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1.
Eur Rev Med Pharmacol Sci ; 24(16): 8537-8550, 2020 08.
Article in English | MEDLINE | ID: mdl-32894559

ABSTRACT

OBJECTIVE: The glutathione S-transferases (GSTs) overexpression in hyperproliferating tumour cells resistant to chemotherapy was demonstrated. An increased GST-π activity weakens the efficacy of anti-cancer drugs by promoting their efflux from cells. MATERIALS AND METHODS: This review summarises available information on the physiological role of GSTs, in particular the role of GST-π, in regulation of signalling pathways mechanisms and cellular homeostasis for understanding and explaining the basis for GST-π application as a target for anticancer therapy and implications for clinical practice. RESULTS: GST-π can weaken the effect of TNF receptor-associated factor 2 (TRAF2) on apoptosis signal-regulating kinase-1 by inactivation of MAP kinase pathways (c-Jun N-terminal, p38 kinases). GST-π is involved in the metabolism of endogenous lipids mediators, such as 15-detoxy-Δ12,14-prostaglandin J2 (15d-PGJ2). Reduced binding of 15d-PGJ2 to peroxisome proliferator-activated receptors accompanied by GST-π can result in the inhibition of apoptosis. GSTP1 RNA is able to increase the phosphorylation of signal transducer and activator of transcription 3, what results in negative regulation as regards transcriptional activity thereof and affects the growth factor signalling. However, the oxidation of GST-π results in inhibition of TRAF2-GST-π complexes formation and unblocks cell apoptosis. The inhibition of multidrug resistance related proteins 1 (MRP-1) promoter activity and impairment of MRP-1 function can also act as a potent non-competitive inhibitor of GST-π. CONCLUSIONS: GST-π is recognised as an important target in designing new anticancer drugs. These drugs are often substrates for GST-π or have the affinity with its structure, what results in weakening its activity and achieving therapeutic goal.


Subject(s)
Antineoplastic Agents/pharmacology , Glutathione Transferase/metabolism , Neoplasms/drug therapy , Apoptosis/drug effects , Glutathione Transferase/antagonists & inhibitors , Glutathione Transferase/chemistry , Humans , Isoenzymes/antagonists & inhibitors , Isoenzymes/chemistry , Isoenzymes/metabolism , Models, Molecular , Multidrug Resistance-Associated Proteins/antagonists & inhibitors , Multidrug Resistance-Associated Proteins/metabolism , Signal Transduction/drug effects
2.
Eur Rev Med Pharmacol Sci ; 21(21): 4755-4761, 2017 Nov.
Article in English | MEDLINE | ID: mdl-29164590

ABSTRACT

OBJECTIVE: Polycystic ovarian syndrome and obesity contribute to the metabolic complications for women of reproductive age. The aim of present study was to analyze the effect of abdominal obesity expressed using waist/hip ratio (WHR) in patients with polycystic ovary syndrome on metabolic parameters. PATIENTS AND METHODS: The study included 659 women with PCOS with WHR <0.8 and ≥0.8 aged between 17 and 44 years. Patients were tested for follicular stimulating hormone, luteinizing hormone, 17-beta-estradiol, dehydroepiandrosterone sulfate, androstenedione, sex hormone binding globulin, and total lipid profile during the follicular phase (within 3 and 5 days of their menstrual cycle). Also, fasting glucose and insulin concentrations, and after, oral-glucose glucose administration, were determinate. De Ritis and Castelli index I and II were calculated. RESULTS: Women with WHR ≥0.8 had higher concentration of glucose and  insulin (both fasting and after 120 min of oral administration of 75 g glucose), as well as HOMA-IR value, than women with WHR value < 0.8. Also, abdominal obesity disorders hormonal parameters. Higher free androgen index and lower concentration of sex hormone binding globulin and dehydroepiandrosterone sulfate were found in female with WHR ≥ 0.8. Follicular stimulating hormone, luteinizing hormone, androstenedione, and 17-beta-estradiol, were on similar level in both groups. Elevation in triglycerides, total cholesterol, and low-density lipoprotein levels, as well as decrease in high density lipoprotein level in serum of women with WHR value ≥ 0.8, were found when compared to women with WHR < 0.8. A statistically significant correlation was found between WHR value and glucose, insulin, sex hormone binding globulin, free androgen index and lipid profile parameters. CONCLUSIONS: Abdominal obesity causes additional disorders in metabolic and hormonal parameters in PCOS women, which confirmed changes in analyzed parameters between PCOS women with WHR < 0.8 and WHR ≥ 0.8 and statistically significant correlations between WHR value and analyzed parameters.


