Enzyme Activities In Different Tissues Research Articles

Galactose epimerase deficiency: lessons from the GalNet registry

BackgroundGalactose epimerase (GALE) deficiency is a rare hereditary disorder of galactose metabolism with only a few cases described in the literature. This study aims to present the data of patients with GALE deficiency from different countries included through the Galactosemia Network to further expand the existing knowledge and review the current diagnostic strategy, treatment and follow-up of this not well characterized entity.MethodsObservational study collecting medical data from December 2014 to April 2022 of 22 not previously reported patients from 14 centers in 9 countries. Patients were classified as generalized or non-generalized based on their genotype, enzyme activities in different tissues and/or clinical picture and professional judgment of the treating physician.ResultsIn total 6 patients were classified as generalized and 16 as non-generalized. In the generalized group, acute neonatal illness was reported in 3, cognitive and developmental delays were present in 5 and hearing problems were reported in 3. Four generalized patients were homozygous for the genetic variant NM_001008216.2:c.280G > A (p.Val94Met). In the non-generalized group, no clearly related symptoms were found. Ten novel genetic variants were reported in this study population.ConclusionThe phenotypic spectrum of GALE deficiency ranges from asymptomatic to severe. The generalized patients have a phenotype that is in line with the 9 described cases in the literature and prescribing dietary interventions is the cornerstone for treatment. In the non-generalized group, treatment advice is more difficult. To be able to offer proper counseling, in addition to red blood cell enzyme activity, genetic studies, transferrin glycoform analysis and enzymatic measurements in fibroblasts are recommended. Due to lack of facilities, additional enzymatic testing is not common practice in many centers nor a tailored long-term follow-up is performed.

ACETYLCHOLINESTERASE, AS A POTENTIAL BIOMARKER OF NAPHTHALENE TOXICITY IN DIFFERENT TISSUES OF FRESHWATER TELEOST, ANABAS TESTUDINEUS

Naphthalene, a Polycyclic Aromatic Hydrocarbon is widely used as a fumigant and disinfectant despite its toxic effect and is ranked as the ninth most threatening compound. The present study was carried out to determine the in vivo effect of naphthalene at different concentrations on acetylcholinesterase (AChE) enzyme activity in different tissues of Anabas testudineus. The fishes were exposed to varying concentrations of naphthalene (4.2 mgL–1, 4.4 mgL–1, 4.6 mgL–1, 4.8 mgL–1 and 5 mgL–1) for a period of 72 hours. Acetylcholinesterase enzyme activity was found to be significantly inhibited, in a dose-response manner. The inhibition percentage of AChE activity varied from 9.34–43.95% in brain tissue, 2.56–35.81% in liver tissue, 5.94–34.15% in muscle tissue and 3.92–33.75% in gills in comparison to the tissues of the control group. Maximum inhibition of acetylcholinesterase enzyme activity in treated fish was observed in the brain followed by liver, muscles, and gills. This study highlights the significance and role of acetylcholinesterase as a potential stress biomarker of naphthalene toxicity.

Zinc-lysine prevents chromium-induced morphological, photosynthetic, and oxidative alterations in spinach irrigated with tannery wastewater.

Anthropogenic activities have resulted in severe environmental degradation. Untreated wastewater from tanneries is hazardous to all kinds of life on earth. Effluent from tanning industries, containing large amount of Cr, is used to irrigate the crops in Pakistan. The current experiment was carried out to study the effects of tannery wastewater on spinach and the role of lysine-Zn in mitigating the severity of stress. The plants were grown in soil and the following treatments were used: irrigation with 0%, 33%, 66%, and 100% wastewater (ww) along with two doses (0 mM, 10 mM) of Zn-lysine. Foliar application of zinc-lysine enhanced the plant growth, biomass, Zn contents, photosynthesis, and enzyme activities in different tissues of plant. Zinc-lysine (10 mM) considerably decreased the Cr content in roots and shoots, along with ameliorating the oxidative stress by enhancing the activities of antioxidant enzymes in plants. Addition of Zn-lys (10 mM) improved the plant height by 19%, root length by 57%, leaf dry weight by 19%, and root dry weight by33% under 100% Cr treatment. Zn-lys significantly reduces the oxidative stress and concentration of Cr as compared with the Cr treatments alone. Application of Zn-lys (10 mM) reduced the Cr contents in roots by 27 and 22 under 33 and 66% Cr treatment, respectively. Taken together, Zn-lys chelates efficiently ameliorated the toxic effects of chromium. Zn-lysine has the extravagant potential of mitigating the heavy metal toxicity without harming the normal growth and development of the plants.

