Osmolality Of Solution Research Articles

Promotion of Germination Using Hydroxamic Acid Inhibitors of 9-cis-Epoxycarotenoid Dioxygenase.

Abscisic acid (ABA) inhibits seed germination and the regulation of ABA biosynthesis has a role in maintenance of seed dormancy. The key rate-limiting step in ABA biosynthesis is catalyzed by 9-cis-epoxycarotenoid dioxygenase (NCED). Two hydroxamic acid inhibitors of carotenoid cleavage dioxygenase (CCD), D4 and D7, previously found to inhibit CCD and NCED in vitro, are shown to have the novel property of decreasing mean germination time of tomato (Solanum lycopersicum L.) seeds constitutively overexpressing LeNCED1. Post-germination, D4 exhibited no negative effects on tomato seedling growth in terms of height, dry weight, and fresh weight. Tobacco (Nicotiana tabacum L.) seeds containing a tetracycline-inducible LeNCED1 transgene were used to show that germination could be negatively and positively controlled through the chemical induction of gene expression and the chemical inhibition of the NCED protein: application of tetracycline increased mean germination time and delayed hypocotyl emergence in a similar manner to that observed when exogenous ABA was applied and this was reversed by D4 when NCED expression was induced at intermediate levels. D4 also improved germination in lettuce (Lactuca sativa L.) seeds under thermoinhibitory temperatures and in tomato seeds imbibed in high osmolarity solutions of polyethylene glycol. D4 reduced ABA and dihydrophaseic acid accumulation in tomato seeds overexpressing LeNCED1 and reduced ABA accumulation in wild type tomato seeds imbibed on polyethylene glycol. The evidence supports a mode of action of D4 through NCED inhibition, and this molecule provides a lead compound for the design of NCED inhibitors with greater specificity and potency.

Low osmolality and shear stress during liposuction impair cell viability in autologous fat grafting

Liposuction and subsequent autologous fat grafting have become essential techniques for fat augmentation in plastic surgery. However, standard harvesting techniques that ensure the survival of adipocytes and stromal vascular fraction (SVF) cells and thus preserve the transplanted fat volume are lacking. In particular, the effect of different parameters of the tumescent solution has not been studied in this context. We hypothesized that the osmolality of the tumescent solution could have a significant effect on the survival of adipocytes and SVF cells. We developed two distinct invitro models based on freshly harvested excision fat from patients undergoing surgical treatment. First, we investigated the effect of osmolality by incubating excision fat in different tumescent solutions and analyzed the total cell survival and the differentiation potential of SVF cells. Vital whole-mount staining, isolation yield of SVF cells, clonogenicity, and osteogenic and adipogenic differentiation capacities were analyzed. Second, we addressed the additional effect of mechanical stress by simulating a liposuction on pieces of excision fat after incubation with the tumescent solutions. Osmolality of the tumescent solution by itself did not have a significant effect on adipocyte and SVF viability or SVF differentiation. However, when osmolality was combined with liposuction, a significant trend toward lower viability and more lipid droplets with lower osmolality was observed. Especially, SVF viability was significantly lower after liposuction with a hypotonic (150mOsm/kg) solution. This study demonstrates the considerable effect of osmolality during liposuction and may lead to the development of "cell-protective" tumescent solutions.

Physico-chemical properties of late-incubation egg amniotic fluid and a potential in ovo feed supplement.

