sodium-acetate--anhydrous and ammonium-acetate

A plasmid preparation is a method used to extract and purify plasmid DNA. Methods developed to purify plasmid DNA from bacteria generally involve harvesting and alkaline lysis of the bacteria and precipitation of chromosomal DNA and protein, followed by purification of the plasmid DNA. Here, we describe the mini-preparation of plasmid DNA by a rapid small-scale method, adapted for Listeria monocytogenes. The quality of plasmid DNA isolated using this method is sufficient for analytical purposes but may be upscaled for further downstream analysis. Electrophoretic separation of the resultant lysate allows conclusions to be made on the presence, number, copy number, and size of the plasmids in the analyzed bacterial strains.

Topics: Acetates; Buffers; DNA; Electrophoresis; Listeria monocytogenes; Listeriosis; Muramidase; Plasmids; Sodium Acetate; Sucrose

When working with RNA, the need often arises to concentrate a sample or purify it from various salts, nucleotides, and proteins. RNA precipitation is an easy and cost-effective method for the concentration of RNA, leaving a pellet that can be resuspended in the buffer of choice.

Topics: Acetates; Animals; Chemical Precipitation; Ethanol; Lithium Chloride; RNA; Sodium Acetate

Solvent-free Claisen-Schmidt reactions of cycloalkanones with various substituted benzaldehydes (aryl aldehydes) using solid NaOH (20 mol%) and applying a grinding technique were studied. Quantitative yields (96-98%) of α,α'-bis-(substituted-benzylidene)cycloalkanones were obtained. Aliphatic aldehydes also provided α,α'-bis-(substituted-alkylidene)cycloalkanones in very good yields with minor amounts of a-(substituted-alkylidene)cycloalkanones. The catalytic performance of solid NaOH was examined. The molar ratio of NaOH was optimized. The catalytic effect of solid NaOH was also evaluated by comparing it with KOH, NaOAc, and NH(4)OAc and it turns out that 20 mol% of solid NaOH was good enough to catalyze the Claisen-Schmidt reactions of cycloalkanones with various substituted benzaldehydes. Additionally, the regioselectivity of the Claisen-Schmidt reaction of acetone with benzaldehyde was examined. Using the same method, we could synthesize the corresponding bis-benzylidene- and mono-benzylideneacetone separately in 98% and 96% yields, respectively.

Topics: Acetates; Benzylidene Compounds; Catalysis; Cycloparaffins; Freezing; Hydroxides; Potassium Compounds; Sodium Acetate; Sodium Hydroxide; Solid-Phase Synthesis Techniques

Exposure of Daphnia in degassed (boiled) culturing water (hypoxia simulation) led to solitary lethal outcomes after more than 24 h. Before this term, hypoxia had no appreciable effect on the toxicity of sodium or ammonium acetate salts. The sensitivity of daphnias to the lethal effects of the tested chemicals did not change under conditions of normal oxygenation and increased sharply (by two orders of magnitude) under conditions of hypoxia, loosing the linear relationship with toxicant concentration. Ammonium acetate toxicity more markedly increased under conditions of hypoxia than sodium acetate toxicity. These data should be taken into consideration when predicting the results of combined effects of toxicants on water ecosystems and on human organism.

Topics: Acetates; Animals; Daphnia; Female; Oxygen; Sodium Acetate

To test the hypothesis that ammonia detoxification in ruminants consumes amino acids to the detriment of milk protein production, we infused four lactating dairy cows with ammonium acetate or sodium acetate in switchback experiments. Plasma ammonia concentrations increased to 411 microm within 1 h of the start of infusion of ammonium acetate at 567 mmol/h. The rate constant for ammonia clearance from plasma was 0 x 054/min and the half-life was 12 x 9 min. Infusion at 567 mmol/h for 1 h followed by 1 h without infusion, repeated four times between am- and pm-milking, caused a decrease in feed intake. Compared with sodium acetate, continuous infusion of ammonium acetate at 360 mmol/h throughout an entire 10-h milking interval increased plasma ammonia concentrations to 193 microm and caused a 20% decrease in milk, protein and lactose production with no effect on percentage composition of milk or the yield of milk fat. Arterial concentrations of glucose and non-esterified fatty acids tended to increase; there was no effect on arterial acetate, beta-hydroxybutyrate or triacylglcerol, and branched-chain amino acids, Lys and Thr decreased. Mammary plasma flow, estimated by assuming 100% uptake/output of Phe+Tyr, was significantly correlated with milk yield. Mammary uptakes of acetate tended to be reduced by hyperammonaemia, but uptakes of other energy metabolites and amino acids were not affected. Thus, while an increase in amino acid consumption during hyperammonaemia was apparent from the drop in circulating concentrations of Leu, Ile, Val, Lys and Thr, there was no evidence to support the hypothesis that milk yield is affected by the lower concentrations. An ammonia-induced depression in feed intake may have caused the decrease in milk synthesis.

