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1.
Effects of treatmentin vivo with the antimalarials:chloroquine (CQ), primaquine (PQ) and quinine(Q) on lysosomal enzymes and lysosomal membrane integrity were examined. Treatment with the three antimalarials showed an apparent increase in the membrane stability. CQ treatment resulted in increase in both the ‘free’ and ‘total’ activities of all the enzymes i.e. acid phosphatase, RNase II, DNase II and cathepsin D. PQ treatment lowered the ‘free’ and ‘total’ activities of acid phosphatase and cathepsin D, but the DNase II activities increased. Treatment with Q resulted in increased ‘free’ and ‘total’ activities of RNase II and DNase II. While ‘free’ activities of acid phosphatase and cathepsin D were low; the ‘total’ activities increased significantly. Our results suggest that a generalized increase in free nucleases activities following prolonged treatment with antimalarials may lead to cell damage and/or necrosis.  相似文献   

2.
The cysteine proteinases like cathepsins B, L and H are main hydrolytic enzymes present in lysosomes and play an important role in intracellular protein degradation. Tuberculosis and leprosy, both are tissue- destructive diseases. Main drugs used in chemotherapy of these diseases may inhibit the main lysosomal cysteine proteinases i.e. cathepsins B, L and H released during tissue destruction and thus prevent the further destruction of tissue by these enzymes. So the aim of this study is to see the effect of antituberculous and antileprotic drugs on these proteolytic enzymes. The effect of commonly used antituberculous and antileprotic drugs was screened on the activities of purified brain lysosomal cysteine proteinases namely cathepsins B [EC 3.4.22.1], L [EC 3.4.22.15] and H [EC 3.4.22.16]. Among the antileprotic drugs, only clofazimine inhibited the enzymic activities whereas dapsone had no effect whatsoever. In antituberculous drugs, rifampicin was the most inhibitory while isoniazid had little inhibitory potency. Streptomycin and pyrazinamide did not effect the activities at all. As regards the mechanism of inhibition, clofazimine and isoniazid inhibited the enzymes in a non-competitive manner with K values of 0.25 mM and 5.0 mM for cathepsin B, 0.071 mM and 0.833 mM for cathepsin L and 1.513 mM and 0.885 mM for cathepsin H, While rifampicin could effect in a competitive manner with Ki values of 0.03 mM, 0.125 mM and 0.027 mM for cathepsin B, L and H respectively.  相似文献   

3.
The changes in the erythrocyte lipid peroxidation products (MDA), levels of glutathione (GSH), ascorbic acid and plasma vitamin E (non enzymatic antioxidant parameters) and activities of antioxidant enzymes superoxide dismutase (SOD), glutathione peroxidase (GPX), catalase in erythrocytes and plasma glutathione - S - transferase (GST) activity were estimated in patients with rheumatoid arthritis. This work was undertaken to assess oxidative stress and anti oxidant status in patients with rheumatoid arthritis. It was observed that there was a significant increase in erythrocyte MDA levels, activities of SOD, GPX, plasma GST and a significant decrease in erythrocyte GSH, ascorbic acid, plasma vitamin E levels and catalse activity in patients with rheumatoid arthritis when compared to controls. The results of our study suggests higher oxygen free radical production, evidenced by increased MDA and decreased GSH, ascorbic acid, vitamin E and Catalase activity, support to the oxidative stress in rheumatoid arthritis. The increased activities of antioxidant enzymes may be a compensatory regulation in response to increased oxidative stress.  相似文献   

4.
Mucopolysaccharidosis are a group of rare metabolic disorders of the lysosomal storage disease family caused by the absence or malfunctioning of lysosomal enzymes responsible for their breakdown. It encompasses disorders in which undegraded or partly degraded glycosaminoglycans accumulate in the lysosomes of many tissues owing to a deficiency of specific lysosomal enzymes. Here we report a case of a 7 years old child displaying the symptoms of Morquio’s disease (Mucopolysaccharidosis type IV). Urine screening tests were performed which gave contrasting results.  相似文献   

