Regulation of alkaline-resistant laccase from Perenniporia tephropora KU-Alk4 was proved to be controlled by several factors. One important factor was the initial pH, which drove the fungus to produce different kinds of ligninolytic enzymes. P. tephropora KU-Alk4 could grow at pH 4.5, 7.0, and 8.0. The fungus produced laccase and MnP at pH 7.0, but only laccase at pH 8.0. The specific activity of laccase in the pH 8.0 culture was higher than that in the pH 7.0 culture. At pH 8.0, glucose was the best carbon source for laccase production but growth was better with lactose. Low concentrations of glucose at 0.1% to 1.0% enhanced laccase production, while concentrations over 1% gave contradictory results. Veratryl alcohol induced the production of laccase. A trace concentration of copper ions was required for laccase production. Biomass increased with an increasing rate of aeration of shaking flasks from 100 to 140 rpm; however, shaking at over 120 rpm decreased laccase quantity. Highest amount of laccase produced by KU-Alk4, 360 U/mL, was at pH 8.0 with 1% glucose and 0.2 mM copper sulfate, unshaken for the first 3 days, followed by addition of 0.85 mM veratryl alcohol and shaking at 120 rpm. The crude enzyme was significantly stable in alkaline pH 8.0~10.0 for 24 hr. After treating the pulp mill effluent with the KU-Alk4 system for 3 days, pH decreased from 9.6 to 6.8, with reduction of color and chemical oxygen demand at 83.2% and 81%, respectively. Laccase was detectable during the biotreatment process.
Biological Oxygen Demand Analysis ; Biomass ; Carbon ; Copper ; Copper Sulfate ; Fungi ; Glucose ; Hydrogen-Ion Concentration ; Ions ; Laccase* ; Lactose ; Physiology

Copper sulfate is a copper compound used widely in the chemical and agriculture industries. Most intoxication occurs in developing countries of Southeast Asia particularly India, but rarely occurs in Western countries. The early symptoms of intoxication are nausea, vomiting, diarrhea, and abdominal cramps, and the most distinguishable clue is bluish vomiting. The clinical signs of copper sulfate intoxication can vary according to the amount ingested. A 75-year old man came to our emergency room because he had taken approximately 250 ml copper sulfate per oral. His Glasgow Coma Scale (GCS) score was 14 and vital signs were blood pressure 173/111 mmHg, pulse rate 24 bpm, respiration rate 24 bpm, and body temperature 36.1degrees.... Arterial blood gas analysis (ABGa) showed mild hypoxemia and just improved after 2 L/min oxygen supply via nasal cannula. Other laboratory tests and chest CT scan showed no clinical significance. Three hours later, the patient's mental status showed sudden deterioration (GCS 11), and ABGa showed hypercarbia. He was arrested and his spontaneous circulation returned after 8 minutes CPR. However, 22 minutes later, he was arrested again and returned after 3 minutes CPR. The family did not want additional resuscitation, so that he died 5 hours after ED visit. In my knowledge, early deaths are the consequence of shock, while late mortality is related to renal and hepatic failure. However, as this case shows, consideration of early definite airway preservation is reasonable in a case of supposed copper sulfate intoxication, because the patients can show rapid deterioration even when serious clinical manifestation are not presented initially.
Agriculture ; Anoxia ; Asia, Southeastern ; Blood Gas Analysis ; Blood Pressure ; Body Temperature ; Cardiopulmonary Resuscitation ; Catheters ; Colic ; Copper ; Copper Sulfate* ; Developing Countries ; Diarrhea ; Emergency Service, Hospital ; Glasgow Coma Scale ; Heart Rate ; Humans ; India ; Liver Failure ; Mortality ; Nausea ; Oxygen ; Poisoning* ; Respiratory Insufficiency* ; Respiratory Rate ; Resuscitation ; Shock ; Tomography, X-Ray Computed ; Vital Signs ; Vomiting