Topics: Acetates; Amino Acids; Ammonia; Animals; Cattle; Cattle Diseases; Dairying; Eating; Female; Hyperammonemia; Lactation; Mammary Glands, Animal; Milk; Sodium Acetate

Fundamental studies on the nanopreparation of neutral analytes in CZE by analyte focusing by micelle collapse (AFMC) are presented. The background solution (BGS) is prepared using an electrolyte salt (i.e. sodium or ammonium acetate). The sample solution of the neutral analytes (S) is prepared using SDS at a concentration above the cmc. To induce AFMC, the conductivity of the S must be greater than the BGS. This was achieved by the addition of the electrolyte salt to the S. Dilution of the micellar carrier from the injected S occurs at the BGS zone closest to the boundary between the S and BGS (micellar dilution zone). The dilution of SDS below the cmc causes the collapse of the micelles with subsequent release of previously bound analyte molecules. The continued transport and release causes the analytes to be accumulated at the micellar dilution zone. This nanopreparative technique is compatible with detection using mass spectrometry and can be utilized as a sample injection step for microfluidic devices. The disadvantage of this technique is that the neutral analytes are not separated after concentration. Here, the effect of retention factor of the analyte, conductivity ratio of the S and BGS, SDS concentration in the S, electrolyte salt (i.e. sodium acetate) concentration in the BGS, and organic modifier content in the BGS were examined. A study on the effect of the sample matrix injection prior to the sample injection to the performance of AFMC-CZE to neutral analytes is also presented.

Topics: Acetates; Electrolytes; Electrophoresis, Capillary; Methanol; Micelles; Nanotechnology; Sodium Acetate; Sodium Dodecyl Sulfate

This study examines a new method for the remote loading of doxorubicin into liposomes. It was shown that doxorubicin can be loaded to a level of up to 98% into large unilamellar vesicles composed of egg phosphatidylcholine/cholesterol (7/3 mol/mol) with a transmembrane phosphate gradient. The different encapsulation efficiencies which were achieved with ammonium salts (citrate 100%, phosphate 98%, sulfate 95%, acetate 77%) were significantly higher as compared to the loading via sodium salts (citrate 54%, phosphate 52%, sulfate 44%, acetate 16%). Various factors, including pH-value, buffer capacity, solubility of doxorubicin in different salt solutions and base counter-flow, which likely has an influence on drug accumulation in the intraliposomal interior are taken into account. In contrast to other methods, the newly developed remote loading method exhibits a pH-dependent drug release property which may be effective in tumor tissues. At physiological pH-value doxorubicin is retained in the liposomes, whereas drug release is achieved by lowering the pH to 5.5 (approximately 25% release at 25 degrees C or 30% at 37 degrees C within two h). The DXR release of liposomes which were loaded via a sulfate gradient showed a maximum of 3% at pH 5.5.

Topics: Acetates; Ammonium Sulfate; Cholesterol; Citrates; Citric Acid; Doxorubicin; Hydrogen-Ion Concentration; Liposomes; Microscopy, Electron, Transmission; Phosphates; Phosphatidylcholines; Quaternary Ammonium Compounds; Sodium; Sodium Acetate; Sodium Citrate; Sulfates

Mass Spectra of charge states of folded proteins were obtained with nanospray and aqueous solution containing 20 microM the protein (ubiquitin, cytochrome c, lysozyme) and one of the NaA salts NaCl, NaI, NaAc (acetate) (1-10 mM). At very low collision activated decomposition (CAD), the mass spectra of a protein with charge z exhibited a replacement of zH+ with zNa+ and also multiple adducts of NaA. Higher CAD converts the NaA adduct peaks to Na minus H peaks. These must be due to loss of HA where the H was provided by the protein. The degree of HA loss with increasing CAD followed the order I < Cl < Ac. Significantly, the intensity of the ions with n (Na minus H) adducts showed a downward break past an n(MAX) which is equal to the number of acidic residues of the protein plus the charge of the protein. All the observations could be rationalized within the framework of the electrospray mechanism and the charge residue model, which predict that due to extensive evaporation of solvent, the solutes will reach very high concentrations in the final charged droplets. At such high concentrations, positive ions such as Na+, NH4+ form ion pairs with ionized acidic residues and the negative A- form ion pairs with ionized basic residues of the protein. Adducts of Na+, and NaA to backbone amide groups occur also. This reaction mechanism fits all the experimental observations and provides predictions that the number of acidic and basic groups at the surface of the gaseous protein that remain ionized can be controlled by the absence or presence of additives to the solution.

