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Ammonium chloride in the prophylaxis of parturient hypocalcaemia.

Twenty Holstein X Friesian cows weighing 642 +/- 29 kg, were used at the 4th or 5th calving. They received a daily ration offering them 80 g calcium, 80 g inorganic phosphate and 60 g magnesium. Ten of these animals were supplemented with NH4Cl (20 g per animal per day, from the 15th day before calving until the 5th day after calving). This treatment had no significant effect on blood pH, ionised calcium, total calcium, inorganic phosphate, magnesium and protein levels. Among control and treated animals, three and four cows, respectively, were afflicted with parturient hypocalcaemia. These results demonstrate that in our experimental conditions NH4Cl was completely ineffective in preventing parturient hypocalcaemia.

Ammonium Chloride↗

Relation between hyaluronan and sulphated glycosaminoglycan synthesis and degradation in cultured embryonic fibroblasts. Effect of concanavalin A and ammonium chloride administration.

In order to evaluate the relationship between glycosaminoglycan (GAG) synthesis and degradation, the effect of NH4Cl, which inhibits lysosomal degradation, on GAG production was analysed in vitro in concanavalin A (Con A) stimulated fibroblasts from 7 and 14-day-old chick embryos. 35SO4 incorporation into total proteoglycan (PG), 3H incorporation into individual GAG classes, beta-N-acetyl-D-glucosaminidase and beta-D-glucuronidase activity were determined. The results indicate a correlation between Con A and NH4Cl effects: NH4Cl induced a reduction principally in the GAG classes most stimulated by Con A. Thus HA and DS are much more stimulated by Con A and inhibited by NH4Cl than are CS and HS.

Acetylglucosaminidase↗

The effect of ammonium chloride and tunicamycin on the glycoprotein content and infectivity of herpes simplex virus type 1.

Infectious virions of MP, a syncytial strain of herpes simplex virus type 1, are formed in the presence of 50 mM NH4Cl. Underglycosylated virion glycoproteins are synthesized in infected cells and are incorporated into virions in the presence of the same concentration of NH4Cl. We conclude that fully glycosylated glycoproteins are not required for viral infectivity. Virus particles, deficient in glycosylated glycoproteins, are assembled in the presence of tunicamycin but they are not infectious. The decrease in infectivity could be due to the decreased amount of the gB or possibly other peptides and/or to the lack of the high-mannose saccharides of precursor glycoproteins.

Ammonium Chloride↗

[The ammonium chloride modification of the effect of sodium nitroprusside on the contractile activity of guinea pig papillary muscles].

The rise of intracellular pH (pH) caused by the replacement of 20 mM NaCl by NH4Cl was shown to enhance guinea-pig papillary muscle contractility while the pH decrease under the NH4Cl abolition from perfusion solution was obtained to reduce the contractile activity. Sodium nitroprusside (SN, 10 microM) does not influence the cardiac muscle contractility in controls but significantly decreases its levels under intracellular alkalinization. SN washout does not restore contractility, hence, the subsequent NH4Cl removal results in contractility increase. In most cases simultaneous action of SN and NH4Cl was accompanied by the development of cardiac mechanical instability. Thus, pHi change can provoke unexpected effects of nitrates.

Ammonium Chloride↗

Comparison of the effects of isotonic and hypertonic sodium bicarbonate solutions on acidemic calves experimentally induced by ammonium chloride administration.

The objectives of this study were to evaluate and compare the effects of intravenously (IV) administered infusion of isotonic solution (ISB) or hypertonic sodium bicarbonate solution (HSB) on acid-base equilibrium and the plasma osmolarity in acidemic calves experimentally induced by 5 M-NH(4)Cl, IV infusion (1.0 ml/kg, over 1 hr). The ISB and HSB infusion induced progressive and significant increases in their HCO(3)(-) and BE levels that persisted throughout the period of fluid administration. The plasma osmolarity in the ISB groups was significantly decreased. The plasma osmolarity in the HSB group was significantly higher than in the calves in the other groups (p<0.05). ISB solution might be safe and effective for treating and reviving conscious calves from experimentally induced metabolic acidosis.

Acidosis↗

The elucidation of the effect of ammonium chloride on pyruvate distribution and pyruvate dehydrogenase interconversion in isolated rat hepatocytes.

The distribution of pyruvate between cell compartments measured in isolated hepatocytes in the presence of lactate was in agreement with delta pH across plasma and mitochondrial membranes. In isolated liver mitochondria NH4Cl decreased the transmembrane potential (delta psi) by about 14 mV, whereas no change of delta pH was observed. In the presence of lactate or alanine NH4Cl increased the mitochondrial pyruvate concentration presumably due to the inhibition of the flux through pyruvate carboxylase. In the presence of lactate or alanine changes in the amount of the active form of pyruvate dehydrogenase (PDHa) were correlated with the mitochondrial pyruvate concentration, NH4Cl increased the amount of PDHa by lowering the mitochondrial ATP/ADP and NADH/NAD+ ratios.

