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Demineralization of bone by ammonium chloride for the pyroantimonate method.

Rat bones were fixed by the pyroantimonate method and demineralized by treatment with ammonium chloride (NH4Cl). Bones from very young rats (neonatal and 2-week-old rats) that had been treated with NH4Cl were easy to cut into ultrathin sections. However, bones of 8-week-old rats needed to be demineralized for a longer time and were not very easy to cut. The extent of demineralization was measured by scintillation counting. After demineralization, precipitates of calcium pyroantimonate (Ca-pyroantimonate) were well retained and the fine structure of the tissue was well preserved. In addition, the distribution of Ca-pyroantimonate in the demineralized bone was similar to that in undecalcified specimens. This demineralization method is used in combination with the pyroantimonate method.

Ammonium Chloride↗

Effect of ammonium chloride on the multiplication of infectious pancreatic necrosis virus.

The multiplication of IPNV in CHSE cells is inhibited by ammonium chloride. This inhibition is complete if NH4Cl is added early in the infective cycle. Immunofluorescence analysis shows that NH4Cl reduces the number of virus producing cells and thus suggests it may act by inhibiting virus internalization. In addition, NH4Cl can produce partial inhibition of IPNV multiplication if added later in the replicative cycle. Analysis of 3H-uridine incorporation into viral RNA shows that this later effect could be an inhibition of viral RNA synthesis.

Ammonium Chloride↗

Inhibition of multiplication of herpes simplex virus type 1 by ammonium chloride and chloroquine.

Multiplication of herpes simplex virus type 1 (HSV-1) strain HF in Vero cells was inhibited by ammonium chloride and chloroquine. A series of one-step growth experiments showed that the maturation of intracellular virus was prevented immediately after the addition of weak bases in the late stage of infection, indicating that the site of inhibition by weak bases is some step in the maturation process of HSV-1.

Ammonium Chloride↗

Characterization of ammonium chloride derivatives by salt clustering in electrospray mass spectrometry.

Clustering of ammonium chloride salts was studied using an electrospray source to characterize the form, mono- vs. dihydrochloride, of organic compounds by mass spectrometry. This new way of taking advantage of cluster formation is applied to aminomethylacridines as their storage requires the synthesis of such derivatives. Both positive and negative cluster mass spectra were obtained and allowed the determination of the nature of the hydrochloride forms as the salt mass is straightforwardly derived from the mass of the cluster monomer, calculated from singly and multiply charged ion distributions. The identity of the counterion is confirmed from the mass of the ionic moieties in the clusters.

Journal Article↗

Short- and long-term effects of ammonium chloride on RNA, DNA and protein concentrations in muscle and serum in the rainbow trout, Oncorhynchus mykiss.

1. Effects of ammonium-induced acidosis on white muscle RNA, DNA and protein concentrations were examined in the rainbow trout, Oncorhynchus mykiss. Serum protein concentrations were also determined. 2. A single administration of ammonium chloride at a dose of sublethal level caused no changes in these parameters after 3 and 6 hr. 3. Multiple administrations at a lesser dose caused significant decreases in RNA and serum protein concentrations 48 and 72 hr after the first administration. 4. DNA and muscle protein concentrations remained unchanged throughout the experimental period. 5. Significant decreases in RNA-DNA ratios and RNA-muscle protein ratios were primarily attributed to decreases in RNA concentrations.

Ammonium Chloride↗

Difference in the effects of acetazolamide and ammonium chloride acidosis on ventilatory responses to CO2 and hypoxia in humans.