The aim of this study was to identify and characterize new Flammulina velutipes laccases from its whole-genome sequence. Of the 15 putative laccase genes detected in the F. velutipes genome, four new laccase genes (fvLac-1, fvLac-2, fvLac3, and fvLac-4) were found to contain four complete copper-binding regions (ten histidine residues and one cysteine residue) and four cysteine residues involved in forming disulfide bridges, fvLac-1, fvLac-2, fvLac3, and fvLac-4, encoding proteins consisting of 516, 518, 515, and 533 amino acid residues, respectively. Potential N-glycosylation sites (Asn-Xaa-Ser/Thr) were identified in the cDNA sequence of fvLac-1 (Asn-454), fvLac-2 (Asn-437 and Asn-455), fvLac-3 (Asn-111 and Asn-237), and fvLac4 (Asn-402 and Asn-457). In addition, the first 19~20 amino acid residues of these proteins were predicted to comprise signal peptides. Laccase activity assays and reverse transcription polymerase chain reaction analyses clearly reveal that CuSO4 affects the induction and the transcription level of these laccase genes.
Copper Sulfate ; Cysteine ; DNA, Complementary ; Flammulina* ; Fungi* ; Genome ; Histidine ; Laccase* ; Polymerase Chain Reaction ; Protein Sorting Signals ; Reverse Transcription

OBJECTIVETo assess various antioxidative activities of Satureja khozestanica essential oil (SKE) and its effect on oxidation of low density lipoprotein (LDL) induced by CuSO4 in vitro by monitoring the formation of conjugated dienes and malondialdehyde (MDA).

METHODSThe formation of conjugated dienes, lag time and MDA were measured. Inhibition of this Cu-induced oxidation was studied in the presence of several concentrations of SKE. Also total antioxidant activity and free radical scavenging of SKE were determinated.

RESULTSIt was demonstrated that SKE was able to inhibit LDL oxidation and decrease the resistance of LDL against oxidation. The inhibitory effects of SKE on LDL oxidation were dose-dependent at concentrations ranging from 50 to 200 µg/mL. Total antioxidant capacity of SKE was (3.20±0.40) nmol of ascorbic acid equivalents/g SKE. The SKE showed remarkable scavenging activity on 2, 2-diphenyl-picrylhydrazyl, IC50 (5.30±0.11) ng/mL.

CONCLUSIONSThis study shows that SKE is a source of potent antioxidants and prevents the oxidation of LDL in vitro and it may be suitable for use in food and pharmaceutical applications.

Antioxidants ; chemistry ; pharmacology ; Copper Sulfate ; chemistry ; Humans ; Lipoproteins, LDL ; chemistry ; metabolism ; Male ; Malondialdehyde ; chemistry ; Oils, Volatile ; chemistry ; pharmacology ; Oxidation-Reduction ; drug effects ; Satureja ; chemistry

Copper sulfate ingestion is a rare cause of corrosive gastrointestinal injury in the Republic of Korea. In developing countries, copper sulfate is chiefly used for agricultural purposes as a pesticide and in the leather industry. It is also used in school science classes in the form of bright blue crystals. Copper sulfate is a powerful oxidizing agent that is corrosive to mucous membranes. Concentrated solutions are acidic, with a pH of 4. We report a case of corrosive gastritis and esophagitis due to accidental copper sulfate ingestion in a 12-year-old boy.
Burns ; Child ; Copper ; Copper Sulfate ; Developing Countries ; Eating ; Esophagitis ; Gastritis ; Humans ; Hydrogen-Ion Concentration ; Mucous Membrane ; Republic of Korea

BACKGROUNDThe protective effects of magnesium sulfate against ischemia-reperfusion injury of the small intestine in Sprague-Dawley (SD) rats have been confirmed in our previous research. However, its exact mechanism is unclear. This study was to evaluate the role of PI3K/Akt signal pathway in the protective effect of magnesium sulfate against ischemia-reperfusion injury of the small intestine in SD rats.

