scholarly journals CONCENTRATION OF ACID PHOSPHATASE, RIBONUCLEASE, DESOXYRIBONUCLEASE, ß-GLUCURONIDASE, AND CATHEPSIN IN "DROPLETS" ISOLATED FROM THE KIDNEY CELLS OF NORMAL RATS

1956 ◽  
Vol 2 (5) ◽  
pp. 513-521 ◽  
Author(s):  
Werner Straus
Keyword(s):  
Acid Phosphatase ◽  
Mitochondrial Fraction ◽  
Liver Cells ◽  
Enzymatic Properties ◽  
Kidney Cells ◽  
Supernatant Fluid ◽  
Fraction I

1. Three fractions of "droplets" having diameters of 1 to 5 µ (fraction I), 0.5 to 1.5 µ (fraction II), and 0.1 to 1.0 µ (fraction III) were isolated from the kidney cells of normal rats. 2. All three "droplet" fractions showed 10 to 15 times higher activities of acid phosphatase, ß-glucuronidase, ribonuclease, desoxyribonuclease, and cathepsin than the total homogenate and the mitochondrial fraction. 3. After a rough fractionation of the total homogenate, approximately 50 per cent of the 5 enzymes was found in the fractions which contained the "droplets" and approximately 30 per cent in the supernatant fluid. 4. The similarities between the enzymatic properties of the "droplets" from kidney cells and of the fractions isolated from liver cells by other investigators have been discussed.

scholarly journals BIOCHEMICAL HETEROGENEITY OF THE CYTOPLASMIC PARTICLES ISOLATED FROM RAT LIVER HOMOGENATE

1953 ◽  
Vol 1 (1) ◽  
pp. 27-46 ◽  
Author(s):  
ALEX B. NOVIKOFF ◽  
ESTELLE PODBER ◽  
JEAN RYAN ◽  
ELSIE NOE
Keyword(s):  
Acid Phosphatase ◽  
Nucleic Acid ◽  
Rat Liver ◽  
Liver Homogenate ◽  
Mitochondrial Fraction ◽  
Nuclear Fraction ◽  
Supernatant Fluid ◽  
Cytoplasmic Granules ◽  
Isolated Mitochondria ◽  
Low Levels

By separating the cytoplasmic granules of 0.88 M sucrose homogenates of rat liver into eight fractions it has been possible to demonstrate marked chemical and enzymatic heterogeneity among the isolated microsomes and less pronounced heterogeneity among the isolated mitochondria. The more readily sedimented microsomes are rich in ribose nucleic acid and show high esterase, adenosine-5’-phosphatase, acid phosphatase and uricase activities, while the less readily sedimented microsomes, although rich in ribose nucleic acid esterase and adenosine-5’-phosphatase have low levels of acid phosphatase and uricase activities. The very small mitochondria differ from the larger ones in the levels of activities of all enzymes studied, with the exception of adenosine-5’-phosphatase; the most striking differences were found in the cases of acid phosphatase and uricase. A centrifugation schedule is given to isolate a "nuclear fraction," a mitochondrial fraction, a mixed fraction of smallest mitochondria and microsomes (of two types differing in optical density), a fraction of optically less dense microsomes, and a "supernatant fluid."

scholarly journals DEVELOPMENT OF AN IMPROVED MEDIUM FOR THE ISOLATION OF LIVER MITOCHONDRIA

1956 ◽  
Vol 2 (5) ◽  
pp. 609-624 ◽  
Author(s):  
M. S. C. Birbeck ◽  
E. Reid
Keyword(s):  
Adverse Effect ◽  
Molecular Weight ◽  
Electron Microscope ◽  
Acid Phosphatase ◽  
Mitochondrial Fraction ◽  
Liver Mitochondria ◽  
High Concentration ◽  
Mitochondrial Fractions ◽  
Sucrose Medium ◽  
Optimal Medium

