Solute concentrations of the pulmonary epithelial lining fluid of anesthetized rats

Uncertainty persists concerning the best method of estimating the volume and solute concentrations of the pulmonary epithelial lining fluid (ELF) recovered during bronchoalveolar lavage (BAL). In the present study, measurements were made of the BAL-to-plasma concentration ratios of a variety of solutes in an anesthetized rat model. One minute after an intravenous injection of labeled Na+ and urea, 5 ml of isotonic mannitol, saline, or glucose were injected into the trachea and an initial aliquot of the BAL was immediately removed. Initial BAL-to-plasma concentration ratios of urea, Na+, Cl-, Ca2+, and total protein were similar (ranging from 0.013 to 0.017) after BAL with mannitol, but albumin and transferrin ratios were approximately 60% lower and K+ ratios were five times greater. Lavage with saline yielded BAL-to-plasma urea concentration ratios similar to those obtained with mannitol lavage. The BAL-to-plasma specific activity of urea was about twice that of Na+, indicating that urea diffused into the ELF more rapidly than Na+ during the 70 s that elapsed between the time the radioactive urea and Na+ were injected into the circulation and the time when lavage was complete. Subsequent lavage samples also indicated that urea rapidly diffuses into the fluid-filled lungs. These experiments suggest that isotonic mannitol may be a useful solution for lavage, because it allows use of Na+ and perhaps Cl- as additional indicators of ELF dilution by BAL.(ABSTRACT TRUNCATED AT 250 WORDS)     

Coupled Solute Fluxes in Toad Skin

Net inward flux of mannitol across toad skin induced by making the outside solution hypertonic with urea has been investigated. No significant relation between net mannitol flux and net Na flux could be detected when both fluxes were measured simultaneously. In addition, the net mannitol flux caused by hypertonic solution was not altered by inhibition of Na transport with ouabain or by replacement of all Na in the bathing solutions by choline. The rate of net mannitol flux was dependent on the magnitude of the urea concentration difference across the skin and the direction of net flux could be reversed by reversing the direction of the urea concentration difference. These observations suggest that the mannitol transfer is the result of a coupling between the flows of urea and mannitol.     

Effect of hyperosmotic solutions on salt excretion and thirst in rats

We investigated urinary changes and thirst induced by infusion of hyperosmotic solutions in freely moving rats. Intracarotid infusions of 0.3 M NaCl (4 ml/20 min, split between both internal carotid arteries) caused a larger increase in excretion of Na(+) and K(+) than intravenous infusions, indicating that cephalic sensors were involved in the response to intracarotid infusions. Intravenous and intracarotid infusions of hyperosmotic glycerol or urea (300 mM in 150 mM NaCl) had little or no effect, suggesting the sensors were outside the blood-brain barrier (BBB). Intracarotid infusion of hypertonic mannitol (300 mM in 150 mM NaCl) was more effective than intravenous infusion, suggesting that cell volume rather than Na(+) concentration of the blood was critical. Similarly, intracarotid infusion (2 ml/20 min, split between both sides), but not intravenous infusion of hypertonic NaCl or mannitol caused thirst. Hyperosmotic glycerol, infused intravenously or into the carotid arteries, did not cause thirst. We conclude that both thirst and electrolyte excretion depend on a cell volume sensor that is located in the head, but outside the BBB.     

Response of the Frog Skin to Steady-State Voltage Clamping : I. The shunt pathway

Properties of the shunt pathway (a pathway in parallel to the Na transport system) in frog skin have been examined. The permeability of this shunt to urea increases markedly when the skin is depolarized to -100 mv (inside negative) but hyperpolarization to +100 mv produces no change in urea permeability compared to short-circuit conditions. The permeability increase at depolarizing potentials is dependent on the external solute concentration and is considerably reduced by the presence of external Ca. Neither urea permeability nor its response to changes in potential difference are affected by complete inhibition of Na transport by ouabain. In ouabain-poisoned skins, movements of Na, K, Cl, and mannitol through the shunt change in parallel with urea movements. Ion fluxes under these conditions and their response to potential can be described by the constant field equation. The selectivity of the shunt is in the order Cl > urea > K > Na > mannitol and this order does not appear to be affected by the absolute magnitude of the shunt permeability. Arguments are presented suggesting that the pathway is mainly between cells and that its permeability may be affected by cell swelling.     