Dynamic responses of antioxidant enzymes in pearl oyster Pinctada martensii exposed to di(2-ethylhexyl) phthalate (DEHP)

Di(2-ethylhexyl) phthalate (DEHP) is recognized as one of the most ubiquitous contaminants in marine environments and causes adverse effects on the health of marine organisms. The purpose of this study was to investigate the toxic effects of DEHP on the pearl oyster Pinctada martensii. The Pinctada martensii was exposed to 0.0, 0.5, 2, 8, or 32mgL−1 DEHP for 7 and 10days using parameters of antioxidant. Antioxidant indicators included levels of superoxide dismutase (SOD), catalase (CAT), glutathione (GSH), peroxidase (POD), and total antioxidant capacity (T-AOC) in the gills and hepatopancreas of Pinctada martensii for 7 and 10days. Besides, we used the lowest observed effect concentration (LOEC) of five enzyme activities in different tissues of Pinctada martensii for 7 and 10days to compare sensitivity. The results showed that the gills were more sensitive than the hepatopancreas of Pinctada martensii and that GSH activity in the gills and CAT activity in the hepatopancreas might be suitable biomarkers after 7days of DEHP exposure. After 10days of DEHP exposure, the GSH activity and CAT activity in the gills and SOD activity in the hepatopancreas could be regarded as biomarkers. Compared to the LOEC, GSH activity in the gills and CAT activity in the hepatopancreas after 7days of DEHP exposure were more sensitive than any other biomarkers. In addition, after 10days of DEHP exposure, GSH activity in the gills and hepatopancreas were much more sensitive than other activities. In conclusion, GSH activity demonstrated its potential to be used as a biomarker for the monitoring of DEHP pollution in the marine environment.

Effect of curcumin and quercetin on lysosomal enzyme activities in streptozotocin-induced diabetic rats

Diabetes causes impairment of various enzyme activities in the physiological system, including lysosomal enzymes. The effect of feeding curcumin, quercetin and aminoguanidine on lysosomal enzyme activities viz., N-acetyl-β-d-glucosaminidase, β-d-glucuronidase, β-d-galactosidase and acid phosphatase were studied in different tissues of streptozotocin-induced diabetic rats. Rats were divided into four control groups and four diabetic groups. Experimental groups were fed with diet supplemented with curcumin (0.5%) or quercetin (0.1%) or aminoguanidine (0.05%). Lysosomal enzyme activities were determined in various tissues. The specific activity of N-acetyl-β-d-glucosaminidase in liver of diabetic rats was decreased when compared to control rats and was ameliorated with curcumin and quercetin treatment by 67% and 78%, respectively. On the other hand, β-d-glucuronidase activity was higher in the brain of diabetic rats (0.90 ± 0.04 nmol/mg protein/min), when compared to control rats (0.45 ± 0.02 nmol/mg protein/min) and was decreased in curcumin (0.75 ± 0.05 nmol/mg protein/min) and quercetin (0.74 ± 0.11 nmol/mg protein/min) treated rats. β-d-galactosidase activity in spleen of curcumin and quercetin fed diabetic group rats was ameliorated by 68% and 58%, respectively, in comparison to diabetic rats. Acid phosphatase activity in diabetic rats decreased in testis when compared to control. Curcumin and quercetin feeding modulated lysosomal enzyme activities in different tissues during diabetes and the effect was comparable to well-known anti-glycative agent - aminoguanidine.