Adenosine Diphosphate↗

[Early action of colchicine, ammonium chloride and prolactin, on secretion of milk lipids in the lactating mammary gland (author's transl)].

The secretion of milk lipids was investigated by incubating mammary tissue fragments from lactating ewes and rabbits. After the lipids were pulse-labelled (3 min with 14C-sodium acetate or 14C-sodium butyrate), the intracellular and extracellular lipid globules, localized in the acinar lumen, were labelled (as shown by electron microscopy), and the labelled lipids were released into the medium. Colchicine, a microtubule modifier inhibiting protein secretion, did not modify lipid secretion. NH4Cl, a lysosomotropic agent, did not decrease lipid secretion. When prolactin was added immediately after the pulse, the amount of labelled lipids secreted for two hours increases significantly. These results show that prolactin stimulates lipid secretion in the same way as protein secretion. However, microtubular integrity, necessary for protein secretion, is not required for lipid secretion.

Ammonium Chloride↗

Inhibition of hemoglobin degradation in Plasmodium falciparum by chloroquine and ammonium chloride.

The effect of chloroquine (CQ) and NH4Cl on hemoglobin degradation within Plasmodium falciparum was studied by SDS gel electrophoresis. CQ inhibited hemoglobin digestion such that accumulation of hemoglobin in the parasites occurred. Quantitative analysis indicated that the content of hemoglobin in isolated CQ-treated parasites was increased from 2.39 +/- 0.47 micrograms hemoglobin (Hb)/mg protein to 8.34 +/- 0.77 micrograms Hb/mg protein (P less than 0.001) within 45 min and further to 18.7 +/- 1.23 micrograms Hb/mg protein for 2 hr compared with the untreated parasites. These results suggest that the inhibition of hemoglobin degradation in malarial parasites might be the primary target of CQ antimalarial action. The CQ-like effect on the hemoglobin digestion in P. falciparum was observed with another lysosomotropic weak base, NH4Cl, suggesting that the CQ effect on hemoglobin degradation and its antimalarial action, as well as the effect of NH4Cl, are related to their properties of lysosomotropic weak base.

Ammonium Chloride↗

Neonatal chorda tympani transection alters adult preference for ammonium chloride in the rat.

The immature gustatory system of the neonatal rat is characterized by sensitivity to disruption by early interventions such as receptor or nerve damage. The present studies examined the effect of chorda tympani transection (neoCTX) of neonates on adult preference for salt and nonsalt stimuli. NeoCTX at 10 days of age led to a striking change in adult rats' preference for NH4Cl solutions but little change in preference for other solutions, including NaCl and KCl. Permanent anatomical effects of neoCTX included failure of the nerve to regenerate and a loss of all fungiform taste buds. Preference for NH4Cl was not due to an inability to discriminate it from NaCl. Following taste aversion conditioning to NaCl, neoCTX rats clearly distinguished between NaCl and NH4Cl. The effects on NH4Cl preference reflect a sensitive period during development because adult rats receiving similar surgery did not show any change in NH4Cl preference.

Ammonium Chloride↗

Comparative effects of ouabain, natriuretic factor and ammonium chloride in the toad urinary bladder.

Electrical changes and direct effects on Na-K ATPase activity induced by an endogenous digitalis-like natriuretic factor (NF), NH4Cl and ouabain were studied in toad bladders. NF inhibited the SCC and the Na-K ATPase activity in a similar manner to ouabain, but induced a greater increase in calculated direct current resistance (R) (p less than 0.05). NH4Cl was a weak inhibitor of Na-K ATPase activity, although it produced steeper SCC inhibition slopes than those observed with ouabain or NF (p less than 0.01). The data suggested the same mechanism of action of NF and ouabain on the sodium pump, with an additional effect of the former on apical sodium permeability of the cells and/or closure of paracellular routes leading to an increased tissue resistance. In contrast, the effects of NH4Cl were mostly compatible with intracellular inhibition of apical sodium entry into the cell.

Ammonium Chloride↗

Biosynthesis and transport of lysosomal enzymes in human monocytes and macrophages. Effects of ammonium chloride, zymosan and tunicamycin.

