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PubMed1988

Two-dimensional flow cytometric analysis of T cell subsets maintained in IL-2 medium after autologous mixed leukocyte reaction activation.

Human T cells are stimulated with an autologous mixed leukocyte reaction (AMLR) and can be propagated in interleukin-2. Staining of the cultured cells with the combination of two monoclonal antibodies was evaluated by two-dimensional flow cytometry at weekly intervals. AMLR activation resulted in an initial preservation of the CD4+ (helper/inducer T) subset predominance over the CD8+ (suppressor/cytotoxic T) cells, noted on normal circulating blood lymphocytes. However, during culture in interleukin-2, there was a progressive increase in the percentages of CD8+ Leu 15- cytotoxic T, CD4+ Leu 8- helper T, and CD3+ HLA-DR+ activated T cells, and a concomitant decrease in those of CD4+ Leu 8+ suppressor inducer T and CD8+ Leu 15+ suppressor T cells if the responder sheep red blood cell (SRBC)-rosetting T cells were made up by tris ammonium chloride, but not by hypotonic shock treatment to lyse SRBC. The significant difference between hypotonic shock-T cells and ammonium chloride-T cells in the phenotypic changes of T cell subsets after long-term culture in an interleukin-2 medium may suggest a regulatory role of the ammonium chloride-sensitive T cells in the AMLR.

Adult↗

PubMed1982

[Results of treatment by rotation in left-sided abomasal displacement (author's transl)].

The results are given of the treatment of thirty cows with left-sided abomasal displacement by rotation, followed by intravenous injection of hypertonic sodium chloride solution and a calcium borogluconate solution. In addition, ammonium chloride was administered orally for a few days. The rations of the animals were gradually restored to normal levels. The purpose was to examine whether recurrences following rotation could be prevented by supplementary measures. Of these thirty animals, five did not respond to treatment because of complications that would otherwise not have reacted favourably to any form of treatment either. Correction of the abomasal displacement was successful in more than 90 per cent (23 animals) of the remaining five cows. However, abomasal displacement recurred in sixteen of these twenty-three animals (70%). Of these fourteen cows, two animals developed an abomasal torsion. It is concluded that the supplementary measures adopted will not prevent recurrences.

Abomasum↗

PubMed1995

Granule swelling in stimulated bovine adrenal chromaffin cells: regulation by internal granule pH.

Adrenal medullary chromaffin cells secrete catecholamines through exocytosis of their intracellular chromaffin granules. Osmotic granule swelling has been implicated to play a role in the generation of membrane stress associated with the fusion of the granule membrane. However, controversy exists as to whether swelling occurs before or after the actual fusion event. Using morphometric methods we have determined the granule diameter distributions in rapidly frozen, freeze-substituted chromaffin cells. Our measurements show that intracellular chromaffin granules increase in size from an average of 234 nm to 274 nm or 277 nm in cells stimulated to secrete with nicotine or high external K+, respectively. Granule swelling occurs before the formation of membrane contact. Ammonium chloride, an agent which inhibits stimulated catecholamine secretion by approximately 50% by altering the intragranular pH, also inhibits granule swelling. In addition, ammonium chloride-treated secreting cells show more granule-plasma membrane contacts than untreated secreting cells. Sodium propionate induces granule swelling in the absence of secretagogue and has been shown to enhance nicotine- and high K(+)- induced catecholamine release. These results indicate that in adrenal chromaffin cells granule swelling is an essential step in exocytosis before fusion pore formation, and is related to the pH of the granule environment.

Adrenal Medulla↗

PubMed1988

Ion fluxes and abscisic Acid-induced proline accumulation in barley leaf segments.

The increase in proline induced by ABA, a process stimulated by NaCl or KCl in barley leaves, did not occur when Na(+) (or K(+)) was present in the external medium as the gluconate salt, namely with an anion unable to permeate the plasma membrane. However, proline increase was restored, to different extents, by the addition of various chloride salts but not by ammonium chloride. Moreover, it was shown that the stimulation of the process by NaCl (or KCl) was variously affected by the presence of different salts; all the ammonium salts (10 millimolar NH(4) (+) concentration) inhibited this stimulation almost completely. Inhibition by NH(4) (+) was accompanied by a decreased Na(+) influx (-40%). Also, in the case of Na-gluconate, Na(+) uptake was reduced and the addition of Cl(-) as the calcium or magnesium salt (but not as ammonium salt) restored both the ion influxes and the increase in proline typical of NaCl treatments. Both 4,4'-diisothiocyano-2,2'-disulfonic acid stilbene (DIDS), an anion transport inhibitor, and tetraethylammonium chloride (TEA), a K(+) channels-blocking agent, caused, as well as with a reduction of ion influxes, an inhibition of the proline accumulation. The inhibition was practically total with 1 millimolar DIDS and about 80% with 20 millimolar TEA. A possible role of ion influxes in the process leading to the increase in proline induced by ABA is proposed.