ObjectiveThis study explored the physico-chemical properties of late-incubation egg amniotic fluid and a potential in ovo feed (IOF) supplement.MethodsAmniotic fluid was collected from broiler breeders (Ross 308, 51 weeks and Cobb 500, 35 weeks) on day 17 after incubation. A mixture of high-quality soy protein supplement – Hamlet Protein AviStart (HPA) was serially diluted in MilliQ water to obtain solutions ranging from 150 to 9.375 mg/mL. The mixtures were heat-treated (0, 30, 60 minutes) in a waterbath (80°C) and then centrifuged to obtain supernatants. The amniotic fluid and HPA supernatants were analysed for their physico-chemical properties.ResultsOnly viscosity and K+ were significantly (p<0.05) different in both strains. Of all essential amino acids, leucine and lysine were in the highest concentration in both strains. The osmolality, viscosity and pCO2 of the supernatants decreased (p<0.05) with decreasing HPA concentration. Heat treatment significantly (p<0.05) affected osmolality, pH, and pCO2, of the supernatants. The interactions between HPA concentration and heat treatment were significant with regards to osmolality (p<0.01), pH (p<0.01), pCO2 (p<0.05), glucose (p<0.05), lactate (p<0.01) and acid-base status (p<0.01) of HPA solutions. The Ca2+, K+, glucose, and lactate increased with increasing concentration of HPA solution. The protein content of HPA solutions decreased (p<0.05) with reduced HPA solution concentrations. The supernatant from 150 mg/mL HPA solution was richest in glutamic acid, aspartic acid, arginine and lysine. Amino acids concentrations were reduced (p<0.05) with each serial dilution but increased with longer heating.ConclusionThe values obtained in the primary solution (highest concentration) are close to the profiles of high-protein ingredients. This supplement, as a solution, hence, may be suitable for use as an IOF supplement and should be tested for this potential.

Comparison of Purification Solutions With Different Osmolality for Porcine Islet Purification.

The osmolality of the purification solution is one of the most critical variables in human islet purification during islet isolation. We previously reported the effectiveness of a combined continuous density/osmolality gradient for the supplemental purification of human islets. We herein applied a combined continuous density/osmolality gradient for regular purification. The islets were purified with a continuous density gradient without osmolality preparation [continuous density/normal osmolality (CD/NO)] or continuous density/osmolality solution with osmolality preparation by 10× Hank's balanced salt solution (HBSS) [continuous density/continuous osmolality (CD/CO)]. The osmolality of the low-density solution was 400 mOsm/kg in both groups and that of the high-density solution was 410 mOsm/kg in the CD/NO group and 500 mOsm/kg in the CD/CO group. Unexpectedly, we noted no significant differences between the two solutions in terms of the islet yield, rate of viability and purity, score, stimulation index, or the attainability and suitability of posttransplantation normoglycemia. Despite reports that the endocrine and exocrine tissues of pancreata have distinct osmotic sensitivities and that high-osmolality solutions result in greater purification efficiency, the isolation and transplant outcomes did not markedly differ between the two purification solutions with different osmolalities in this study.

Randomized, masked, in vitro comparison of three commercially available tear film osmometers.

PurposeThe purpose of this study was to compare the precision and accuracy of commercially available tear film osmometers.MethodsContrived tear solution target values representing the physiological range of tear osmolarity (normal eyes 297 mOsm/L, moderately dry eyes 342 mOsm/L, and severe dry eyes 383 mOsm/L) were constructed using a mix of mono- and divalent electrolytes, metabolites, serum albumin, and pH balanced to 7.4. Solution values were randomized and masked from the investigators during testing. Osmometers (Wescor 5520 Vapro Pressure Osmometer: device A, TearLab Osmolarity System: device B, and i-Med Pharma i-Pen: device C) were calibrated according to manufacturer instructions. Each level was tested 64× on each osmometer across two sites. Accuracy was reported as a correlation coefficient against expected linear dilutions, precision was calculated as percent coefficient of variation.ResultsDevice A reported a correlation with known solutions of r2=0.98, with averages of 305.6±4.0, 352.2±5.5, and 389.8±4.0 mOsm/L, and coefficient of variations (CVs) of 1.3%, 1.6%, and 1.0%, respectively. Device B reported an r2=0.96, with averages of 300.6±3.7, 341.4±7.9, and 376.8±5.1 mOsm/L, and CVs of 1.2%, 2.3%, and 1.4%, respectively. Device C reported an r2=0.03, with averages of 336.4±21.5, 342.0±20.7, and 345.7±22.0 mOsm/L, and CVs of 6.4%, 6.1%, and 6.4%, respectively.ConclusionIn this randomized, masked, in vitro study, device A and device B had significantly better accuracy and precision in measuring osmolarity of contrived tear solutions of known target values compared to device C. Device C showed insufficient performance to accurately and precisely delineate osmolarity levels in the physiological range. Furthermore, in vivo studies would be required to compare performance in human subjects.