Association of mitochondrial function with feed efficiency in rainbow trout: Diets and family effects

A 2 × 4 factorial experiment was conducted to evaluate the effects of commercial diets (40/10 or 45/19% protein/fat, Melick Aquafeed, Inc.) and rainbow trout families (designated as 076, 254, 256 or 257) on the performance, mitochondrial gene expression, and mitochondrial respiratory chain enzyme activities in the liver, muscle and intestine. Four families of rainbow trout (average weight = 100, 97, 82 and 72 g for 076, 254, 256 and 257, respectively) were stocked into twenty four 152-L aquaria at a density of 10 fish/aquarium. Three aquaria were used for each treatment. Fish were fed twice daily to apparent satiation for 16 weeks. Results indicated that family type had significant (P < 0.05) main effects on weight gain, feed consumption, feed efficiency and specific growth rate. Diet composition had no significant (P > 0.05) main effect on weight gain, absolute feed consumption, feed efficiency, specific growth rate, and condition factor, regardless of the family type. There was no significant (P > 0.05) interaction between family and diet composition for weight gain, feed consumption, feed efficiency, specific growth rate, and condition factor. Family with the least weight gain and feed effciency had significantly (P < 0.05) low nutrient utilization efficiencies in terms of protein efficiency ratio, protein productive value, energy efficiency ratio, energy productive value, and lipid productive value. There were variations in the expression levels of some mitochondrial encoded genes and mitochondrial complex enzyme activities in different tissues. Family and diet caused significant down-regulation of complex I gene (ND1) in the liver and its up-regulation in the intestine and muscle tissues. Generally, family with high feed efficiency and better nutrient utilization had higher numerical values for respiratory chain enzyme activities, down-regulation of hepatic complex I ND1 gene and its up-regulation in the intestine and muscle compared to the low feed efficiency family. The mitochondrial respiratory chain complex activity was significantly affected by family and diets in the liver, and muscle for complex I; liver and intestine for complex II; liver, intestine and muscle for complex III; liver and muscle for complex IV. Regardless of family, fish fed diet containing 45% protein and 19% fat had significantly higher (P < 0.0001 to P = 0.0426) levels of activities in the liver and muscle for complex I, intestine for complex II, muscle for complex III, and liver for complex IV. These data demonstrate that family 257of rainbow trout appears to possess superior traits in growth performance compared with the family 256. Our data indicate that genetic factor has important impact on rainbow trout production and that a commercial diet containing 40% dietary protein and 10% dietary fat is as good as diet with higher protein and fat levels for enhanced growth performance, gene expression, and mitochondrial enzyme activities.

Molecular characterization of a glutathione peroxidase gene and its expression in the selected Vibrio-resistant population of the clam Meretrix meretrix

Glutathione peroxidase (GPx) is an important member of cellular enzymatic antioxidant system, which may be involved in pathogen defense of host. In the present study, a selenium-dependent glutathione peroxidase (MmeGPx) gene from clam Meretrix meretrix was cloned and analyzed. The MmeGPx gene was composed of two introns of 723 bp and 238 bp and an open reading frame (ORF) of 711 bp. The ORF encodes a protein of 237 amino acids with a putative selenocysteine residue encoded by an unusual stop codon. MmeGPx shares a higher level of similarity with human GPx 3 than with other human GPx isozymes. The level of MmeGPx mRNA roughly paralleled GPx enzyme activity in different tissues except in gills, with the highest mRNA expression and enzyme activity occurring in hepatopancreas. MmeGPx mRNA expressions were detected in different larval stages and the results showed that MmeGPx mRNA increased significantly in pediveliger stage, which may be a response to oxidative stress. After challenge of clam with a Vibrio parahaemolyticus-related bacterium (MM21), the expression of MmeGPx was significantly up-regulated at 6 h and 12 h in hepatopancreas, which suggested that MmeGPx may be involved in the immune response to MM21 infection. To better understand its role in the immunity of clam, the expression of MmeGPx in hepatopancreas was compared between a selected Vibrio-resistant population and a control population after immersion challenge with MM21. Early up-regulation of MmeGPx was observed in the resistant population. These results suggested that MmeGPx might be involved in maintaining the redox state of immune system, and the early immune response to pathogen infection may help the clam against pathogen infection.

