OSMOTIC RELATIONSHIPS IN THE HEN'S EGG,AS DETERMINED BY COLLIGATIVE PROPERTIES OF YOLK AND WHITE

The osmotic pressure of the yolk and white of the hen''s egg have been shown to be identical, by means of direct freezing point determinations, dialyses, and vapor pressure measurements. Dialysates of egg yolk slow the rate of ice formation compared with NaCl solutions. They also show a marked change of freezing rate as the freezing point is approached. The anomalous freezing behavior of this material may lead to errors in the determination of the true freezing point which would tend to make the value for the yolk erroneously low. The postulate of a vital activity at the yolk membrane maintaining an osmotic pressure difference is thus shown to be unnecessary, since a simple osmotic equilibrium exists between the yolk and the white.     

Measuring Osmotic Pressure of Sap within Live Cells by Means of a Visual Melting Point Apparatus

A freezing slide apparatus is described for visual observation of freezing water and melting ice within plant cells. The slide consists of an ordinary microscope slide glued into a Plexiglass jacket, through which cold 90% ethyl alcohol is pumped at varying rates for temperature control. Temperature is recorded by means of an iron-constantan thermocouple wire (25-micron diameter) connected to a recording potentiometer. Tissue strips were quick frozen (at a cooling rate of 33 C per ½ minute) and then warmed very slowly (at a rate of 2 C per minute) for observation of melting points. This apparatus has been used to determine osmotic pressures of cell sap of guard and adjoining epidermal cells of Chrysanthemum morifolium and Pelargonium hortorum. An accuracy of ± 1.2 atmospheres is possible. Wide variations among osmotic pressures of both guard and epidermal cells were found at any one stomatal aperture in both species.     

Is Aboriginal Food Less Allergenic? Comparing IgE-Reactivity of Eggs from Modern and Ancient Chicken Breeds in a Cohort of Allergic Children

Background

Hen''s egg allergy ranks among the most frequent primary food allergies in children. We aimed to investigate sensitization profiles of egg allergic patients and compare in vitro IgE reactivities of eggs from ancient chicken breeds (Araucana and Maran) with those from conventional laying hen hybrids.

Methodology

Egg allergic children (n = 25) were subjected to skin prick test, double blind placebo controlled food challenge, and sensitization profiles to Gal d 1–5 were determined by allergen microarray. IgE binding and biological activity of eggs from different chicken breeds were investigated by immunoblot, ELISA, and mediator release assays.

Principal Findings

We found that Gal d 1 and Gal d 2 are generally major egg allergens, whereas Gal d 3–5 displayed high sensitization prevalence only in patients reacting to both, egg white and yolk. It seems that the onset of egg allergy is mediated by egg white allergens expanding to yolk sensitization in later stages of disease. Of note, egg white/yolk weight ratios were reduced in eggs from Auraucana and Maran chicken. As determined in IgE immunoblots and mass analysis, eggs from ancient chicken breeds did not differ in their protein composition. Similar IgE-binding was observed for all egg white preparations, while an elevated allergenicity was detected in egg yolk from Araucana chicken.

Conclusion/Significance

Our results on allergenicity and biological activity do not confirm the common assumption that aboriginal food might be less allergenic. Comprehensive diagnosis of egg allergy should distinguish between reactivity to hen''s egg white and yolk fractions to avoid unnecessary dietary restrictions to improve life quality of the allergic child and its family.     

OSMOTIC RELATIONSHIPS IN THE HEN'S EGG AS DETERMINED BY RELATIVE VAPOR PRESSURES

It has been shown by a comparison of the relative vapor pressures of egg yolk and egg white before and after the addition of sodium chloride to the white that the osmotic pressure of the yolk is greater than that of the white.     

Theoretical and experimental studies on freezing point depression and vapor pressure deficit as methods to measure osmotic pressure of aqueous polyethylene glycol and bovine serum albumin solutions