Subject(s)
Obesity, Abdominal/diagnosis , Polycystic Ovary Syndrome/diagnosis , Adolescent , Adult , Blood Glucose/analysis , Body Mass Index , Dehydroepiandrosterone Sulfate/blood , Female , Follicle Stimulating Hormone/blood , Humans , Insulin/blood , Luteinizing Hormone/blood , Obesity, Abdominal/complications , Polycystic Ovary Syndrome/complications , Sex Hormone-Binding Globulin/analysis , Waist-Hip Ratio , Young Adult
3.
Environ Toxicol Pharmacol ; 44: 99-106, 2016 Jun.
Article in English | MEDLINE | ID: mdl-27137107

ABSTRACT

Arsenic, lead and cadmium, potent environmental toxicants have been reported to induce diabetes mellitus, but their potential biological mechanism(s) have not been much investigated. The present study was designed to correlate parameters of pro/antioxidant balance with occupational exposure on heavy metals and smoking in smelters with diabetes compared on control group. The results showed a significant increase in the concentration of arsenic, cadmium and lead in the blood and urine of smelters, while smoking caused a further increase in the concentration of these metals. Increasing γ-glutamyltransferase activity and lead concentration due to occupational exposure in copper foundry, tobacco smoke and co-existing diabetes were observed. Also these factors have synergistic effects on metallothionein and glutathione concentrations as well as glutathione dependent enzymes activities. Our data suggests that sub-chronic arsenic, lead and cadmium exposure induces diabetic condition which may be mediated due to increased oxidative stress in blood.


Subject(s)
Diabetes Mellitus/blood , Metallurgy , Occupational Exposure/analysis , Smoking/blood , Adult , Air Pollutants/blood , Air Pollutants/urine , Arsenic/urine , Cadmium/blood , Diabetes Mellitus/urine , Environmental Monitoring , Glutathione/blood , Glutathione Peroxidase/blood , Glutathione Transferase/blood , Humans , Lead/blood , Male , Metallothionein/blood , Middle Aged , Smoking/urine , Superoxide Dismutase/blood , Young Adult , gamma-Glutamyltransferase/blood
4.
Neoplasma ; 53(6): 495-9, 2006.
Article in English | MEDLINE | ID: mdl-17167718

ABSTRACT

The clinical and histochemical examination of hormone-producing serous cystadenomas of the pancreas are presented. The study material was obtained from five female patients. The patients underwent diagnostic examinations, including ultrasonography, computer tomography (CT), magnetic resonance imaging (MRI) and Doppler ultrasonography examination of abdomen. In all cases the presence of serous cystadenoma of pancreas was detected in the histopathologically verified sections. The test applied to immunohistochemically localize paraffin-embedded sections of neoplastic tissues of the pancreas was the LSAB2-HRP test using monoclonal antibodies against epithelial membrane antigen (EMA), carcinoembryonic antigen (CEA), synaptophysin, p53 and polyclonal antibodies against insulin, glucagon, somatostatin and pancreatic polypeptide. In one patient, ultrasonography revealed an irregular space filled with fluid resembling a multicellular cystic lesion. The Doppler ultrasonography examination showed a pathologically vascularized focus in the pancreatic head. In the adenoma sections of this patient, the immunohistochemical techniques revealed a strong positive somatostatin, pancreatic polypeptide and synaptophysin expression in the lining epithelium of neoplastic cysts.