Osmotic and Heat Stress Effects on Segmentation.

During vertebrate embryonic development, early skin, muscle, and bone progenitor populations organize into segments known as somites. Defects in this conserved process of segmentation lead to skeletal and muscular deformities, such as congenital scoliosis, a curvature of the spine caused by vertebral defects. Environmental stresses such as hypoxia or heat shock produce segmentation defects, and significantly increase the penetrance and severity of vertebral defects in genetically susceptible individuals. Here we show that a brief exposure to a high osmolarity solution causes reproducible segmentation defects in developing zebrafish (Danio rerio) embryos. Both osmotic shock and heat shock produce border defects in a dose-dependent manner, with an increase in both frequency and severity of defects. We also show that osmotic treatment has a delayed effect on somite development, similar to that observed in heat shocked embryos. Our results establish osmotic shock as an alternate experimental model for stress, affecting segmentation in a manner comparable to other known environmental stressors. The similar effects of these two distinct environmental stressors support a model in which a variety of cellular stresses act through a related response pathway that leads to disturbances in the segmentation process.

Osmoregulated Chloride Currents in Hemocytes from Mytilus galloprovincialis.

We investigated the biophysical properties of the transport mediated by ion channels in hemocytes from the hemolymph of the bivalve Mytilus galloprovincialis. Besides other transporters, mytilus hemocytes possess a specialized channel sensitive to the osmotic pressure with functional properties similar to those of other transport proteins present in vertebrates. As chloride fluxes may play an important role in the regulation of cell volume in case of modifications of the ionic composition of the external medium, we focused our attention on an inwardly-rectifying voltage-dependent, chloride-selective channel activated by negative membrane potentials and potentiated by the low osmolality of the external solution. The chloride channel was slightly inhibited by micromolar concentrations of zinc chloride in the bath solution, while the antifouling agent zinc pyrithione did not affect the channel conductance at all. This is the first direct electrophysiological characterization of a functional ion channel in ancestral immunocytes of mytilus, which may bring a contribution to the understanding of the response of bivalves to salt and contaminant stresses.

Osmolality of antisense oligonucleotide parenteral formulations: Implications on counterion dissociation and recommended osmometry techniques

The intrinsic osmolality of aqueous solutions of sodium salt antisense oligonucleotides (ASOs) has been studied to inform formulation practices, understand the molecular basis underlying the difference between theoretical and empirical results, and determine suitable measurement methods. It was found that regardless of nucleotide sequence, ASO concentration of ∼140mg/mL has isotonic osmolality of ∼290mOsm/kg water (SI unit: mmol osmotically-active particles/kg water), such that lower concentration formulations require excipients for tonicity adjustment. The range of osmolality values at a given active ingredient concentration can be ascribed to drug substance lot-to-lot purity differences impacting total oligonucleotide content (i.e., including oligonucleotide-related impurities). Empirical osmolality measurements were found to be ∼70% of theoretical values, which corresponds to an osmotic coefficient value of ∼0.7, thus inferring incomplete counterion dissociation. When comparing theoretical (ideal) osmolality of multiple sequences with various nucleotide compositions and chemistries at the same w/v concentration, the “average osmolar mass” (molar mass of the oligonucleotide, including the sodium counterions, divided by the ideal Van’t Hoff factor, iid) appears to be the strongest factor governing theoretical osmolality values. Other factors examined were the sequence length, backbone chemistry, 2′ sugar chemistry, and nucleotide composition. A head-to-head comparison between two osmolality techniques showed that vapor pressure osmometry is generally more suitable than freezing point osmometry for oligonucleotide solutions greater than ∼150mg/mL due to viscosity effects, but the two techniques are comparable otherwise.