Topics: Acetates; Acetic Acid; Amino Acids, Acidic; Amino Acids, Basic; Cytochromes c; Ions; Muramidase; Particle Size; Proteomics; Sodium Acetate; Sodium Chloride; Sodium Iodide; Spectrometry, Mass, Electrospray Ionization; Ubiquitin

Paclitaxel, a unique antimitotic chemotherapy agent that inhibits cell division by binding to microtubules and prevents them from "depolymerizing," has received widespread interest because of its efficacy in fighting certain types of cancer, including breast and ovarian cancer. Paclitaxel undergoes aggregation at millimolar concentrations in both aqueous media and solvents of low polarity (mimicking hydrophobic environments). Its aggregation may have impact on its aqueous stability and its ability to stabilize microtubules. Here, we investigated the dimerization phenomenon of paclitaxel by electrospray ionization mass spectrometry (ESI-MS). Paclitaxel dimers were stable in solutions of acetonitrile/aqueous ammonium acetate (80/20) and aqueous sodium acetate/acetonitrile (92/8 or 95/5) at various pH values. Additional experiments using solution-phase hydrogen/deuterium exchange were employed to ascertain whether or not the observed dimers were formed in solution or as an artifact of the ESI process by ion-molecule reaction. The evidence supports formation of the dimer in solution, and the approach used can be extended to investigation of other types of drug-drug interactions.

Topics: Acetates; Acetonitriles; Antineoplastic Agents, Phytogenic; Dimerization; Drug Stability; Hydrogen-Ion Concentration; Paclitaxel; Sodium Acetate; Solutions; Solvents; Spectrometry, Mass, Electrospray Ionization

Caenorhabditis elegans has two heterotrimeric G-protein gamma subunits, gpc-1 and gpc-2. Although GPC-1 is specifically expressed in sensory neurons, it is not essential for the detection of odorants or salts. To test whether GPC-1 is involved in sensory plasticity, we developed a water soluble compound adaptation assay. The behaviour of wild-type animals in this assay confirms that prolonged exposure to salts can abolish chemo-attraction to these compounds. This process is time and concentration dependent, partly salt specific and reversible. In contrast, gpc-1 mutant animals show clear deficits in their ability to adapt to NaAc, NaCl and NH4Cl, but normal wild-type adaptation to odorants. Two other loci previously implicated in odorant adaptation, adp-1 and osm-9, are also involved in adaptation to salts. Our finding that G proteins, OSM-9 and ADP-1 are involved in taste adaptation offer the first molecular insight into this process.

Topics: Acetates; Adaptation, Physiological; Ammonium Chloride; Animals; Caenorhabditis elegans; Caenorhabditis elegans Proteins; Chemotaxis; GTP-Binding Protein gamma Subunits; Heterotrimeric GTP-Binding Proteins; Ion Channels; Nerve Tissue Proteins; Osmolar Concentration; Smell; Sodium Acetate; Sodium Chloride; Taste; Transient Receptor Potential Channels; TRPV Cation Channels

Glutamine is purported to inhibit recycling of citrulline to arginine and to limit nitric oxide release in vitro. However, vasoactive effects of glutamine have not been clearly demonstrated in vivo. During hyperammonemia, impaired cerebrovascular reactivity to CO(2) is related to glutamine accumulation. We tested the hypotheses that 1) glutamine infusion in the absence of hyperammonemia impairs cerebrovascular CO(2) reactivity and 2) arginine infusion preserves CO(2) reactivity during glutamine infusion and during hyperammonemia. Pentobarbital sodium-anesthetized rats were equipped with a closed cranial window for measuring pial arteriolar diameter. Intravenous infusion of 3 mmol. kg(-1). h(-1) of L-glutamine for 6 h produced threefold increases in plasma and cerebrospinal fluid concentrations. Dilation to hypercapnia was reduced by 45% compared with that of a time control group at 6 h but not at 3 h of glutamine infusion. Coinfusion of 2 mmol. kg(-1). h(-1) of L-arginine with glutamine maintained the hypercapnic vasodilation at the control value. Infusion of ammonium acetate at a rate known to produce threefold increases in cortical tissue glutamine concentration resulted in no significant hypercapnic vasodilation. Coinfusion of arginine with ammonium acetate maintained hypercapnic vasodilation at 60% of the control value. Arginine infusion did not augment hypercapnic vasodilation in a control group. We conclude that glutamine modulates cerebrovascular CO(2) reactivity in vivo. Glutamine probably acts by limiting arginine availability because the vascular inhibitory effect required >3 h to develop and because arginine infusion counteracted the vascular effect of both endogenously and exogenously produced increases in glutamine.

