scholarly journals The distributions of some granule-associated enzymes in guinea-pig polymorphonuclear leucocytes

1970 ◽  
Vol 116 (2) ◽  
pp. 207-216 ◽  
Author(s):  
Robert H. Michell ◽  
Morris J. Karnovsky ◽  
Manfred L. Karnovsky
Keyword(s):  
Alkaline Phosphatase ◽  
Acid Phosphatase ◽  
Guinea Pig ◽  
Soluble Fraction ◽  
Maximum Activity ◽  
Polymorphonuclear Leucocyte ◽  
Urate Oxidase ◽  
Polymorphonuclear Leucocytes ◽  
Granule Fraction ◽  
Two Populations

1. Homogenates of guinea-pig polymorphonuclear leucocytes were separated by differential centrifugation into six particulate fractions and a soluble fraction. 2. The distributions in these fractions of protein, DNA, succinate dehydrogenase, β-glucuronidase, peroxidase, alkaline phosphatase, acid phosphatase (against p-nitrophenyl phosphate and β-glycerophosphate), cathepsin, and catalase were compared. 3. Almost all of the DNA sedimented in the first two pellets, indicating that the nuclei were relatively intact. 4. The four hydrolases and peroxidase showed different distribution patterns, although these activities were previously reported to be localized mainly in the single ‘granule’ fraction isolated from leucocytes. 5. The particles containing peroxidase, acid phosphatase and alkaline phosphatase all exhibited latency. Maximum activity for each enzyme was obtained at roughly similar concentrations of Triton X-100. 6. The acid phosphatase of these cells was distributed between two populations of particles that differed in both sedimentation characteristics and density. The acid phosphatase(s) of the two populations showed slightly different substrate specificities. This bimodal distribution was not an artifact of the procedure used to elicit the cells. 7. Catalase was recovered almost entirely in the soluble fraction and showed no latency in freshly prepared homogenates. No urate oxidase was detected. 8. We conclude that the ‘granule’ fraction of the polymorphonuclear leucocyte, as isolated by previous workers, contains at least three, probably more, populations of particles with different enzyme contents, and that these cells probably do not contain peroxisomes.

PHOSPHATASE OF RABBIT POLYMORPHONUCLEAR LEUCOCYTES

1949 ◽  
Vol 27e (5) ◽  
pp. 290-307 ◽  
Author(s):  
D. M. Cram ◽  
R. J. Rossiter
Keyword(s):  
Alkaline Phosphatase ◽  
Enzyme Activity ◽  
Acid Phosphatase ◽  
Bile Salts ◽  
Polymorphonuclear Leucocytes ◽  
Alkyl Sulphate ◽  
Low Concentrations ◽  
Activity Curve ◽  
Surface Active ◽  
Active Substances

Rabbit polymorphonuclear leucocytes contain an active phosphatase that readily hydrolyzes disodium phenyl phosphate. The pH activity curve of the enzyme was found to have two maxima, one in the region of pH 10 and the other in the region of pH 5. The alkaline phosphatase was much more active than the acid phosphatase. The concentration of alkaline phosphatase in rabbit white cells was approximately one thousand times that of the enzyme in the serum. Under the conditions of study, the alkaline phosphatase activity was proportional to the concentration of the enzyme. The effect of substrate concentration on the enzyme activity was studied and the Michaelis constant (Ks) determined. An excess of substrate inhibited the enzyme. The course of the reaction was linear with time for the first 60 min.; after 90 min. the activity fell off faster than would be expected if the reaction were of the first order.Magnesium and glycine, in low concentrations, caused an increase in the enzyme activity, whereas zinc, cyanide, borate, phosphate, bile salts, and glycine, in higher concentrations, were inhibitory. Fluoride had no demonstrable effect. Surface-active substances, such as saponin, bile salts, or alkyl sulphate, liberated the enzyme from the cells. Similar results were obtained when α-glycerophosphate or β-glycerophosphate was used as the substrate.The alkaline phosphatase can be considered to belong to Class AI of Folley and Kay (22) and the acid phosphatase to Class AII. The alkaline phosphatase can also be considered to be a Phosphatase II of Cloetens (9).

