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PubMed1983

Ultrastructure of normal human T cell subpopulations. Parallel tubular arrays in T gamma lymphocytes and clustered dense bodies in T mu lymphocytes.

Ultrastructures of normal T-cell subpopulations, T gamma and T mu cells, were studied. T gamma cells were isolated and identified by repeating the rosetting method; firstly, by E rosette formation with neuraminidase-treated sheep red blood cells (SRBC), and next by EA gamma-rosette formation with ox red blood cells coated with IgG antibody (EAox). Before EAox rosetting, SRBC on isolated T cells were lysed by autologous plasma instead of ammonium chloride solution. Normal T gamma cells were heterogeneous with regard to their granules; the majority of T gamma cells had parallel tubular arrays (PTA) and a few had electron-dense granules. When ammonium chloride solution was employed to lyse SRBC, PTA were never observed; PTA in normal T gamma cells and in chronic lymphocytic leukemia cells with T gamma character both seemed to change into electron-dense granules after ammonium chloride treatment. In contrast to T gamma cells, T mu cells were characterized by clustered dense bodies, i.e. focal aggregates of electron-dense granules.

Ammonium Chloride↗
PubMed1974

The metabolic basis of portasystemic encephalopathy and the effect of selective vs nonselective shunts.

Portasystemic encephalopathy has been a major deterent to the utilization of total or non-selective shunts. A procedure to determine the maximum rate of urea synthesis (MRUS) has been developed and a depression in the ability to synthesize urea has been shown to correlate closely with the development of encephalopathy. Utilizing this procedure and a modified ammonium tolerance curve, a controlled comparison was instituted between selective and non-selective shunts. Following a non-selective or total shunt, there was a definite deterioration in both the MRUS and the ammonium chloride tolerance curve which was accompanied by a high rate of clinical encephalopathy. In marked contrast, the selective shunt, which maintains portal venous perfusion of the liver, showed little or no change in the MRUS and the ammonium chloride tolerance curve following the selective shunt and there was a very low incidence of encephalopathy. The difference between the non-selective and selective shunt in maintenance of urea synthesis, metabolism of ammonium chloride, and the development of clinical encephalopathy show the selective shunt procedure to be definitively superior in this regard.

Ammonium Chloride↗
PubMed1999

MHC class I-restricted presentation of maleylated protein binding to scavenger receptors.

Pathways for loading exogenous protein-derived peptides on MHC class I are thought to be present mainly in monocyte-lineage cells and to involve phagocytosis- or macropinocytosis-mediated antigenic leakage into either cytosol or extracellular milieu to give peptide access to MHC class I. We show that maleylation of OVA enhanced its presentation to an OVA-specific MHC class I-restricted T cell line by both macrophages and B cells. This enhanced presentation involved uptake through receptors of scavenger receptor (SR)-like ligand specificity, was TAP-1-independent, and was inhibited by low levels (2 mM) of ammonium chloride. No peptide loading of bystander APCs by maleylated (maleyl) OVA-pulsed macrophages was detected. Demaleylated maleyl-OVA showed enhanced MHC class I-restricted presentation through receptor-mediated uptake and remained highly sensitive to 2 mM ammonium chloride. However, if receptor binding of maleyl-OVA was inhibited by maleylated BSA, the residual presentation was relatively resistant to 2 mM ammonium chloride. Maleyl-OVA directly introduced into the cytosol via osmotic lysis of pinosomes was poorly presented, confirming that receptor-mediated presentation of exogenous maleyl-OVA was unlikely to involve a cytosolic pathway. Demaleylated maleyl-OVA was well presented as a cytosolic Ag, consistent with the dependence of cytosolic processing on protein ubiquitination. Thus, receptor-specific delivery of exogenous protein Ags to APCs can result in enhanced MHC class I-restricted presentation, suggesting that the exogenous pathway of peptide loading for MHC class I may be a constitutive property dependent mainly on the quantity of Ag taken up by APCs.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1979

Urinary excretion of chlorpheniramine and pseudoephedrine in humans.