Topics: Acetates; Ammonia; Analysis of Variance; Animals; Arginine; Arterioles; Blood Pressure; Carbon Dioxide; Cerebrovascular Circulation; Drug Synergism; Glutamic Acid; Glutamine; Hypercapnia; Infusions, Intravenous; Male; Pia Mater; Rats; Rats, Wistar; Sodium Acetate

Non-aqueous capillary electrophoresis was used to study the separation selectivity of positively charged drug substances and negatively charged diuretics. Study was made of the effects of organic solvent composition and the background electrolyte on the separation. The separation selectivity could be altered considerably by varying the methanol/acetonitrile composition. In addition, the migration order and the resolution of the pharmaceuticals could be altered merely by changing the electrolyte cation or the anion. The electrolytes tested were alkali metal acetates, ammonium acetate, ammonium chloride and ammonium bromide. As with aqueous background electrolyte solutions, the electroosmotic flow was decreased with increasing size of the alkali metal cation of the electrolyte in methanol/acetonitrile 50:50 (v/v).

Topics: Acetates; Acetonitriles; Ammonium Chloride; Bromides; Dextromoramide; Dose-Response Relationship, Drug; Electrolytes; Electrophoresis, Capillary; Ephedrine; Hydrogen-Ion Concentration; Levorphanol; Lithium Compounds; Methadone; Methanol; Morphine; Potassium Acetate; Quaternary Ammonium Compounds; Sodium Acetate; Time Factors; Viscosity

The effects of different types of salts and salt concentrations on the selectivity in the adsorption of serum proteins have been compared for the amphiphilic agarose-based adsorbents Phenyl-Sepharose, Octyl-Sepharose, butyl-agarose and mercaptopyridine-derivatized agarose. By use of multivariate analysis, the complex interrelationships for the different combined effects were evaluated. From these analyses conclusions about similarities and/or dissimilarities in the mechanisms involved in adsorption of proteins on respective adsorbent were made.

Topics: Acetates; Adsorption; Blood Proteins; Chromatography, Agarose; Osmolar Concentration; Potassium Chloride; Salts; Sodium Acetate; Sodium Chloride; Sulfates

The effects of adaptation/stimulus temperature (25 degrees C vs. 35 degrees C) on taste nerve responses to salt stimulation and amiloride suppression were assessed in rats. We measured the integrated responses of the chorda tympani nerve to 500 mM concentrations of NaCl, Na2SO4, sodium acetate (NaAc), KCl, K2SO4, potassium acetate (KAc), NH4Cl, (NH4)2SO4, and ammonium acetate (NH4Ac) mixed with or without 100 microM amiloride hydrochloride at 35 degrees C. Taste nerve responses to all Na+ and NH4+ salts, but not K/ salts, were significantly smaller at 25 degrees C than at 35 degrees C. Amiloride significantly suppressed taste nerve responses to all salts (Na+ salts > K+ salts > NH4+ salts); amiloride suppression of Na+ and NH4+ salts was significantly greater at 25 degrees C than at 35 degrees C. Benzamil-HCl, a more potent Na+ channel blocker compared to amiloride, strongly suppressed taste nerve responses to NaCl and KCl, but not to NH4Cl. Amiloride and benzamil suppression of NaCl responses were similar; however, amiloride suppressed KCl responses more than did benzamil. The results suggest that: (1) amiloride-sensitive Na+ channels are involved to varying degrees in the transduction of sodium and potassium salt taste, and (2) amiloride may inhibit membrane proteins other than passive Na+ channels during stimulation with potassium and ammonium salts.

Topics: Acetates; Amiloride; Animals; Chorda Tympani Nerve; Male; Potassium Acetate; Rats; Rats, Sprague-Dawley; Salts; Sodium Acetate; Taste; Temperature