Subject(s)
Biomarkers, Tumor/metabolism , Cystadenoma, Serous/metabolism , Glucagon/metabolism , Insulin/metabolism , Pancreatic Neoplasms/metabolism , Somatostatin/metabolism , Adult , Aged , Carcinoembryonic Antigen/metabolism , Cystadenoma, Serous/pathology , Female , Humans , Magnetic Resonance Imaging , Middle Aged , Mucin-1/metabolism , Pancreatic Neoplasms/pathology , Synaptophysin/metabolism
5.
Int J Occup Med Environ Health ; 14(3): 275-85, 2001.
Article in English | MEDLINE | ID: mdl-11764857

ABSTRACT

The aim of the present study was to assess the activity of membrane enzymes: alanine aminopeptidase (AAP), gamma-glutamyltransferase (GGT) and trehalase in amniotic fluid of women who smoke cigarettes during pregnancy complicated by idiopathic oligohydramnios or premature rupture of the membranes (PROM). The enzyme activity was measured between 22 and 31 (group A) and between 32 and 39 (group B) weeks of gestation. In the women of group A with idiopathic oligohydramnios, AAP activity was five times higher than in PROM women. AAP activity was declining with the progression of gestation, and in the B group women with oligohydramnios, it was over eight times lower than in group A. A threefold increase in GGT activity was found in women of group A with oligohydramnios as compared to women of group A with PROM. No statistically significant differences in trehalase activity were found in amniotic fluid of women with oligohydramnios and PROM, AAP, GGT and trehalase activity in women with idiopathic oligohydramnios correlated with the cadmium ion concentration, and AAP and GGT activity with the lead ion concentration in amniotic fluid which confirms toxical properties of these heavy metals present in cigarette smoke. It has already been confirmed that measurements of the brush border enzyme activity in amniotic fluid are very useful in prenatal diagnosis and detection of the placenta disorders.


Subject(s)
Amniotic Fluid/chemistry , CD13 Antigens/metabolism , Fetal Membranes, Premature Rupture/enzymology , Oligohydramnios/enzymology , Smoking/adverse effects , Trehalase/metabolism , gamma-Glutamyltransferase/metabolism , Adult , CD13 Antigens/analysis , Enzyme Activation , Female , Fetal Membranes, Premature Rupture/diagnosis , Fetal Membranes, Premature Rupture/etiology , Gestational Age , Humans , Oligohydramnios/diagnosis , Oligohydramnios/etiology , Pregnancy , Probability , Risk Assessment , Sensitivity and Specificity , Trehalase/analysis , gamma-Glutamyltransferase/analysis
6.
Int J Occup Med Environ Health ; 13(3): 185-93, 2000.
Article in English | MEDLINE | ID: mdl-11109742

ABSTRACT

To assess the exposure to tobacco smoke in pregnant women with oligohydramnios, idiopathic or caused by premature rupture of the membranes (PROM), cotinine concentrations were measured, using enzyme-like immunosorbent assay (ELISA). In women with idiopathic oligohydramnios (22-31 weeks of gestation), serum cotinine concentration was 1010 +/- 445 micrograms/L which provides evidence that women of this group were heavy smokers. In these women, significantly higher Cd concentrations in blood and amniotic fluid were found as compared to other pregnant women. A positive correlation between Cd concentrations in blood and amniotic fluid was observed (PROM r = 0.784; p < 0.001; idiopathic oligohydramnios r = 0.7118; p < 0.02). In oligohydramnios cases of both types, Cd concentration in amniotic fluid was over two times and Pb concentration ten times lower than blood concentrations of these metals, whereas amniotic fluid Zn concentration was two times lower than that found earlier in women with normal pregnancy. In the group of women with idiopathic oligohydramnios who were mostly exposed to tobacco smoke, a considerably larger number of still births and new-borns with CNS disorders than in PROM cases, were observed. Zn deficiency at increased exposure to Cd and Pb could play a significant role in etiology of these abnormalities. A positive correlation was found between Zn and Cu concentrations (r = 0.862; p < 0.05) in PROM cases which indicates regular transport of trace metals to the fetal ovum. The condition of infants born to this group of women was much better, and prematurity was the only complication of pregnancy.


