Behavior of plant tissue in osmotic solutions

The effect of the concentration of sucrose solutions on the cellular structure of potato tissue in equilibrium at 27 degreesC was Studied. Two different methods of investigation were used to determine the volume of the different phases composing the cellular tissue of the potato when in equilibrium with the solutions. one based on data of the concentration itself and the overall volume of 2 mm slices after 48 h at equilibrium, and the other on microscopic images of cells in thin slices of fresh tissue stained with neutral red after an hour in equilibrium to show protoplasts, vacuoles and plasmolysis spaces. The results of these methods were compared with those obtained by a predictive thermodynamic approach considering the semipermeability of cell membranes. Phase volume data obtained from microscopic analysis were more similar to what was predicted by the theoretical model than those obtained by means of composition measurement. where the long equilibrium time apparently led to the loss of semi permeability of the cell membranes, since total volumes calculated without consideration of the cell membranes were similar to those measured. This suggests that the length of time of osmotic dehydration brings about a change in cell structure and the consequent involvement of a different mechanism in mass transfer. (C) 2002 Elsevier B.V. Ltd. All rights reserved.

A comparison between diuretics and angiotensin-receptor blocker agents in patients with stage I hypertension (PREVER-treatment trial): Study protocol for a randomized double-blind controlled trial

Background: Cardiovascular disease is the leading cause of death in Brazil, and hypertension is its major risk factor. The benefit of its drug treatment to prevent major cardiovascular events was consistently demonstrated. Angiotensin-receptor blockers (ARB) have been the preferential drugs in the management of hypertension worldwide, despite the absence of any consistent evidence of advantage over older agents, and the concern that they may be associated with lower renal protection and risk for cancer. Diuretics are as efficacious as other agents, are well tolerated, have longer duration of action and low cost, but have been scarcely compared with ARBs. A study comparing diuretic and ARB is therefore warranted.Methods/design: This is a randomized, double-blind, clinical trial, comparing the association of chlorthalidone and amiloride with losartan as first drug option in patients aged 30 to 70 years, with stage I hypertension. The primary outcomes will be variation of blood pressure by time, adverse events and development or worsening of microalbuminuria and of left ventricular hypertrophy in the EKG. The secondary outcomes will be fatal or non-fatal cardiovascular events: myocardial infarction, stroke, heart failure, evidence of new subclinical atherosclerosis and sudden death. The study will last 18 months. The sample size will be of 1200 participants for group in order to confer enough power to test for all primary outcomes. The project was approved by the Ethics committee of each participating institution.Discussion: The putative pleiotropic effects of ARB agents, particularly renal protection, have been disputed, and they have been scarcely compared with diuretics in large clinical trials, despite that they have been at least as efficacious as newer agents in managing hypertension. Even if the null hypothesis is not rejected, the information will be useful for health care policy to treat hypertension in Brazil. Clinical trials registration number: ClinicalTrials.gov: NCT00971165. © 2011 Fuchs et al; licensee BioMed Central Ltd.

Osmotic Dehydration of Pineapple with Impregnation of Sucrose, Calcium, and Ascorbic Acid

Mass transfer was evaluated during osmotic dehydration of pineapple in solutions with until four components aiming to investigate the solutes concentration influence on impregnation. In the first step, the experimental trials for optimization of solution concentration were based on 23 factorial design. In the second step, effective diffusion coefficients were determined. Equations representing the influence of the concentration of sucrose, calcium lactate, and ascorbic acid in osmotic solutions on water loss and gains of sucrose, calcium, and vitamin C were found. Results showed that both calcium lactate and sucrose concentration affected calcium and sucrose gain. On the other hand, only vitamin C gain was significantly affected by the ascorbic acid concentration in the studied concentration range. However, when comparing diffusivities in pineapple immersed in sucrose solutions, with and without calcium lactate, with and without ascorbic acid, it was possible to verify that diffusivities of water, sugar, and calcium increased in presence of ascorbic acid in solution. Calcium in solution diminished the water and sucrose diffusivities. High calcium and vitamin C contents were obtained in 1 h immersion in the solutions studied. © 2013 Springer Science+Business Media New York.