A specific high-pressure liquid chromatographic method for the determination of chlorpheniramine and pseudoephedrine in urine was developed and applied in a urinary excretion study of normal healthy subjects who received a sustained-release dosage form contianing 8 mgof chlorpheniramine maleate and 120 mg of pseudoephedrine hydrochloride. Five subjects received one dose on Day 1, followed by multiple dosing every 12 hr for 7 days without ammonium chloride administration. Four subjects received one dose of the sustained-release dosage form together with ammonium chloride. Urine samples were collected during the 1st day and at steady state. The method is specific and simultaneously determines choorpheniramine, two metabolites (mono- and di-desmethylchlorpheniramine), pseudoephedrine, and norpseudoephedrine. The assay recovery was less than 97% (0.06-3 microgram/ml) for chlorpheniramine maleate and less than 98% (1.5-75 microgram/ml) for pseudoephedrine hydrochloride. Excretion of chlorpheniramine and its two metabolites in urine was enhanced after ammonium chloride administration. At steady state, a change in urine pH from 5.69 to 6.46 resulted in more than a 25% decrease in chlorpheniramine and monodesmethylchlorpheniramine excretion. In spite of expected changes in its biological half-life, the overall amount of unchanged pseudoephedrine excreted in urine was not affected by urine pH, presumably because it is primarily excreted in urine as intact drug.

Chlorpheniramine↗
PubMed1988

Thermoregulatory responses in mice following acute administration of principal nitrogenous excretory substances.

This study was designed to assess the effects of some key excretory nitrogenous substances on body temperature and selected ambient temperature (Ta) in the mouse. In the first experiment, a dosage-response curve was developed to assess the effects of urea, creatinine, and ammonium chloride on colonic temperature at a Ta of 20 degrees C. All three substances elicited a drop in body temperature at a critical dosage. The threshold dosages were 3280 mg/kg for urea, 1279 mg/kg for creatinine, and 365 mg/kg for ammonium chloride. In a second experiment the selected Ta was monitored using a temperature gradient system. Mice were injected with dosages of the nitrogenous substances that had previously been shown to cause hypothermia at a Ta of 20 degrees C. Urea and ammonium chloride had no significant effect on the selected Ta nor on the colonic temperature after 90 min in the temperature gradient. Creatinine elicited a slight lowering of the selected Ta but had no effect on colonic temperature. The thermoregulatory responses to extremely toxic dosages of the nitrogenous substances appear to be quite dissimilar to that when animals are treated with xenobiotic compounds.

Ammonium Chloride↗
PubMed1992

Phagolysosomal alkalinization and the bactericidal effect of antibiotics: the Coxiella burnetii paradigm.

Most infections due to intracellular bacteria respond poorly to antibiotic treatment. The chemical conditions within the subcellular site of bacteria may change antibiotic activity. Coxiella burnetii multiplies within phagolysosomes. The antimicrobial activity of antibiotics combined with the lysosomotropic agents amantadine (1 microgram/mL), chloroquine (1 microgram/mL), and ammonium chloride (1 mg/mL), which alkalinized Coxiella burnetii-containing phagolysosomes from pH 4.8 to 5.3, 5.7, and 6.8, respectively, was evaluated. Percentages of residual viable bacteria (RVB) in cell cultures were significantly reduced after exposure to combinations of doxycycline (4 micrograms/mL) with amantadine (RVB = 18.2% +/- 8.7%, P < .05), chloroquine (RVB = 0.64% +/- 0.38%, P < .01), or ammonium chloride (RVB = 0.29% +/- 0.17%, P < .01); the same was seen with pefloxacin (1 microgram/mL) with chloroquine (RVB = 27.6% +/- 10.8%, P < .05) or ammonium chloride (RVB = 3.72% +/- 1.1%, P < .05). Such bactericidal activity correlated with increased phagolysosomal pH, as determined by Pearson's correlation coefficient, suggesting that phagolysosomal alkalinization is critical for the bactericidal effect of antibiotics.

Amantadine↗
PubMed1992

Intracellular pH on protein kinase C and ionomycin potentiation of isoproterenol-stimulated cyclic AMP and cyclic GMP production in rat pinealocytes.