For survival in adverse environments where there is drought, high salt concentration or low temperature, some plants seem to be able to synthesize biochemical compounds, including proteins, in response to changes in water activity or osmotic pressure. Measurement of the water activity or osmotic pressure of simple aqueous solutions has been based on freezing point depression or vapor pressure deficit. Measurement of the osmotic pressure of plants under water stress has been mainly based on vapor pressure deficit. However, differences have been noted for osmotic pressure values of aqueous polyethylene glycol (PEG) solutions measured by freezing point depression and vapor pressure deficit. For this paper, the physicochemical basis of freezing point depression and vapor pressure deficit were first examined theoretically and then, the osmotic pressure of aqueous ethylene glycol and of PEG solutions were measured by both freezing point depression and vapor pressure deficit in comparison with other aqueous solutions such as NaCl, KCl, CaCl(2), glucose, sucrose, raffinose, and bovine serum albumin (BSA) solutions. The results showed that: (1) freezing point depression and vapor pressure deficit share theoretically the same physicochemical basis; (2) theoretically, they are proportional to the molal concentration of the aqueous solutions to be measured; (3) in practice, the osmotic pressure levels of aqueous NaCl, KCl, CaCl(2), glucose, sucrose, and raffinose solutions increase in proportion to their molal concentrations and there is little inconsistency between those measured by freezing point depression and vapor pressure deficit; (4) the osmotic pressure levels of aqueous ethylene glycol and PEG solutions measured by freezing point depression differed from the values measured by vapor pressure deficit; (5) the osmotic pressure of aqueous BSA solution measured by freezing point depression differed slightly from that measured by vapor pressure deficit.     

Isotopic Labeling of Embryo,Yolk Sac,and Intracellular Parasites of the Embryonated Hen's Egg by Yolk Replacement

A technique is described which allows the replacement of 50% of the yolk of the embryonated hen''s egg with large volumes of diverse but chemically defined solutions. By using an electrosurgical unit and a polyethylene tunnel, the procedure was performed on eggs from days 3 through 7 with greater than 90% surgical success and viability for the short term. More than 50% of the eggs replaced showed viability for 2 weeks, and a significant proportion went full term. ³²PO₄ and amino acids (³H and ¹⁴C) added to the replaced eggs were incorporated into the macromolecules of the embryo and yolk sac as well as into parasitic rickettsiae cultivated in the replaced eggs. The incorporated ³²PO₄ was shown to be assimilated into a variety of biochemical species.     

THE VAPOR PRESSURE OF DOG'S BLOOD AT BODY TEMPERATURE

The vapor pressures of dog''s blood and blood plasma were determined at 37.5° by the dynamic method and the osmotic pressures calculated from the experimental data. The vapor pressures calculated from experimentally determined freezing point data agreed, within the experimental error, with the values obtained from direct measurement. The vapor pressure lowering produced by the colloid constituents of the blood was also determined and found to be minimal compared to that of the other constituents.     

The freezing point depression of mammalian tissues in relation to the question of osmotic activity of cell fluid

The freezing point depression of freshly excised frozen tissues, pulverized in a hydraulic press or in a mortar, is greater than that of plasma. Even at 0°C. the freezing point depression of such homogenates increases significantly with time. Dilution data indicate that such freezing point data are valid. The presence of intact cells has been shown in smears of tissues pulverized in a mortar, but not in smears of those crushed in a hydraulic press. The osmolarity of various diluent solutions affects the calculated osmotic activity of tissue homogenates presumably because of delayed diffusion between the diluent and cell fluid. With a hypertonic NaCl diluent, spuriously low values of tissue osmotic activity are found from calculations assuming instantaneous mixing between homogenates and diluents. The limitations of data from cryoscopic experiments and from tissue-swelling experiments are discussed in relation to the basic question of whether or not cell fluid is isotonic to extracellular fluid.     

Separation of lipid-free egg yolk proteins by high-pressure liquid chromatography using solvents containing formic acid

A method for obtaining total protein patterns from lipid-containing systems, in particular egg yolk, is described. After dispersion of the yolk in 8 M guanidine hydrochloride solution, lipid is removed by extraction with chloroform-methanol and petrol. The protein solution is applied to a high-pressure liquid chromatograph and eluted with a gradient of formic acid, isopropanol, and acetonitrile. In measurements on a known yolk protein, duck apovitellenin I, the method did not cause irreversible formylation of N-terminal or other residues. The method was used (1) to compare protein patterns of whole yolk from hen and quail eggs; (2) to isolate and partially sequence quail apovitellenin I; and (3) to compare protein patterns of "white yolk" and "yellow yolk."     

