Combination Of Mannitol Research Articles

The effects of formulation variables on the stability of freeze-dried human growth hormone.

Formulation often has a dramatic effect on degradation of proteins during the freeze-drying process as well as impacting on the "shelf-life" stability of the freeze-dried product. This research presents the results of a formulation optimization study of the "in-process" and shelf-life stability of freeze-dried human growth hormone (hGH). Chemical decomposition via methionine oxidation and deamidation of asparagine residues as well as irreversible aggregation were characterized by HPLC assay methodology. In-process degradation and stability of low moisture freeze-dried solids were studied at 25 and 40 degrees C in a nominal nitrogen headspace (approximately 0.5% O2). Formulation variables included pH, level of salts, and the nature of the lyoprotectant. Studies of the effect of shear on aggregation in solutions indicated that shear comparable to that experienced during filtration does not induce aggregation. Irreversible changes in hGH during the freeze-drying process were minimal, but chemical decomposition via methionine oxidation and asparagine deamidation and aggregation did occur on storage of the freeze-dried solid. Decomposition via methionine oxidation was significant. A combination of mannitol and glycine, where the glycine remains amorphous, provided the greatest protection against decomposition and aggregation. It is postulated that an excipient system that remains at least partially amorphous is necessary for stabilization. However, the observation that dextran 40 formulations showed poor stability toward aggregation demonstrates that an amorphous excipient system is not a sufficient condition for stability. Stability of the solid was optimal when produced from solutions in the pH range, 7-7.5, with severe aggregation being observed at high pH. The level of sodium phosphate buffer affected stability of the solid, although this relationship was complex. Freeze-drying in the presence of NaCl produced severe aggregation and precipitation during the freeze-drying process as well as acceleration of oxidation and/or deamidation.

Maturation and greenhouse planting of alfalfa artificial seeds

Conversion (plant production) was obtained from direct-planting alfalfa somatic embryos and encapsulated somatic embryos (artificial seeds) of alfalfa into a growth chamber and greenhouse. The embryos were planted in a commercial soil potting mix under nonsterile conditions in a manner similar to zygotic seed. Embryo maturation with abscisic acid (ABA), prior to planting, gave 48% conversion in soil under growth chamber conditions. Under greenhouse conditions, 64% conversion was obtained when humidified air was used to prevent soil surface drying. Previously, conversion in soil was between 0–6% without the ABA maturation treatment. The replacement of ABA with mannitol or combinations of mannitol and ABA during maturation resulted in lower conversion in the growth chamber than with ABA alone. ABA may be promoting the accumulation of embryo storage reserves such as proteins and carbohydrates for growth after planting in the soil environment.

Proline Accumulation and Its Implication in Cold Tolerance of Regenerable Maize Callus

Embryogenic callus of maize (Zea mays L.) inbreds B37wx, H99, H99(3)H95, Mo17, and Pa91 accumulated proline to levels 2.1 to 2.5 times that of control callus when subjected to mannitol-induced water stress, cool temperatures (19 degrees C) and abscisic acid (ABA). A combination of 0.53 molar mannitol plus 0.1 millimolar ABA induced a proline accumulation to about 4.5 times that of control callus, equivalent to approximately 0.18 millimoles proline per gram fresh weight of callus. Proline accumulation was directly related to the level of mannitol in the medium. Levels of ABA greater than 1.0 micromolar were required in the medium to induce proline accumulation comparable to that induced by mannitol. Mannitol and ABA levels that induced maximum accumulation of proline also inhibited callus growth and increased tolerance to cold. Proline (12 millimolar) added to the culture media also increased the tolerance of callus to 4 degrees C. The increased cold tolerance induced by the combination of mannitol and ABA has permitted the storage of the maize inbreds A632, A634Ht, B37wx, C103DTrf, Fr27rhm, H99, Pa91, Va35, and W117Ht at 4 degrees C for 90 days which is more than double the typical survival time of callus. These studies show that proline and conditions which induce proline accumulation increase the cold tolerance of regenerable maize callus.

The protective effect of combined administration of anti-oxidants and perfluorochemicals on cerebral ischemia.

We previously published results of investigations which indicated that the combination of mannitol, which acts as a free radical scavenger, and perfluorochemicals (PFC), which have a strong oxygen-carrying capacity, can be therapeutic in cases of brain infarction. The present experiment tested the hypothesis that the effectiveness of such treatment could be increased by an optimal combination of such scavengers and other chemicals. Fifty-two dogs were used, employing the "canine model of a completely ischemic brain regulated with the perfusion method." A total of six drugs with free radical scavenger capacities were tested: mannitol, vitamin E, vitamin C, Nizofenone (Y-9197), dexamethasone (DEXA) and suloctidil (MY-103). These drugs were administered intravenously 15 minutes prior to the production of ischemia, when cerebral blood flow was reduced to one-tenth its normal volume. After one hour, recirculation was allowed and the recovery of electrical activity of the brain observed for three hours. Judged by the degree of recovery of brain electrical activity, five drugs were considered to have protective effect against brain ischemia: mannitol, vitamin E, MY-103, DEXA and Y-9197. Among these five drugs, mannitol, vitamin E and DEXA are known to be safe and easily used clinically. The combined administration of these three drugs, together with PFC, was also investigated. It was found that the speed and degree of recovery of brain electrical activity were greater when these drugs were given together than when one was administered alone.(ABSTRACT TRUNCATED AT 250 WORDS)

In Vitro and Invivo Evaluation of Some Fast Release Dosage Forms of Hydrochlorothiazide

AbstractThe utilization of ternary sugar solid dispersion and solvent deposition systems for increasing the dissolution rate of hydrochlorothiazide (hot) were investigated. The dispersion systems were prepared by the fusion method using various combinations of mannitol and sorbitol, and urea and polyethylene glycol 4000 (peg 4000) were used for comparison. An 1:2 mixture of sorbitol-mannitol was found to be an excellent carrier. The dissolution rate of this sample was closely comparable to that of hot-peg 4000 solid dispersions. Drug-urea eutectic mixtures were inferior to both the sugar and polymer dispersions. Solvent deposition systems of hot with microfine cellulose and potato starch gave higher dissolution rates at the initial sampling times. It is proposed that solid dispersion systems of this drug may prove to be valuable. Tablets fabricated from fast-release hot granules showed better in vivo results than a marketed tablet. A linear relationship was observed between in vitro-in vivo data of some o...