In view of the unsatisfactory appearance, under the electron microscope, of liver mitochondria isolated in isotonic sucrose medium, alternative media have been examined. It was found to be advantageous to replace sucrose by raffinose, and to add levan or, preferably, dextran, together with heparin in suitable concentration. With the optimal medium, the constituents of which are raffinose, versene (optional), dextran of high molecular weight, heparin, and AMP (optional), most of the mitochondria in the osmium-fixed pellet are apparently intact, and show the membranes characteristic of mitochondria as seen in cell sections. The optimal medium has no adverse effect on the activity of the several tissue enzymes which have been studied, except that Mg++-activated ATPase is partially inhibited if the medium is present in high concentration in the assay system. Mitochondrial fractions isolated in the new medium have, in common with sucrose fractions, appreciable "free" ATPase activity, this activity being evidently a poor criterion of mitochondrial integrity. Use of the new medium does not decrease the proportion of cytoplasmic ATPase which fails to sediment with the mitochondria, but does give a mitochondrial fraction low in RNA and in acid phosphatase activity and little contaminated with microsomal material. Particles tentatively identified as "lysosomes" have been seen in certain sections.

scholarly journals BETA GLUCURONIDASE-RICH CYTOPLASMIC PARTICLES IN ANDROGEN-STIMULATED MOUSE KIDNEY

1963 ◽  
Vol 16 (2) ◽  
pp. 253-258 ◽  
Author(s):  
Andrew G. Plaut ◽  
William H. Fishman
Keyword(s):  
Alkaline Phosphatase ◽  
Acid Phosphatase ◽  
Specific Activity ◽  
Mouse Kidney ◽  
The Other ◽  
Mouse Testis ◽  
Glucuronidase Activity ◽  
Other Hand ◽  
Fraction I ◽  
Gonadotropic Hormones

Androgens produced by stimulating mouse testis with gonadotropic hormones cause a rise in renal ß-glucuronidase but not an increase in acid or alkaline phosphatase. All subcellular components increase in ß-glucuronidase activity, with a relatively greater increment in particulate enzyme as compared with that free in the cytoplasm (non-sedimentable). A small percentage of recovered ß-glucuronidase, acid phosphatase, and alkaline phosphatase is found in material which rises to the surface during centrifugation in sucrose media (fraction I). The specific activity of ß-glucuronidase and acid phosphatase in this fraction is normally quite high with respect to the homogenate, while that of alkaline phosphatase is not. On the other hand, the fraction I material from androgen-stimulated mice exhibits a further increase in specific activity with respect to ß-glucuronidase and not acid phosphatase. It thus appears that there is an independence in the behavior of individual enzymes in response to physiologic stimuli in spite of obvious morphologic proximity.

EFFECT OF CORTISONE ON THE INTRACELLULAR DISTRIBUTION OF PHOSPHATASES AND RIBONUCLEASES IN RAT LIVER

1956 ◽  
Vol 34 (2) ◽  
pp. 170-179 ◽  
Author(s):  
Claude Allard ◽  
Gaston De Lamirande ◽  
George Weber ◽  
Antonio Cantero
Keyword(s):  
Body Weight ◽  
Weight Ratio ◽  
Mitochondrial Fraction ◽  
Intracellular Distribution ◽  
Cortisone Acetate ◽  
Supernatant Fluid ◽  
Body Weight Ratio ◽  
Average Cell ◽  
Mitochondrial Population ◽  
The One

The effect of cortisone acetate administration on the mitochondrial population and intracellular distribution of phosphatases and ribonucleases was investigated. Cortisone acetate was administrated at a dosage of 25 mgm. per 100 gm. body weight, daily, during five days and sacrificed on the sixth day. The livers of these animals were removed and homogenized in 0.25 M sucrose and subjected to differential centrifugation to prepare nuclear, mitochondrial, microsomal, and supernatant fractions. Nuclear and mitochondrial counts were made for each homogenate prepared. From these the number of mitochondria per average cell was computed. The intracellular distribution of RNAase and PO4ase activities was determined and expressed per average cell and per liver/body weight ratio. The activity of the mitochondrial fraction was also expressed per average mitochondrion. Results of this investigation showed that cortisone lowered the number of large granules per liver cell without influencing the acid PO4ase and acid and alkaline RNAase activities per cell. This suggested that mitochondria isolated from cortisone treated rats possessed abnormally high acid PO4ase and RNAase activities or/and that the remaining population of granules was different from the one in liver cells of untreated animals. The investigation of alkaline phosphatase revealed that cortisone increased the Mg++ sensitive alkaline PO4ase localized in the supernatant fluid without influencing the Mg++ insensitive enzyme.