The effects of acetazolamide, a potent carbonic anhydrase inhibitor, and ammonium chloride (NH4Cl) on arterial blood gas tension, resting ventilation, and ventilatory responses to CO2 (HCVR) and hypoxia (HVR) were studied in healthy male subjects. Both drugs induced chronic metabolic acidosis with the reduction in plasma bicarbonate by a mean of 7.0 +/- 2.0 (S.D.) mM after acetazolamide and by 5.6 +/- 1.8 mM after NH4Cl. The ratio in the decrement of PaCO2 to that of plasma bicarbonate (delta PaCO2/delta [HCO3-]) was 1.51 in the former and 0.98 in the latter. Both drugs increased inspiratory minute ventilation (VI) predominantly due to increased tidal volume (VT) with acetazolamide and to increased respiratory frequency (f) with NH4Cl. In HCVR, the increments in CO2- ventilation slope and in ventilation at PETCO2 60 mmHg after drug administration were 0.77 +/- 0.51 l X min-1 X mmHg-1 and 20.0 +/- 11.2 l/min with acetazolamide and 0.59 +/- 0.40 l X min-1 X mmHg-1 and 8.0 +/- 2.8 l/min with NH4Cl, respectively. On the other hand, HVR both in terms of delta VI/delta SaO2 slope and of ventilation at SaO2 75% significantly increased after NH4Cl but not after acetazolamide administration. Thus, augmented VT and HCVR in the acetazolamide group and increased f and HVR in the NH4Cl group suggested that the central chemosensitive mechanism in the former and the peripheral chemosensitive mechanism in the latter may predominantly be responsible for the elevated ventilatory activities.

Acetazolamide↗

Evaluation of the effect of ammonium chloride treatment on the activity of human effector cells in antibody-dependent and spontaneous cell-mediated cytotoxicity.

The effect of ammonium chloride treatment, used to eliminate contaminating red blood cells from lymphoid cell preparations, on the cytotoxic activity of human effector cells in antibody-dependent and spontaneous cell-mediated cytotoxicity was quantitatively determined. No significant difference between the cytotoxic efficiency of treated and untreated cell preparations was observed.

Ammonium Chloride↗

Effect of supplementation of dry cat food with D,L-methionine and ammonium chloride on struvite activity product and sediment in urine.

Feeding dry foods supplemented with urine acidifier (D,L-methionine (Met) or ammonium chloride) decreased urinary pH and struvite activity product in clinically normal cats. As a result, the number of struvite crystals in urine was greatly reduced. Supplementation with 3% Met but not 1% Met caused decrease in the urinary concentration of sediment, which resulted from a reduction in the HCl-soluble fraction. The concentration of HCl-insoluble sediment was not affected by supplementation with the urine acidifier.

Ammonium Chloride↗

Aggregation and chimney formation during the solidification of ammonium chloride.

Experiments study large-scale pattern formation during the growth of ammonium chloride (NH4Cl) from solution in a thin (Hele-Shaw) geometry. In particular a solid-liquid mixture ("mushy layer") forms in which growing solid NH4Cl crystals form a solid network interspersed with liquid. There are different ways that the mushy layer can be formed, however. If the cell is heated from below and cooled from above, thermal convection generates large-scale recirculating flows that carry seed crystals from the upper (cold) boundary to the (warmer) side and bottom boundaries. Ballistic deposition of these seed crystals leads to aggregation patterns with significant voids (filled with liquid) with a wide range of length scales. If the cell is cooled from below with a warm environment, the solid NH4Cl grows dendritically without deposition, resulting in a compact mushy layer. Plume convection within this mushy layer produces one or two well-defined "chimneys." If the environment is cool (comparable to the liquidus temperature of the solution), the mushy layer forms by a combination of dendritic growth and ballistic deposition, resulting in a more permeable mushy layer and enhanced chimney formation. The effects of ballistic deposition are enhanced if the cell is tipped, in which case the voids reappear. Plume convection and chimney formation are dramatically enhanced in this case. Additional experiments are done in which fluid flows in the system are enhanced artificially to verify that enhancements in chimney formation are due primarily to the aggregation process, and not to the increases in fluid flows due to thermal and compositional convection.

Journal Article↗

Isolation of Babesia bigemina and babesia bovis merozoites by ammonium chloride lysis of infected erythrocytes.

1. We describe the isolation of viable merozoites from erythrocytes infected with Babesia bovis or Babesia bigemina organisms by ammonium chloride lysis. 2. Parasite morphology was examined by both light and transmission electron microscopy. Erythrocyte-free parasites maintain their viability and infectivity, retain their antigenicity and are suitable for use in the indirect fluorescent antibody assay.

Ammonium Chloride↗

Effect of ammonium chloride and dietary phosphorus in the azotaemic rat. I. Renal function and biochemical changes.