METHODSRat model of intestinal ischemia-reperfusion injury was used. The SD rats were divided into four groups randomly: sham operation group, ischemia-reperfusion group, magnesium sulfate group and magnesium sulfate plus LY294002 (an inhibitor of PI3K) group. The pathological changes of intestinal mucosa were examined; the activity of diamine oxidase (DAO) in plasma, the plasma contents of malondialdehyde (MDA), and apoptosis rate of the intestinal mucosal cells were determined and compared. The expression of p-Akt was detected by Western blotting.

RESULTSThere were more evident pathological changes of the intestinal mucosa (higher Chiu's score, P < 0.05), enhanced DAO activity (P < 0.05), elevated contents of MDA (P < 0.05), higher apoptosis rate (P < 0.05), and lower level of p-Akt (P < 0.05) in the ischemia-reperfusion group compared with the sham operation group. There were less evident pathological changes of the intestinal mucosa (lower Chiu's score, P < 0.05), lower DAO activity (P < 0.05), lower contents of MDA (P < 0.05), and lower apoptosis rate (P < 0.05), but higher level of p-Akt (P < 0.05) in the magnesium sulfate group compared with the ischemia-reperfusion group. There were more evident pathological changes of the intestinal mucosa (higher Chiu's score, P < 0.05), higher contents of MDA (P < 0.05), higher DAO activity (P < 0.05) and higher apoptosis rate (P < 0.05), and lower level of p-Akt (P < 0.05) in the magnesium sulfate plus LY294002 group compared with the magnesium sulfate group.

CONCLUSIONSActivation of PI3K/Akt signal pathway results in the reduction of cell apoptosis, which likely accounts for the protective effect of magnesium sulfate against intestinal ischemia-reperfusion injury.

Amine Oxidase (Copper-Containing) ; metabolism ; Animals ; Apoptosis ; drug effects ; Blotting, Western ; Disease Models, Animal ; Intestinal Mucosa ; cytology ; drug effects ; Intestine, Small ; drug effects ; Magnesium Sulfate ; therapeutic use ; Malondialdehyde ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; prevention & control ; Signal Transduction ; drug effects

Culture medium and fermentation conditions for lipid production by Rhodosporidium toruloides were optimized with single factor and uniform design experiment. The best medium recipe was found with 70 g/L glucose, 0.1 g/L (NH4)2SO4, 0.75 g/L yeast extract, 1.5 g/L MgSO4. 7H2O, 0.4g/L KH2PO4, sterilized at 121 degrees C for 15 min, and then supplemented with ZnSO4 1.91 x 10(-6) mmol/L, CaCl2 1.50 mmol/L, MnCl2 1.22 x 10(-4) mmol/L and CuSO4 1.00 x 10(-4) mmol/L. The optimal fermentation conditions were as follows: 50 mL of medium (pH 6.0) in 250 mL Erlenmeyer flask with 10% inoculum (28h) under orbital shaking at 200 r/min for 120h at 30 degrees C. Under these conditions, yeast biomass accumulated lipids up to 76.1%.
Basidiomycota ; growth & development ; metabolism ; Copper ; pharmacology ; Culture Media ; Fermentation ; Hydrogen-Ion Concentration ; Lipids ; biosynthesis ; Magnesium Sulfate ; pharmacology ; Zinc ; pharmacology

AIMTo establish a new, reliable vomiting model in minks.

METHODSAdult male minks (Mustela vison) were randomly divided into groups (n = 6). Cisplatin, apomorphine, copper sulfate and X-radiation were used to establish vomiting model. Retching and vomiting were observed after the vomiting models were given anti-vomiting agents. After the behavioral experiment, assay of 5-HT in the ileum was performed by immunohistologic method.

RESULTSCisplatin 7.5 mg.kg-1 i.p., apomorphine 1.6 mg.kg-1 s.c. and copper sulfate 40 mg.kg-1 ig were shown to evoke vomiting. Retching and vomiting were significantly inhibited in ondansetron and metoclopramide pretreated minks (P < 0.05, P < 0.01).

CONCLUSIONAs a new vomiting model, minks may be of great value in studying vomiting mechanism and screening new antiemetic drugs.