Mannitol production from glycerol by resting cells of Candida magnoliae

Production of mannitol from glycerol by resting cells of Candida magnoliae under aerobic condition was investigated. The resting cells were suspended in aqueous solution of glycerol in Erlenmeyer flasks and incubated on rotary shaker. The samples were analyzed by ion exclusion–HPLC equipped with refractive index and UV detector. The resting cells of C. magnoliae produced mannitol from fructose, sucrose and glycerol but not from glucose. Addition of yeast extract and/or potassium phosphate to the glycerol solution adversely affected its conversion to mannitol. The conversion of glycerol to mannitol was dependent on oxygen availability. Using resting cells, the yield of mannitol was as high as 45%. This is probably the first report of conversion of glycerol to mannitol by osmophilic yeast.     

Osmoregulation in red blood cells of Bufo viridis

The effect of hyperosmotic solution of NaCl, urea and mannitol on Bufo viridis red blood cells were studied. The percentage of water content in B. viridis red blood cells decreased significantly in NaCl and mannitol hypertonic solutions compared to urea hypertonic solution. The urea concentration found in red blood cells in a urea hypertonic solution was significantly higher than in red blood cells acclimated to NaCl and mannitol hypertonic solutions. The Na+ concentration was significantly lower in red blood cells immersed in urea hypertonic solution than in red blood cells immersed in hypertonic NaCl and mannitol solutions. However, the K+ concentration increased at a similar rate in three different hypertonic solutions.     

Influence of carbon source on α-amylase production by Aspergillus oryzae

The influence of the carbon source on alpha-amylase production by Aspergillus oryzae was quantified in carbon-limited chemostat cultures. The following carbon sources were investigated: maltose, maltodextrin (different chain lengths), glucose, fructose, galactose, sucrose, glycerol, mannitol and acetate. A. oryzae did not grow on galactose as the sole carbon source, but galactose was co-metabolized together with glucose. Relative to that on low glucose concentration (below 10 mg/l), productivity was found to be higher during growth on maltose and maltodextrins, whereas it was lower during growth on sucrose, fructose, glycerol, mannitol and acetate. During growth on acetate there was no production of alpha-amylase, whereas addition of small amounts of glucose resulted in alpha-amylase production. A possible induction by alpha-methyl-D-glucoside during growth on glucose was also investigated, but this compound was not found to be a better inducer of a-amylase production than glucose. The results strongly indicate that besides acting as a repressor via the CreA protein, glucose acts as an inducer.     

Co-fermentation of carbon sources by Enterobacter aerogenes ATCC 29007 to enhance the production of bioethanol

We investigated the enhancement of bioethanol production in Enterobacter aerogenes ATCC 29007 by co-fermentation of carbon sources such as glycerol, glucose, galactose, sucrose, fructose, xylose, starch, mannitol and citric acid. Biofuel production increases with increasing growth rate of microorganisms; that is why we investigated the optimal growth rate of E. aerogenes ATCC 29007, using mixtures of different carbon sources with glycerol. E. aerogenes ATCC 29007 was incubated in media containing each carbon source and glycerol; growth rate and bioethanol production improved in all cases compared to those in medium containing glycerol alone. The growth rate and bioethanol production were highest with mannitol. Fermentation was carried out at 37 °C for 18 h, pH 7, using 50 mL defined production medium in 100 mL serum bottles at 200 rpm. Bioethanol production under optimized conditions in medium containing 16 g/L mannitol and 20 g/L glycerol increased sixfold (32.10 g/L) than that containing glycerol alone (5.23 g/L) as the carbon source in anaerobic conditions. Similarly, bioethanol production using free cells in continuous co-fermentation also improved (27.28 g/L) when 90.37 % of 16 g/L mannitol and 67.15 % of 20 g/L glycerol were used. Although naturally existing or engineered microorganisms can ferment mixed sugars sequentially, the preferential utilization of glucose to non-glucose sugars often results in lower overall yield and productivity of ethanol. Here, we present new findings in E. aerogenes ATCC 29007 that can be used to improve bioethanol production by simultaneous co-fermentation of glycerol and mannitol.     

