Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.1.21.3 (deoxyribonuclease)
1,528 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A folate-binding protein (binder) from human choroid plexus was solubilized with Triton X-100 and partially purified in three steps: (1) affinity chromatography, (2) Sephadex G-200 column chromatography, and (3) polyacrylamide gel electrophoresis. When the partially purified binder was subjected to sodium dodecyl sulfate--polyacrylamide gel electrophoresis, the binding activity was located in the region of the gel with a molecular weight between 45,000 and 60,000. The specific activity of the binder after the three purification steps was 1.2 mu g folic acid/mg protein, a 316-fold purification. Binding activity of the partially purified binder decreased below pH 6.0 and above pH 8.0, was unaffected by treatment with ribonuclease or deoxyribonuclease, but was abolished with trypsin, chymotrypsin, or protease (Streptomyces griesus). The binding of folic acid to the human binder was inhibited by folate Greater Than H4-folate Greater Than methyl-H4-folate approximately dihydrofolate approximately pteroic acid Greater Than methotrexate approximately aminopterin.
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PMID:Partial purification and characterization of a folate-binding protein from human choroid plexus. 727 9

An ATP-dependent deoxyribonuclease was isolated from lymphocyte nuclei. The enzyme preparation sediments with about 4 S through sucrose gradients and shows one stainable band after sodium dodecyl sulfate gel electrophoresis. We find three, possibly four, activities associated with the enzyme: a DNA-independent ATPase activity; an ATP-independent endonuclease; an ATP-dependent nuclease which degrades nicked DNA to acid-soluble material; and an unwinding activity producing single-stranded regions in nicked DNA.
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PMID:A lymphocyte ATP-dependent deoxyribonuclease. Isolation and properties. 730 58

Cystic fibrosis (CF), a lethal disease common to Caucasians, is characterized by a defect in the CF transmembrane conductance regulator and the resulting defective cAMP-regulated Cl- secretion by epithelial cells. Clinical manifestations include both pancreatic and pulmonary insufficiency. Traditional therapeutic modalities address these problems with pancreatic enzyme replacement, vitamins and nutritional supplementation, antibiotics, and respiratory therapy. However, newer therapies directed at the specific underlying defects have emerged. In this review, we discuss agents that increase Cl- secretion via preserved Cl- secretory pathways, such as uridine triphosphate, or that enhance Na+ resorption, such as amiloride, thereby correcting altered airway secretions. We also discuss agents, including deoxyribonuclease (DNase), that directly reduce sputum viscosity. CF is an early target for in vivo gene therapy, since it is a monogenic autosomal recessive disease in which restoration of normal cAMP-regulated Cl- conductance can be achieved by complementation with a normal gene. The early clinical gene therapy therapy work, with gene introduction by both viral and nonviral vectors, is discussed.
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PMID:Molecular strategies for therapy of cystic fibrosis. 759 94

Patients in the intensive care unit (ICU) with compromised lung function often have excessive pulmonary secretions and have difficulty clearing mucus. A variety of pharmacotherapeutics have been used in an attempt to change the rate, amount, and viscosity of respiratory secretions. These drugs are known as mucokinetic or mucolytic drugs, but their efficacy remains controversial in the research literature. Most commonly used methods of mucolysis/mucokinesis include: bland aerosols (saline), pH adjustment (sodium bicarbonate), and disulfide disruption (N-acetylcysteine, MUCOMYST). Clinical research needs to be done on the use of recombinant human deoxyribonuclease (rhDNase) in ICU patients because recombinant human deoxyribonuclease affects only purulent secretions, which makes it a safe, easily tolerated compound. Nursing interventions in the control of mucus in the ICU patient needs to be re-examined in light of recent research where the efficacy of chest percussion was questioned, as was the use of saline bolus instillation before suctioning and the potential for dornase alfa.
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PMID:Mucolytics and the critically ill patient: help or hindrance? 774 33

Deoxyribonuclease activity was detected in E. granulosus protoscoleces from sheep hydatid cysts by electrophoresis in agarose gels of DNA fragments obtained after incubation of integral DNA with a protoscoleces preparation. Preliminary characterization showed that deoxyribonuclease activity was optimal at neutral-alkaline pH, magnesium ions were required, and it was able to digest different types of DNA, making random cuts. Electrophoresis in DNA-containing sodium dodecylsulfate (SDS)-polyacrylamide gels indicated a relative molecular mass, under non-reducing conditions, of 32 kDa. Deoxyribonuclease activity was also found in sheep hydatid fluid. It shared optimal pH, ionic and substrate requirements with the enzyme from protoscoleces but had a higher relative molecular mass (40 kDa), the same as that of normal sheep serum deoxyribonuclease.
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PMID:Deoxyribonuclease activity in Echinococcus granulosus protoscoleces and hydatid fluid. 808 90