Development of sperm cryopreservation protocol of endangered spiny eel, Mastacembelus armatus (Lacepede 1800) for ex-situ conservation

The study was conducted to develop sperm cryopreservation protocol for the endangered spiny eel, Mastacembelus armatus as well as to use the cryopreserved sperm in breeding. Activation of sperm was evaluated by observing motility in different osmolalities of NaCl solution (48–319 mOsmol/kg) and found that sperm motility decreased with increasing osmolality of the NaCl solution and it was totally inhibited at 319 mOsmol/kg. The highest motility and swimming duration of sperm were observed as 14.5 ± 0.5 min and 4.75 ± 0.25 min at 128 mOsmol/kg and 67 mOsmol/kg, respectively. The concentration of sperm was found to be 1.75 × 1010 to 2.25 × 1010 cells/ml. Evaluation of toxicity of cryoprotectants of dimethyl sulfoxide (Me2SO) and methanol to sperm was performed at 5, 10 and 15% concentrations during 5–30 min incubation period and found that cryoprotectants with 5 and 10% concentrations produced better motility up to 10 min incubation. In the selection of suitable diluent for cryopreservation of sperm, Alsever's solution with 10% Me2SO performed best producing 91.67 ± 1.67% and 65 ± 2.89% equilibration and post-thaw motility, respectively. In breeding trials, sperm preserved with Alsever's solution plus Me2SO produced more or less similar fertilization (82.25 ± 4.46%) and hatching (12 ± 0.65%) to those preserved with 0.9% NaCl solution plus Me2SO as 79.88 ± 4.67% and 11 ± 0.46% fertilization and hatching respectively. Fresh sperm used as control produced 87.63 ± 3.6% and 14 ± 0.5% fertilization and hatching respectively. Both cryopreserved and fresh sperm-derived fry performed well but the control fry showed a significantly better (P < 0.05) growth. Fry derived from four diluents did not show any significant (P > 0.05) growth difference but Alsever's solution plus Me2SO produced better results.

Osmolarity and spectrophotometric property of brilliant blue green define the degree of toxicity on retinal pigment epithelial cells exposed to surgical endoilluminator.

ObjectiveTo evaluate the effect of varying concentrations of brilliant blue green (BBG) and their different biochemical characteristics on retinal pigment epithelial (RPE) cells under xenon light source illumination at varying distances to identify safe parameters for intraoperative use.MethodsHuman retinal RPE cells (ARPE-19) were exposed to two concentrations (0.25 and 0.50 mg/mL) of BBG and illuminated with a xenon surgical illuminator at varying distances (10 and 25 mm), intensity levels, and time intervals (1, 5, and 15 minutes). Additionally, the effect of osmolarity was examined by diluting BBG in different concentrations of glucose. Cytotoxicity of BBG and osmolarity effects on cell viability were evaluated using a WST-1 assay. Light absorption and emission characteristic of BBG in different solvents were measured using a plate reader at different wavelengths. Lastly, the activity of caspase-3 was also studied.ResultsCell viability of ARPE-19 cells was 77.4%±12.7%, 78.7%±17.0%, and 65.0%±19.7% at 1, 5, and 15 minutes to exposure of high illumination xenon light at 10 mm (P<0.05) compared to controls. At both distances of illumination (10 and 25 mm), similar cell viabilities were seen between 1 and 5 minutes of exposure. However, there was a decline in viability when the illumination was carried out to 15 minutes in all groups (P<0.05). There was no significant reduction in cell viability in presence or absence of xenon light in different osmolar solutions concentrations of glucose (P>0.05). Maximal light absorption of BBG was noted between 540 and 680 nm. Activated caspase-3 level was not significant in both the concentrations of BBG (P>0.05).ConclusionOur findings suggest that BBG at 0.25 mg/mL during vitreoretinal surgery is safe and not toxic to RPE cells up to 5 minutes under focal high illumination (10 mm) and up to 15 minutes under medium diffuse illumination (25 mm). BBG was safe to be mixed with isotonic glucose solution at the concentration range of 2.5%–10%, regardless of the illumination status.