BACKGROUND: Both dietary phosphorus restriction and the ingestion of ammonium chloride (NH(4)Cl) given to rats on a high-phosphorus diet have been shown to preserve renal function in the azotaemic rat. Parathyroidectomy also has been reported to preserve renal function and, in addition, to prevent kidney hypertrophy in the remnant kidney model. Our goals were (i) to evaluate in azotaemic rats the effect of dietary phosphorus on renal function in a shorter time frame than previously studied and (ii) to determine whether NH(4)Cl administration (a) enhances the renoprotective effect of dietary phosphorus restriction and (b) improves renal function in the absence of parathyroid hormone (PTH). METHODS: High (H; 1.2%), normal (N; 0.6%) and low (L; <0.05%) phosphorus diets (PD) were given for 30 days to 5/6 nephrectomized rats. In each dietary group, one-half of the rats were given NH(4)Cl in the drinking water. The six groups were HPD + NH(4)Cl, HPD, NPD + NH(4)Cl, NPD, LPD + NH(4)Cl and LPD. The effect of NH(4)Cl administration was also evaluated in 5/6 nephrectomized, parathyroidectomized (PTX) rats on NPD. RESULTS: In each of the three dietary phosphorus groups, creatinine and urea clearances were greater (P<0.01) in rats receiving NH(4)Cl. Neither creatinine nor urea clearance was reduced by high dietary phosphorus. Urine calcium excretion was greatest in the LPD group and was increased (P < or = 0.001) in all three groups by NH(4)Cl ingestion. An inverse correlation was present between plasma calcium and phosphorus in the parathyroid intact (r = -0.79, P<0.001) and PTX groups (r = -0.46, P = 0.02). In PTX rats, NH(4)Cl ingestion increased (P < or = 0.01) creatinine and urea clearances and both an increasing plasma calcium concentration (r = 0.67, P<0.001) and urine calcium excretion (r = 0.73, P<0.001) increased urine phosphorus excretion. CONCLUSIONS: At 30 days of renal failure (i) NH(4)Cl ingestion increased creatinine and urea clearances, irrespective of dietary phosphorus; (ii) high urine calcium excretion, induced by dietary phosphorus restriction and NH(4)Cl ingestion, did not adversely affect renal function; (iii) high dietary phosphorus did not decrease renal function; (iv) the absence of PTH did not preserve renal function or prevent NH(4)Cl from improving renal function; and (v) both an increasing plasma calcium concentration and urine calcium excretion resulted in an increase in urine phosphorus excretion in PTX rats.

Ammonium Chloride↗

Responses in primary astrocytes and C6-glioma cells to ammonium chloride and dibutyryl cyclic-AMP.

Elevated brain ammonia levels are a major factor in the genesis of hepatic encephalopathy (HE). The mechanism of ammonium chloride (NH4Cl) neurotoxicity involves interruption of oxidative metabolism. This leads to decreased levels of ATP concentration and subsequent glial fibrillary acidic protein (GFAP) degradation of astrocytes and fibrous C6-glioma cells. Our study investigates NH4Cl toxicity by evaluating changes in ATP concentration and mitochondrial function as well as by evaluating alterations in GFAP expression. NH4Cl induced decreases in ATP were detected after 15 minutes in C6-glioma cells and 24 hours in both cell types. Mitochondrial function, assessed by MTT (2-4,5-dimethylthiazol A-yl)-2, 5-diphenyltetrazolium bromide) assay, was impaired in both cell types at 24 hours following NH4Cl treatment. GFAP was also significantly decreased in both cell types. Morphologic and metabolic toxicities were greater in C6-glioma cells. The data clearly indicate that NH4Cl interrupts oxidative metabolism. The greater toxicity seen in C6-glioma cells may be due to their greater dependence on oxidative metabolism. Lastly, the decrease in GFAP is probably a consequence of diminished ATP.

Ammonium Chloride↗

The effect of ammonium chloride and sodium bicarbonate ingestion on the physical working capacity at the fatigue threshold.