EFFECT OF CORTISONE ON THE INTRACELLULAR DISTRIBUTION OF PHOSPHATASES AND RIBONUCLEASES IN RAT LIVER

1956 ◽  
Vol 34 (1) ◽  
pp. 170-179 ◽  
Author(s):  
Claude Allard ◽  
Gaston De Lamirande ◽  
George Weber ◽  
Antonio Cantero
Keyword(s):  
Body Weight ◽  
Weight Ratio ◽  
Mitochondrial Fraction ◽  
Intracellular Distribution ◽  
Cortisone Acetate ◽  
Supernatant Fluid ◽  
Body Weight Ratio ◽  
Average Cell ◽  
Mitochondrial Population ◽  
The One

The effect of cortisone acetate administration on the mitochondrial population and intracellular distribution of phosphatases and ribonucleases was investigated. Cortisone acetate was administrated at a dosage of 25 mgm. per 100 gm. body weight, daily, during five days and sacrificed on the sixth day. The livers of these animals were removed and homogenized in 0.25 M sucrose and subjected to differential centrifugation to prepare nuclear, mitochondrial, microsomal, and supernatant fractions. Nuclear and mitochondrial counts were made for each homogenate prepared. From these the number of mitochondria per average cell was computed. The intracellular distribution of RNAase and PO4ase activities was determined and expressed per average cell and per liver/body weight ratio. The activity of the mitochondrial fraction was also expressed per average mitochondrion. Results of this investigation showed that cortisone lowered the number of large granules per liver cell without influencing the acid PO4ase and acid and alkaline RNAase activities per cell. This suggested that mitochondria isolated from cortisone treated rats possessed abnormally high acid PO4ase and RNAase activities or/and that the remaining population of granules was different from the one in liver cells of untreated animals. The investigation of alkaline phosphatase revealed that cortisone increased the Mg++ sensitive alkaline PO4ase localized in the supernatant fluid without influencing the Mg++ insensitive enzyme.

scholarly journals CHANGES IN "DROPLET" FRACTIONS FROM RAT KIDNEY CELLS AFTER INTRAPERITONEAL INJECTION OF EGG WHITE

1957 ◽  
Vol 3 (6) ◽  
pp. 933-947 ◽  
Author(s):  
Werner Straus
Keyword(s):  
Acid Phosphatase ◽  
Intraperitoneal Injection ◽  
Rat Kidney ◽  
Hydrolytic Enzymes ◽  
Bottom Layer ◽  
Egg White ◽  
Supernatant Fluid ◽  
Enzyme Content ◽  
Osmotic Effects

1. Kidney homogenates from rats injected with egg white and from control rats were fractionated simultaneously into six fractions and the content of acid phosphatase, ribonuclease, desoxyribonuclease, cathepsin, and ß-glucuronidase in corresponding fractions from treated and untreated animals was compared. These observations were correlated with the amount of dark brown bottom sediments in fractions NDrI, DrII, and DrIII, and with the number of droplets in fraction NDrI. 2. It was found that after injection of egg white the amount of small droplets decreased as indicated by the decrease of the dark brown bottom layer in the sediment of fraction DrIII and by the concomitant decrease of hydrolytic enzymes in the same fraction, and that the number of large droplets increased as indicated by the increase of brown sediment in fraction NDrI and the increase in the number of droplets counted in a bacterial counting chamber in the same fraction. It was concluded that the treatment with egg white induced the transformation of small droplets into large droplets. 3. The decrease of hydrolytic enzymes in the fractions containing the small droplets was accompanied by a marked increase of these enzymes in the supernatant fluid. The enzyme content of fraction NDrI was not increased after treatment, although it contained greatly increased numbers of large droplets. Counting of the droplets in this fraction showed decreased enzymatic activity of the average large droplet after treatment with egg white. It was suggested that during the transformation of small into large droplets, a portion of the hydrolytic enzymes was released into the surrounding cytoplasm, and that this was partly responsible for the increased enzyme content of the supernatant fluid after fractionation of the kidney homogenate. In contrast to the four other hydrolytic enzymes, ß-glucuronidase was not increased in the supernatant fluid. 4. Eighteen hours after intraperitoneal injection of egg white, the specific enzymatic activities of kidney homogenates showed a 25 to 35 per cent increase for cathepsin, ribonuclease, and desoxyribonuclease, no change for acid phosphatase and ß-glucuronidase, and approximately a 7 per cent decrease for cytochrome oxidase. The increase of cathepsin, ribonuclease, and desoxyribonuclease in the total homogenate was interpreted as an indication of the formation of new enzymes, and it was suggested that this partly accounted for the increase of these enzymes in the supernatant fluid. 5. The activation of the enzymes by osmotic effects was investigated in vitro by incubation of droplet fractions in the presence of different concentrations of sucrose.

scholarly journals Function of mannan chains of yeast repressible acid phosphatase on its enzymatic properties.