The tonoplast intrinsic aquaporin (TIP) subfamily of Eucalyptus grandis: Characterization of EgTIP2, a root-specific and osmotic stress-responsive gene

Aquaporins have important roles in various physiological processes in plants, including growth, development and adaptation to stress. In this study, a gene encoding a root-specific tonoplast intrinsic aquaporin (TIP) from Eucalyptus grandis (named EgTIP2) was investigated. The root-specific expression of EgTIP2 was validated over a panel of five eucalyptus organ/tissues. In eucalyptus roots, EgTIP2 expression was significantly induced by osmotic stress imposed by PEG treatment. Histochemical analysis of transgenic tobacco lines (Nicotiana tabacum SR1) harboring an EgTIP2 promoter:GUS reporter cassette revealed major GUS staining in the vasculature and in root tips. Consistent with its osmotic-stress inducible expression in eucalyptus, EgTIP2 promoter activity was up-regulated by mannitol treatment, but was down-regulated by abscisic acid. Taken together, these results suggest that EgTIP2 might be involved in eucalyptus response to drought. Additional searches in the eucalyptus genome revealed the presence of four additional putative TIP coding genes, which could be individually assigned to the classical TIP1-5 groups. © 2013 Elsevier B.V.

Effect of calcium on the osmotic dehydration kinetics and quality of pineapple

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Osmotic dehydration of mango with ascorbic acid impregnation: influence of process variables

The present study investigated the osmotic dehydration (OD) of mangoes cv.Palmer, dehydrated in a sucrose solution with added ascorbic acid (AA), aiming toevaluate the influence of the process variables (sucrose concentration, AA concen-tration and time) on vitamin C, water, sugars and total phenolic compound (TPC)contents by way of an analysis of the effects and the variance of the factors. Thesucrose concentration did not affect the vitamin C content, but the AA concentra-tion in the solution and the process time exerted a positive influence on thisresponse. The addition of AA also presented a significant positive effect on theTPC contents because this vitamin interferes with the quantification of the TPC.On discounting the interferences, no losses of phenolic compounds were found.Impregnation with vitamin C during OD was shown to be effective in obtainingminimally processed mangoes with high nutritional value.

Improvement of water transport and carotenoid retention during drying of papaya by applying ultrasonic osmotic pretreatment

The effect of ultrasound and osmotic dehydration pretreatments on papaya drying kinetics was investigated. The ultrasound pretreatment was carried out in an ultrasonic bath at 30 A degrees C. The osmotic pretreatment in sucrose solution was carried out in an incubator at 34 A degrees C and agitation of 80 rpm for 210 min. The drying process was conducted in a fixed bed dryer at 70 A degrees C. Experimental data were fitted successfully using the Page model for dried fresh and pretreated fruits, with coefficient of determination greater than 0.9992 and average relative error lower that 14.4 %. The diffusional model was used to describe the moisture transfer, and the effective water diffusivity was identified in the order of 10(-9) m(2) s(-1). It was found that drying rates of osmosed fruits were the lowest due to the presence of infused solutes, while the ultrasound pretreatment contributed to faster drying rates. Evaluation of the dried fruit was performed by means of total carotenoids retention. Ultrasound treatments in distilled water prior to air-drying gave rise to dried papayas with retention of carotenoids in the range 30.4-39.8 % and the ultrasonic-assisted osmotic dehydration of papayas showed carotenoids retention values up to 64.9 %, whereas the dried fruit without pretreatment showed carotenoids retention lower than 24 %.

Osmotic potentials on water uptake and germination of Guazuma ulmifolia Lam. (Sterculiaceae) seeds

Osmotic potentials on water uptake and germination of Guazuma Ulmifolia Lam. (Sterculiaceae) seeds. This work was carried out in the Germination Lab. of the Department of Botany, Institute of Biosciences, São Paulo State University (UNESP), Botucatu, São Paulo State, Brazil. The aims of this work were to determine the water uptake curve and to evaluate the germination of Guazuma ulmifolia seeds subjected to different water potentials. For the water uptake curve, seven replicates of 50 pre-scarified seeds were placed onto paper moistened with 15 mL PEG 6000 solution under the potentials 0 (control), -0.3 and -0.6 MPa at 25o C in the darkness. For the germination assay, four replicates of 50 seeds were subjected to the same above-described conditions; however, one lot of seeds was modified when there was variation in the refractometric index, whereas the remaining ones were kept in the same solutions until the end of the experiment. All three phases of water uptake were detected under 0 and -0.3 MPa; however, phase II was prolonged under -0.6 MPa and germination was not observed. For 0 and -0.3 MPa, the adopted statistical models consisted of asymptotic (phases I and II) and exponential (phase III) functions, y = a*[1 - b*exp (-c*t) + exp (-d + e*(t - t0)]. For -0.6MPa, only the asymptotic function y = a* [1 - b* exp (-c*t)] was used since there was no evidence of germination. The germination final percentage and speed index were lower under -0.3 MPa, mainly when solutions were not replaced; besides, germination was not detected under -0.6 MPa, with or without solution replacement.