Hyperosmolarity and the Net Transport of Nonelectrolytes in Frog Skin

The permeability of frog skin to a series of nonelectrolytes (thiourea, urea, mannitol, and sucrose) under the influence of 2.5 times normal osmolarity in the outer bathing solution has been investigated. Although the flux of the tracer nonelectrolytes across the skin in either direction is greatly increased by hyperosmolarity, the influx is found to be increased to a significantly greater extent than the outflux. Flux ratios as high as 3:1 can be observed. The net inward movement of the nonelectrolyte proceeds in spite of a sizeable bulk flow of water in the opposite direction. Possible driving forces for this phenomenon are discussed.     

Chronotropic response to hyperosmotic solutions of isolated rat atria

The changes in spontaneous rate of isolated rat atria in response to increased extracellular osmotic pressure were examined using sucrose, urea and several polyhydroxyalcohols (mannitol, glycerol and ethylene glycol) as test solutes. Sucrose, mannitol and urea induced a fall in atrial rate, which was transient with the last compound. On the other hand, media made hyperosomotic by addition of glycerol or ethylene glycol increased the beating frequency. Sucrose effect was not affected by low extracellular calcium, nifedipine or atropine. Glycerol-induced increase in atrial rate was a calcium-dependent mechanism sensitive to nifedipine. Thus, positive chronotropic effect occurs in the rat atria only with certain diffusible solutes which probably promote calcium entry. The response to pure osmotic change, resulting from changes in concentration of ions within the cell as water moves out, is a negative chronotropic effect.     

Effects of a Single Therapeutic Dose of Glycerol on Cerebral Metabolism in the Brains of Young Mice: Possible Increase in Brain Glucose Transport and Glucose Utilization

Abstract: This is a study of the effects of a single “therapeutic” dose of glycerol [2 g(22 mmol)/kg i.p.] on brain carbohydrate and energy metabolism in normal nursing weanling mice. Findings were correlated with brain water and electrolyte content and with metabolite changes in plasma, red blood cells, and liver. Plasma glycerol levels peaked at 21 mM 7.5 min after injection and returned to the control value, 0.16 mM, by 2 h. Plasma Na⁺ concentration decreased and plasma protein increased for as long as 2 h after injection. Although red blood cells were freely permeable to glycerol, there was no evidence for glycerol metabolism in these cells. Glycerol levels in liver paralleled those in plasma. Glycerol injection increased liver glucose concentration 23% and doubled hepatic glycerol-1-phosphate levels. Liver ATP levels were reduced 24% after glycerol injection. Brain water concentration was significantly reduced from 7.5 min to 30 min after glycerol injection; brain Na⁺ and K⁺ levels were unchanged. There was no evidence for glycerol entry into brain (the amount detected in brain tissue could be explained by the glycerol content in the blood of the brain). While plasma glucose increased 33%, brain glucose increased 87%. Concomitantly there were statistically significant increases in fructose-1,6-diphosphate, lactate, α-ketoglutarate, and malate levels. The disproportionately high brain glucose value suggests increased transport of glucose from the blood to the brain. Increases in fructose-1,6-diphosphate, lactate, α-ketoglutarate, and malate are compatible with an increased metabolic flux in the glycolytic pathway and Krebs citric acid cycle. As has been previously shown for urea and/or mannitol, these changes may result from the effects of the hyperosmolar glycerol solution on the blood-brain barrier and on cerebral glucose utilization. The sustained lowering of plasma Na⁺ concentration after a single “therapeutic” glycerol injection suggests a need for monitoring plasma Na⁺ levels in the clinical situation. Possible lowering of hepatic ATP levels by the use of glycerol in humans is another concern.     