Effect of carbonic anhydrase inhibition on superficial and deep nephron bicarbonate reabsorption in the rat.

The nephron segment responsible for the acetazolamide-insensitive fraction of renal bicarbonate reabsorption has not been clearly delineated. This study compares superficial and deep nephron bicarbonate reabsorption before and after acetazolamide at two dose levels (20 and 50 mg/kg per h) in mutant Munich-Wistar rats employing both cortical and papillary micropuncture and microcalorimetry. Systemic acid-base balance and right whole kidney glomerular filtration rate were similar in all groups examined. The effects of the two doses of acetazolamide were indistinguishable and resulted in a significant increase in whole kidney bicarbonate excretion that compared favorably with the fraction delivered out of the left papillary tip. Acetazolamide inhibited superficial proximal bicarbonate reabsorption by 80.0%, whereas reabsorption up to the deep loop of Henle was decreased by only 52% (P less than 0.001). Bicarbonate reabsorption that was insensitive to acetazolamide occurred in the superficial and deep loop of Henle and between the distal tubule and base collecting duct. Because water reabsorption in these segments could serve to generate transepithelial bicarbonate concentration gradients favorable for reabsorption, we attempted to minimize water abstraction by combined administration of mannitol and acetazolamide. During this condition a significant increase in bicarbonate delivery up to the deep loop of Henle was noted (52 vs. 65%), whereas superficial nephron reabsorption was not altered. Furthermore, an outwardly directed bicarbonate concentration gradient from the deep loop of Henle to vasa recta was demonstrated during acetazolamide (delta tCO2 = 20.9 +/- 3.3 mM), but was abolished during combined mannitol and acetazolamide administration (delta tCO2 = 3.5 +/- 0.9 mM). It is concluded that carbonic anhydrase inhibition results in a disparate effect on nephron bicarbonate reabsorption when juxtamedullary and superficial nephron segments are compared. Our findings suggest that a mechanism for residual bicarbonate reabsorption during acetazolamide administration may be passive reabsorption driven by favorable transepithelial concentration gradients.

Modification of acute focal ischemia by treatment with mannitol and high-dose dexamethasone.

A simple implanted device was used to occlude the left middle cerebral artery (MCA) of 50 conscious cats. In the acute experiments, 10 cats were given mannitol (1.2 gm/kg intravenously) and 10 cats were untreated. The neurological status of the treated cats improved. Perfusion with a mixture of colloidal carbon and buffered paraformaldehyde was carried out from 30 minutes to 12 hours following MCA occlusion. Results of the morphological examination of the brains demonstrated that mannitol delayed the development of neuronal alterations and edema. Measurement of capillary luminal diameters in the ischemic cortex of the untreated cats revealed progressive narrowing with microcirculatory obstruction occurring at 6 hours. Capillary narrowing was absent or minimal in the treated cats during the initial 6 hours. Breakdown of the blood-brain barrier also was delayed by treatment. In the 30 subacute experiments, 10 cats were untreated, 10 cats received mannitol (1.2 gm/kg intravenously), and 10 cats were given a combination of mannitol and high-dose dexamethasone. Although early transient clinical improvement occurred in the treated animals, morphological findings in the brains of the treated and untreated cats following 48 hours of ischemia were not significantly different. Despite the finding that the beneficial effects of mannitol were relatively short-lived (6 to 12 hours), the delay in the development of irreversible changes probably is important as it would allow a longer period of time for the institution of a more definitive form of therapy.

The influence of combined intra-aortic balloon counterpulsation and hyperosomotic mannitol on regional myocardial blood flow in ischemic myocardium in the dog.

We investigated the combined effectiveness of intra-aortic balloon counterpulsation and hyperosmotic mannitol (25%) on regional myocardial blood flow during acute coronary insufficiency. Cardiac output and paced heart rate were held constant in chloralose-anesthetized dogs during right heart bypass. Acute coronary insufficiency was produced by ligation of the proximal left anterior descending coronary artery (LAD). Regional myocardial blood flow was measured using radioactive microspheres. Left ventricular end-diastolic pressure, mean aortic pressure, maximum left ventricular dp/dt, and hematocrit were unchanged by combined mannitol infusion and balloon pumping. Studies of combined treatment with balloon pumping and mannitol immediately after the second of two 13-minute consecutive reversible ligations of the LAD demonstrated that (1) collateral coronary blood flow increased 46% (P less than 0.02) in ischemic myocardium compared with mannitol infusion along during the first LAD ligation, and (2) collateral coronary blood flow increased 27% (P less than 0.05) in ischemic myocardium compared with balloon pumping along during the first LAD ligation. Studies in which combined treatment was delayed until 20 minutes after LAD ligation demonstrated that collateral coronary blood flow was elevated by 33% (P les than 0.05) in ischemic myocardium compared to control studies in which balloon pumping alone had no effect. The results suggest that the increase in collateral coronary blood flow was in part a result of an increased transmural pressure gradient produced by balloon diastolic augmentation and the ability of mannitol to reduce coronary vascular resistance in ischemic myocardium.