An inhibitor of leukotriene synthesis affects vasopressin secretion following osmotic stimulus in rats

Previous studies revealed the presence of LTC4 synthase in paraventricular vasopressinergic neurons, suggesting a role for leukotrienes (LTs) in certain neuroendocrine system functions. Our aim was to study the effect of an inhibitor of LT synthesis in the release of arginine vasopressin (AVP) following an osmotic stimulus in rats. Male Wistar rats received an intra-cerebroventricular injection of 2 mu l of the LT synthesis inhibitor MK-886 (1, 2, or 4 mu g/kg), or vehicle (DMSO 5%), 1 h before an intraperitoneal injection of hypertonic saline (NaCl 2 M) or isotonic saline (NaCl 0.01 M) in a volume corresponding to 1% of body weight. Thirty minutes after the osmotic stimulus, the animals were decapitated and blood was collected for determining hematocrit, plasma osmolality and plasma AVP levels. As expected, the injection of hypertonic saline significantly increased (P<0.05) the hematocrit, plasma osmolality and plasma AVP levels. While inhibiting LT synthesis by central administration of MK-886 did not cause any additional increase in hematocrit or osmolality, plasma AVP levels were augmented (P<0.05). We conclude that central leukotrienes may have a modulatory role in AVP secretion following an osmotic stimulus, this deserving future studies. (C) 2012 Elsevier B.V. All rights reserved.

Cannabinoid CB1 receptor mediates glucocorticoid effects on hormone secretion induced by volume and osmotic changes

1. The present study provides the first in vivo evidence that the cannabinoid CB1 receptor mediates the effects of dexamethasone on hormone release induced by changes in circulating volume and osmolality. Male adult rats were administered with the CB1 receptor antagonist rimonabant (10 mg/Kg, p.o.), followed or not in 1 hour by dexamethasone (1 mg/Kg, i.p.). Extracellular volume expansion (EVE, 2 mL/100 g of body weight, i.v.) was performed 2 hours after dexamethasone or vehicle treatment using either isotonic (I-EVE, 0.15 mol/L) or hypertonic (H-EVE, 0.30 mol/L) NaCl solution. Five minutes after EVE, animals were decapitated and trunk blood was collected for all plasma measurements. 2. Rimonabant potentiated oxytocin (OT) secretion induced by H-EVE and completely reversed the inhibitory effects of dexamethasone in response to the same stimulus. These data suggest that glucocorticoid modulation of OT release is mediated by the CB1 receptor. 3. Although dexamethasone did not affect vasopressin (AVP) secretion induced by H-EVE, the administration of rimonabant potentiated AVP release in response to the same stimulus, supporting the hypothesis that the CB1 receptor regulates AVP secretion independently of glucocorticoid-mediated signalling. 4. Dexamethasone alone did not affect atrial natriuretic peptide (ANP) release stimulated by I-EVE or H-EVE. However, pretreatment with rimonabant potentiated ANP secretion induced by H-EVE, suggesting a possible role for the CB1 receptor in the control of peripheral factors that modulate cardiovascular function. 5. Rimonabant also reversed the inhibitory effects of dexamethasone on H-EVE-induced corticosterone secretion, reinforcing the hypothesis that the CB1 receptor may be involved in the negative feedback exerted by glucocorticoids on the activity of the hypothalamicpituitaryadrenal axis. 6. Collectively, the results of the present study indicate that the CB1 receptor modulates neurohypophyseal hormone secretion and systemic factors, such as corticosterone and ANP, thus participating in homeostatic responses to altered extracellular volume and plasma tonicity.

Influence of Osmotic Dehydration on Ascorbic Acid Loss in Pickled Dry Peppers (Capsicum chinense)

The objective of this work was (1) to develop a dehydrated pepper with 45% humidity, determining the drying curves for pepper, with and without osmotic pre-treatment and (2) to evaluate the influence of both drying and osmotic treatment on the content ascorbic acid (vitamin C) in fresh pepper and pepper with 45% humidity. The experiments were carried out using the peppers cut in half, with and without osmotic pre-treatment, followed by drying in an oven at 70 degrees C. The results showed that the osmotic pretreatment did not influence the retention of ascorbic acid during the drying of pepper. The sensory analysis regarding the color, flavor, and texture attributes revealed that there was no difference in the acceptability.