The Influence of Short Ether Anesthesia on Some Liver Enzyme Activities in the Rat

1973 ◽  
Vol 51 (8) ◽  
pp. 604-607 ◽  
Author(s):  
E. Katona
Keyword(s):  
Alkaline Phosphatase ◽  
Acid Phosphatase ◽  
Malate Dehydrogenase ◽  
Enzyme Activities ◽  
Urate Oxidase ◽  
Arylsulfatase A ◽  
Ether Anesthesia ◽  
Arylsulfatase B ◽  
Thiamine Pyrophosphatase ◽  
Liver Homogenates

Rats were anesthetized by ether inhalation for 4–5 min and sacrificed 1–48 h after anesthesia. From their liver homogenates, the activities of nine enzymes were determined. Activities of urate oxidase and arylsulfatase-A did not change significantly but arylsulfatase-B was slightly decreased. Malate dehydrogenase, arylsulfatase-B, and thiamine pyrophosphatase reached their highest and "malic enzymes" their lowest activities at the same time, 5 h after anesthesia. Alkaline phosphatase first decreased, later increased. Acid phosphatase and glucose-6-phosphatase activities decreased following ether anesthesia. Thesechanges in the enzyme activities generally agree and partly explain previously reported effects of ether anesthesia observed in the serum.

scholarly journals Distribution of basal lysosomes in exocrine acinar cells.

1987 ◽  
Vol 35 (5) ◽  
pp. 565-570 ◽  
Author(s):  
C Oliver ◽  
Y Yuasa
Keyword(s):  
Acid Phosphatase ◽  
Golgi Apparatus ◽  
Guinea Pig ◽  
Secretory Granules ◽  
Acinar Cells ◽  
The Other ◽  
Basal Region ◽  
Golgi Region ◽  
Basal Portion ◽  
Two Populations

We examined the distribution of trimetaphosphatase (TMPase)-positive basal lysosomes in pancreas, parotid, submandibular, sublingual, and exorbital lacrimal glands from rats, rabbits, and guinea pigs. The location of the basal lysosomes was compared to that of the acid phosphatase (AcPase)-positive lysosomes. In all of the tissues examined from rat and rabbit, AcPase activity was localized primarily to the Golgi region. Reaction product was localized in GERL, immature secretory granules, and lysosomes lying adjacent to the Golgi apparatus. TMPase activity was found in basal lysosomes and in occasional elongated lysosomes adjacent to the Golgi apparatus. In guinea pig, the distribution of TMPase activity was identical to that seen in the other two species, but a significant number of lysosomes in the basal region of the cells also contained AcPase activity. These results confirm and extend our previous finding (J Histochem Cytochem 31:1209, 1983) that exocrine acinar cells possess two distinct populations of lysosomes. The lysosomes in the Golgi region contain both AcPase and TMPase activity, whereas those in the basal portion of the cells are reactive predominantly for TMPase. The functional significance of the two populations of lysosomes is not understood at present.

ALKALINE AND ACID PHOSPHATASE IN CEREBROSPINAL FLUID: DATA FOR NORMAL FLUIDS AND FLUIDS FROM PATIENTS WITH MENINGITIS, POLIOMYELITIS, OR SYPHILIS

1950 ◽  
Vol 28e (2) ◽  
pp. 56-68 ◽  
Author(s):  
K. G. Colling ◽  
R. J. Rossiter
Keyword(s):  
Alkaline Phosphatase ◽  
Acid Phosphatase ◽  
Blood Plasma ◽  
Cell Count ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Protein Concentration ◽  
White Cell Count ◽  
Polymorphonuclear Leucocytes ◽  
Alkaline And Acid Phosphatase

Many normal cerebrospinal fluids contain an alkaline (pH 9.8) and an acid (pH 4.9) phosphatase. Both the alkaline and the acid phosphatase were significantly increased in the spinal fluids from patients with meningitis or poliomyelitis, but not in the fluids from patients with syphilis. The alkaline phosphatase activity was correlated with both the concentration of protein in the spinal fluid and with the white cell count, whereas the acid phosphatase was correlated with neither. When correction was made for the significant correlation between cell count and protein concentration, the partial correlation between alkaline phosphatase activity and both protein concentration and cell count remained significant statistically. In pathological conditions it appears likely that the alkaline phosphatase is derived partly from the polymorphonuclear leucocytes in the fluid and partly from the blood plasma. The acid phosphatase is probably derived from the lymphocytes of the fluid and possibly also from the blood plasma. It is unlikely that either of these enzymes comes from the substance of the brain or spinal cord. Acid phosphatase would be of more value than alkaline phosphatase as a diagnostic aid, since normal fluids contain much less of this enzyme.

scholarly journals PHAGOCYTIN: A BACTERICIDAL SUBSTANCE FROM POLYMORPHONUCLEAR LEUCOCYTES