تقييم ممارسات الأمهات في حالات الاسهال عند الأطفال وقياس الأسمولية وPH لبعض محاليل الجفاف والمشروبات الغازية

The aims of this study were to evaluate the mothers’ practice in case of children diarrhea and measure osmolality and PH of some oral rehydration solutions (ORS) and carbonated beverages. The study consisted of two parts; the first part was a face to face questionnaire that was conducted at a group of community pharmacies between July and November 2013. In the second part, Vapor Pressure Osmometer (VAPRO) was used for osmolality determination, while 3520 pH meter was used for the pH determination of some ORS and carbonated beverages. A total of 200 mothers were surveyed in the study. Among them, 85.0% told that they keep breastfeeding during diarrhea. During diarrhea, around half of the mothers (102; 51.0%) told that they give their children potato and (129; 64.5%) told that they avoid sweets, 42.5% told that they use ORS and 47.5% told that they use carbonated beverages. The osmolality of all carbonated beverages tested (except for diet coca cola) and two ORS was higher than the level recommended by the World Health Organization for rehydration solutions (210-268 mOsm/L). A wide range of variability in PH was found. The values ranged from 3.98 to 8.86. It can be concluded that mothers’ practice in case of diarrhea is good in general; however, there is a room for improvement. Continuation of breast feeding and the use of suitable ORS should be encouraged. Use of carbonated beverages should be discouraged. كانت أهداف هذه الدراسة تقييم ممارسات الأمهات في حالات الاسهال عند الأطفال وقياس الأسمولية وPH لبعض محاليل الجفاف والمشروبات الغازية. وتكونت الدراسة من جزأين؛ كان الجزء الأول عبارة عن استبيان وجها لوجه والذي أجري على مجموعة من صيدليات المجتمع بين تموز وتشرين الثاني عام 2013. وفي الجزء الثاني، تم قياس الاسموزية و درجة الحموضة لعدد من محاليل الجفاف والمشروبات الغازية. تم مسح ما مجموعه 200 الأمهات في الدراسة. وقال 85٪ من بينهن أنها تبقي الرضاعة الطبيعية خلال الإسهال. ما يقرب من نصف الأمهات قلن 51.0٪ أنهن يعطين أطفالهن البطاطا اثناء الاسهال و64.5٪ قلن أنهن يتجنبن الحلويات، وقال 42.5٪ أنهن يستخدمن محاليل الجفاف لاطفالهن بينما ذكر 47.5٪ أنهن يستخدمن المشروبات الغازية. وكانت الأسموزية لجميع المشروبات الغازية (باستثناء كوكا كولا دايت) واثنين من محاليل الجفاف أعلى من المستوى الموصى به من قبل منظمة الصحة العالمية (210-268 ميلي أسمول/ لتر) و كان هناك تباين في درجة الحموضة وتراوحت القيم بين 3.98-8.86. ويمكن أن نخلص إلى أن ممارسة الأمهات في حالة الإسهال جيدة بصفة عامة؛ ومع ذلك، هناك مجال للتحسين. وينبغي تشجيع استمرار الرضاعة الطبيعية واستخدام محاليل جفاف مناسبة. وينبغي الامتناع عن استخدام المشروبات الغازية.