5.
Recent acquisitions on the early detection and monitoring of the progression of diabetic complications (nephropathy) using the techniques of enzymology (lysosomal enzymes) are reviewed. it appears that the kidney is the principal source of urinary lysosomal enzymes. Urinary samples for lysosomal enzyme determination can be either 24-hour or spot-collection. The use of synthetic substrates (4-methylumbelliferyl substrates) provides an easy, inexpensive, sensitive and highly reproducible method of lysosomal enzyme assay. It is recommended that more than one enzyme be assayed in the process. The use of fractional enzyme excretion (FEE) ratios is further recommended. The urinary lysosomal glycosidases investigated and found to be of particular diagnostic value in the early detection of diabetic nephropathy include N-acetyl-β-D-glucosaminidase (β-hexosaminidase, NAG), β-glucuronidase and β-galactosidase, with NAG being the most useful indicator. Urinary NAG can be used in monitoring the progression of diabetic nephropathy. The fluorimetric assay of lysosomal glycosidases is particuarly recommended in developing countries since it is simple, sensitive and inexpensive.  相似文献   

6.
Early, accurate diagnosis of lysosomal storage disorders is a major challenge, even for trained specialists. Finding innovative, accurate diagnostic methods, and high throughput, cost-effective tools are crucial to medical progress and will contribute to improved quality of life. The goal of this work was to improve currently used protocols to determine activity of acid β-galactosidase, and discuss the possibility analysing lysosomal enzymes with microfluidic systems. A principle of the determination of β-galactosidase activity was fluorometric measurement of a deprotonated form of 4-methylumbelliferone released in the enzymatic reaction. Measurements were performed using Jurkat T cells as a source of the enzyme. We observed the temperature-dependent substrate inhibition effect and determined the substrate (4-MU-β-d-galactopyranoside) concentration which should be used to determine acid β-galactosidase activity at 37 °C (0.8 mM) and at room temperature (0.6 mM). We proved that the sample incubation time may be significantly reduced to only a few minutes. We also showed that the amount of alkaline buffer used to stop the enzymatic reaction may be minimized and even, in some cases, eliminated. The presented results show how the sensitivity of the available methods to diagnose patients suffer from gangliosidosis GM1 or Morquio B disease can be improved. The proposed method may be easily implemented with microfluidic systems, which currently are promising tools for point-of-care applications.  相似文献   

7.
The microbicidal capacity of polymorphonuclear leucocytes of diabetic and control subjects was evaluated by estimating the release of lysosomal enzymes viz beta-glucuronidase, lysozyme, acid phosphatase, alkaline phosphatase, in response to a particulate stimulus-serum treated zymosan (STZ). The cells untreated and pretreated with cytochalasin B were exposed to STZ The total enzyme activities were estimated after cell lysis. The total enzyme activities were not altered in diabetic subjects as compared to control subjects. The release of lysosomal enzymes by cells pretreated with cytochalasin B was high as compared to untreated cells. The release of lysosomal enzymes from the cells isolated from diabetic patients, untreated and pretreated with cytochalasin B was reduced as compared to controls. The findings of the present study that the total lysosomal enzyme activities are normal while the release of these enzymes in response to stimulus is impaired in diabetics, suggest that the bactericidal capacity of these cells which involves phagocytosis is impaired in these patients.  相似文献   