Subject(s)
Amniotic Fluid/chemistry , Fetal Membranes, Premature Rupture/metabolism , Metals, Heavy/blood , Oligohydramnios/metabolism , Tobacco Smoke Pollution/adverse effects , Adult , Central Nervous System Diseases/congenital , Cotinine/analysis , Enzyme-Linked Immunosorbent Assay , Female , Fetal Death , Humans , Oligohydramnios/etiology , Pregnancy , Pregnancy Outcome
7.
Ginekol Pol ; 71(4): 274-8, 2000 Apr.
Article in Polish | MEDLINE | ID: mdl-10860258

ABSTRACT

OBJECTIVES: The aim of the study was analysis of neonatal status from pregnancies complicated oligohydramnios and from pregnancies complicated premature rupture of the membranes (PROM). MATERIALS AND METHODS: Authors analyzed 15 newborns from pregnancies with oligohydramnios diagnosed and 15- from pregnancies complicated PROM. Mothers of these newborns smoked cigarettes or were exposed to tobacco smoke. The features taken into consideration was prematurity, hypotrophy, malformations, birth weight, Apgar scale gained by newborns in first minutes after delivery. RESULTS AND CONCLUSIONS: The higher perinatal mortality, more frequent occurrence of hypotrophy and malformations were shown in newborns from pregnancies complicated oligohydramnios. The most frequent developmental anomaly was defect of nervosal system. Status of newborns from PROM pregnancies was getting better in according to lengthening duration of gestation. It wasn't stated in case of pregnancies with oligohydramnios complications.


Subject(s)
Fetal Membranes, Premature Rupture/diagnosis , Health Status , Infant, Newborn/physiology , Nicotiana , Oligohydramnios/diagnosis , Plants, Toxic , Pregnancy Complications , Smoking/adverse effects , Apgar Score , Female , Humans , Pregnancy
8.
Ginekol Pol ; 71(4): 268-73, 2000 Apr.
Article in Polish | MEDLINE | ID: mdl-10860257

ABSTRACT

OBJECTIVES: Morphological lesions in placenta and changed metal distribution through placenta, which were observed in pregnancy exposed to tobacco smoke, could cause alternations in lysosomal enzymes secretion to amniotic fluid. DESIGN: Assessment of total activity and molecular forms of N-beta-D-acetylglucosaminidase in amniotic fluid in pregnancy with oligohydramnios or PROM. MATERIALS AND METHODS: The materials for studies were amniotic fluids collected from pregnancy with oligohydramnios and from pregnancy with PROM. Determination of N-beta-D-acetylglucosaminidase activity was performed with use of sodium salt of 3-crezolosulfthaleinyl-N-beta-D-acetylglucosamide as a substrate (test Boehringer, Niemcy). Creatinine was determined by kinetic method (test of Analco-GBG firm). Protein was determine by Bredford method. NAG-B was assessed after previous thermal NAG-A inactivation in 56 degrees C, for 2 hours and the difference between activity of total form and termostabile form of NAG were assessed. The microsomal and cytosol fractions were separated by ultracentrifugation. RESULTS AND CONCLUSIONS: The statistically significant growth of NAG activity in amniotic fluid in pregnant with oligohydramnios was observed. The cytosol form took over 90% of whole activity. The activity of NAG microsomal fraction was lower in amniotic fluid in women with PROM diagnosis. The positive coleration between NAG-A activity and the Pb level was observed in this group of women as well. In the second group of women with oligohydramnios, the positive coleration between NAG and Cd level, NAG-A and level, NAG-B and Pb level were demonstrated. Obtained results show on damage of placental and foetal membranes cell structure. It could be caused by metal ions cumulation and releasing of molecular N-beta-D-acetylglucosaminase form to amniotic fluid.