Cryoprotectant kinetic analysis of a human articular cartilage vitrification protocol

We recently published a protocol to vitrify human articular cartilage and a method of cryoprotectant removal in preparation for transplantation. The current study’s goal was to perform a cryoprotectant kinetic analysis and theoretically shorten the procedure used to vitrify human articular cartilage. First, the loading of the cryoprotectants was modeled using Fick’s law of diffusion, and this information was used to predict the kinetics of cryoprotectant efflux after the cartilage sample had been warmed. We hypothesized that diffusion coefficients obtained from the permeation of individual cryoprotectants into porcine articular cartilage could be used to provide a reasonable prediction of the cryoprotectant loading and of the combined cryoprotectant efflux from vitrified human articular cartilage. We tested this hypothesis with experimental efflux measurements. Osteochondral dowels from three patients were vitrified, and after warming, the articular cartilage was immersed in 3 mL X-VIVO at 4 °C in two consecutive solutions, each for 24 h, with the solution osmolality recorded at various times. Measured equilibrium values agreed with theoretical values within a maximum of 15% for all three samples. The results showed that diffusion coefficients for individual cryoprotectants determined from experiments with 2-mm thick porcine cartilage can be used to approximate the rate of efflux of the combined cryoprotectants from vitrified human articular cartilage of similar thickness. Finally, Fick’s law of diffusion was used in a computational optimization to shorten the protocol with the constraint of maintaining the theoretical minimum cryoprotectant concentration needed to achieve vitrification. The learning provided by this study will enable future improvements in tissue vitrification.

Effect of the osmolarity change in multipurpose solutions induced by an improper contact lens case cleaning procedure.

To determine whether variations in multipurpose disinfecting solution (MPDS) storage osmolarity from inappropriate contact lens (CL) case cleaning affect ocular surface integrity and wearer comfort. There were twenty contact lens cases (study CLCs) in the study group. Ten were filled with ReNu Multiplus(®) and 10 with SoloCare Aqua™ (MPDS-1 and -2, respectively) and kept closed for 8h; the cases were then emptied and kept open for air-drying for 16h. This procedure was carried out every day for two months. Storage solution osmolarity was measured on days 0, 15, 30, 45 and 60. Ten subjects were then fitted with both month-old lenses stored in the study CLCs and with new lenses stored in new cases with fresh solution for 24h (control CLCs). Symptoms, tear osmolarity and percentage of subjects whose conjunctival hyperaemia and ocular surface staining scores changed were determined after 1h of wear. Study CLC osmolarity increased in both solutions after two months (p<0.05). For MPDS-1 there were differences in stinging between study CLCs and control CLCs after 10min of CL wear (p=0.04), and in comfort after 10 (p=0.035) and 60min wear (p=0.042). Significant (p<0.05) differences between study CLC and control CLC groups were also found for MPDS-2 in limbal hyperaemia (study: 50% change; control: 0% change) and bulbar and corneal staining (study: 80% change; control: 20% change). The stored-MPDS osmolarity increase caused by air-drying the CLCs could affect the ocular surface. This increase might reduce lens wear comfort.

Investigation of the Effects of Extracellular Osmotic Pressure on Morphology and Mechanical Properties of Individual Chondrocyte.

It has been demonstrated that most cells of the body respond to osmotic pressure in a systematic manner. The disruption of the collagen network in the early stages of osteoarthritis causes an increase in water content of cartilage which leads to a reduction of pericellular osmolality in chondrocytes distributed within the extracellular environment. It is therefore arguable that an insight into the mechanical properties of chondrocytes under varying osmotic pressure would provide a better understanding of chondrocyte mechanotransduction and potentially contribute to knowledge on cartilage degeneration. In this present study, the chondrocyte cells were exposed to solutions with different osmolality. Changes in their dimensions and mechanical properties were measured over time. Atomic force microscopy (AFM) was used to apply load at various strain-rates and the force-time curves were logged. The thin-layer elastic model was used to extract the elastic stiffness of chondrocytes at different strain-rates and at different solution osmolality. In addition, the porohyperelastic (PHE) model was used to investigate the strain-rate-dependent responses under the loading and osmotic pressure conditions. The results revealed that the hypo-osmotic external environment increased chondrocyte dimensions and reduced Young's modulus of the cells at all strain-rates tested. In contrast, the hyper-osmotic external environment reduced dimensions and increased Young's modulus. Moreover, using the PHE model coupled with inverse FEA simulation, we established that the hydraulic permeability of chondrocytes increased with decreasing extracellular osmolality which is consistent with previous work in the literature. This could be due to a higher intracellular fluid volume fraction with lower osmolality.