8.
Changes in plasma fatty acid (FA) composition and desaturase activities are observed in metabolic syndrome (MS). However, whether these changes are a reflection of dietary intakes of fats and FAs is not well established. The current study was aimed at assessing plasma FA composition and desaturase enzyme activities as biomarkers of dietary intakes in subjects with MS. Case control study was done on 41 MS patients and was compared with age matched 45 controls. Dietary intakes, anthropometric and clinical parameters were measured. FA composition was analysed using gas chromatography-flame ionisation detector and desaturase enzyme activities were estimated as ratios of product to precursor FAs. Higher levels of 14:0, 16:0, 16:1, 18:1, D9D-18 activity and lower levels of 18:0 and 18:2 n-6 were seen in MS group when compared to controls (p < 0.05). Strong positive correlations were seen between plasma triglyceride (TG) levels and 14:0, 16:0, 16:1, 18:1, total saturated fatty acid, total monounsaturated fatty acid, and D9D activities, while 18:0, 18:2 n-6 and total polyunsaturated fatty acid were negatively correlated with TG. Positive correlations were seen between plasma 14:0, 18:1 and D9D-18 activity with total energy intake and carbohydrate (CHO) intakes but not with fat intake. Plasma FA profile appears to be a better index of total energy intake and CHO intake than fat intake, suggesting it might be a good reflection of endogenous FA metabolism. Changes in FA composition may therefore serve as an early index of dysregulation of FA metabolism, resulting in increased risk of MS.  相似文献   

9.
There was increase in erythrocyte lipid peroxidation and adenosine deaminase (ADA) activity in red blood cells (RBC's), white blood cells (WBC's) and plasma, in 30 patients of confirmed diabetes mellitus, 10 each of insulin dependent diabetes mellitus (IDDM or type I), non-insulin dependent diabetes mellitus (NIDDM or type II) and diabetes with ketoacidosis when compared with 20 healthy individuals (controls). Glycosylated hemoglobin A1c%, plasma free fatty acid (FFA) and glycerol levels were also elevated in all the cases when compared with controls. The increase was more in diabetic ketoacidosis group. Significant positive correlations were seen between erythrocyte lipid peroxidation and ADA activities in RBC's, WBC's and plasma. It is suggested that decreased tissue adenosine levels due to increase in ADA activity, is related to the severity of hyperglycemia and lipid peroxidation in diabetes mellitus.  相似文献   

10.
The microbicidal capacity of polymorphonuclear (PMN) leukocytes of normal human subjects was evaluated by estimating the release of lysosomal enzymes from normal cells and cells rendered incapable of ingesting particles by treatment with cytochalasin B, on exposure to a particulate stimulus-serum treated zymosan (STZ). The release of lysosomal enzymes namely beta-glucuronidase, lysozyme, acid phosphatase, alkaline phosphatase was found to be 16.28%, 27.84%, 20.54%, 15.40% respectively. The cells rendered incapable of ingesting particles showed enhanced response. The percentage release of beta-glucuronidase, lysozyme, acid and alkaline phosphatase was increased to 20.58%, 37.95%, 25.87% and 33.04% respectively. This type of response in cytochalasin B treated PMN leukocytes is due to the conversion of these cells into “secretory cells”, which release lysosomal enzymes outside the cell, by the process of reverse endocytosis.  相似文献   

11.
Changes in the levels of glycoprotein (GP) and glycosminoglycans (GAG) were studied in diabetes associated with myocardial infarction in rats. Diabetes was induced by alloxan while myocardial infarction was induced using isoproterenol. Most of the GAG fraction increased in the heart and aorta in diabetes superimposed with myocardial infarction. The carbohydrate of GP also increased in heart while in serum only sialic acid showed an increase. The activity of lysosomal enzymes showed an increase in the case of β-glucuronidase and β-hexosaminidase while in the case of β-galactosidase and β-fucosidase it showed a decrease in the heart.  相似文献   

12.
The antiulcerogenic effects of the methanolic extract ofSolanum nigrum berries (SBE) on aspirin induced ulceration in rats with respect to antioxidant status in the gastric mucosa have been investigated. Oxygen free radicals are considered to be important factors in the pathogenesis of gastric ulcer. The level of lipid peroxides, which were elevated highly in rats with acute gastric mucosal injury was taken as an index of oxidative stress. The activities of antioxidant defense enzymes were also decreased considerably by oral gastric administration of aspirin. The decreased levels of antioxidant enzymes and increased mucosal injury were altered to near normal status upon pretreatment with (SBE) when compared to the ulcer induced rats. The results indicate that (SBE) may exert its gastroprotective effect by a free radical scavenging action. Our observations suggest that (SBE) may have considerable therapeutic potential in the treatment of gastric diseases.  相似文献   