Biochemical Enzyme Activity in Different Tissues of Rats Exposed to a Novel Phosphorothionate (RPR‐V)

The effects of a novel phosphorothionate (RPR‐V) synthesized at Indian Institute of Chemical Technology, Hyderabad, was studied using three sub‐chronic doses of 0.033 (low), 0.066 (medium) and 0.099 (high) mg kg−1 in male and female rats for a period of 90 days. This long term and repeated treatment of RPR‐V revealed that the membrane bound target enzymes Aspartate aminotransferase and Alanine aminotransferase increased significantly in serum and kidney, whereas these enzymes significantly decreased in liver and lung tissues when measured after 45 and 90 days of treatment. This compound also caused significant inhibition of RBC Acetylcholinesterase, target enzyme of organophosphorus compounds revealing its effect on normal synaptic transmission. Two way Anova studies disclosed that the alterations were mostly dose and time dependent, sexual dimorphism was not observed when the activities of male rats were compared with female rats. Enzyme recoveries were recorded after 28 days of post treatment, high degree positive correlation was observed with regard to these enzymes between serum versus kidney, whereas in case of serum versus liver and lung tissues high degree negative correlation was recorded. These enzyme profiles elucidates that they increased in serum but they decreased significantly in liver and lung indicating necrosis of these tissues. However, in case of kidney the level of these enzymes increased significantly with parallel to serum, which is suggestive of an increase synthesis of these enzymes, may be an adaptive mechanism due to the stress of the toxicant. These biomarker enzymes can be detected rapidly and hence may be used for the prediction and diagnosis of pesticide insults.

Glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) enzyme activities in different tissues ofSarotherodon mossambicus(Peters) exposed to a carbamate pesticide, carbaryl

The activity levels of glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) enzymes were estimated in liver, muscle and brain tissues of the fish, Sarotherodon mossambicus (Peters), which had been exposed to sub-lethal (3 mg litre−1) and lethal (25 mg litre−1) concentrations of the carbamate insecticide carbaryl. Based on the results obtained, the changes in GOT and GPT levels in liver, muscle and brain following different periods of sub-lethal and lethal carbaryl exposures suggested that S mossambicus showed adaptive elevation in the activity levels of the two aminotransferase enzymes in the tissues, thereby probably aiding gluconeogenesis through transamination of glucogenic amino acids to meet the energy demand under carbaryl toxicity. © 1999 Society of Chemical Industry

Immunological analysis of acetyl-CoA carboxylase mass, tissue distribution and subunit composition

Changes in the mass and subunit structure of liver acetyl-CoA carboxylase (ACC) accompany altered nutrition in vivo. Enzyme activity in different tissues and cell lines is also, in part, determined by variations in both total mass and ACC isoenzyme composition. ACC isoenzyme mass and hetero/homo-isoenzyme association were quantified by three sandwich e.l.i.s.a. assays, i.e. an avidin-based assay that measured total isoenzyme mass and two antibody-sandwich assays which measure polypeptide association. Results from the avidin-based assay reveal that the two major isoenzymes, of molecular mass 265 kDa (ACC 265) and 280 kDa (ACC 280), are present in markedly variable concentration in several rat and mouse tissues and in cell lines of rat and mouse origin. Hepatic ACC mass has been reported to be distributed between mitochondrial and cytosolic fractions and to undergo only a change in subcellular distribution without alteration in total mass on induction/repression of activity in vivo [Roman-Lopez, Shriver, Joseph & Alfred (1989) Biochem. J. 260, 927-930]. However, in the present study, immunoblotting and e.l.i.s.a. analysis reveals that, in rat liver, the mass of both isoenzymes is predominantly cytosolic in distribution, is markedly diminished on fasting and rises 6-8-fold on refeeding of a high-carbohydrate diet. These data support the results of several other investigations of hepatic ACC mass, and are consistent with known nutritionally altered changes in ACC mRNA content. By the two antibody-sandwich e.l.i.s.a. assays, isoenzyme complexes either composed of both ACC 280 and 265 or with multiple copies of ACC 265 are detectable in rat liver enzyme; their concentration varies independently of total ACC mass with the nutritional state of the rat, being lowest in fasting and highest on fasting/refeeding. E.l.i.s.a. analysis, applicable to crude tissue/cell extracts, provides a simple, sensitive and quantitative measurement of ACC mass and subunit composition. Its use may permit needed quantitative insight into the role of variable total ACC and isoenzyme mass and of alterations in ACC subunit composition that occur in vivo or in isolated cells in response to a variety of hormonal and nutritional influences.