1984 ◽  
Vol 48 (6) ◽  
pp. 1445-1450 ◽  
Author(s):  
Takemitsu MIZUNAGA ◽  
Tadahiro OSHIDA ◽  
Akiko TAKASAKI ◽  
Yoshiharu MARUYAMA
Keyword(s):  
Acid Phosphatase ◽  
Enzymatic Properties

Spectrochemical Behavior of Carcinogenic Polynuclear Aromatic Hydrocarbons in Biological Systems. Part I: Steady-State Fluorometry of BaP and BeP in Living Cells

1996 ◽  
Vol 50 (11) ◽  
pp. 1345-1351 ◽  
Author(s):  
Ping Chiang ◽  
Kuang-Pang Li ◽  
Tong-Ming Hseu
Keyword(s):  
Aromatic Hydrocarbons ◽  
Normal Liver ◽  
Liver Cells ◽  
Polynuclear Aromatic Hydrocarbons ◽  
Kidney Cells ◽  
Excimer Emission ◽  
First Order ◽  
Polycyclic Aromatic ◽  
Different Origins ◽  
Time Plot

The rates of interactions of polycyclic aromatic hydrocarbons (PAHs) with biological cells studied with fluorescence appear to be different for cells from different origins. The monomer emission of benzo[ a]pyrene (BaP) is enhanced as in liposomes, but more significantly in normal liver cells than in liver cancer cells or kidney cells, and that enhancement is proportional to the amount of cells added. When PAHs are allowed to interact with cells for a certain period of time, metabolism appears to occur. The excimer emission is seen to dissipate continuously as reactions proceed, whereas the monomer emission increases, passes through a maximum, then starts to decrease when excimer emission becomes exhausted. The time plot of the BaP excimer emission in semilogarithmic coordinates indicates that the decrease or the mass transfer of microcrystalline BaP to the cell membrane is a first-order process. Metabolism has been investigated by monitoring the monomer emission. Liver cells have higher monomer emission than the kidney cells in the early stages of interaction, indicating that these cell membranes are more amenable to PAH absorption. Effects of vitamin K3 and radiation have also been investigated.

scholarly journals The effects of added purines on urate and purine synthesis de novo by isolated chick liver, kidney and lymphoid cells

1976 ◽  
Vol 158 (3) ◽  
pp. 549-556 ◽  
Author(s):  
P Badenoch-Jones ◽  
P J Buttery
Keyword(s):  
De Novo ◽  
Cell Types ◽  
Liver Cells ◽  
Lymphoid Cells ◽  
Kidney Cells ◽  
Purine Synthesis ◽  
Liver And Kidney ◽  
Stimulation Of

1. Isolated chick lymphoid cells, together with isolated chick liver and kidney cells, incorporate [1-14C]glycine or [14C]formate into urate. 2. Of the cell types used, bursal cells incorporate 14C into urate at the fastest rate, although the output of total urate by bursal cells is only 10% that of liver cells. 3. When suspended in Eagle's medium the incorporation of 14C into urate is inhibited by adenine and guanine up to 1 mM. In contrast, the addition of 1 mM-AMP or -GMP results in a relatively large stimulation of this incorporation. 4. Added adenine is rapidly taken up by liver cells and then released in an unmetabolized form; AMP is taken up more slowly and is rapidly metabolized. The metabolites (possibly including adenine) are then released. 5. Intracellular liver 5-phosphoribosyl 1-pyrophosphate is approx. 0.7mM and remains constant or falls slightly during a 3 h incubation of the cells. 6. The addition of adenine or guanine, AMP or GMP, does not alter liver intracellular 5-phosphoribosyl 1-pyrophosphate concentrations. Added 5-phosphoribosyl 1-pyrophosphate is not taken up by liver cells. 7. The results are discussed in the context of the control of urate and purine synthesis de novo in the chick.

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