1956 ◽  
Vol 103 (5) ◽  
pp. 589-611 ◽  
Author(s):  
James G. Hirsch
Keyword(s):  
Bactericidal Activity ◽  
Soluble Fraction ◽  
Proteolytic Enzymes ◽  
Cell Types ◽  
Rabbit Heart ◽  
Polymorphonuclear Leucocyte ◽  
Polymorphonuclear Leucocytes ◽  
Large Numbers ◽  
Aqueous Salt Solutions ◽  
Salt Fractionation

A technique has been developed for collecting large numbers of polymorphonuclear leucocytes from peritoneal exudates in rabbits. These cells are obtained essentially free from other cell types and from debris. When microphages so procured are disrupted by physical methods and extracted with aqueous salt solutions, the soluble fraction manifests striking bactericidal activity, especially on Gram-negative enteric bacilli. The susceptible microorganisms are not lysed. This bactericidal substance, which has been called phagocytin, appears to be limited in distribution mainly to the polymorphonuclear leucocyte. No phagocytin is present in extracts of rabbit heart, kidney, or skeletal muscle, and rabbit liver and spleen contain much less than do packed leucocytes. Extracts of human and of guinea pig microphages show less bactericidal activity than rabbit cell preparations. Similar extracts of rat and mouse polymorphonuclear leucocytes contain no demonstrable phagocytin. As indicated by its behavior on dialysis, on exposure to proteolytic enzymes, and on salt fractionation, phagocytin appears to be a protein with general properties characteristic of a globulin. It is clearly different from lysozyme and from properdin. Although phagocytin is reasonably stable at temperatures of 65°C. and lower for several hours, solutions of it gradually lose bactericidal activity on standing for prolonged periods at 4°C. This instability, and also the ease with which phagocytin is inactivated, presumably by adsorption, on exposure to a variety of materials, have thus far rendered fruitless efforts to isolate it.

scholarly journals THE ISOLATION AND PROPERTIES OF THE SPECIFIC CYTOPLASMIC GRANULES OF RABBIT POLYMORPHONUCLEAR LEUCOCYTES

1960 ◽  
Vol 112 (6) ◽  
pp. 983-1004 ◽  
Author(s):  
Zanvil A. Cohn ◽  
James G. Hirsch
Keyword(s):  
Polymorphonuclear Leucocyte ◽  
The Other ◽  
Considerable Proportion ◽  
Polymorphonuclear Leucocytes ◽  
Cytoplasmic Granules ◽  
Surface Active Agents ◽  
Granule Fraction ◽  
Liver Lysosomes ◽  
Surface Active ◽  
Phosphatase Alkaline

A method has been described for isolation of the specific cytoplasmic granules of rabbit polymorphonuclear leucocytes. Homogeneous suspensions of leucocytes were disrupted by lysis in 0.34 M sucrose. This procedure liberated the cytoplasmic contents of the cell and dissolved a considerable proportion of the nuclei. Following disruption, the sucrose lysate was separated into three fractions by differential centrifugation, i.e. 400 g or nuclear pellet, 8,200 g or granule pellet and the postgranule supernate. Microscopic examination revealed that the 8,200 g pellet was composed of intact granules as well as occasional mitochondria. The other two fractions were morphologically heterogeneous. Studies with isolated granules demonstrated their lysis by a variety of weak acids and surface-active agents. When buffered solutions were employed between the ranges of pH 2.0 and 9.0, granule lysis began at pH 5.5 and was complete at pH 4.0. Chemical analysis disclosed that the granule pellet contained protein and phospholipid with only traces of nucleic acids. Approximately 70 to 80 per cent of the total cellular antimicrobial agent phagocytin was present in the granule fraction. This material was liberated from the granules by acid (pH 5.0 or lower). Studies on selected enzymes showed that acid phosphatase, alkaline phosphatase, nucleotidase, ribonuclease, deoxyribonuclease, and beta glucuronidase were predominantly localized in the granule fraction. Approximately 50 per cent of total cellular lysozyme and cathepsin were also present in the 8,200 g pellet. Disruption of the granules was associated with the release of the majority of granule protein and enzymes in a non-sedimentable form. The properties and composition of rabbit polymorphonuclear leucocyte granules seem to be analogous to those of liver lysosomes.

scholarly journals Collagenolytic cathepsin activity in rabbit peritoneal polymorphonuclear leucocyte granules

1978 ◽  
Vol 172 (1) ◽  
pp. 83-89 ◽  
Author(s):  
W T Gibson ◽  
D W Milsom ◽  
F S Steven ◽  
J S Lowe
Keyword(s):  
Density Gradient ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Polymorphonuclear Leucocyte ◽  
Polymorphonuclear Leucocytes ◽  
Differential Centrifugation ◽  
Ph Optimum ◽  
Cathepsin Activity ◽  
Granule Fraction ◽  
Alpha Beta