Journal Article↗

PubMed1999

[Ionophore characteristics of heliomycin].

A highly purified product, identified as a well-known antibiotic geliomycin, has been derived by gradient extraction of the mycelium from Streptomyces robefuscus isolated from soil specimens as a result of purposeful search for a producer of hydrophobic ionophore antibiotics. The ionophore properties of geliomycin are highly labile and inducible. Cationic or anionic nature and the angle of the function performed depend on the concentration, type of electrolyte, protocol of measurements, and some additives used for the preparation of geliomycin electrodes to measurements. Stable development of the cationic function was detected for potassium chloride. The lability of ionophore properties was the highest with ammonium chloride as the electrolyte. Ammonium acetate and potassium and sodium chlorides notably increased the stability of geliomycin electrodes. Selective activity of geliomycin towards potassium ion is a more permanent characteristic. The results permit a conclusion about the inducibility of ionophore properties of geliomycin, which may play an important role in the vital activity of the producer and manifest as a regulatory factor in maintenance of water and acid base balance.

Anti-Bacterial Agents↗

PubMed1989

Effect of sulfate on calcium and magnesium homeostasis following urinary diversion.

The present investigation was undertaken to elucidate the mechanisms of renal calcium and magnesium wastage in patients with urinary diversion through intestinal segments. Patients with urinary intestinal diversions demonstrated a hyperchloremic metabolic acidosis, increased serum sulfate and phosphate, and increased urinary excretion of calcium, phosphate, sulfate, and magnesium. A study was conducted in rats utilizing oral or intravenous loads of various solutes to investigate the possible mechanisms of these findings in patients. Ammonium enhances renal sulfate excretion and intestinal sulfate absorption. Increased filtered loads of sulfate inhibit renal tubule sodium and calcium reabsorption. Dietary ammonium and sulfate result in increased serum phosphorous concentrations. Ammonium appears to directly inhibit renal magnesium reabsorption. Renal tubule dysfunction resulting from relieved obstruction appears to enhance the inhibitory effect of sulfate on renal sodium and calcium reabsorption but does not alter the effect of ammonium on renal magnesium excretion. Taken together, these findings suggest that patients with hyperchloremic metabolic acidosis from the chronic reabsorption of ammonium chloride by intestinal urinary diversions may also reabsorb urinary sulfate. This load of ammonium and sulfate inhibits renal calcium and magnesium reabsorption.

Acidosis, Renal Tubular↗

PubMed2006

Confounding effects of platelets on flow cytometric analysis and cell-sorting experiments using blood-derived cells.

BACKGROUND: Flow cytometric analysis and cell-sorting of peripheral blood leukocytes is commonplace; however, platelet contamination is typically ignored during immunophenotypic analysis and sorting of blood-derived cells. METHODS: Red blood cells, platelets, T & B lymphocytes, monocytes, and granulocytes were sorted from rat blood preparations. Presort enrichment was performed by differential centrifugation for all cell types. Additionally, leukocyte samples were prepared by ammonium chloride lysis of red blood cells. RESULTS: Unless proper precautions were taken, significant numbers of platelets were sorted along with (nonplatelet) cells of interest. The amount of platelet contamination varied greatly from experiment to experiment with the highest level of leukocyte-platelet association observed in the neutrophil/granulocyte population in samples prepared using ammonium chloride-based red blood cell-lysing solution. CONCLUSIONS: Addition of an immunophenotypic marker for platelet identification is a simple, yet prudent, measure to help evaluate the impact of platelets on immunophenotypic staining when performing flow cytometric analysis or sorting of blood-derived cells and should become a routine practice. Platelet presence in postsort fractions can be due to free platelets as well as target cell-associated platelets and both sources of contamination must be addressed.

Ammonium Chloride↗

PubMed2005

Rapid fixation of methylene chloride by a macrocyclic amine.

