PURPOSE: To investigate the effects of Helicobacter pylori (H. pylori)-CagA and the urease metabolite NH₄⁺ on mucin expression in AGS cells. MATERIALS AND METHODS: AGS cells were transfected with CagA and/or treated with different concentrations of NH₄⁺CL. Mucin gene and protein expression was assessed by qPCR and immunofluorescence assays, respectively. RESULTS: CagA significantly upregulated MUC5AC, MUC2, and MUC5B expression in AGS cells, but did not affect E-cadherin and MUC6 expression. MUC5AC, MUC6, and MUC2 expression in AGS cells increased with increasing NH₄⁺ concentrations until reaching a peak level at 15 mM. MUC5B mRNA expression in AGS cells (NH₄⁺ concentration of 15 mM) was significantly higher than that at 0, 5, and 10 mM NH₄⁺. No changes in E-cadherin expression in AGS cells treated with NH₄⁺ were noted, except at 20 mM. The expression of MUC5AC, MUC2, and MUC6 mRNA in CagA-transfected AGS cells at an NH₄⁺ concentration of 15 mM was significantly NH₄⁺ concentration, and was significantly higher compared to that in untreated cells. No significant change in the expression of E-cadherin mRNA in CagA-transfected AGS cells was observed. Immunofluorescence assays confirmed the observed changes. CONCLUSION: H. pylori may affect the expression of MUC5AC, MUC2, MUC5B, and MUC6 in AGS cells via CagA and/or NH₄⁺, but not E-cadherin.
Ammonium Chloride ; Ammonium Compounds* ; Cadherins ; Fluorescent Antibody Technique ; Helicobacter pylori* ; Helicobacter* ; Mucins* ; RNA, Messenger ; Stomach ; Urease ; Virulence*

BACKGROUND: We evaluated the reliability and accuracy of the combined use of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) bacterial identification and Vitek 2 antimicrobial susceptibility testing (AST) for bacteria from positive blood culture bottles. METHODS: Direct identification and AST were performed in parallel to the standard methods in monomicrobial positive blood culture bottles. In total, 254 isolates grown on aerobic and/or anaerobic bottles were identified with MALDI-TOF Vitek MS (bioMerieux, France), and 1,978 microorganism/antimicrobial agent combinations were assessed. For isolates from anaerobic bottles, an aliquot of the culture broth was centrifuged, washed, and filtered through a nylon mesh. For isolates from aerobic/pediatric bottles, a lysis step using 9.26% ammonium chloride solution and 2% saponin solution was included. RESULTS: The overall correct identification rate was 81.8% (208/254) and that for gram-positive/gram-negative isolates was 73.9%/92.6%, respectively, and it was 81.8%, 87.6%, and 57.9% for isolates from aerobic, anaerobic, and pediatric bottles, respectively. Identification was not possible in 45 cases, and most of these isolates were streptococci (N=14) and coagulase-negative staphylococci (N=11). Misidentification occurred only in one case. Compared with standard methods, direct AST showed 97.9% (1,936/1,978) agreement with very major error of 0.25%, major error of 0.05%, and minor error of 1.8%. CONCLUSIONS: This simple and cost-effective sample preparation method gives reliable results for the direct identification and AST of bacteria. For the identification of streptococci and coagulase-negative staphylococci, the method should be further improved.
Adult ; Ammonium Chloride/chemistry ; Anti-Infective Agents/*pharmacology ; Child ; Gram-Negative Bacteria/drug effects/*isolation & purification/metabolism ; Gram-Positive Bacteria/drug effects/*isolation & purification/metabolism ; Humans ; Reagent Kits, Diagnostic ; Saponins/chemistry ; *Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

The influences of temperature and nutritional conditions on the mycelium growth of oyster mushroom Pleurotus ostreatus (PO) and Pleurotus cystidiosus (PC) were investigated in laboratory experiment during the summer season of 2014. The results of the experiment indicated that potato dextrose agar (PDA) and yam dextrose agar (YDA) were the most suitable media for the mycelium growth of oyster mushroom PO while four media (PDA, YDA, sweet potato dextrose agar, and malt extract agar medium) were not significantly different in supporting mycelium growth of oyster mushroom PC. The optimal temperature for mycelium growth of both oyster mushroom species was obtained at 28degrees C. Mycelium growth of oyster mushroom PO was improved by carbon sources such as glucose, molasses, and at 1~5% sucrose concentration, mycelium colony diameter of mushroom PO was achieved the highest value. Whereas glucose, dextrose, and sucrose as carbon sources gave the good mycelium growth of oyster mushroom PC, and at 1~3% sucrose concentration, mycelium colony diameter of PC was achieved the maximum value. Ammonium chloride concentrations at 0.03~0.09% and 0.03~0.05% also gave the greatest values in mycelium colony diameter of mushroom PO and PC. Brown rice was found to be the most favourable for mycelium growth of two oyster mushroom species. In addition, sugarcane residue, acasia sawdust and corn cob were selected as favourable lignocellulosic substrate sources for mycelium growth of both oyster mushrooms.
Agar ; Agaricales ; Ammonium Chloride ; Carbon ; Dioscorea ; Glucose ; Ipomoea batatas ; Molasses ; Mycelium* ; Pleurotus* ; Saccharum ; Seasons ; Solanum tuberosum ; Sucrose ; Zea mays