Subject(s)
Acetylglucosaminidase/analysis , Acetylglucosaminidase/metabolism , Amniotic Fluid/chemistry , Fetal Membranes, Premature Rupture/diagnosis , Fetal Membranes, Premature Rupture/enzymology , Nicotiana , Oligohydramnios/diagnosis , Oligohydramnios/enzymology , Plants, Toxic , Pregnancy Complications , Smoking/adverse effects , Adult , Female , Humans , Middle Aged , Pregnancy
9.
Ginekol Pol ; 71(4): 311-6, 2000 Apr.
Article in Polish | MEDLINE | ID: mdl-10860265

ABSTRACT

OBJECTIVES: The ingredients of tobacco smoke have the impact on uterine blood vessels. They caused vascular insufficiency of placenta during development of gestation and changes in placental tissue and fetal membranes. It manifest changeable metal permeability and others symptoms. DESIGN: Assessment of metal contents in blood and amniotic fluid in pregnancies with oligohydramnios and PROM. MATERIALS AND METHODS: The materials for studies were blood and amniotic fluid of 30 pregnancies. All of them had made amino-punction between 20 and 38 week of pregnancy before procedure of amniotic fluid supplementation. 15 patients had diagnosed oligohydramnios and the next 15 women had diagnosed premature rupture of membranes. Metals were determined by method of electrothermal atomical absorbic spectrophotometry (ET-ASA). RESULTS AND CONCLUSIONS: Twice lower concentration of Zn and Cd and ten times lower concentration of Pb in amniotic fluids in examined women than women in normal pregnancy were observed. The women, with oligohydramnios in earlier period of gestation, smoked many more cigarettes and cotinine++ and Cd. were much higher. Both these facts had relevance with each other certainly. The stillborns were many more in this group. The different distribution of Cd, Pb, Zn in pregnant women with PROM and oligohydramnios, comparing with women in normal pregnancy was demonstrated. It is probable to be effect of placental tissue and fetal membranes disfunction.


Subject(s)
Amniotic Fluid/chemistry , Cadmium/blood , Copper/analysis , Fetal Membranes, Premature Rupture/blood , Fetal Membranes, Premature Rupture/diagnosis , Lead/blood , Oligohydramnios/blood , Oligohydramnios/diagnosis , Placenta/blood supply , Zinc/analysis , Adult , Female , Humans , Middle Aged , Pregnancy , Pregnancy Complications
10.
Int J Occup Med Environ Health ; 10(2): 187-202, 1997.
Article in English | MEDLINE | ID: mdl-9278131

ABSTRACT

Concentrations of metals (Zn, Cu, Se, Cd), metallotionein (MT) and ceruloplasmin (Cp), and the activity of N-acetyl-beta-D-glucosaminidase (NAG) and gamma-glutamyltransferase (gammaGT) in pregnant women who smoke and in passive smokers as well as in their newborns were measured. In smoking women a many-fold increase in cotinine concentrations in maternal and infant urine, amniotic fluid, placenta and milk was found as compared to passive smokers. Smoking lowered serum Se concentration in mothers and infants, increased serum ceruloplasmin in mothers and decreased serum Cu concentration in infants. Serum Zn level in infants was higher than in mothers, and smoking furthered its increase. Statistically significant increase in concentrations of Cd in placenta, amniotic fluid and milk, of Cu in placentae and amniotic fluid and of Zn in placenta and milk was revealed. Higher concentrations of metals were accompanied by an elevated MT level in placenta and amniotic fluid. In smoking women a significant increase in NAG activity in urine, amniotic fluid and milk, increase in gammaGT activity in urine and amniotic fluid and decrease in gammaGT activity in milk and placenta were observed. There was high, positive correlation between MT/NAG and MT/gammaGT in amniotic fluid of passive and active smokers, while similar correlation in milk was indicated only in the group of active smokers. The biochemical parameters analysed proved their usefulness in the evaluation of effects of exposure to cigarette smoke on pregnant woman and her fetus.