A simple macrocyclic amine is alkylated by methylene chloride to give a quaternary ammonium chloride salt. When methylene chloride is the solvent, the reaction exhibits pseudo-first-order kinetics, and the reaction half-life at 25.0 degrees C is 2.0 min. The reaction half-life for a structurally related, acyclic amine is approximately 50 000 times longer. Detailed calculations favor a mechanism where the methylene chloride associates with the macrocycle to form an activated prereaction complex. The macrocyclic nitrogen subsequently attacks the methylene chloride with a classic SN2 trajectory, and although the carbon-chlorine bond breaks, the chloride leaving group does not separate from the newly formed cationic macrocycle, such that the product is a tightly associated ion-pair. X-ray crystal structures of the starting amine and the product salt, as well as kinetic data, support this mechanism.

Journal Article↗

PubMed1961

Algicidal and sanitizing properties of Armazide.

Algicidal and sanitizing properties of Armazide, a new swimming pool additive, consisting of 12% w/v each of dodecylamine hydrochloride, trimethyl alkyl ammonium chloride, and methyl alkyl dipolyoxypropylene ammonium methyl sulfate in solution, were evaluated by laboratory techniques against algae and sewage. Results indicated it to be highly effective in low concentrations especially in conjunction with low concentrations of chlorine. Swimming pool field tests were found to confirm the laboratory findings. Various treatment levels and methods are described for swimming pools using treatments based upon the actual condition of the pool and water. The use of the product permitted a reduction in chlorine residual in pools resulting in greatly reduced requirements for chlorinating chemicals along with absence of irritation, odor, and other undesirable results usually associated with standard pool chlorination methods.

Chlorine↗

PubMed1988

Microbial inhibition on hospital garments treated with Dow Corning 5700 antimicrobial agent.

We evaluated the efficacy of the antimicrobial activity of cotton-polyester fabric treated with 3-(trimethoxysilyl)propyldimethyloctadecyl ammonium chloride (DC 5700), a quaternary ammonium compound bound irreversibly to the material. Significant antimicrobial activity was observed with 58 of 61 gram-positive cocci but with only 1 of 35 gram-negative bacilli and 0 of 5 yeasts. No inhibition of bacilli or yeasts was observed when the DC 5700 concentration ranged from 0.05 to 1.0% or when antimicrobial activity was assayed by the agar overlay bioassay or broth agitation methods.

Anti-Bacterial Agents↗

PubMed1990

[Microbiological evaluation of chemical disinfectants for household use].

Five disinfectants for household use were advertised on television during 1988 and the first half of 1989. The products were tested by a qualitative (Use-Dilution with 10 carriers, a conventional and simplified, adapted method) and a qualitative, adapted method with a view to evaluating their antimicrobial activity. The active compounds of the products, according to their respective labels were: 1--Parachlorophenol (0-Benzil) 0.1%; 2--Eter 2.4.4' Chloro (III) 2' hydroxiphenylic 0.1%; 3--N-alkyl dimethylbenzyl ammonium chlorides, N-alkyl dimethylethybenzyl ammonium 50%-1.6%; 4--Formaldehyde 37% (0.3% solution); 5--No information. The microorganisms used were: Staphylococcus aureus ATCC 6538, Pseudomonas aeruginosa ATCC 15442 and Salmonella choleraesuis ATCC 10708. In the qualitative method the pseudomonas strain was recovered from disinfectants 1, 2, and 3 and the salmonella strain from disinfectants 2 and 3. All disinfectants showed germicidal effect 5.0 (99.999%) of reduction) in 15 seconds against all strains. Disinfectant 3 was contaminated with Enterobacter sp to the order of 10(4) cells/ml. This contaminant was sensitive against disinfectants 1, 4 and 5, in the qualitative method and had a relative resistance to disinfectant 2 in the quantitative method.

Disinfectants↗

PubMed1975

Duodenal control of gastric emptying in the milk-fed calf.

1. It has been ascertained experimentally that an open duodenal cannula does not cause any marked variation in emptying of the abomasum when compared to the rate of emptying of the normal viscus or when the cannulae are closed. 2. Duodenal re-entrant cannulae provide a preparation whereby it is possible to infuse the duodenum via the distal cannula and to collect simultaneously the effluent fluid of the abomasum from the proximal arm of the cannula (Fig. 1). 3. With this preparation the duodenal infusate is not contaminated by the gastric effluent and thus the duodenal stimulus can be restricted to the single non-varying effect of the infusate. 4. In the milk-fed calf using this preparation the following facts have been established. (a) On infusion into the duodenum, hypotonic and isotonic solutions of sodium chloride and sodium bicarbonate increase abomasal emptying; bicarbonate being the more effective stimulus. (b) Potassium chloride, calcium chloride, glucose and hydrochloric acid inhibit gastric emptying when infused into the duodenum. (c) Ammonium chloride, urea, lactose and acetic acid have little effect. (d) Hypertonic solutions of all substances tested were inhibitory to gastric emptying when infused into the duodenum. 5. It has been demonstrated unequivocally that it is the effect of the infusate in the duodenum which controls gastric emptying for when the stomach is isolated from the duodenum, gastric emptying can be manipulated from the duodenum whatever the stomach contents may be. 6. Our results bring firm experimental confirmation of the views discussed by Hunt & Knox (1968) that gastric outflow is regulated by mechanisms initiated from receptors situated in the duodenum.