Lung cell culture may be useful as an in vitro alternative to study the susceptibility of the lung to various toxic agents. Lungs from female Wistar rats were enzymatically digested by recirculating perfusion through the pulmonary artery with a sequence of solutions containing deoxyribonuclease, chymopapain, pronase, collagenase, and elastase. Lung tissue was microdissected and resuspended and the cells obtained were washed by centrifugation. By this isolation method, 2 x 10(8) cells per rat lung were obtained with an average viability of 97%. Lung cells cultured in medium containing antibiotics and serum maintained a viability of > 70% for 5 d. Rat primary lung cells were exposed to various toxic agents and their viability was assessed by formazan production capacity after 18 h of incubation. Compared to rat and mouse hepatocyte cultures (EC50 = 5.8 mM), rat primary lung cells were much more susceptible to hydrogen peroxide (EC50 = 0.6 mM). All cell types were equally sensitive to the more potent toxicant tert-butylhydroperoxide (EC50 = 0.1 mM). Paraquat was more toxic to lung cells (EC50 = 0.03 mM) than to rat (EC50 = 2.8 mM) and mouse (EC50 = 0.2 mM) hepatocytes. In contrast, rat lung cells were less sensitive to sodium nitroprusside (EC50 = 2.6 mM) compared to rat (EC50 = 0.2 mM) and mouse (EC50 = 0.03 mM) hepatocytes. Nitrofurantoin and menadione (at EC50 = 0.04 mM and 0.006 mM, respectively) were more toxic to rat lung and liver cells than to murine hepatocytes (EC50 = 0.2 mM and 0.04 mM, respectively). Our findings demonstrate the applicability of this rat primary lung cell culture for studying the effects of lung toxicants.
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PMID:Isolation and characterization of rat primary lung cells. 856 79

During purification of fungal deoxyribonuclease (DNase) from Syncephalastrum racemosum, a protein which was functionally unknown and persistently existed in the DNase-containing fractions through chromatography over DEAE-cellulose, hydroxylapatite, and phenyl-Sepharose was identified. The protein was finally separated from DNase after affinity chromatography on a cibacron blue-Sepharose column and purified to apparent homogeneity after gel chromatography on a Superdex 200 HR column. Ten tryptic peptides of this protein were isolated and sequenced. Searching in the sequence data bank with the aid of the computer program PC/Gene, we found that this protein was highly homologous to aspartic proteinases, such as pepsin and rhizopuspepsin. Because of its fungal origin and because the protein indeed showed catalytic cleavage on peptide bonds of bovine serum albumin, RNase, and carbonic anhydrase, we termed this protein syncephapepsin. The molecular weight of syncephapepsin is 38,000 daltons, based on gel filtration and sodium dodecyl sulfate-polyacrylamide electrophoresis.
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PMID:Identification of a fungal protein of Syncephalastrum racemosum as aspartic proteinase. 883 44

The acute pancreatitis is still serious diagnostic and therapeutic problem. In spite of many experiments there are no satisfactory methods of the treatment of this disease. Usually, it is diagnosed clinically by acute abdominal pain and increased pancreatic enzymes in blood and urine. The main disorder of this disease is the necrosis of the pancreatic gland. The aim of our investigations was the evaluation of effect of necrolytic enzymes on the course of acute experimental pancreatitis. Fibrolan was used. Fibrolan (Parke-Davis) is a preparation, which has necrolytic properties. It consists of two bovine enzymes: fibrinolysin and deoxyribonuclease. The examinations were carried out on 380 Wistar rats. The acute pancreatitis was induced by retrograde injection of 0.1 ml/0.1 kg body weight of 3% sodium taurocholate solution into the pancreatic duct as previously described by other authors. The experimental animals were divided into four groups: K0-control group (healthy animals), KP-animals with acute experimental pancreatitis without medication, S-rats with acute experimental pancreatitis, which were treated with 0.9% NaCl solution, and F-animals with acute experimental pancreatitis treated with Fibrolan. Fibrolan and 0.9% NaCl were injected into the peritoneal cavity three times a day with eight hours intervals from the 24th experimental hour. The serum amylase and lipase activities and the amylase activity in the urine were determined in each animal group in the 24th, 48th, 72nd, 96th, 120th, 144th experimental hour. For histopathological analysis pancreatic tissue samples were taken from the pancreatic gland. The intensification of the histological changes of these tissue samples was judged using a point score as described by Spormann et al. The results were statistically analysed. The animals of the KP group showed significant histological changes of the pancreas during the whole examination time. Point score: 75-100. Microscopically, tissue samples taken from the animals treated with Fibrolan showed less changes from the 72nd examination hour. There were observed regenerative processes and the improvement of the histological state was evident (point score: 0-50). Changes were less in rats treated with 0.9% NaCl solution than in KP group but the regenerative processes were slower than in F group (point score: 50-75). Enzymes levels were significant lower in F group than in others from the 48th experimental hour.
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PMID:[The effect of intraperitoneally administered necrolytic enzymes on the course of experimental acute pancreatitis]. 942 54