A carbon source-dependent antimetabolite sensitivity test in Bacillus megaterium B71 for isolation of ethionine resistant mutants

Alteration of carbon sources significantly altered the analogue sensitivity of Bacillus megaterium B71. DL-Ethionine (ETN) was highly inhibitory with glucose, mannitol, sucrose, citrate, glycerol and arabinose. DL-Norleucine, L-homoserine and S(2'-aminoethyl)-L-cysteine were either highly inhibitory, slightly inhibitory or non-inhibitory depending on the carbon sources used. Maltose markedly overcame the inhibitory effect of ETN in liquid culture. Uninhibited growth was poor on citrate and arabinose when compared with other carbon sources. Six carbon sources showing comparable growth were used to determine minimum inhibitory concentrations (MIC) of the analogues. The MIC of ETN was highest (450 μg/ml) with maltose and lowest (4 μg/ml) with mannitol. ETN sensitivity was inversely related to the endogenous L-methionine pool size, and was relatively low with mannitol which was used to isolate ETN resistant mutants of B. megaterium B71. The best mutant BUE-118 produced 435 μg/ml of L-methionine.     

耐高渗压高压甘油的一个假丝酵母新种：产甘油假丝酵母

从自然标本中分离获得一高产甘油的菌株ＷＬ２００２－５,仅发酵葡萄糖及微发酵蔗糖,能利用葡萄糖、蔗糖、乙醇生产、微利用甘油和柠檬酸,不利用肌醇,硝酸盐、与ＤＢＢ显色反应为阴性。     

Comparison between somatic and zygotic embryo development in Quercus robur L.

ABSTRACT

Somatic embryogenesis from juvenile explants as an efficient way for oak clonal propagation is drastically limited by the low rate of embryo germination. A comparison of the development of immature somatic and zygotic embryos, and a study of the changes in sugar content and lignin accumulation during somatic versus zygotic embryo development were conducted in view of understanding the effect of reserve substance deficiency upon somatic embryo maturation. A morphological comparison of somatic and zygotic embryos led to the identification of 4 to 7 similar developmental stages in both types of embryos, thus indicating that the accumulation phase in both zygotic and somatic embryos occurs at the same stage, when the cotyledons became thicker and opaque. Carbohydrate analysis showed the presence of glycerol, inositol, mannitol, galactose, trehalose, xylose, arabinose, glucose, fructose and sucrose in all stages of zygotic and somatic embryo development, but in different amounts. The amount of glycerol, inositol, glucose and sucrose during the early stages is larger in zygotic embryos than in somatic ones, but the time course of their accumulation is similar in both types of embryos. Lignin content, which increased continuously during development, showed a similar behaviour in zygotic and somatic embryos. In somatic embryos which were able to germinate, lignin content was higher than in nongerminating embryos at the same stage.     

Na+ Dependence of Tyrosine Transport Across the Synaptosomal Membrane Reflects Changes in the Morphology of Synaptosomes

The Na+ dependence of tyrosine uptake into rat brain synaptosomes and synaptosomal plasma membrane vesicles (SPMV) was examined in the present study. At low tyrosine concentrations, the isoosmotic substitution of Na+ by sucrose in the incubation medium led to an increase of tyrosine uptake in synaptosomes and to a decrease in SPMV. The removal of extracellular Ca2+ and Mg2+ and addition of isoosmotic sucrose completely prevented the augmented tyrosine uptake in Na+-free incubated synaptosomes. Morphological differences were found at the electron-microscopic level when synaptosomes were incubated in Na+-free and Na+-containing media. The internal volume measured for synaptosomes incubated in a Na+-free medium was almost half of that obtained in a Na+-containing medium, in good agreement with the observations made with the electron microscope. Also, the omission of Ca2+ and Mg2+ resulted in a specific swelling of only the synaptosomes incubated in Na+-free medium. When synaptosomes and SPMV were preloaded with several neutral amino acids, the tyrosine uptake rate was greatly increased, indicating fully operational exchange mechanisms for these amino acids. We propose that the enhancement of high-affinity synaptosomal tyrosine uptake observed in Na+-free medium is a consequence of a specific shrinkage of the synaptosomes and a parallel increase of the exchange rate with endogenous neutral amino acids.     