Photosynthetic carbon metabolism in potato leaves under changes in light intensity

Photosynthetic assimilation of 14CO2 was examined in leaves of potato (Solanum tuberosum L.) plants that were grown under direct sunlight and then transferred to 50% irradiance for various periods. The rate of 14CO2 assimilation correlated with light intensity: the photosynthetic rate reduced by 52% after 5-day shading and by 70% after 30-min shading. In all shaded and shade-adapted plants, the sucrose/hexose ratio decreased by a factor of 3.5–4.1; furthermore, the radioactivity of glycolate cycle metabolites and the serine/glycine ratio were lowered. In plants shaded for 5 days or 30 min, the radioactivity of aspartate and malate was higher than at continuous high irradiance, especially in plants shaded for 30 min, whereas a sudden illumination of the shaded plants reduced the radioactivity of these substances. We suppose that low irradiance averted the reentry of glycolate path carbon into the Calvin cycle and redirected this carbon source for the production of four-carbon acids that acidified the apoplast. This acidification activated the apoplastic invertase, which enhanced sucrose hydrolysis and hindered the sucrose export from the leaf. Hydrolysis of sucrose promoted the increase in osmolarity of the apoplastic solution, this increase being stronger at close distances to the stomatal pores where water is intensely evaporated. The increase in osmolarity of extracellular medium led to closing of stomata and the suppression of photosynthesis.

AGREEMENT BETWEEN DIFFERENT EQUATIONS TO ESTIMATE OSMOLARITY OF PARENTERAL NUTRITION SOLUTIONS.

our aim was to measure the osmolality of several PN formulas at different component concentrations to determine if equations described in literature to calculate osmolarity accurately predict osmolalality in other experimental conditions different than these used to develop them. osmolality of 12 different types of PN solutions, 9 for central and 3 for peripheral perfusion were measured by using freezing point depression in cross-sectional study. We evaluated the agreement (Pearson correlation test) and differential bias between measured osmolality and calculated osmolarity for three different equations described in the literature: Pereira Da Silva, ASPEN Practice Manual and ASPEN guidelines. mean ± SD osmolality of PN solutions was 1789 ± 256 (range 1 540 - 2 372) and 751 ± 64 mOsm/kg (range 689 - 817) for central and peripheral infusion, respectively. The osmolality of PN formulations was mainly due to glucose (r = 0.975) and amino acids (r = 0.948). All studied equations had a good correlation in the bivariate analysis (p = 0.000). All equations had a trend to underestimate the osmolality compared with the measured value. However, ASPEN guidelines equation overestimated the osmolality for peripheral PN. measurement of osmolality of peripheral PN solutions is important to reduce the risk of phlebitis. The different equations described previously show a good correlation between them although in general underestimate the osmolality.

Concept of relative variability of cardiac action potential duration and its test under various experimental conditions.

Beat-to-beat variability of action potential duration (short-term variability, SV) is an intrinsic property of mammalian myocardium. Since the majority of agents and interventions affecting SV may modify also action potential duration (APD), we propose here the concept of relative SV (RSV), where changes in SV are normalized to changes in APD and these data are compared to the control SV-APD relationship obtained by lengthening or shortening of action potentials by inward and outward current injections. Based on this concept the influence of the several experimental conditions like stimulation frequency, temperature, pH, redox-state and osmolarity were examined on RSV in canine ventricular myocytes using sharp microelectrodes. RSV was increased by high stimulation frequency (cycle lengths <0.7 s), high temperature (above 37ºC), oxidative agents (H2O2), while it was decreased by reductive environment. RSV was not affected by changes in pH (within the range of 6.4-8.4) and osmolarity of the solution (between 250-350 mOsm). The results indicate that changes in beat-to-beat variability of APD must be evaluated exclusively in terms of RSV; furthermore, some experimental conditions, including the stimulation frequency, redox-state and temperature have to be controlled strictly when analyzing alterations in the short-term variability of APD.