Animals ; Antiemetics ; therapeutic use ; Apomorphine ; Cisplatin ; Copper Sulfate ; Disease Models, Animal ; Male ; Metoclopramide ; therapeutic use ; Mink ; Ondansetron ; therapeutic use ; Vomiting ; chemically induced ; drug therapy

BACKGROUND: Modified lipoproteins may be involved in nephro- and glomerulosclerosis. Diabetic nephropathy-like lesions have also been induced in a rat model by glycated and glycoxidized albumin. In cultured rat or human mesangial cells, enhanced cell proliferation and production of mesangial matrix in response to lipoproteins and their modified forms have been demonstrated by [3H]-thymidine incorporation and cell counting assays. But these methods are relatively complex and most of them have used only one or two of the lipoprotein, albumin and their modified forms. METHODS: We investigated the effects of native and modifed lipoproteins, and albumin on cultured human mesangial cell proliferation using non-radioactive colorimetric method by MTS/PMS assay. Lipoproteins added were low density lipoprotein(LDL), high density lipoprotein(HDL), very low density lipoprotein(VLDL), oxidized LDL(oxidation with copper sulfate in vitro) and glycated LDL and we also used albumin, glycated albumin, and interleukin-1beta as a positive control. RESULTS: Interleukin-1beta promoted the proliferation of cultured human mesangial cells up to concentration 20 ng/mL. LDL induced the proliferation of mesangial cells in a concentration-dependent manner up to concentration 100 microgram/mL. HDL and VLDL had no significant proliferative effect. Oxidized LDL caused the proliferation of mesangial cells at low concentration up to concentration 25 microgram/mL. Addition of glycated LDL resulted in a concentration- dependent inhibition of mesangial cells. Albumin and glycated albumin inhibited the proliferation of mesangial cells at low concentration of 100 microgram/mL, but cell growth was increased at higher concentrations. CONCLUSION: We demonstrated the effects of the single and modified proteins on the proliferation of cultured human mesangial cell by relatively simple colorimetric method. Results were almostly identical to those of previous studies obtained by radioactive method or cell counting assay.
Animals ; Cell Count ; Cell Proliferation ; Copper Sulfate ; Humans* ; Interleukin-1beta ; Lipoproteins* ; Mesangial Cells* ; Models, Animal ; Rats

Oxidized low-density lipoprotein (oxLDL) induces a wide range of cellular responses to produce atherosclerotic lesion, but key factors determining the response are not understood. In this study, purified LDL was oxidized with copper sulfate, and its physical properties and the related biological responses were investigated. The average hydrodynamic diameter of the lightly oxidized LDL was approximately 25 nm and its Rf value relative to nLDL on agarose gel was between 1.0 and 1.25. The diameter of the extensively oxidized LDL was over 30 nm, the Rf value was over 2.0. A 24 h-exposure of resting RAW264.7 macrophage cells to 100 microg/ml of the lightly oxidized LDL induced proliferation and macrophage activation whereas the extensively oxidized LDL induced cell death at the same concentration. In contrast, 200 microg/ml of oxLDL caused cell death regardless of oxidation degree. Short incubation (4-6 h) of the highly oxidized LDL (100 microg/ml) also resulted in cell proliferation. OxLDL-induced cell death showed mixed characteristics of apoptosis and/or necrosis depending on the strength and duration of the insult. These results suggest that cellular responses induced by oxLDL be dependent on the oxidation degree, the duration of exposure, and the concentration of oxLDL. Copyright 2000 Academic Press.
Animal ; Apolipoproteins B/metabolism ; Apoptosis/physiology ; Apoptosis/drug effects ; Cell Death/physiology* ; Cell Division/physiology ; Copper Sulfate/metabolism ; Dose-Response Relationship, Drug ; Human ; Lipid Peroxidation ; Lipids/metabolism ; Lipoproteins, LDL/pharmacology ; Lipoproteins, LDL/metabolism* ; Macrophages/pathology ; Macrophages/drug effects ; Macrophages/cytology* ; Mice ; Necrosis ; Oxidation-Reduction ; Thiobarbituric Acid Reactive Substances/metabolism