In rat pinealocytes, alpha 1-adrenergic activation, which leads to cytoplasmic alkalinization, also potentiates the beta-adrenergic stimulated cyclic AMP (cAMP) and cyclic GMP (cGMP) responses. Both elevation of intracellular calcium ([Ca2+]i) and activation of protein kinase C are involved in the potentiation mechanism. Recently, intracellular pH has also been found to modulate the adrenergic-stimulated cyclic nucleotide responses, suggesting intracellular pH may also affect the potentiation mechanism. This possibility was examined in the present study. Cytoplasmic alkalinization by ammonium chloride had an enhancing effect on the isoproterenol and ionomycin-stimulated cAMP and cGMP accumulation. In comparison, cytoplasmic acidification by sodium propionate reduced the isoproterenol and ionomycin-stimulated cAMP and cGMP responses. Direct measurement of [Ca2+]i indicated that neither ammonium chloride nor sodium propionate had an effect on the ionomycin-stimulated elevation of [Ca2+]i, suggesting their effects on cyclic nucleotide responses may be independent of [Ca2+]i. In cells stimulated by isoproterenol and an activator of protein kinase C, ammonium chloride had an enhancing effect on both cAMP and cGMP responses, whereas sodium propionate had no effect. Taken together, these results suggest that a site distal to elevation of [Ca2+]i and activation of protein kinase C, of importance to the potentiation mechanism, is modulated by intracellular pH.

Ammonium Chloride↗
PubMed1999

Studies on semduramicin and nutritional responses: 3. Electrolyte balance.

Four experiments were conducted to determine whether fluctuations in dietary electrolyte level (milliequivalents Na+K-Cl per kilogram) or different dietary sodium chloride levels would affect performance, water intake, or excreta moisture of chickens fed semduramicin vs nonmedicated controls. In all experiments, male commercial broiler chicks were used, and all diets were fed with and without 25 mg semduramicin/kg diet. The basal diets were based on corn, soybean meal, and poultry oil. Experiments 1, 3, and 4 were conducted using male broiler chicks in battery brooders to 18 d of age. In Experiment 1, six electrolyte levels were fed [basal (0.2% Na, 0.33% Cl, 1.10% K); basal plus 0.1% Na (from sodium chloride); basal plus 0.1% K (from potassium chloride); basal plus 0.2% Na (from sodium carbonate); basal plus 0.34% K (from potassium chloride); basal plus 0.15% Cl (from ammonium chloride)]. There were no significant changes in BW gain, feed consumption, or feed conversion ratio caused by any dietary treatment. Water consumption was not affected by the inclusion of Na, Cl, K, or electrolyte levels, but excreta moisture was affected. The highest and lowest excreta moisture levels came from chicks fed the lowest chloride levels. There were no significant dietary effects on serum Na, Cl, or K by dietary electrolytes or semduramicin. Experiment 2 was conducted with triplicate floor pens of 33 male broilers each for 42 d with four electrolyte levels [basal (0.2% Na, 0.34% Cl, 1.03% K); basal plus 0.1% Na (from sodium carbonate); basal plus 0.1% Cl (from ammonium chloride); basal plus 0.18% K (from potassium carbonate)]. Increasing electrolyte level had a significant effect on BW gain at 35 d but not at 42 d [mainly because of differences in K (1.943 kg at 35 d) vs Cl (2.013 kg at 35 d)]. At 42 d, there were no differences in growth because of N, K, Cl, or semduramicin. Potassium supplementation caused a significant increase in litter moisture (P = 0.031). Semduramicin did not affect litter moisture (P = 0.892), nor were there significant semduramicin interactions with Na, K, Cl, or semduramicin. The basal diets in Experiments 3 and 4 were identical to the basal diet in Experiment 1 except there was no sodium chloride added. The diets fed in Experiment 3 contained 0, 0.1, 0.2, 0.3, or 0.4% added sodium chloride. The diets fed in Experiment 4 contained 0.1, 0.3, 0.5, 0.7, and 0.9% added sodium chloride. The results of Experiments 3 and 4 show that about 0.4% added sodium chloride is necessary to achieve maximum growth and feed conversion. It is clear that semduramicin had no significant effect upon the variation observed in any of the variables measured.

Aging↗
PubMed2002

Effect of elevated systemic concentrations of ammonia and urea on the metabolite and ionic composition of oviductal fluid in cattle.

