Concentration dependent effects of dextran on the physical properties of acid milk gels

The effect of dextran from Leuconostoc mesenteroides (DEX₅₀₀), added to milk prior to acidification with glucono-δ-lactone (GDL) or Streptococcus thermophilus DSM20259, was studied with respect to polysaccharide concentration. The incorporation of 5–30 g/kg DEX₅₀₀ significantly affected gelation behavior. Increasing DEX₅₀₀ concentrations resulted in a linear increase of gel stiffness (GDL gels: R² = 0.96; microbial acidification: R² = 0.94; P < 0.05) and 30 g/kg DEX₅₀₀ resulted in a 2-fold higher stiffness compared to gels without polysaccharide. The respective stirred gels depicted a significant reduction in syneresis, which decreased from 30.4% (0 g/kg DEX₅₀₀) to 22.0% (30 g/kg DEX₅₀₀) for chemically acidified gels after 1 d of storage. Physical characteristics of DEX₅₀₀ in aqueous solution were helpful to explain its behavior in the complex system milk.     

Cell Wall Changes in Nectarines (Prunus persica) : Solubilization and Depolymerization of Pectic and Neutral Polymers during Ripening and in Mealy Fruit

Nectarine fruit (Prunus persica L. Batsch var nectarina [Ait] maxim) cultivar Fantasia were either ripened immediately after harvest at 20°C or stored for 5 weeks at 2°C prior to ripening. Fruit ripened after 5 weeks of storage did not soften to the same extent as normally ripened fruit, they lacked juice, and had a dry, mealy texture. Pectic and hemicellulosic polysaccharides were solubilized from the mesocarp of the fruit using phenol:acetic acid:water (PAW) treatment to yield PAW-soluble material and cell wall material (CWM). The carbohydrate composition and relative molecular weight (M_r) of polysaccharide fractions released from the CWM by sequential treatment with cyclohexane-trans-1,2-diamine tetra-acetate, 0.05 m Na₂CO₃, 6 m guanidinium thiocyanate, and 4 m KOH were determined. Normal ripening of nectarines resulted in solubilization of pectic polymers of high M_r from CWM during the first 2 d at ripening temperatures. Concurrently, galactan side chains were removed from pectic polymers. Solubilized pectic polymers were depolymerized to lower M_r species during the latter stages of ripening. Upon removal from cool storage, fruit had undergone some pectic polymer solubilization, and after ripening, pectins were not depolymerized and were of high M_r. Side chains did not appear to be removed from insoluble pectic polymers and branched pectins accumulated in the CWM. The molecular weight profiles obtained by gel filtration of the hemicellulosic fractions from normally ripening and mealy fruit were similar. The results suggest that mealiness results as a consequence of altered pectic polymer breakdown, including that associated with neutral side chains.     

Pectic polysaccharides of cabbage (Brassica oleracea)

Pectic substances extracted from cabbage cell walls with water, at 80°, and (NH₄)₂C₂O₄, at 80°, accounted for 45%(w/w) of the purified cell wall material. Only a small amount of neutral arabinan was isolated. Partial acid hydrolysis and methylation analysis revealed that the major pectic polysaccharide had a rhamnogalacturonan backbone to which a highly branched arabinan was linked, at C-4 of the rhamnose units, mainly through short chains of (1→4)-linked galactopyranose residues. The bulk of the soluble pectic substances had only small amounts of proteins associated with them. After further extraction of the depectinated material with 1M and 4M KOH, to remove the hemicelluloses, the cellulose residue was found to contain a pectic polysaccharide which was solubilized by treatment with cellulase. The general structural features of the pectic polymers are discussed in the light of these results.     

Structure of a pectic polysaccharide fraction from zea shoots

A pectic fraction, accounting for about 0.3% of the total cell wall polysaccharide, was derived from the hot water extract of an insoluble fraction of the buffer-homogenate of Zea shoots. The pectic polysaccharide fraction was characterized by fragmentation analysis after hydrolysis with acid and Erwinia carotovora pectate lyase. The results suggest that the fraction consists of mostly a linear homopolygalacturonan with neutral sugar components or a homogalacturonan and a rhamnogalacturonan with neutral sugar components.     

Polysaccharides from the pulp of cupuassu (Theobroma grandiflorum): Structural characterization of a pectic fraction

Cupuassu (Theobroma grandiflorum, Schumann) is a Brazilian Amazonian fruit whose pulp contains volatile compounds that have been extensively studied. In this work, the pulp from fruits of cupuassu was ground, treated with MeOH–H₂O, and defatted with p-Tol–EtOH. The residue (4% in relation to the fresh pulp) was submitted to sequential extractions with water, aqueous citric acid and aqueous NaOH, resulting in polysaccharide fractions with 0.3–15% yield. The main pectic fraction (7% yield) was obtained with water at 25 °C (W-1 fraction) and was chosen to be better characterized. Chemical and spectroscopic analyses showed that W-1 is composed mainly of a homogalacturonan highly esterified (DE 53%; DA 1.7%) with some rhamnogalacturonan insertions, carrying side chains containing galactose and arabinose.     