Subject(s)
Body Fluids/chemistry , Pregnancy Complications/metabolism , Prenatal Exposure Delayed Effects , Smoking/metabolism , Tobacco Smoke Pollution/adverse effects , Acetylglucosaminidase/analysis , Adult , Amniotic Fluid/chemistry , Ceruloplasmin/analysis , Cotinine/analysis , Environmental Exposure/analysis , Environmental Monitoring/methods , Female , Humans , Infant, Newborn , Metallothionein/analysis , Metals/analysis , Milk, Human/chemistry , Placenta/chemistry , Pregnancy , Smoking/adverse effects , Spectrophotometry, Atomic , Tobacco Smoke Pollution/analysis , gamma-Glutamyltransferase/analysis
11.
Int J Occup Med Environ Health ; 10(3): 273-82, 1997.
Article in English | MEDLINE | ID: mdl-9524405

ABSTRACT

Concentration of metals (Zn, Cu, Cd), Metallothionein (MT) and the activity of n-acetyl¿beta-glucosaminidase (NAG) in amniotic fluids and milk from smoking mothers and passive smokers, living in the area of higher environmental pollution with heavy metals, were measured. In active smokers a three-fold increase in Cd concentration, higher Cu and lower Zn concentrations, higher MT level and enhanced NAG activity in amniotic fluids as well as higher Cd, Cu and Zn concentrations and increased MT and NAG in milk were indicated, when compared to passive smokers. Some dynamics in metal and metallothionein secretion with mother's milk was noted on successive days after delivery. The highest MT level was observed during first 24 hrs postpartum, whereas Cd concentration was found to be highest on the third day. There were no significant differences in concentrations of Cu and Zn secreted with milk on those days. In environmentally exposed women higher Cd and Cu concentrations, enhanced MT level and Nag activity, higher amniotic fluid and lower milk Zn concentration were observed in comparison to women living in the Cieplice region.


Subject(s)
Amniotic Fluid/metabolism , Maternal Exposure , Metals, Heavy , Milk, Human/chemistry , Smoking/metabolism , Ceruloplasmin/metabolism , Copper/metabolism , Female , Hexosaminidases/metabolism , Humans , Metallothionein/metabolism , Pregnancy , Tobacco Smoke Pollution
12.
Int J Occup Med Environ Health ; 10(4): 395-403, 1997.
Article in English | MEDLINE | ID: mdl-9575665

ABSTRACT

SDS-PAGE was used for separation of heat-stable proteins in placenta, amniotic fluid and milk of women who smoke actively and those exposed to tobacco smoke (passive smokers). In the number of protein bands stained with Coomassie brilliant blue R-250, several bands were stained with silver nitrate. The presence of low-molecular band with molecular weight of 6.5 kDa, corresponding to the mobility of electrophoretic MT-1 was detected in placenta, amniotic fluid and milk. The involvement of MT-2 isoform was much lower, although more evident in amniotic fluid and milk of active smokers. In this group of women an enhanced concentration of the band with molecular weight of 25-30 kDa was observed in placenta, colostrum and milk on the second day after delivery; several protein bands emerged in the area of the same mobility in amniotic fluid; the presence of the band with molecular weight of 12.5 kDa and the absence of protein bands of 25-30 kDa were found in milk excreted on the third day post partum. Our results show the apparent differences between proteingrams of placenta, amniotic fluid and milk of active and passive smokers.


Subject(s)
Amniotic Fluid/chemistry , Environmental Exposure/analysis , Metallothionein/analysis , Milk, Human/chemistry , Placenta/chemistry , Smoking/adverse effects , Tobacco Smoke Pollution/adverse effects , Adult , Cotinine/analysis , Culture Techniques , Electrophoresis , Environmental Monitoring/methods , Female , Humans , Metallothionein/biosynthesis , Molecular Weight , Pregnancy
13.
Chem Biol Interact ; 93(3): 197-219, 1994 Dec.
Article in English | MEDLINE | ID: mdl-7923440