High dietary protein leads to elevated systemic concentrations of ammonia and urea, and these, in turn, have been associated with reduced fertility in cattle. The effect of elevating systemic concentrations of ammonia and urea on the concentrations of electrolytes and nonelectrolytes in bovine oviductal fluid were studied using estrus-synchronized, nulliparous heifers (n = 25). Heifers were randomly assigned to 1 of 3 treatments consisting of jugular vein infusion with either ammonium chloride (n = 8), urea (n = 8), or saline (n = 9). Oviducts were catheterized, and fluid was recovered over a 3-h period on either Day 2 or 8 of the estrous cycle. No difference (P > 0.05) was found in the concentrations of any electrolyte or nonelectrolyte between oviducts ipsi- or contralateral to the corpus luteum. Plasma and oviductal concentrations of urea were increased by infusion with urea (P < 0.001) and ammonium chloride (P < 0.05) but not by saline (P > 0.05). Plasma and oviductal concentrations of ammonia were elevated by infusion with ammonium chloride (P < 0.001) but not by infusion with urea or saline (P > 0.05). No effect (P > 0.05) of treatment was found on oviductal or plasma concentrations of glucose, lactate, magnesium, potassium, or sodium or on plasma concentrations of insulin or progesterone. The concentration of calcium in oviductal fluid was reduced by urea infusion and was negatively associated with systemic and oviductal concentrations of urea. Oviductal concentrations of sodium were higher on Day 8 than on Day 2 (P < 0.05). No effect of sample day was found on any of the other electrolytes or nonelectrolytes measured (P > 0.05). Elevated systemic concentrations of ammonia and urea are unlikely to reduce embryo survival through disruptions in the oviductal environment.

Ammonia↗
PubMed1997

Mis-sorting of procathepsin D in metastogenic tumor cells is not due to impaired synthesis of the phosphomannosyl signal.

Synthesis, processing and sorting of pro-cathepsin D (proCD), the precursor of a lysosomal protease involved in tumor-cell proliferation and invasion, were compared in various human tumor cell lines. In cultures of HepG2, HT29 and MCF7 cells the extracellular CD activity was up to 10 times higher than in cultures of U937 and MG63 cells. Ammonium chloride, which disrupts the pH-dependent receptor-mediated transport of lysosomal enzymes, exerted a differential effect on the secretion of CD activity in these cells. As judged by metabolic labeling and immunoprecipitation, the secretion of CD synthesized in 24 hr in MG63 and U937 cells was enhanced 4- to 10-fold by ammonium chloride, while it was only slightly increased in HepG2, HT29 and MCF7 cells. In all tumor cells examined, a portion of proCD was segregated into the endosomal-lysosomal pathway in the presence of ammonium chloride. However, the intralysosomal maturation of CD was only inhibited by this drug in HT29 and MCF7 cells. The Golgi-associated processing of proCD, leading to the synthesis of uncovered phosphomannosyl groups, proceeded with comparable efficiency in all tumor cells examined. These results suggest that hypersecretion of proCD in HepG2, HT29 and MCF7 cells is linked to transport mechanisms, as yet unidentified, which are independent of mannose-6-phosphate.

Acetylglucosamine↗
PubMed1985

The alloimmunogenicity of heated blood and lymphoid cells.

Murine lymph node cells heated to 45 degrees C for 1 h or 56 degrees C for 15 min lost their ability to provoke a primary cytotoxic alloantibody response, though they were able to provoke a secondary response in animals primed with normal lymph node cells two months previously. The primary immunogenicity of whole blood and spleen cell preparations was destroyed by 56 degrees C but not by 45 degrees C treatment. Treatment of spleen cells with 45 degrees C heat, and ammonium chloride to remove red cells, destroyed their immunogenicity, whereas ammonium chloride treatment alone did not, suggesting that the red cells were the immunogenic component of heated spleen cells and, by implication, of blood. Further evidence for a difference in the immunogenicity of 45 degrees C heated blood and normal blood was provided by the finding that heated blood did not prime for a response to 45 degrees C lymph node cells given two months later. Preliminary investigations of the tolerogenicity of heated cells were unsuccessful, indicating, in view of the published data, that the precise protocol for tolerance induction is very critical.