A comparison between diuretics and angiotensin-receptor blocker agents in patients with stage I hypertension (PREVER-treatment trial): study protocol for a randomized double-blind controlled trial

Abstract Background Cardiovascular disease is the leading cause of death in Brazil, and hypertension is its major risk factor. The benefit of its drug treatment to prevent major cardiovascular events was consistently demonstrated. Angiotensin-receptor blockers (ARB) have been the preferential drugs in the management of hypertension worldwide, despite the absence of any consistent evidence of advantage over older agents, and the concern that they may be associated with lower renal protection and risk for cancer. Diuretics are as efficacious as other agents, are well tolerated, have longer duration of action and low cost, but have been scarcely compared with ARBs. A study comparing diuretic and ARB is therefore warranted. Methods/design This is a randomized, double-blind, clinical trial, comparing the association of chlorthalidone and amiloride with losartan as first drug option in patients aged 30 to 70 years, with stage I hypertension. The primary outcomes will be variation of blood pressure by time, adverse events and development or worsening of microalbuminuria and of left ventricular hypertrophy in the EKG. The secondary outcomes will be fatal or non-fatal cardiovascular events: myocardial infarction, stroke, heart failure, evidence of new subclinical atherosclerosis and sudden death. The study will last 18 months. The sample size will be of 1200 participants for group in order to confer enough power to test for all primary outcomes. The project was approved by the Ethics committee of each participating institution. Discussion The putative pleiotropic effects of ARB agents, particularly renal protection, have been disputed, and they have been scarcely compared with diuretics in large clinical trials, despite that they have been at least as efficacious as newer agents in managing hypertension. Even if the null hypothesis is not rejected, the information will be useful for health care policy to treat hypertension in Brazil. Clinical trials registration number ClinicalTrials.gov: NCT00971165

Influence of osmotic dehydration on ascorbic acid loss in pickled dry peppers (Capsicum chinense)

The objective of this work was (1) to develop a dehydrated pepper with 45% humidity, determining the drying curves for pepper, with and without osmotic pre-treatment and (2) to evaluate the influence of both drying and osmotic treatment on the content ascorbic acid (vitamin C) in fresh pepper and pepper with 45% humidity. The experiments were carried out using the peppers cut in half, with and without osmotic pre-treatment, followed by drying in an oven at 70 ºC. The results showed that the osmotic pretreatment did not influence the retention of ascorbic acid during the drying of pepper. The sensory analysis regarding the color, flavor, and texture attributes revealed that there was no difference in the acceptability.

Advances in understanding of osmotic dehydration and vacuum impregnation of fruits

Osmotic Dehydration and Vacuum Impregnation are interesting operations in the food industry with applications in minimal fruit processing and/or freezing, allowing to develop new products with specific innovative characteristics. Osmotic dehydration is widely used for the partial removal of water from cellular tissue by immersion in hypertonic (osmotic) solution. The driving force for the diffusion of water from the tissue is provided by the differences in water chemical potential between the external solution and the internal liquid phase of the cells. Vacuum Impregnation of porous products immersed in a liquid phase consist of reduction of pressure in a solid-liquid system (vacuum step) followed by the restoration of atmospheric pressure (atmospheric step). During the vacuum step the internal gas in the product pores is expanded and partially flows out while during the atmospheric step, there is a compression of residual gas and the external liquid flows into the pores (Fito, 1994). This process is also a very useful unit operation in food engineering as it allows to introduce specific solutes in the tissue which can play different functions (antioxidants, pH regulators, preservatives, cryoprotectants etc.). The present study attempts to enhance our understanding and knowledge of fruit as living organism, interacting dynamically with the environment, and to explore metabolic, structural, physico-chemical changes during fruit processing. The use of innovative approaches and/or technologies such as SAFES (Systematic Approach to Food Engineering System), LF-NMR (Low Frequency Nuclear Magnetic Resonance), GASMAS (Gas in Scattering Media Absorption Spectroscopy) are very promising to deeply study these phenomena. SAFES methodology was applied in order to study irreversibility of the structural changes of kiwifruit during short time of osmotic treatment. The results showed that the deformed tissue can recover its initial state 300 min after osmotic dehydration at 25 °C. The LF-NMR resulted very useful in water status and compartmentalization study, permitting to separate observation of three different water population presented in vacuole, cytoplasm plus extracellular space and cell wall. GASMAS techniques was able to study the pressure equilibration after Vacuum Impregnation showing that after restoration of atmospheric pressure in the solid-liquid system, there was a reminding internal low pressure in the apple tissue that slowly increases until reaching the atmospheric pressure, in a time scale that depends on the vacuum applied during the vacuum step. The physiological response of apple tissue on Vacuum Impregnation process was studied indicating the possibility of vesicular transport within the cells. Finally, the possibility to extend the freezing tolerance of strawberry fruits impregnated with cryoprotectants was proven.