ABSTRACT

The knowledge of brain metallothionein (MT) regulation and especially of MT presence in specific cell types is scarce. Therefore, the effect of several well-known MT inducers, measured by radioimmunoassays using antibodies that cross-react with MT-I and MT-II or specific for MT-I and which do not cross-react with human growth inhibitory factor (GIF or MT-III), has been studied in primary cultures of neurons or astrocytes obtained from rat cerebrum. MT-I levels in glial cells were about ten times higher than those in neuronal cells (538 +/- 194 vs. 49 +/- 16 pg MT-I/micrograms protein, mean +/- S.D. from three separate cell preparations). Increasing the concentration of Zn in the bovine serum albumin (BSA)-containing culture medium up to 50 microM significantly increased MT-I levels by up to 3.5-fold in neurons and 2.5-fold in astrocytes. In contrast, Cu up to 50 microM increased MT-I levels in a saturable manner in both neurons (up to 5-fold) and astrocytes (up to 1.5-fold), the maximum effect occurring at 5 microM Cu. In general, the combination of Zn and Cu further increased MT-I levels. The effect of the metals on MT-I appeared to reflect metal uptake, since MT-I induction was less marked when the BSA concentration in the medium was increased from 2 to 10 mg/ml. Dexamethasone increased MT-I levels in both neurons and astrocytes in vitro in a concentration-dependent manner. Endotoxin, IL-1 and IL-6 did not have a significant effect on glial MT levels at the concentrations studied. The administration of dexamethasone to rats increased MT-I levels in non-frontal cortex, cerebellum, pons+medulla, midbrain and hippocampus, but not in hypothalamus, frontal cortex and striatum. Endotoxin increased liver but not brain MT-I levels. Immunocytochemical studies in adult rat brain preparations with a polyclonal antibody that cross-reacts with MT-I and MT-II indicated that immunostaining was always nuclear in glial cells, whereas in neurons it was nuclear in the cerebral cortex, hippocampus and the granular layer of the cerebellum, and nuclear plus cytoplasmic in Purkinje cells in the cerebellum, hypothalamic nuclei and gigantocellular reticular nucleus in the brain stem. Meninges, choroidal plexus, ependymal and endothelial cells were also MT-immunoreactive.


Subject(s)
Astrocytes/drug effects , Copper/pharmacology , Glucocorticoids/pharmacology , Metallothionein/metabolism , Neurons/drug effects , Zinc/pharmacology , Analysis of Variance , Animals , Astrocytes/cytology , Brain/cytology , Brain/embryology , Cells, Cultured , Corticosterone/pharmacology , Cross Reactions , Dexamethasone/pharmacology , Endotoxins/pharmacology , Female , Immunohistochemistry , Male , Metallothionein/drug effects , Neurons/cytology , Radioimmunoassay , Rats , Rats, Sprague-Dawley
16.
Biomed Biochim Acta ; 45(3): 305-13, 1986.
Article in English | MEDLINE | ID: mdl-3085660

ABSTRACT

Applying molecular filtration, the presence of two zinc-binding protein fractions in the human placenta was found. Their molecular weights were 47000 and 7900, respectively. A method for isolation of the high-molecular weight fraction from the placenta was elaborated. It consisted in the fractionation with acetone, Sephadex G-75 filtration, DEAE-Sephadex A-25 chromatography and preparative electrophoresis in polyacrylamide gel. It was calculated that 3.93 g-atoms of Zn are bound to one molecule of the protein. Rabbit antibodies against the zinc-containing protein formed an immunoprecipitate with the whole human amniotic fluid and with two protein fractions obtained following filtration of the amniotic fluids on Bio-Gel P-2.