The purpose of this investigation was to examine the effect of ammonium chloride (NH4Cl) and sodium bicarbonate (NaHCO3) ingestion on the physical working capacity at the fatigue threshold (PWCFT). Eighteen adult males (mean age, SD = 23, 2 years) volunteered for two experiments (experiment 1, n = 9; experiment 2, n = 9). In both experiments, the subjects orally ingested 0.3 g.kg-1 body weight of NH4Cl and NaHCO3 over a 3-h period in random order on days separated by 72 h or more. In experiment 1, following ingestion of the substance, the subjects performed a discontinuous incremental cycle ergometer test to the onset of PWCFT which was estimated from integrated electromyography voltages at the vastus lateralis muscle. In experiment 2, the subjects performed a continuous PWCFT test. The results of these experiments indicated that NH4Cl and NaHCO3 ingestion had no significant (P greater than 0.05) effect on PWCFT (experiment 1: NH4Cl = 257, SD 26 W; NaHCO3 = 256, SD 22 W; t = 0.06; r = 0.866; experiment 2: NH4Cl = 231, 14 W; NaHCO3 = 216, 16 W; t = 1.78; r = 0.857).

Administration, Oral↗

Effects of ammonium chloride on membrane currents of acinar cells dispersed from the rat parotid gland.

In acinar cells freshly dispersed from rat parotid glands, the effects of ammonium chloride (NH4Cl) on membrane currents were studied using the whole-cell clamp method. When membrane currents were recorded with command pulses to 0 mV, applied at 2-s intervals from a holding potential of -70 mV, NH4Cl (5-20 mM) transiently decreased outward currents and then slowly increased both outward and inward currents. After reaching a peak in about 40-50 s, both outward and inward currents gradually decreased in the presence of NH4Cl and, on its wash-out, the currents returned to the control level. Butyrate (5-20 mM) had little effect on the resting membrane currents, but markedly inhibited the response to NH4Cl. Tetraethylammonium (5 mM) strongly reduced both the resting and NH4Cl-induced outward currents, whereas it slightly potentiated the NH4Cl-induced inward current without affecting the membrane current at the holding potential. Without ATP in the patch pipettes, carbachol-induced membrane currents were relatively resistant to Ca2+ removal from the external medium, but NH4Cl-induced currents were quickly abolished in the absence of Ca2+. We conclude that intracellular alkalinization with NH4Cl increases Ca2+ influx and activates Ca(2+)-dependent outward K+ and inward Cl- currents.

Ammonium Chloride↗

Antibody-dependent cellular cytotoxicity and natural cytotoxicity: effect of pre-treatment of human lymphocytes with deionised water and ammonium chloride.

Deionised water is an efficient red blood cell lytic agent with no demonstrable deleterious effects on ADCC and natural cytotoxicity (blood and tonsil). Treatment with isotonic ammonium chloride, however, produces a significant reduction of both ADCC and natural cytotoxicity. This reduction is less pronounced if the treatment is carried out at 4 degrees C and is temporary, recovery occurring over a 24 h period of incubation at 37 degrees C.

Ammonium Chloride↗

Effects of a high-protein diet versus dietary supplementation with ammonium chloride on struvite crystal formation in urine of clinically normal cats.

OBJECTIVE: To evaluate the effects of a high-protein diet versus dietary supplementation with ammonium chloride (NH4Cl) on struvite crystal formation in the urine of clinically normal cats by measuring the urine concentration of hydrochloric acid (HCl)-insoluble sediment, urine pH, struvite activity product (SAP), number of struvite crystals in urine, and urine volume. ANIMALS: 23 healthy adult cats. PROCEDURE: Urine was fractionated by centrifugation with subsequent extraction of the sediment with 1 N HCl (study 1). Diets containing either 29% crude protein or 55% crude protein were fed to cats in a crossover trial of 3 weeks/period (study 2). Diets supplemented with either sodium chloride (NaCl) or NH4Cl were fed, by use of a 3 x 3 Latin-square design with 3 wk/period (study 3). In studies 2 and 3, urine samples were collected for the last 7 days of each period. RESULTS: The HCl-insoluble sediment contained Tamm-Horsfall glycoprotein (THP; study 1). The high-protein diet (study 2) and dietary supplementation with NH4Cl (study 3) resulted in a decrease in urine pH, SAP, and the number of struvite crystals in urine. However, the high-protein diet decreased urine concentrations of HCl-insoluble sediment containing THP (study 2), in contrast to the NH4Cl supplementation that increased urine volume without a significant effect on the urine concentration of the HCl-insoluble sediment (study 3). CONCLUSIONS AND CLINICAL RELEVANCE: Our results indicate that compared with dietary supplementation with NH4Cl, the high-protein diet is preferable as a urine acidifier for the prevention of struvite crystal formation in clinically normal cats.

Ammonium Chloride↗