Mucociliary clearance (MCC), the process in which airway mucus together with substances trapped within are moved out of the lungs, is an important defence mechanism of the human body. Drugs may alter this process, such that it is necessary to know the effect of the drugs on MCC. Indeed, agents stimulating MCC may be used therapeutically in respiratory medicine, especially in patients suspected of having an impairment of their mucociliary transport system. In contrast, caution should be taken with drugs depressing MCC as an undesired side-effect, independently of their therapeutic indication. Since cough clearance (CC) serves as a back-up system when MCC fails, the influence of drugs must be examined not only on MCC but also on CC. Ultimately, the clinical repercussions of alterations in mucus transport induced by drug administration must be studied. Tertiary ammonium compounds (anticholinergics), aspirin, anaesthetic agents and benzodiazepines have been shown to be capable of depressing the mucociliary transport system. Cholinergics, methylxanthines, sodium cromoglycate, hypertonic saline, saline as well as water aerosol have been shown to increase MCC. Adrenergic antagonists, guaifenesin, S-carboxymethylcysteine, sodium 2-mercapto-ethane sulphonate and frusemide have been reported not to alter the mucociliary transport significantly. Amiloride, uridine 5'-triphosphate (UTP), quaternary ammonium compounds (anticholinergics), adrenergic agonists, corticosteroids, recombinant human deoxyribonuclease (rhDNase), N-acetylcysteine, bromhexine and ambroxol have been reported either not to change or to augment MCC. Indirect data suggest that surfactant as well as antibiotics may improve the mucociliary transport system. As for the influence of drugs on CC, amiloride and rhDNase have been demonstrated to increase the effectiveness of cough. A trend towards an improved CC was noted after treatment with adrenergic agonists. The anticholinergic agent ipratropium bromide, which is a quaternary ammonium compound, has been suggested to decrease CC significantly. Bromhexine, ambroxol and neutral saline seemed not to alter CC, either positively or negatively. Finally, treatment with either amiloride, recombinant human deoxyribonuclease, bromhexine, ambroxol, N-acetylcysteine, S-carboxymethylcysteine or hypertonic saline has been suggested as a possible cause of clinical improvement in patients, such as the experience of dyspnoea, the case of expectoration or the frequency of infective exacerbations. Other agents did not show a clinical benefit.
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PMID:Effects of drugs on mucus clearance. 1051 29

A protease designated pleureryn, with an N-terminal sequence dissimilar from previously reported mushroom metalloendopeptidases and showing only limited resemblance to aspartic proteinases, albeit considerable homology to DNA replication licensing factor, was isolated from fresh fruiting bodies of the edible mushroom Pleurotus eryngii. The purification protocol entailed ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel Blue gel, ion exchange chromatography on CM-Sepharose, and FPLC-gel filtration on Superdex 75. The protease was unadsorbed on DEAE-cellulose but adsorbed on Affi-gel Blue gel and CM-Sepharose. It demonstrated a single band with a molecular weight of 11.5 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Pleureryn demonstrated a protease activity of 9364 U/mg toward casein. It exhibited a pH optimum of 5.0 and a temperature optimum of 45 degrees C, with substantial activity remaining at high temperatures and pH 4 and 12. The activity of the protease was adversely affected by pepstatin A, indicating that it is an aspartic protease. PMSF, trypsin inhibitor, and EDTA exerted no striking effect, suggesting that it is neither a serine protease nor a metalloprotease. It inhibited translation in a rabbit reticulocyte lysate system with an IC(50) of 20 nM. Pleureryn also exhibited some inhibitory activity against HIV-1 reverse transcriptase, reminiscent of a suppressive action of HIV-1 protease on its homologous reverse transcriptase but was devoid of ribonuclease, deoxyribonuclease, and antifungal activities.
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PMID:Pleureryn, a novel protease from fresh fruiting bodies of the edible mushroom Pleurotus eryngii. 1172 12


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