13.
The activity of six hydrolytic enzymes-carboxyl esterase, acid phosphatase, alkaline phosphatase, β-galactosidase, β-glucosidase and β-hexosaminidase, were studied in different regions of the normal human brain tissue obtained at autopsy. Protein estimation and activities of the hydrolytic enzymes with respective substrates were assayed by spectrophotometric and spectroflourometric methods. Amongst the eight regions of the brain-frontal, parietal, occipital, temporal, thalamus, cerebellum and hippocampus, the pineal gland showed highest activity for all hydrolytic enzymes studied except for carboxyl esterase. Among six hydrolases studied, hexosaminidase exhibited highest activity in all regions of the human brain while alkaline phosphatase activity was the least amongst all regions studied. A majority of the enzymes studied showed higher activity in gray matter as compared to the white matter except acid phosphatase and β-glucosidase which exhibited higher activity in the white matter. The most significant finding in the present study was the high activity of all hydrolytic enzymes noted in the pineal gland as compared to all other regions of the human brain. Such a finding has not been hitherto reported earlier in human brain tissue samples. If the specific activities of these enzymes are to be considered as any functional index, then pineal gland may be more metabolically active tissue with respect to the hydrolytic function as compared to the other regions of the brain.  相似文献   

14.
The effect of oral administration of vitamin E for twenty-eight consecutive days on blood glucose, reduced glutathione levels, antioxidant enzymes (superoxide dismutase and catalase activities), and levels of malondialdehyde (as an index of free radical-mediated lipid peroxidation) was observed in the whole blood and liver of normal and alloxan-induced diabetic rats. It was found that oral administration of vitamin E significantly (p<0.05) lowered the blood glucose level and increased the body weight of the diabetic rats. The activities of superoxide dismutase and levels of reduced glutathione increased significantly (p<0.05) while the level of lipid peroxidation decreased.  相似文献   

15.
Serum Adenosine deaminase (ADA) activity in normal healthy control subjects increases upto 30 years, remains steady between 31–60 years of age and shows a steep increase in the age group of 61–70 years. This was compared with serum aspartate transaminase (AST) and alanine transaminase (ALT) activity which also showed a gradual increase upto 40 years of age and decreased thereafter. The activities of serum ADA, AST and ALT increased in patients with hepatitis of all age groups compared to their respective controls. The degree of increase in the activities of the above enzymes in hepatitis, decreased with age. The present study also shows that while studying serum ADA activity in hepatitis for diagnostic purposes, the value obtained in a particular age group should be compared with normal range of values for the respective age group only.  相似文献   

16.
Activities of antioxidant enzymes involved oxygen free radical metabolism in the liver and lung were investigated in 2,4, and 30 months old Wistar albino rats. The levels of SOD, GSH-Px, GSH and protein-SH were decreased whereas MDA increased, in the liver as well as the lung of the old rats (30 months old) compared to the young rats (2 and 4 months old). The data suggests that the failure of the free radical protective mechanism may be one of the important contributors to the process of ageing.  相似文献   

17.
Psoriasis is a chronic inflammatory, proliferative skin disease characterized by pathological skin lesions due to various exogenous and endogenous factors. It is associated with a number of biochemical and immunological disturbances. Recently, it has been suggested that increased reactive oxygen species (ROS) production and compromised function of antioxidant system may be involved in the pathogenesis of this disease. In the present study, 90 psoriasis patients were selected. Disease severity was assessed by psoriasis area severity index score and grouped as mild, moderate and severe (each group consists of 30 subjects) and compared with 30 healthy controls. Serum levels of malondialdehyde, nitric oxide end products and the activities of antioxidant enzymes such as erythrocyte-superoxide dismutase, catalase and total antioxidant status were investigated in these groups/subjects. As compared to controls, we found severitywise significantly increased serum malondialdehyde, nitric oxide end products with decrease in erythrocyte-superoxide dismutase activity, catalase activity and total antioxidant status in patients with psoriasis suggesting worsening of the disease. It seems to be linked with the enhancement of Reactive Oxygen Species production and decreased antioxidant potential in psoriasis.  相似文献   