Human monocytes and macrophages synthesize lysosomal enzymes as larger precursors. The polypeptide patterns of several lysosomal-enzyme precursors and their mature forms are similar to those observed in human fibroblasts. Like fibroblasts, the monocytes and macrophages release small amounts of lysosomal-enzyme precursors. The lysosomotropic NH4+ cation enhances this release. In contrast, zymosan, a degranulating agent, causes release of both the mature and the precursor forms of the lysosomal enzymes. Both NH4Cl and zymosan inhibit maturation of the precursors. The fractional amounts of mature cathepsin D and beta-hexosaminidase released in the presence of zymosan are strikingly different. Probably, in the macrophages several lysosomal organelles are packaged with different relative contents of lysosomal enzymes. The transport of the precursors of cathepsin D into lysosomes is inhibited by tunicamycin. Therefore oligosaccharide side chains are likely to function as signals in packaging of lysosomal enzymes in macrophages also.

Ammonium Chloride↗

Acute response of urine pH following ammonium chloride administration to dogs.

To test the acidifying ability of the distal portion of the nephrons in healthy dogs, 0.2 g of NH4Cl/kg of body weight was given PO. Samples for venous blood gas analysis and urine pH were taken hourly for 6 hours. Systemic acidemia developed, as evidenced by a statistically significant (P less than 0.05) decrease in blood pH 1 hour after NH4Cl administration. Four hours after administration, mean urine pH decreased to a low of 5.16 +/- 0.1 and was less than 5.5 3 hours after administration. Changes in urine pH 2 hours after administration were statistically significant (P less than 0.05). In human beings, NH4Cl loading is used to detect patients with distal renal tubular acidosis (defective hydrogen ion secretion by the distal nephrons) and normal acid/base values. Distal renal tubular acidosis is diagnosed if urine pH fails to decrease to less than 5.5 after NH4Cl administration. On the basis of the findings of this study, a similar value would be valid for dogs.

Ammonium Chloride↗

Spontaneous firing of NG108-15 cells induced by transient exposure to ammonium chloride.

1. We report that NG108-15 (neuroblastoma x glioma) cells differentiated in defined serum-free media are capable of exhibiting stable automaticity (the spontaneous occurrence of regenerative action potentials) following exposure to extracellular perfusates containing NH4Cl. 2. Membrane depolarization (4-5 mV) concomitant with an increased pHi during NH4Cl exposure are followed by hyperpolarization (5-7 mV), sub-threshold oscillations, and spontaneous firing after the removal of NH4Cl. 3. Cells cultured in 10% serum did not exhibit automaticity. Cells cultured in serum-free media are twice as likely to show automaticity as those cultured in reduced (1.5%) serum media. 4. We have examined factors that contribute to the events following NH4Cl exposure, namely, membrane depolarization and hyperpolarization, subthreshold oscillations, and automaticity. The inward currents activated at more negative potentials and the ionic currents associated with pronounced afterhyperpolarization in NG108-15 cells cultured in serum-free media provide a basis for the repetitive activity in general and automaticity in particular.

Amiloride↗

[Effect of the segregation of neutral red, acridine orange and ammonium chloride by L cells (subline LSM) on lysosomal hydrolase activity].

Kinetics of Neutral red (NR) and Acridine orange (AO) uptake by cultured L cells (subline LSM) has been studied. It was found that the uptake of both NR and AO, with their constant concentrations in the medium was characterized as a two-phase process. During 2 hours, these cells concentrated as much as 90% of the total amount of NR and AO taken up during the whole incubation period. The segregation and accumulation of NR, AO as well as NH4Cl took place in lysosomes. NR and AO concentrations within the cells exceed by 600 and 400 times, respectively, those in the medium. NR, AO and NH4+ accumulation in cells resulted in inhibition of the activity of the following lysosomal hydrolases: cathepsins B and D, acid lipase, N-acetyl-beta,D-glucosaminidase, beta-galactosidase, acid phosphatase and galactosyltransferase, the latter being a marker of Golgi apparatus. The effect of lysosomal enzyme activity inhibition on the cell economy, and a possible role of lysosomotropic agents as regulators of the lysosomal apparatus functional activity are discussed.

Acridine Orange↗

Processing and intracellular transport of cathepsin G and neutrophil elastase in the leukemic myeloid cell line U-937-modulation by brefeldin A, ammonium chloride, and monensin.