Thermodynamic modeling and cryomicroscopy of cell-size, unilamellar, and paucilamellar liposomes

Cryomicroscope studies of large unilamellar liposomes indicate that liposomes are an excellent model for studying membrane response to freezing and thawing. Liposomes are attractive for such use because they can be custom-manufactured for a particular investigation. In addition, liposome responses to freezing and thawing mimic real cell behavior in a number of significant ways. Analogous behavior includes osmotic shrinkage at slow cooling rates, internal ice formation at fast cooling rates, comparable nucleation temperatures, and a variety of comparable thawing responses. Experimental determination has been made of the equilibrium osmotic properties and the nonequilibrium water transport properties of the egg lecithin liposomes used in the freezing studies. These properties have been used in a computer model to simulate volume changes resulting from water transport during freezing and thawing. Comparison between computer model predictions and experimental data for the liposome volume response during freezing indicates reasonable agreement whereas computer simulations of volume response during thawing do not match experimental data well.     

Effect of egg yolk concentration on cryopreserving Spanish ibex (Capra pyrenaica) epididymal spermatozoa

Tris-egg yolk based diluents provide adequate cryoprotection for the sperm of most wild species in which they have been tested. The objective of the current study was to evaluate various Tris-based diluents containing different concentrations of egg yolk, for the fertilizing ability of epididymal spermatozoa of the Spanish ibex (Capra pyrenaica) after freezing and thawing. For this purpose, we used heterologous in vivo fertilization by intrauterine insemination of domestic goats (Capra hircus). In Experiment 1, a Tris-citric acid-glucose (TCG) diluent containing 6% (v/v) egg yolk and a TCG extender containing 20% egg yolk were compared. In Experiment 2, a TCG-6% egg yolk extender was compared with Triladyl-20% egg yolk. Diluted samples were cooled slowly to 5 degrees C over 1 h and equilibrated at that temperature for 2 h. At that point, aliquots of samples were loaded into 0.25 ml straws, and frozen in nitrogen vapor for 10 min. The fertility of spermatozoa frozen in TCG-6% egg yolk was higher (P<0.05) than for those extended with TCG-20% egg yolk, and tended to be higher than for those frozen with Triladyl-20% egg yolk. From the results of this study, the use of Tris-based extenders containing low concentrations of egg yolk (6%) is recommended for cryopreserving Spanish ibex epididymal spermatozoa.     

Protection against lethal freezing temperatures by mucus in an Antarctic limpet

The antarctic limpet, Patinigera polaris, is sometimes caught in near-shore ice and exposed to temperatures substantially below ?2 °C. In-frozen animals always secrete an envelope of mucus which prevents extracellular ice propagation down to ?10 °C. Survival in limpets without mucus protection is significantly lower. Ice propagation through limpet mucus is retarded below its equilibrium freezing point in a manner similar to polar fishes. The capacity of mucus as a cryoprotectant has not previously been described.     

The freezing point depression of mammalian tissues after sudden heating in boiling distilled water

The calculated freezing point depression of freshly excised boiled mammalian tissue is approximately the same as that of plasma. The boiling procedure was chosen to eliminate the influence of metabolism on the level of the freezing point depression. Problems created by the boiling, such as equilibrium between tissue and diluent, change in activity coefficient by dilution, and loss of CO(2) content, are discussed. A frozen crushed tissue homogenate is hypertonic to plasma. Boiling and dilution of such hypertonic homogenate exposed to room temperature for 5 to 15 minutes did not produce significant or unexplicable decreases in its osmotic activity. Moreover, freezing and crushing of a boiled diluted tissue did not produce any increase of the isoosmotic level of freezing point depression. It is possible to explain these data either with the hypothesis of hypertonic cell fluid or with that of isotonic cell fluid. In the case of an assumed isotonic cell fluid, data can be explained with one assumption, experimentally backed. In the case of an assumed hypertonic theory data can be explained only with the help of at least three ad hoc postulates. The data support the validity of the classical concept which holds that cell fluid is isotonic to extracellular fluid.     

Further comments on negative solvent pressure in osmosis

A previously proposed gedanken experiment on osmotic pressure and flow is supplemented to illustrate the circumstances in which equilibrium is achieved. The concept that solvent pressure can be negative and can be described as a partial pressure is thus given additional credence.     

Graphical analysis of nonideal monomer N-mer, isodesmic, and type II indefinite self-associating systems by equilibrium ultracentrifugation.