Collagenolytic cathepsin activity was detected in lysed rabbit peritoneal polymorphonuclear leucocytes. The pH optimum was around 3, and activity was greatly enhanced by the presence of cysteine and EDTA. Digestion of polymeric collagen resulted in the release of alpha, beta, and gamma-chains. Collagenolytic cathepsin activity was associated mainly with the granule fraction isolated from homogenates by differential centrifugation. The granule fraction was further fractionated by isopycnic density-gradient centrifugation, and the collagenolytic cathepsin activity was shown to be associated with the azurophil and tertiary granules, both lysosome-like organelles.

scholarly journals The determination and localization of sialic acid in guinea-pig granulocytes

1980 ◽  
Vol 192 (2) ◽  
pp. 543-550 ◽  
Author(s):  
J W DePierre ◽  
J Lazdins ◽  
M L Karnovsky
Keyword(s):  
Plasma Membrane ◽  
Sialic Acid ◽  
Guinea Pig ◽  
Total Cell ◽  
Polymorphonuclear Leucocytes ◽  
Intact Cells ◽  
Granule Fraction ◽  
Specific Determination ◽  
Chromatographic Procedure

When intact guinea-pig granulocytes (polymorphonuclear leucocytes) disrupted by sonication or with detergent were treated with neuraminidase from Vibrio cholerae, 3.1–3.2 nmol of sialic acid/10(7) cells was released. By using a chromatographic procedure for the specific determination of total cell sialic acid, this releasable portion was found to constitute 70% of the total sialate. All of the neuraminidase-releasable sialic acid of the cells could be removed by enzymic treatment of intact cells with neuraminidase. It thus seemed likely that the neuraminidase-releasable sialic acid is all on the cell surface. To make sure that the result was not due to entry of neuraminidase into the cells, the enzyme was bound covalently to Sepharose 6B, and intact polymorphonuclear leucocytes were treated with the bound enzyme. All of the neuraminidase-releasable sialic acid could still be removed, though more slowly. The cells remained intact and only 1.5–2% of the bound enzyme was released from the Sepharose during incubation. Freed enzyme could have been responsible, at the very most, for release of 18% of the sialic acid. Fractionation studies showed that the nucleus and cytoplasm contain low amounts of sialic acid and that the neuraminidase-releasable sialic acid distributes in a manner similar to the distribution of 5′-nucleotidase, an unambiguous marker for the plasma membrane in these cells. Thus neuraminidase-releasable sialate constitutes a clear marker for the membrane of polymorphonuclear leucocytes. Most of the neuraminidase-insensitive sialate was present in the granule fraction. Removal of sialic acid from intact polymorphonuclear leucocytes did not affect their ecto-AMPase, -ATPase and -p-nitrophenyl phosphatase activities.

Effects of Kanwa on Rat Gastrointestinal Phosphatases

2010 ◽  
Vol 3 (3) ◽  
pp. 1147-1152
Author(s):  
Jacob Bamaiyi ◽  
Omajali ◽  
Sanni Momoh
Keyword(s):  
Alkaline Phosphatase ◽  
Small Intestine ◽  
Acid Phosphatase ◽  
Sodium Carbonate ◽  
Elevated Level ◽  
Food Additive ◽  
Alkaline Phosphatases ◽  
Acid And Alkaline Phosphatases ◽  
Diagnostic Indices ◽  
High Level

This study investigates the effects of kanwa on rat gastrointestinal phosphatases. The rats were administered 7% w/v concentration of  trona (Kanwa) orally for a period of two weeks in order to investigate how this compound is being used as food additive in some homes in Nigeria. The Kanwa used in this study was the handpicked variety obtained from sellers from Anyigba market in eastern part of Kogi State, Nigeria. Kanwa, a hydrated sodium carbonate (Na2CO3NaHCO3.2H2O) was obtained as a dried lake salt. Acid phosphatase has the ability to dephosphorylate molecules containing phosphate group. The decreased and elevated level in serum or plasma acid and alkaline phosphatases serves as diagnostic indices for various diseases. Results showed that there was increase and decrease of acid phosphatase (ACP) activities in both the stomach and small intestine. The activities of alkaline phosphatase (ALP) fluctuated in the small intestine. However, in the stomach, an increase activity of ALP was noticed throughout the period of ‘Kanwa’ administration. We concluded that although the level of ‘Kanwa’ consumed in most homes may not be toxic if not taken continuously or repeatedly. Thus, continuous consumption should be discouraged as accumulation of high level of ‘Kanwa’ may cause damages or injuries to the various organs/tissues and may disrupt normal body function.

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