Glutamate Dehydrogenase in Cerebellar Mutant Mice: Gene Localization and Enzyme Activity in Different Tissues

Many similarities of both the inheritance pattern and the neuropathology can be observed between olivopontocerebellar atrophies, or so-called multiple system atrophies (MSAs), and murine cerebellar mutations like Purkinje cell degeneration, nervous, staggerer, weaver, and reeler. Our study aimed to test whether the glutamate dehydrogenase (GDH) deficiency observed in some MSA patients could be found also in any of the murine mutants. GDH activity was assayed in several organs of these mutants, and no general deficiency was detected. By contrast, the level was found to be elevated in the cerebellum. The GDH gene was localized on mouse chromosome 14 and does not map close to any known neurological mutation in the mouse. We conclude, for the moment, that none of these cerebellar mutant mice can be considered as an animal model for GDH-deficient MSA.

Patterns of proteolytic enzyme activities in different tissues of germinating corn (Zea mays L.)

Profiles of pH dependence and activities of live proteolytic enzymes, amino- and carboxypeptidase and endopeptidases active at pH 3.8, 5.4 and 7.5, with casein as substrate, were determined in crude extracts from the various organs of corn seedlings during germination and early development (30°C, dark, 8 d). With respect to the endopeptidases, caseolytic activities at pH 3.8, 5.4 and 7.5 in extracts from endosperm increased concurrently with loss of endosperm N during germination; however, the relative amounts of the pH 7.5 activity were very small. In scutellum extracts, caseolytic activities at both pH 5.4 and 7.5 increased during the initial stages of development but only the increase at pH 5.4 was concurrent with loss of scutellar N. In shoot extracts, caseolytic activities at pH 5.4 and 7.5 were very low and remained relatively constant. There was a progressive increase in shoot N with time. In root extracts, caseolytic activities at pH 5.4 and 7.5 were higher (3-fold) than in shoot extracts. The activity at pH 5.4 remained constant while the activity at pH 7.5 increased during germination. The rate of accumulation of N by the root was low after day 5. The pattern and ratio but not the amounts of the pH 5.4 and 7.5 caseolytic activities of the root were similar to those observed in senescing leaves of field-grown corn. Addition of mercaptoethanol increased (several-fold) the caseolytic activities at pH 3.8 and 5.4, especially the latter, but not the pH 7.5 activity in endosperm extracts and increased the pH 5.4 activity in extracts from scutellum (30%) and roots (30%) while the effect in shoot extracts was negligible. Carboxypeptidase activity was relatively low in young tissue (root tip, 3-d-old shoots) and increased with development of the various organs except the roots (whole) where the activity remained relatively constant. The increases in carboxypeptidase activities were concurrent with decreases in N from endosperm and scutellum; this result indicates that this enzyme in these tissues may be involved (cooperatively with endopeptidases) in the mobilization of reserve protein.Of all the enzymes tested, only carboxypeptidase activity was markedly (in excess of 50%) inhibited by phenylmethylsulfonylfluoride. Only aminopeptidase activity was found in appreciable amounts in endosperm and scutellum of dry kernels. Aminopeptidase activity was highest in organs with high metabolic activity (scutella, shoot, root tips) and decreased in plant parts undergoing rapid loss of nitrogen (endosperm, senescing leaves).

Distribution of homocarnosine-carnosine synthetase in tissues of rat, mouse, chick and frog

1. 1. The distribution of homocarnosine-carnosine synthetase in brain, heart, liver and muscle of rat, mouse, chick and frog was studied. 2. 2. These studies reveal differences in enzyme activity in different tissues within a species but similarities among the four species. 3. 3. With the exception of chick brain, NAD does not seem to be required by the enzymes from frog and chick tissues. However, it is required by the enzymes from mouse and rat tissues.

Activity stain for the detection of cyclic nucleotide phosphodiesterase separated by polyacrylamide gel electrophoresis and its application to the cyclic nucleotide phosphodiesterase of beef heart

A rapid method for detecting cyclic 3′,5′-nucleotide phosphodiesterases in a gel matrix after electrophoresis has been described. The method can be employed for identifying cyclic nucleotide phosphodiesterase activity in partially purified preparations as well as for screening crude tissue extracts for this activity. When combined with density gel scanning, this technique permits the estimation of the relative levels of enzyme activity in different tissues or preparations. Beef heart contains at least two electrophoretically distinct forms of cyclic nucleotide phosphodiesterase activity, both of which can hydrolyze cyclic 3′,5′-AMP and cyclic 3′,5′-GMP.