A graphical procedure is described by which one can obtain in principle the monomer molecular weight, stoichiometry, equilibrium constant, and second virial coefficient of nonideal monomer N-mer, isodesmic, and type II indefinite self-associating systems. In addition, a method is presented for obtaining both the equilibrium constant and the second virial coefficient from the maximum in a plot of apparent molecular weight vs. concentration if the monomer molecular weight and stoichiometry are known. The usefulness and limitations of the methods are discussed, as well as the quality and range of data required for determination of the relevant parameters. The techniques described are applicable to analysis of self-associating systems by osmotic pressure and light scattering, as well as equilibrium ultracentrifugation measurements.     

Comment on “Determination of the quaternary phase diagram of the water–ethylene glycol–sucrose–NaCl system and a comparison between two theoretical methods for synthetic phase diagrams” Cryobiology 61 (2010) 52–57

Recently, measurements of a considerable portion of the phase diagram for the quaternary system water–ethylene glycol–sucrose–NaCl were published (Han et al., 2010). In that article, the data were used to evaluate the accuracy of two non-ideal multi-solute solution theories: the Elliott et al. form of the multi-solute osmotic virial equation and the Kleinhans and Mazur freezing point summation model. Based on this evaluation, it was concluded that the freezing point summation model provides more accurate predictions for the water–ethylene glycol–sucrose–NaCl system than the multi-solute osmotic virial equation. However, this analysis suffered from a number of issues, notably including the use of inconsistent solute-specific coefficients for the multi-solute osmotic virial equation. Herein, we reanalyse the data using a recently-updated and consistent set of solute-specific coefficients (Zielinski et al., 2014). Our results indicate that the two models have very similar performance, and, in fact, the multi-solute osmotic virial equation can provide more accurate predictions than the freezing point summation model depending on the concentration units used.     

A modified theory of the potential and inhomogeneous ion distributions near charged surfaces

A new method is proposed to derive ion distributions and the electric potential near charged surfaces. The diffuse double layer is described by a proposed thermodynamic equilibrium condition between electrostatic forces and osmotic pressure. Problems related with the osmotic pressure and the integration of Coulomb forces are investigated in this paper. A numeric example is given based on erythrocyte data.     

A simple moder for estimating the ice content of freezing ectotherms

1. 1.Although body ice content is an important variable affecting freeze tolerance, present calorimetric methods for its measurement necessarily require the termination of a freezing protocol.

2. 2.A simple iterative model, based on the colligative properties of solutions and requiring precise measurements of only equilibrium freezing point (of the unfrozen organism) and of core body temperature, allows estimation of the percentage of body water frozen at any time during a freezing episode.

3. 3.This model can also predict the lethal temperature for a freezing ectotherm, assuming that death occurs due to osmotic dehydration when 67% (of any other known lethal fraction) of the body water is frozen.

4. 4.The basic model is easily extended to evaluate the effects of variables such as: body mass, initial body water content, initial osmotic concentration, and test chamber microenvironment.

5. 5.This model is not intended to supplant existing more exact biophysical models of freezing kinetics. Rather it is proposed as a first approximation which is generally supported by published data and which should be of significant practical value for investigators of freeze tolerant organisms.

Equilibrium,quasi-equilibrium,and nonequilibrium freezing of mammalian embryos

The first successful freezing of early embryos to −196°C in 1972 required that they be cooled slowly at ∼1°C/min to about −70°C. Subsequent observations and physical/chemical analyses indicate that embryos cooled at that rate dehydrate sufficiently to maintain the chemical potential of their intracellular water close to that of the water in the partly frozen extracellular solution. Consequently, such slow freezing is referred to as equilibrium freezing. In 1972 and since, a number of investigators have studied the responses of embryos to departures from equilibrium freezing. When disequilibrium is achieved by the use of higher constant cooling rates to −70°C, the result is usually intracellular ice formation and embryo death. That result is quantitatively in accord with the predictions of the physical/chemical analysis of the kinetics of water loss as a function of cooling rate. However, other procedures involving rapid nonequilibrium cooling do not result in high mortality. One common element in these other nonequilibrium procedures is that, before the temperature has dropped to a level that permits intracellular ice formation, the embryo water content is reduced to the point at which the subsequent rapid nonequilibrium cooling results in either the formation of small innocuous intracellular ice crystals or the conversion of the intracellular solution into a glass. In both cases, high survival requires that subsequent warming be rapid, to prevent recrystallization or devitrification. The physical/ chemical analysis developed for initially nondehydrated cells appears generally applicable to these other nonequilibrium procedures as well.