A simple, rapid and selective liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) is developed and validated for quantification of venlafaxine in human plasma with simple liquid-liquid extraction step consisted of extraction with ether and dichloromethane for 10 min and mixing with 1 M sodium acetate in human plasma using fluoxetine as an internal standard (IS). The analyte are separated using an isocratic mobile phase consisted of acetonitrile and 5 mM ammonium formate (4/3, v/v) on a isocratic YMC hydrosphere C18 (2.0x50.0 mm, 3.0 microm) column and analyzed by MS/MS in the multiple reaction monitoring (MRM) mode using the transitions of respective [M+H](+) ions, m/z 278.2-->260.3 and m/z 310.1-->148.1 for quantification of venlafaxine and IS, respectively. The standard calibration curves showed good linearity within the range of 1.0-200.0 ng/mL (r2=0.9986, 1/chi2 weighting). The lower limit of quantification (LLOQ) was 1.0 ng/mL. The retention times of venlafaxine and IS were 0.6 min and 0.7 min that means the potential for the high-throughput potential of the proposed method. In addition, no significant metabolic compounds were found to interfere with the analysis. Acceptable precision and accuracy were obtained for the concentrations over the standard curve range. The validated method was successfully applied to bioequivalence study after 75-mg of venlafaxine sustained-release (SR) capsule in 24 healthy Korean subjects.
Ammonium Compounds ; Calibration ; Chromatography, Liquid ; Ether ; Fluoxetine ; Humans ; Ions ; Liquid-Liquid Extraction ; Methylene Chloride ; Pharmacokinetics ; Plasma* ; Sodium Acetate ; Tandem Mass Spectrometry ; Therapeutic Equivalency* ; Venlafaxine Hydrochloride

BACKGROUND: The present study was attempted to compare enalapril, an angiotensin-converting enzyme inhibitor with losartan an angiotensin II (Ang II) receptor blocker in the inhibitory effects on the secretion of catecholamines (CA) from the perfused model of the rat adrenal gland. METHODS: The adrenal gland was isolated and perfused with Krebs-bicarbonate. CA was measured directly by using the fluorospectrophotometer. RESULTS: Both enalapril and losartan during perfusion into an adrenal vein for 90 minutes inhibited the CA release evoked by acetylcholine (ACh), 1.1-dimethyl-4-phenyl piperazinium (DMPP, a selective Nn agonist), high K+ (a direct membrane-depolarizer), 3-(m-chloro-phenyl-carbamoyl-oxy-2-butynyl-trimethyl ammonium (McN-A-343, a selective M1 agonist), and Ang II in a time-dependent manner. Also, in the presence of enalapril or losartan, the CA release evoked by veratridine (an activator of voltage-dependent Na+ channels), 6-dimethyl-3-nitro-4-(2-trifluoromethyl-phenyl)-pyridine-5-carboxylate (BAY-K-8644, an L-type Ca2+ channel activator), and cyclopiazonic acid (a cytoplasmic Ca2+-ATPase inhibitor) were significantly reduced. Based on the same concentration of enalapril and losartan, for the CA release evoked by ACh, high K+, DMPP, McN-A-343, Ang II, veratridine, BAY-K-8644, and cyclopiazonic acid, the following rank order of inhibitory potency was obtained: losartan > enalapril. In the simultaneous presence of enalapril and losartan, ACh-evoked CA secretion was more strongly inhibited compared with that of enalapril- or losartan-treated alone. CONCLUSIONS: Collectively, these results demonstrate that both enalapril and losartan inhibit the CA secretion evoked by activation of both cholinergic and Ang II type-1 receptors stimulation in the perfused rat adrenal medulla. When these two drugs were used in combination, their effects were enhanced, which may also be of clinical benefit. Based on concentration used in this study, the inhibitory effect of losartan on the CA secretion seems to be more potent than that of enalapril.
(4-(m-Chlorophenylcarbamoyloxy)-2-butynyl)trimethylammonium Chloride ; 3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester ; Acetylcholine ; Adrenal Glands ; Adrenal Medulla ; Ammonium Compounds ; Angiotensin II ; Animals ; Catecholamines ; Cytoplasm ; Dimethylphenylpiperazinium Iodide ; Enalapril* ; Losartan* ; Perfusion ; Rats ; Veins ; Veratridine