Subject(s)
Metalloproteins/analysis , Placenta/analysis , Zinc/analysis , Amniotic Fluid/analysis , Chromatography, Gel , Chromatography, Ion Exchange , Electrophoresis, Polyacrylamide Gel , Female , Humans , Immunodiffusion , Isoelectric Point , Molecular Weight , Pregnancy
17.
Enzyme ; 32(4): 208-17, 1984.
Article in English | MEDLINE | ID: mdl-6151897

ABSTRACT

By digestion of detergent-solubilized gamma-glutamyltransferase (GGT), isolated from bovine kidney with bromelain, the liberation of 4 protein fragments was demonstrated. The fragment migrating most quickly in gel electrophoresis showed gamma-glutamyltranspeptidase activity and the most slowly migrating fragment showed peptidase activity. Protease-solubilized GGT is a sialoprotein with a molecular weight of 95,000. After treatment with sodium dodecylsulfate it was separated into two unequal subunits with molecular weights of 26,000 and 69,000. Sugar components were found only in the heavy subunit. Some catalytic differences were found between the two solubilized GGT forms. The immunoprecipitate obtained from detergent-solubilized GGT retained about 50% of the initial enzyme activity. The enzyme is inactivated with phenylmethanesulfonyl fluoride in the presence of maleate and with 6-diazo-5-oxo-L-norleucine.


Subject(s)
Kidney/enzymology , gamma-Glutamyltransferase/analysis , Animals , Cattle , Electrophoresis, Polyacrylamide Gel , Immunodiffusion , Immunoelectrophoresis, Two-Dimensional , Molecular Weight , Peptide Fragments/analysis , gamma-Glutamyltransferase/antagonists & inhibitors
18.
Folia Histochem Cytochem (Krakow) ; 20(1-2): 25-33, 1982.
Article in English | MEDLINE | ID: mdl-6183174

ABSTRACT

Using solubilization with bromelain, the light form of gamma-glutamyltransferase was purified from Morris hepatoma 5123D. Some properties of this enzyme were compared to those of the light form rat kidney GGT. Anthglutin and its isomer inhibit competitively the former enzyme but non-competitively the latter. On zymograms of rat control sera, five GGT fractions were noted, but in sera of rats with hepatoma 5123D also the light form and the increase of GGT activity ain region of fraction II were observed. Only these two enzyme fractions react with antibody anti heavy form of Morris hepatoma GGT.


Subject(s)
Liver Neoplasms, Experimental/enzymology , gamma-Glutamyltransferase/metabolism , Animals , Epitopes , Glutamates/pharmacology , Kinetics , Molecular Weight , Rats , Rats, Inbred BUF , gamma-Glutamyltransferase/blood , gamma-Glutamyltransferase/immunology
19.
Arch Immunol Ther Exp (Warsz) ; 29(5): 543-9, 1981.
Article in English | MEDLINE | ID: mdl-6127063

ABSTRACT

Light form of bovine kidney gamma-glutamyl transferase was isolated from heavy form of the enzyme after digestion with bromelain. Its apparent molecular weight was 95,000 and in SDS solution it dissociated into two non-identical subunits with molecular weights 26,000 and 69,000. No substantial differences between both forms in activation, kinetic parameters and inhibition with anthglutin and its isomers were noted. Using enzyme immunoassay it was possible to determine one enzyme form in the presence of the other. This was applied for studies of gamma-glutamyltransferase forms in cow serum and colostrum.


Subject(s)
gamma-Glutamyltransferase/immunology , Animals , Cattle , Enzyme-Linked Immunosorbent Assay , Kidney/enzymology , Kinetics , Molecular Weight , gamma-Glutamyltransferase/metabolism
20.
Neoplasma ; 27(3): 241-5, 1980.
Article in English | MEDLINE | ID: mdl-6109259

ABSTRACT

Comparative biochemical and immunological investigations were performed on purified gamma-glutamyltransferase from the Morris hepatoma 5123D and from the 5123D/AS variant passaged in F1 (Buffalo x Wistar) rats. The enzyme was also localized in hepatoma sections by immunofluorescent technique. The obtained results confirm the hypothesis that low gamma-glutamyltransferase activity in Morris hepatoma 5123D/AS is caused by the enzymatic protein defficiency and not by the inhibition of the enzyme.


Subject(s)
Liver Neoplasms, Experimental/enzymology , gamma-Glutamyltransferase/analysis , Animals , Fluorescent Antibody Technique , Neoplasm Transplantation , Rats , Transplantation, Homologous , gamma-Glutamyltransferase/immunology
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