Sodium induces oxytocin release from superfused rat pituitary

The effects of various osmotic agents on the release of oxytocin were examined in a superfusion system. Oxytocin was released significantly from the rat pituitary by superfusion with medium of an osmolality of 350 mOsm/kg H2O adjusted with NaCl, regardless of the presence of the rat hypothalamus. Media adjusted to an osmolality of 350 mOsm/kg H2O with sucrose, glucose, urea or mannitol had no effect on oxytocin release from the hypothalamo-pituitary complex. Medium containing excess Na2SO4 induced significant release of oxytocin from the pituitary without the hypothalamus. The administration of tetraethylammonium chloride had no oxytocin secretion. These data suggest that oxytocin release from the pituitary is influenced by the level of sodium ion rather than the osmotic pressure.     

Production of extracellular polyols in Aureobasidium pullulans

Aureobasidium pullulans produced extracellularly considerable amounts of polyols in the media with sucrose, glucose, fructose and mannose as sole carbon source during the late exponential and stationary phase of growth. The maximum yield of polyol was about 23% in the 20%(w/v) sucrose medium, of which mannitol was the main polyol associated with minute quantities of glycerol. Stress solutes such as NaCl and KCl did not promote polyol production.     

Effect of stress and sampling site on metabolite concentration in rat plasma

The effect of mild stress on various plasma metabolites in the rat has been studied. Mild stress resulted in significant decreases in liver size and glycogen content, as well as in an increase of blood glucose. In addition, plasma lactate, insulin, glycerol and urea, as well as a number of amino acids were altered by stress. These data indicate that minimal stress can have major effects upon the composition of blood, and suggest the need for strict precautions on the handling of animals during blood sampling. The site of blood extraction--tail tip vs. neck--was also found to have a significant effect on plasma lactate, glucose and urea concentrations. In stressed animals the differences between tail- and neck blood composition were increased.     

Effects of D2O and Osmotic Gradients on Potential and Resistance of the Isolated Frog Skin

Exposure of the outside surface of isolated frog skin (R. pipiens and R. catesbeiana) to sulfate solution made up with D₂O decreased skin potential and resistance. Exposure of the inside surface to D₂O solution decreased the potential slightly but increased the resistance. The changes were linearly related to the D₂O concentration. Since D₂O acts like a hyperosmotic solution, the skin potential and resistance were studied upon exposure to solution made hyperosmotic by addition of sucrose, mannitol, acetamide, urea, thiourea, Na₂SO₄, or K₂SO₄. Skin potential and resistance decreased when the outside solution was made hyperosmotic. The changes depended upon the concentration and the nature of the solute. Thiourea and urea solutions were the most effective. Treatment of the inside surface gave relatively small decreases in potential; the resistance either increased or remained unchanged. These effects appeared to depend upon the direction of the osmotic gradient across the skin rather than upon the value of the osmolarity compared to normal body fluids. Experiments with a series of six polyhydric alcohols from methanol to mannitol and the polysaccharides, sucrose and raffinose, showed adonitol with 5 carbons to decrease the potential the most. Smaller and larger compounds of this set gave lesser effects. As yet no consistent explanation of the effects is forthcoming, but their demonstration calls for caution in the indiscriminate use of solutes such as mannitol or sucrose "to make up the osmolality" and in the neglect of urea because "it penetrates freely."