PURPOSE: Kidney stones (nephrolithiasis) are a widespread disease. Thus, blocking stone formation and finding new therapeutic methods is an important area of study. Diosmin (a major component of the bile) is known to have antioxidant as well as renoprotective effects. The present investigation aimed to evaluate the effect of diosmin on renal tissue protection in rats with ethylene glycol-induced nephrolithiasis. MATERIALS AND METHODS: The rats were randomly divided into three groups. Group one (control) did not receive any treatments. In groups two and three, nephrolithiasis was induced by 2.5% (V/V) ethylene glycol + 2.5% (W/V) ammonium chloride (2 mL/d). The second and the third groups received distilled water or diosmin (80 mg/kg/d) by gavage for 21 days. RESULTS: Stereological estimation of the renal structures revealed that the average volume of calcium oxalate (CaOx) in the nephrolithiasis+diosmin rats was -63% less than in the rats with untreated nephrolithiasis (p<0.01). The volume of the glomeruli, proximal and distal convoluted tubules, Henle's loop, collecting ducts, and vessels was reduced -32% to 58% after the induction of nephrolithiasis (p<0.001). In the nephrolithiasis+diosmin rats, on average, -70% to 96% of the glomeruli, proximal convoluted tubules, Henle's loop collecting ducts, and vessels remained intact (p<0.01). Degeneration of the cortical tissue was 5-fold that of the medulla. In the nephrolithiasis+diosmin rats, degeneration in the renal cortical tissue and medulla was reduced -70% and 44%, respectively, compared with that in the untreated nephrolithiasis group (p<0.01). CONCLUSIONS: Diosmin reduces CaOx deposition and the degeneration of glomeruli and tubules in a rat model of nephrolithiasis.
Ammonium Chloride ; Animals ; Calcium ; Calcium Oxalate ; Diosmin ; Ethylene Glycol ; Ethylenes ; Kidney Calculi ; Nephrolithiasis ; Rats ; Water

The role of the kidney in combating metabolic acidosis has been a subject of considerable interest for many years. The present study was aimed to determine whether there is an altered regulation of renal acid base transporters in acute and chronic acid loading. Male Sprague-Dawley rats were used. Metabolic acidosis was induced by administration of NH4Cl for 2 days (acute) and for 7days (chronic). The serum and urinary pH and bicarbonate were measured. The protein expression of renal acid base transporters [type 3 Na+/H+ exchanger (NHE3), type 1 Na+/HCO3- cotransporter (NBC1), Na-K+ ATPase, H(+)-ATPase, anion exchanger-1 (AE-1)] was measured by semiquantitative immunoblotting. Serum bicarbonate and pH were decreased in acute acid loading rats compared with controls. Accordingly, urinary pH decreased. The protein expression of NHE3, H(+)-ATPase, AE-1 and NBC1 was not changed. In chronic acid loading rats, serum bicarbonate and pH were not changed, while urinary pH was decreased compared with controls. The protein expression of NHE3, H(+)-ATPase was increased in the renal cortex of chronic acid loading rats. These results suggest that unaltered expression of acid transporters combined with acute acid loading may contribute to the development of acidosis. The subsequent increased expression of NHE3, H(+)-ATPase in the kidney may play a role in promoting acid excretion in the later stage of acid loading, which counteract the development of metabolic acidosis.
Acidosis ; Adenosine Triphosphatases ; Ammonium Chloride ; Animals ; Humans ; Hydrogen-Ion Concentration ; Immunoblotting ; Kidney ; Male ; Proton-Translocating ATPases ; Quaternary Ammonium Compounds ; Rats ; Rats, Sprague-Dawley ; Sodium-Hydrogen Antiporter

BACKGROUNDAstrocyte swelling is an important consequence of hepatic encephalopathy, and aquaporin-4 has been reported to play a vital role in this swelling. Ammonia causes astrocyte swelling and is also known to modulate aquaporin-4 expression in the astrocyte foot processes. The purpose of this study was to explore the mechanism of ammonia-induced aquaporin-4 expression, which has been suggested to involve the p38 mitogen-activated protein kinase pathway.