Ammonium Chloride↗
PubMed1985

Mass spectrometry of oligosaccharides by chloride-attachment reactions: the origin of fragment loss.

The direct exposure, negative chemical ionisation, chloride-attachment mass spectrometry of trehalose and sucrose gave abundant chloride-attached molecular ions. The same feature was observed when these sugars were subjected to fast-atom bombardment (f.a.b.) in a glycerol matrix containing ammonium chloride. No characteristic fragment ion was found when trehalose was analysed by either method. In contrast, sucrose gave intense chloride-containing fragments, arising by glycosidic cleavage, when analysed by the first method, whereas such cleavage was not detectable by f.a.b.-ammonium chloride analysis. However, the mass-analysed ion kinetic energy (m.i.k.e.) spectra of the (M + Cl)- ions from either trehalose and sucrose, generated under f.a.b.-ammonium chloride conditions, showed glycosidic cleavage reactions in addition to a large loss of HCl. These cleavage reactions might be attributed to SN2-like reactions on the acetal carbon atom and to base-induced eliminations, and they were enhanced by collision-induced dissociations. However, the relative abundance of such glycosidic cleavages from the ionic state would be too weak to explain the presence of the large chloride-containing fragments in the direct exposure spectra of sucrose. Thus, these ions were mainly produced by a thermal cleavage followed by chloride-attachment reactions.

Chlorides↗
PubMed1998

Sparfloxacin pharmacokinetics in healthy volunteers: the influence of acidification and alkalinization.

OBJECTIVE: To investigate the effect of acidification and alkalinization on the pharmacokinetics of sparfloxacin in healthy subjects. METHODS: A single 200-mg oral dose of sparfloxacin was given to nine healthy Japanese volunteers on three separate occasions under different conditions of urinary pH. Acidic and alkaline conditions were achieved by repeated oral doses of ammonium chloride and sodium bicarbonate, respectively. The concentrations of sparfloxacin and its metabolite in plasma and urine were determined by high-performance liquid chromatography assays. RESULTS: The difference between treatments for Cmax, AUCinfinity, and CL x f(-1) were found to be significant. The relative bioavailability of sparfloxacin was 84.4% and 122.3% after ammonium chloride and sodium bicarbonate treatments, respectively. The amount of unchanged sparfloxacin in urine samples collected 0-48 h after sparfloxacin administration represented 10.1% of the dose in the control, 14.3% of the dose in urine acidification and 8.4% of the dose with alkalinization of urine. Renal clearance was found to depend on urinary pH. However, the plasma elimination and the metabolism of sparfloxacin were not significantly altered by acidification or alkalinization of the urine. CONCLUSION: The urinary pH dependence of the renal clearance of sparfloxacin will be of minor clinical importance with regard to the low contribution of renal excretion to the overall elimination of sparfloxacin. On the other hand, the alteration in the environmental pH in the gastrointestinal tract, produced by the concomitant ingestion of ammonium chloride or sodium bicarbonate, influences the absorption and bioavailability of sparfloxacin. This effect is likely to be clinically significant.

Administration, Oral↗
PubMed1994

First morning urine pH in the diagnosis of renal tubular acidosis with nephrolithiasis.

Few reports of patients with renal stones compare fasting first morning urine pH with the lowest urine pH after a single oral dose of 0.1 g/kg ammonium chloride. We studied these values in 110 individuals, including 96 stone formers and 14 non-stone forming controls. We hypothesized that fasting first morning urine pH > or = 6.10 is abnormal. Nine of ten patients who had urine pH > or = 5.25 after NH4Cl, had pH > or = 6.10 for both a single and a second first morning urine, giving a sensitivity of 90%. In contrast, a single fasting first morning urine pH > or = 6.10 was found in 19 of 100 individuals with pH < 5.25 after ammonium chloride. However, only 4 of these 100 individuals had abnormal fasting first morning pH > or = 6.10 on two occasions, giving a specificity of 96% for the two urines. Useful prediction of urine pH > or = 5.25 after ammonium chloride occurs when two fasting first morning urines have pH > or = 6.10.

Acidosis, Renal Tubular↗