Abomasum↗

PubMed1964

BIOSYNTHESIS OF NITRO COMPOUNDS. I. NITROGEN AND CARBON REQUIREMENTS FOR THE BIOSYNTHESIS OF BETA-NITROPROPIONIC ACID BY PENICILLIUM ATROVENETUM.

Shaw, Paul D. (University of Illinois, Urbana), and Nancy Wang. Biosynthesis of nitro compounds. I. Nitrogen and carbon requirements for the biosynthesis of beta-nitropropionic acid by Penicillium atrovenetum. J. Bacteriol. 88:1629-1635. 1964.-beta-Nitropropionic acid was produced by Penicillium atrovenetum when this fungus was grown on a Raulin-Thom medium in shake flasks. The nitro compound was formed in the early stages of growth, and the total amount in the medium decreased when the fungus reached the end of the log phase. When increasing amounts of nitrate were substituted for the ammonia in the growth medium, production of beta-nitropropionic acid decreased. Aspartic acid did not promote the synthesis of the nitro compound unless either ammonium chloride or sodium tartrate was also added to the medium. The addition of small amounts of hydroxylamine or sodium nitrite to the Raulin-Thom medium stimulated beta-nitropropionic acid production to a greater degree on a molar basis than the amount of hydroxylamine or nitrite added. The nature of possible precursors to the nitro group of beta-nitropropionic acid is discussed.

Ammonia↗

PubMed1965

INFLUENCE OF AERATION AND OF PANTOTHENATE ON GROWTH YIELDS OF ZYMOMONAS MOBILIS.

Belaïch, Jean-Pierre (Centre National de la Recherche Scientifique, Marseille, France), and Jacques C. Senez. Influence of aeration and of pantothenate on growth yields of Zymomonas mobilis. J. Bacteriol. 89:1195-1200. 1965.-The growth yields and rates of Zymomonas mobilis were measured in aerobic and anaerobic cultures on glucose medium containing yeast extract, amino acids, or ammonium chloride as the nitrogen source. In the absence of yeast extract, pantothenate was required. The growth yield and rate of the cultures in synthetic (amino acids) or minimal (NH(4)Cl) medium supplemented with pantothenate corresponded only to about one-half the "normal" values obtained in the presence of yeast extract, suggesting a situation of energetically uncoupled growth. Attempts to restore normal growth by the addition of various compounds were unsuccessful. Aeration of the cultures resulted in a partial oxidation of ethyl alcohol to acetate, but did not modify the growth yield nor the division time. Both aerobic and anaerobic cells, however, contained cytochrome c and a cytochrome oxidase of the a(2) type, which was completely inhibited by 10(-4)m cyanide. In anaerobically grown cells, an additional cytochrome of the b type was present. The absence of a Pasteur effect suggests that the transfer of electrons by the respiratory chain of Z. mobilis may not be coupled with oxidative phosphorylation. Aeration had no effect on the catalase content of the cells. As shown by C(14)-glucose incorporation, 2 to 3% of the glucose metabolized was assimilated by the cells in both synthetic and rich complex medium. No intracellular glycogen nor poly-beta-hydroxybutyrate was accumulated when growth was limited by nitrogen or by phosphate in the presence of excess glucose.

Amino Acids↗

PubMed1982

Ammonium handling by superficial and juxtamedullary nephrons in the rat. Evidence for an ammonia shunt between the loop of Henle and the collecting duct.