Na +-coupled betaine symporters involved in osmotic stress response in prokaryotes and eukaryotes

The betaine/GABA transporter BGT1 is one of the most important osmolyte transporters in the kidney. BGT1 is a member of the neurotransmitter sodium symporter (NSS) family, facilitates Na+/Cl--coupled betaine uptake to cope with hyperosmotic stress. Betaine transport in kidney cells is upregulated under hypertonic conditions by a yet unknown mechanism when increasing amounts of intracellular BGT1 are inserted into the plasma membrane. Re-establishing isotonicity results in ensuing depletion of BGT1 from the membrane. BGT1 phosphorylation on serines and threonines might be a regulation mechanism. In the present study, four potential PKC phosphorylation sites were mutated to alanines and the responses to PKC activators, phorbol 12-myristate acetate (PMA) and dioctanoyl-sn-glycerol (DOG) were determined. GABA-sensitive currents were diminished after 30 min preincubation with these PKC activators. Staurosporine blocked the response to DOG. Three mutants evoked normal GABA-sensitive currents but currents in oocytes expressing the mutant T40A were greatly diminished. [3H]GABA uptake was also determined in HEK-293 cells expressing EGFP-tagged BGT1 with the same mutations. Three mutants showed normal upregulation of GABA uptake after hypertonic stress, and downregulation by PMA was normal compared to EGFP-BGT1. In contrast, GABA uptake by the T40A mutant showed no response to hypertonicity or PMA. Confocal microscopy of the EGFP-BGT1 mutants expressed in MDCK cells, grown on glass or filters, revealed that T40A was present in the cytoplasm after 24 h hypertonic stress while the other mutants and EGFP-BGT1 were predominantely present in the plasma membrane. All four mutants co-migrated with EGFP-BGT1 on Western blots suggesting they are full-length proteins. In conclusion, T235, S428, and S564 are not involved in downregulation of BGT1 due to phosphorylation by PKC. However, T40 near the N-terminus may be part of a hot spot important for normal trafficking or insertion of BGT1 into the plasma membrane. Additionally, a link between substrate transport regulation, insertion of BGT1 into the plasma membrane and N-glycosylation in the extracellular loop 2 (EL2) could be revealed. The functional importance of two predicted N-glycosylation sites, which are conserved in EL2 within the NSS family were investigated for trafficking, transport and regulated plasma membrane insertion by immunogold-labelling, electron microscopy, mutagenesis, two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radioactive-labelled substrate into MDCK cells. Trafficking and plasma membrane insertion of BGT1 was clearly promoted by proper N-glycosylation in both, oocytes and MDCK cells. De-glycosylation with PNGase F or tunicamycin led to a decrease in substrate affinity and transport rate. Mutagenesis studies revealed that in BGT1 N183 is the major N-glycosylation site responsible for full protein activity. Replacement of N183 with aspartate resulted in a mutant, which was not able to bind N-glycans suggesting that N171 is a non-glycosylated site in BGT1. N183D exhibited close to WT transport properties in oocytes. Surprisingly, in MDCK cells plasma membrane insertion of the N183D mutant was no longer regulated by osmotic stress indicating unambiguously that association with N-glycans at this position is linked to osmotic stress-induced transport regulation in BGT1. The molecular transport mechanism of BGT1 remains largely unknown in the absence of a crystal structure. Therefore investigating the structure-function relationship of BGT1 by a combination of structural biology (2D and 3D crystallization) and membrane protein biochemistry (cell culture, substrate transport by radioactive labeled GABA uptake into cells and proteoliposomes) was the aim of this work. While the functional assays are well established, structure determination of eukaryotic membrane transporters is still a challenge. Therefore, a suitable heterologous expression system could be defined, starting with cloning and overexpression of an optimized gene. The achieved expression levels in P. pastoris were high enough to proceed with isolation of BGT1. Furthermore, purification protocols could be established and resulted in pure protein, which could even be reconstituted in an active form. The quality and homogeneity of the protein allowed already 2D and 3D crystallization, in which initial crystals could be obtained. Interestingly, the striking structural similarity of BGT1 to the bacterial betaine transporter BetP, which became a paradigm for osmoregulated betaine transport, provided information on substrate coordination in BGT1. The structure of a BetP mutant that showed activity for GABA was solved to 3.2Å in complex with GABA in an inward facing open state. This structure shed some light into the molecular transport mechanisms in BGT1 and might help in future to design conformationally locked BGT1 to enforce the on-going structure determination.