The effects of brefeldin A, monensin, and the weak base NH4Cl on the biosynthesis and processing of cathepsin G and neutrophil elastase of myeloid cells were investigated. Monoblast-like U-937 cells were biosynthetically labeled with [35S]methionine, followed by subcellular fractionation, immunoprecipitation, and analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography. Brefeldin A inhibited proteolytic processing, intracellular transport, and secretion. The effects were reversible inasmuch as removal of brefeldin A resulted in a normal pattern of processing and transfer to high-density organelles, corresponding to lysosomes, and restitution of constitutive secretion of precursor forms. Both cathepsin G and neutrophil elastase acquired resistance to endoglycosidase-H, suggesting that conversion to complex oligosaccharide side chains also occurs in the presence of brefeldin A. Monensin and NH4Cl inhibited final proteolytic processing, indicating either that acidification is necessary for directing cathepsin G and neutrophil elastase to lysosomal-like organelles or that the protease(s) responsible for processing requires an acid pH. We conclude that pH-dependent proteolytic processing of cathepsin G and neutrophil elastase occurs in post-Golgi structures and that transfer to lysosomes or an immediately prelysosomal compartment is mandatory for complete processing.

Ammonium Chloride↗

Calcium status of the growing rat as affected by diet acidity from ammonium chloride, phosphate and protein.

Two trials of 4 and 8 weeks' duration were conducted on 64 male Long-Evans growing rats to study effect(s) of diet protein (20 and 40%), phosphate source (less or more acidic) and NH4Cl (none or 1% in water) on Ca status and acid-base balance. Treatments were arranged in a 2 X 2 X 2 factorial plan in four randomized blocks. All diets were equal in Ca (0.52%) and P (0.41%). Criteria were blood pH and bicarbonate, plasma hydroxyproline and Ca, urinary cAMP and Ca, fecal Ca, and mineral analyses and gravimetric measurement of femurs. Blood pH was lowered due to NH4Cl ingestion at 4 weeks (7.11 and 7.22, P less than 0.05) and 8 weeks (7.27 and 7.36, P less than 0.02). Protein level and phosphate source influenced blood bicarbonate. With the ingestion of more acid there were increases in urinary cAMP and Ca. Fecal Ca increased twofold with more acid. Femur specific gravity was higher with the less acidic phosphate source. In conclusion, acid phosphate, NH4Cl and protein influenced blood pH and loss of Ca via urine and feces with a resultant lowered bone density. The less acidic phosphate source reduced or prevented a lowered bone density.

Acid-Base Equilibrium↗

Inhibition of nitrogenase activity by ammonium chloride in Azotobacter vinelandii.

In Azotobacter vinelandii cells, the short-term inhibition of nitrogenase activity by NH4Cl was found to depend on several factors. The first factor is the dissolved oxygen concentration during the assay of nitrogenase. When cells are incubated with low concentrations of oxygen, nitrogenase activity is low and ammonia inhibits strongly. With more oxygen, nitrogenase activity increases. Cells incubated with an optimum amount of oxygen have maximum nitrogenase activity, and the extent of inhibition by ammonia is small. With higher amounts of oxygen, the nitrogenase activity of the cells is decreased and strongly inhibited by ammonia. The second factor found to be important for the inhibition of nitrogenase activity by NH4Cl was the pH of the medium. At a low pH, NH4+ inhibits more strongly than at a higher pH. The third factor that influenced the extent of ammonia inhibition was the respiration rate of the cells. When cells are grown with excess oxygen, the respiration rate of the cells is high and inhibition of nitrogenase activity by ammonia is small. Cells grown under oxygen-limited conditions have a low respiration rate and NH4Cl inhibition of nitrogenase activity is strong. Our results explain the contradictory reports described in the literature for the NH4Cl inhibition of nitrogenase in A. vinelandii.

Ammonium Chloride↗

The effect of ammonium chloride on metabolism of primary neurons and neuroblastoma cells in vitro.

Hyperammonemia is a consistent finding in many metabolic disorders. The excess ammonia (NH4Cl) interferes with brain energy metabolism possibly in part by inhibiting the tricarboxylic acid (TCA) cycle. Inhibition of the TCA cycle may result in depletion of ATP in the brain cells. In this study, the acute and chronic effect of NH4Cl (7.5 mM and 15 mM) on the metabolism of isolated neurons and neuroblastoma cells was examined. These cells were treated with NH4Cl for 15 minutes and 24 hours. Morphologic and metabolic toxicity were greater in neuroblastoma cells than in primary neurons. Following 15 minutes treatment, concentration of lactate increased significantly in neuroblastoma cells but, the concentration of other metabolites did not change significantly in neuroblastoma cells and in primary neurons. Following 24 hours treatment, the glucose utilization increased in both cell types. This high utilization of glucose in neuroblastoma cells was in concert with an increase in lactate and decrease in glutamate and ATP. In primary neurons, following 24 hours treatment, the glucose utilization significantly increased, but the concentration of the other metabolites did not change significantly. Neuroblastoma cells consumed more glucose than primary neurons in absence of NH4Cl, but generated the same amount of lactate as neurons.

Adenosine Triphosphate↗