Oxygen permeability of soft contact lenses in different pH, osmolality and buffering solution.

To determine the effect of pH, osmolality, and buffering system on the oxygen permeability (Dk) of soft contact lenses. Two hydrogel lenses (nelfilcon A and etafilcon A) and 2 silicone hydrogel lenses (lotrafilcon A and balafilcon A) were used in the study. These lenses were incubated in phosphate-buffered saline (PBS) and borate-buffered saline (BBS) solutions adjusted by 0.8 pH increments to a pH in the range of 5.8-9.0 or in hypotonic (280 mOsmol/kg), isotonic (310 mOsmol/kg) and hypertonic (380 mOsmol/kg) PBS solutions. Polarographic method was used for measuring the Dk and lenses were stacked as 4 layers to correct the boundary effect. Dk values of all contact lenses measured in BBS solutions were more stable than those in PBS solutions. Especially the etafilcon A lens showed a relative big change compared with other types of contact lenses at the same conditions. When the osmolality of PBS solution increased from hypotonic to hypertonic, Dk of all contact lenses decreased. Variations in Dk existed depending on lens materials, etafilcon A lens was the most affected and nelfilcon A was the least affected by osmolality. From the result obtained, it is revealed that Dk of contact lenses is changed by the pH, osmolality, and buffering condition of tear. Thus, Dk of contact lens can be varied by the lens wearers' physiological and/or pathological conditions.

Impact of Luminal Fluid Volume on the Drug Absorption After Oral Administration: Analysis Based on In Vivo Drug Concentration–Time Profile in the Gastrointestinal Tract

The objective of this study is to clarify the influence of fluid volume in the gastrointestinal (GI) tract on the oral drug absorption. In vivo rat luminal concentrations of FITC-dextran (FD-4), a nonabsorbable marker, and drugs (metoprolol and atenolol) after oral coadministration as solutions with different osmolarity were determined by direct sampling of residual water in each segment of the GI tract. The luminal FD-4 concentration after oral administration as hyposmotic solution was significantly higher than that after administration as isosmotic or hyperosmotic solution. As the change in FD-4 concentration reflects the change in the volume of luminal fluid, it indicated that the luminal volume was greatly influenced by osmolality of solution ingested orally. Then, fraction of drug absorbed (Fa) in these segments was calculated by comparing the area under the luminal concentration-time curve of FD-4 with those of drugs. Fa values of two model drugs in each GI segment decreased with increase in luminal fluid volume, and the impact of the fluid volume was marked for Fa of atenolol (a low permeable drug) than for that of metoprolol (a high permeable drug). These findings should be beneficial to assure the effectiveness and safety of oral drug therapy.

Quantifying Hydrostatic Pressure in Plant Cells by Using Indentation with an Atomic Force Microscope

Plant cell growth depends on a delicate balance between an inner drive—the hydrostatic pressure known as turgor—and an outer restraint—the polymeric wall that surrounds a cell. The classical technique to measure turgor in a single cell, the pressure probe, is intrusive and cannot be applied to small cells. In order to overcome these limitations, we developed a method that combines quantification of topography, nanoindentation force measurements, and an interpretation using a published mechanical model for the pointlike loading of thin elastic shells. We used atomic force microscopy to estimate the elastic properties of the cell wall and turgor pressure from a single force-depth curve. We applied this method to onion epidermal peels and quantified the response to changes in osmolality of the bathing solution. Overall our approach is accessible and enables a straightforward estimation of the hydrostatic pressure inside a walled cell.