18.
The activity of enzymes of glycolysis has been studied in erythrocytes from type-II diabetic patients in comparison with control. RBC lysate was the source of enzymes. In the diabetics the hexokinase (HK) activity increased 50 % while activities of phosphoglucoisomerase (PGI), phosphofructokinase (PFK) and aldolase (ALD) decreased by 37, 75 and 64 % respectively but were still several folds higher than that of HK. Hence, it is possible that in the diabetic erythrocytes the process of glycolysis could proceed in an unimpaired or in fact may be augmented due to increased levels of G6P. The lactate dehydrogenase (LDH) activity was comparatively high in both the groups; the diabetic group showed 85 % increase. In control group the HK, PFK and ALD activities showed strong positive correlation with blood sugar level while PGI activity did not show any correlation. In the diabetic group only PFK activity showed positive correlation. The LDH activity only in the control group showed positive correlation with marginal increase with increasing concentrations of glucose.  相似文献   

19.
The present study conducted on twenty-five uncomplicated cases of acute myocardial infarction diagnosed by clinical and electrocardiographic findings indicated significantly increased level of cardiac Troponin-T and increased activities of the enzymes total creatine kinase, creatine kinase-MB, aspartate transaminase and lactate dehydrogenase as compared to the twenty-five healthy control subjects. The level of cardiac Troponin-T and the activities of the enzymes total creatine kinase, creatine kinase-MB, aspartate transaminase and lactate dehydrogenase was found to be higher in “Q” wave myocardial infarction patients as compared to the non-“Q” wave and the controls. Since cardiac Troponin-T has been shown to increase in unstable angina and renal failure without cardiac disease and creatine kinase-MB activity has been found to be normal in patients with unstable angina and increase very slightly in patients with renal failure, it was concluded that a combination of cardiac Troponin-T and creatine kinase-MB activity was sufficient for the early diagnosis of acute myocardial infarction.  相似文献   

20.
Mitochondrial diseases are a heterogeneous group of disorders in which a primary mitochondrial dysfunction is proven by morphological, biochemical, and genetic examinations. The mitral valve has important function in the regulation of blood flow from one chamber to another. Often, the mitral valve becomes abnormal with age, in Rheumatic fever or it is abnormal from birth (Congenital) or it can be destroyed by infection i.e. bacterial endocarditis and needs replacement. Myocardial function depends on energy produced by mitochondria and in any of these disease conditions, mitochondrial functions and enzyme activities may be impaired. With this in view, we analyzed the relationship between preoperative clinical conditions (as per New York heart Association) and extent of mitochondrial enzyme activities in damaged Human mitral valve in two types of heart disease such as Rheumatic Heart Disease (RHD) and Bacterial Endocarditis (BE). Thirty nine Patients undergoing cardiopulmonary bypass (CPB) for routine valvular heart surgery were included in the study. Controls included 11 normal porcine mitral valve samples without any evidence of heart disease. Mitochondrial enzymes like cytochrome oxidase (COX), succinate dehydrogenase (SDH), malate dehydrogenase (MDH), citrate synthase (CS) and ATPase were determined. Mitochondrial COX, SDH, CS and Total ATPase activities were significantly decreased in disease condition like BE and RHD when compared with control (P<0.001). On the other hand as per New York Heart Association (NYHA) preoperative clinical classification, all the mitochondrial enzymes were significantly (p<0.05) impaired in class IV as compared with NYHA class I, II and III. Present study shows that impairment in the mitochondrial enzymes activities are more pronounced in bacterial endocarditis (BE). It also indicates that damage to mitochondrial enzymes are most pronounced in NYHA class IV.  相似文献   

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