Papillary and surface micropuncture was used to assess the effects of a chronic metabolic acidosis on the renal tubular handling of ammonium by surface nephrons, juxtamedullary nephrons, and the terminal segment of collecting duct. Rats chronically fed ammonium chloride had an expected decline in arterial pH and bicarbonate concentration associated with a doubling in the amount of ammonium excreted and a decline in urine pH. The glomerular filtration rate and absolute delivery of water and sodium to micropuncture sites of surface and deep nephrons was not measurably altered. Ammonium delivery to the end of the proximal tubule increased from 853+/-102% to 1,197+/-142% (SE) of the filtered load of ammonium after the induction of metabolic acidosis. This increase was due to a rise in tubular fluid ammonium content from 2.31+/-0.23 to 4.06+/-0.28 mM/liter. After the induction of acidosis, absolute and fractional delivery of ammonium ion to the end of the distal tubule was less than to the end of the accessible portion of the proximal tubule. These findings indicate that ammonium is lost in the intervening segment.Ammonium handling by deep nephrons was profoundly affected by acid loading. Absolute delivery to the bend of the loop of Henle increased twofold while fractional delivery rose from 1,222+/-108% to 1,780+/-132% of the filtered ammonium. This was due to a marked increase in ammonia entry. During acidosis, ammonium delivery to the terminal segment of the collecting duct was doubled (709+/-137% in controls vs. 1,415+/-150% in acidosis, P < 0.005) but did not change between proximal and tip collecting duct sites. In both groups of animals delivery of ammonium to the terminal segment of the collecting duct was greater than to end distal tubular micropuncture sites suggesting that ammonia entry occurred between these two sites. The differences in delivery was greater after the induction of a metabolic acidosis (887+/-140% vs. 384+/-144%, P < 0.05). Thus, the present study indicates that deep nephrons contribute to the adaptive increase in ammonium excretion seen during the induction of metabolic acidosis. The data also suggest that ammonia leaves the nephrons at a site(s) along the loop of Henle to enter the collecting duct and that the induction of a metabolic acidosis enhances this reentry.

Acidosis, Renal Tubular↗

PubMed1991

The effect of permeant and impermeant osmoticants on exocytosis in guinea pig sperm.

Guinea pig sperm were suspended in calcium-containing medium supplemented with various concentrations of the tetrasaccharide, stachyose. At concentrations up to and including 0.6 M, stachyose was without effect on the A23187-induced acrosome reaction. At 1.0 M stachyose, greater than 97% of sperm retained their acrosome after exposure to A23187, as judged by light microscopy. Electron microscopy demonstrated, however, that exocytotic membrane fusion had occurred, although with substantial retention of the acrosomal matrix. Sperm incubated in 1.0 M stachyose solutions also underwent exocytotic membrane fusion in the absence of A23187 and external calcium. Sperm suspended in 0.175 M ammonium chloride solution progressively lost motility over 30 min, but without acrosomal swelling. By contrast, sperm in 0.19 M ammonium acetate underwent substantial swelling of the acrosome within 2-5 min. 70-80% of these sperm were able to exclude the vital dye propidium iodide with their acrosomes swollen. These sperm underwent acrosomal shrinkage if resuspended in normal medium within 5-10 min, and the majority (60-70%) recovered some motility. These sperm could undergo an A23187-induced acrosome reaction. Electron microscopy indicated that swelling in ammonium acetate solution solubilizes much of the acrosomal matrix and causes internal fusion between adjacent regions of the outer acrosomal membrane. There was no exocytotic membrane fusion in ammonium acetate solution, however. The evidence suggests that there is no stachyose osmolality for guinea pig sperm which will suppress the membrane fusion associated with exocytosis, and that sufficiently high osmolalities cause exocytotic membrane fusion in the absence of calcium.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetates↗

PubMed1998

Intracellular pH and chemoresponse to NH4+ in Paramecium.

Paramecium are attracted to ammonium chloride solutions relative to sodium chloride control solutions, but little is known about the mechanisms by which attraction is evoked. A known effect of ammonium solutions in other cell types is an alteration of intracellular pH. We show here that intracellular pH is elevated upon initial exposure to 5 mM NH4Cl, but appears to decline within 10 minutes, both in wild type cells and in two mutants which do not show sustained attraction to NH4Cl using the standard behavioral assay, the T-maze. We also present quantitative values of swimming parameters that underlie the response to NH4Cl.

Ammonium Chloride↗

PubMedDate not supplied

Influence of ammonium sulfate and sodium chloride on ethyl methanesulfonate-induced chromosomal aberrations and HPRT mutations in V79 hamster cells.

V79 hamster cells were mutagenized with ethyl methanesulfonate (EMS) and immediately afterwards posttreated with hypertonic solutions of sodium chloride or ammonium sulfate. The posttreatment had a clear effect on chromosomal aberrations, but TGr mutations were only enhanced by ammonium sulfate, but not by sodium chloride posttreatment. It is suggested that hypertonic salt posttreatment leads to conformational changes in the DNA, resulting in an increase in TGr mutations and chromosomal aberrations.

Ammonium Sulfate↗