroid dehydrogenase activity in intact erythrocytes of Nigerian patients, in particular with regard to haemoglobin genotypes and G6PD* activity was studied. The G6PD activity of the erythrocyte did not affect the oxidative transformation of testosterone to androstenedione and of oestradiol to oestrone. The reduction (reverse transformation) was inhibited in G6PD-deficient erythrocytes but this inhibition was offset by the addition of 0.025 m glucose to the incubation medium. The per cent oxidation transformation of testosterone was higher in Hb-AA than in Hb-SS erythrocytes. It is suggested that the differences may be a result of either lower enzyme activity in the Hb-SS erythrocytes or of differences in the uptake and possibly binding of sex steroids by intact Hb-SS and Hb-AA erythrocytes.

Antioxidant Enzyme Levels Inversely Covary with Hearing Loss After Amikacin Treatment

2003 ◽  
Vol 14 (03) ◽  
pp. 134-143 ◽  
Author(s):  
James J. Klemens ◽  
Robert P. Meech ◽  
Larry F. Hughes ◽  
Satu Somani ◽  
Kathleen C.M. Campbell
Keyword(s):  
Hearing Loss ◽  
Enzyme Activity ◽  
Antioxidant Enzyme ◽  
Guinea Pigs ◽  
Auditory Brainstem ◽  
Antioxidant Enzyme Activity ◽  
Control Group ◽  
Glutathione S Transferase ◽  
Brainstem Response ◽  
The Difference

This study's purpose was to determine if a correlation exists between cochlear antioxidant activity changes and auditory function after induction of aminoglycoside (AG) ototoxicity. Two groups of five 250-350 g albino guinea pigs served as subjects. For 28 days, albino guinea pigs were administered either 200 mg/kg/day amikacin, or saline subcutaneously. Auditory brainstem response testing was performed prior to the first injection and again before sacrifice, 28 days later. Cochleae were harvested and superoxide dismutase, catalase, glutathione peroxidase, glutathione-S-transferase, glutathione reductase activities and malondialdehyde levels were measured. All antioxidant enzymes had significantly lower activity in the amikacin group (p ≤ 0.05) than in the control group. The difference in cochlear antioxidant enzyme activity between groups inversely correlated significantly with the change in ABR thresholds. The greatest correlation was for the high frequencies, which are most affected by aminoglycosides. This study demonstrates that antioxidant enzyme activity and amikacin-induced hearing loss significantly covary.

Changes in 3β-hydroxy-Δ5-steroid dehydrogenase activity in granulosa tissue during the ovulatory cycle of the laying hen (Gallus domesticus)

1985 ◽  
Vol 106 (3) ◽  
pp. 269-273 ◽  
Author(s):  
D. G. Armstrong
Keyword(s):  
Enzyme Activity ◽  
Dehydrogenase Activity ◽  
Domestic Fowl ◽  
Ovarian Follicle ◽  
Laying Hen ◽  
Gallus Domesticus ◽  
Ovulatory Cycle ◽  
Lh Surge ◽  
Maximal Plasma ◽  
Steroid Dehydrogenase

ABSTRACT The object of this study was to examine changes in the activity of granulosa 3β-hydroxy-Δ5-steroid dehydrogenase during the ovulatory cycle of the domestic fowl. The enzyme activity in granulosa tissue from the largest follicle increased significantly during the period 8–14 h before an expected ovulation. The increase in activity occurs before the preovulatory surge of LH and near the time of lights off. During the 4–8 h period before an ovulation, i.e. the time of maximal plasma LH concentrations, 3β-hydroxy-Δ5-steroid dehydrogenase activity decreased in granulosa tissue from the largest follicle. This observation is explained by proposing that the enzyme is inhibited by the large amounts of progesterone found in the tissue at this time. The results indicate that important biochemical changes are taking place within granulosa tissue of the largest ovarian follicle before the preovulatory LH surge. J. Endocr. (1985) 106, 269–273

A Study of the Histochemical Demonstration of Cytochrome Oxidase

1964 ◽  
Vol s3-105 (72) ◽  
pp. 497-502
Author(s):  
R. G. BUTCHER ◽  
J. V. DIENGDOH ◽  
J. CHAYEN
Keyword(s):  
Enzyme Activity ◽  
Cardiac Muscle ◽  
Cytochrome Oxidase ◽  
Histochemical Demonstration ◽  
Lugol’S Iodine ◽  
Liver And Kidney

Burstone's procedure for the histochemical demonstration of cytochrome oxidase has been studied and applied to sections prepared by freezing in hexane and cutting by the controlled-temperature freezing-sectioning technique. The method has been modified by the inclusion of a treatment with Lugol's iodine to yield a stronger colour, which is stable for at least several weeks and which localizes the enzyme activity more precisely. The variants of the reaction have been compared in their effect on cardiac muscle, liver, and kidney of the rat.

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