METHODSWe exposed cultured astrocytes to ammonium chloride, an in vitro model of hepatic encephalopathy. The purity of cultured astrocytes was evaluated by fluorescent glial fibrillary acidic protein labeling; cell morphology was assessed by light microscopy; the expression of aquaporin-4, phospho-p38, and p38 were detected by Western blotting analysis. Statistical analysis was performed by one-way factorial analysis of variance, and the relationship between variables was calculated by linear regression using SPSS version 13.0 program for Windows (SPSS, Chicago, IL, USA).

RESULTSThe purity of cultured astrocytes was (96.6 +/- 1.4)%. Astrocytes swelled significantly when exposed to 5 mmol/L ammonium chloride for 24 hours as compared to non-exposed astrocytes. Co-treatment with 10 micromol/L SB203580 (an inhibitor of p38) attenuated the degree of ammonium chloride induced astrocyte swelling. Western blotting analysis revealed that the expression levels of phospho-p38 and aquaporin-4 in ammonium chloride treated cells were significantly increased relative to the control group (P < 0.001); SB203580 co-treatment inhibited the increased expression of phospho-p38 and aquaporin-4 relative to the ammonium chloride treated group (P = 0.002 and P = 0.015 respectively). The phosphorylation of p38 and upregulation of aquaporin-4 were highly correlated (r = 0.909). There were no significant differences in total p38 expression among the groups (P = 0.341).

CONCLUSIONSAmmonium chloride induced upregulation of aquaporin-4 in astrocytes is regulated by the p38 mitogen-activated protein kinase pathway. Inhibiting p38 activation prevented ammonium chloride induced aquaporin-4 protein upregulation.

Ammonium Chloride ; pharmacology ; Animals ; Aquaporin 4 ; genetics ; metabolism ; Astrocytes ; drug effects ; metabolism ; Blotting, Western ; Cells, Cultured ; Enzyme Inhibitors ; pharmacology ; Imidazoles ; pharmacology ; Phosphorylation ; drug effects ; Pyridines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; metabolism

BACKGROUND: Hand eczema is a common skin disease in the general population. The etiology of hand eczema is obscure and many causative factors have been proposed. However, there are only a few reported studies of the relevance of contact allergy in hand eczema. Objective: The purpose of this study was to evaluate the diagnostic value of the patch test for patients with hand eczema. METHODS: We analyzed the clinical characteristics and the results of the patch tests of the 37 patients with hand eczema and we then compared these with the clinical subtypes. RESULTS: 26 patients (70.3%) showed a positive test to one or more allergens. The common allergens were nickel sulfate (35.1%), mercury ammonium chloride (21.6%), and cobalt chloride (18.9%). The positive rates for a patch test were 82.4% for the vesicular form, 77.8% for the fissured form, 20.0% for the hyperkeratotic form and 66.7% for pompholyx, respectively. We found more significant improvement of the clinical symptoms in the vesicular group (57.1%) than that in the non-vesicular group (9.3%) after avoiding the verified allergens. CONCLUSION: This study shows that the patch test is a useful tool for the detection of contact allergens and it must be performed for the patients with hand eczema, and especially for those patients with the vesicular type.
Allergens ; Ammonia ; Ammonium Chloride ; Cobalt ; Dermatitis, Allergic Contact ; Eczema ; Eczema, Dyshidrotic ; Hand ; Humans ; Hypersensitivity ; Mercuric Chloride ; Nickel ; Occupations ; Patch Tests ; Skin Diseases

BACKGROUND: Hand eczema is a common skin disease in the general population. The etiology of hand eczema is obscure and many causative factors have been proposed. However, there are only a few reported studies of the relevance of contact allergy in hand eczema. Objective: The purpose of this study was to evaluate the diagnostic value of the patch test for patients with hand eczema. METHODS: We analyzed the clinical characteristics and the results of the patch tests of the 37 patients with hand eczema and we then compared these with the clinical subtypes. RESULTS: 26 patients (70.3%) showed a positive test to one or more allergens. The common allergens were nickel sulfate (35.1%), mercury ammonium chloride (21.6%), and cobalt chloride (18.9%). The positive rates for a patch test were 82.4% for the vesicular form, 77.8% for the fissured form, 20.0% for the hyperkeratotic form and 66.7% for pompholyx, respectively. We found more significant improvement of the clinical symptoms in the vesicular group (57.1%) than that in the non-vesicular group (9.3%) after avoiding the verified allergens. CONCLUSION: This study shows that the patch test is a useful tool for the detection of contact allergens and it must be performed for the patients with hand eczema, and especially for those patients with the vesicular type.
Allergens ; Ammonia ; Ammonium Chloride ; Cobalt ; Dermatitis, Allergic Contact ; Eczema ; Eczema, Dyshidrotic ; Hand ; Humans ; Hypersensitivity ; Mercuric Chloride ; Nickel ; Occupations ; Patch Tests ; Skin Diseases