Recrystallization of Ice Crystals in Trehalose Solution at Isothermal Condition

An isothermal ice recrystallization behavior in trehalose solution was investigated. The isothermal recrystallization rate constants of ice crystals in trehalose solution were obtained at ?5 °C, ?7 °C, and ?10 °C. Then the results were compared to those of a sucrose solution used as a control sample. Simultaneous estimation of water mobility in the freeze-concentrated matrix was conducted by ¹H spin–spin relaxation time T₂ to investigate mechanisms causing the different ice crystal recrystallization behaviors of sucrose and trehalose. At lower temperatures, lower recrystallization rates were obtained for both trehalose and sucrose solutions. The ice crystallization rate constants in trahalose solution tended to be smaller than those in sucrose solution at the same temperature. Although different ice contents (less than 3.6%) were observed between trehalose and sucrose solutions at the same temperature, the recrystallization behaviors of ice crystals were not markedly different. The ¹H spin–spin relaxation time T₂ of water components in a freeze-concentrated matrix for trehalose solution was shorter than in a sucrose solution at the same temperature. Results show that the water mobility of trehalose solutions in freeze-concentrated matrix was less than that of sucrose solutions, which was suggested as the reason for retarded ice crystal growth in a trehalose solution. Results of this study suggest that the replacement of sucrose with trehalose will not negatively affect deterioration caused by ice crystal recrystallization in frozen foods and cryobiological materials.     

The Effectiveness of Cryoprotectants in Inhibiting Multiple Freeze-Thaw-Induced Functional and Rheological Changes in the Myofibrillar Proteins of Common Carp (Cyprinus carpio) Surimi

The purpose of the present study was to investigate the effects of cryoprotectants (4 % sucrose?+?4 % sorbitol) on functional and rheological changes, including thermal stability, dynamic rheological and emulsifying properties, gel texture profile, gel whiteness, gel water-binding capacity, and gel microstructure, in the myofibrillar proteins (MP) of common carp (Cyprinus carpio) surimi subjected to multiple freeze-thaw cycles (FT, 0, 1, 3 and 5 times). Increased numbers of FT cycles were accompanied by reduced thermal transition temperature (T _max), reduced enthalpy of denaturation (ΔH) (P?<?0.05), and enhanced susceptibility to thermal aggregation. The observation of the gel microstructure indicated that the FT processes produced empty spaces and changed the aggregate gel structure into a coarser-stranded network. The addition of cryoprotectants significantly prevented the reduction of MP thermal stability (P?<?0.05). The protective effect on protein functionality was demonstrated by the cryoprotectant’s efficacy in maintaining the three-dimensional gel network by small-strain oscillatory rheological testing and increasing the emulsifying capacity and emulsion stability of the MP. The FT process caused a decrease (P?<?0.05) in whiteness, water-binding capacity, and texture (hardness, springiness, and gumminess) of the MP gels, which were reduced by the addition of cryoprotectants. Overall, the FT-induced loss of the abovementioned functional and rheological properties was significantly reduced by the presence of cryoprotectants.     

The State of Water in Muscle Tissue as Determined by Proton Nuclear Magnetic Resonance

The nuclear magnetic resonance (NMR) of water protons in live and glycerinated muscle, suspensions of glycerinated myofibrils, and solutions of several muscle proteins has been studied. T₁ and T₂, measured on partially hydrated proteins by pulsed spin-echo techniques, decreased as the ratio of water to protein decreased, showing that the water which is tightly bound by the protein has short relaxation times. In live muscle fibers the pulse techniques showed that, after either a 180 or a 90° pulse, the relaxation of the magnetization is described by a single exponential. This is direct evidence that a fast exchange of protons occurs among the phases of the intracellular water. The data can be fitted with a model in which the bulk of the muscle water is in a phase which has properties similar to those of a dilute salt solution, while less than 4-5% of the total water is bound to the protein surface and has short relaxation times. Measurements of T₁ and T₂ in protein solutions showed that no change in the proton relaxation times occurred when heavy meromyosin was bound to actin, when myofibrils were contracted with adenosine triphosphate (ATP), or when globular actin was polymerized.     

Polysaccharide and oligosaccharide changes in germinating lupin cotyledons

In germinating lupin cotyledons, there was a rapid depletion of raffinose series oligosaccharides, a temporary increase in sucrose and constant low levels of reducing monosaccharides. The major polysaccharide fraction was extracted with hot NH₄ oxalate—EDTA solution and had the constitution of intercellular/cell wall polysaccharide. GLC examination of component sugars showed that as cotyledons expanded this fraction was depleted and that there was selective hydrolysis of arabinose and galactose, so that the uronic acid proportion increased. Gel and DEAE-cellulose chromatography showed that this fraction became more heterogeneous. The neutral and acidic fractions were separated and the component sugars, viscosities, gel chromatographic behaviour and sedimentation constants of these determined. The results indicated that in the later phase of plant cell wall expansion in germinating lupin cotyledons the arabinogalactan side chains of the pectic polysaccharide fraction are selectively hydrolysed leaving a primary wall with a high uronic acid content.     

Conductance behavior of symmetrical tetraalkylammonium halides in aqueous sucrose solutions

The conductance behavior of some tetraalkylammonium halides (R₄NX) in saturated, aqueous solutions of sucrose has been investigated, and data on the conductance of these salts in water saturated by sucrose at 50° are reported at several temperatures within the range 25 to 70°. In these homogeneous, ternary systems, plots of —log K versus 1/T show a break at the saturation temperature, where two straight lines intersect one another. Divergence of the pair of straight lines has been found to decrease with increasing chain-length of the R₄N⁺ ions, in contrast to the structural behavior of common, alkali-metal ions. The results are interpreted in terms of the hydrophobic nature of the tetraalkylammonium halides, as well as the salting-in behavior of these salts towards sucrose molecules.     

Phytantriol-Based In Situ Liquid Crystals with Long-Term Release for Intra-articular Administration

The purpose of this study was to develop an injectable in situ liquid crystal formulation for intra-articular (IA) administration, and in situ forming a viscous liquid crystalline gel with long-term release of sinomenine hydrochloride (SMH) upon water absorption. The pseudo-ternary phase diagram of phytantriol (PT)-ethanol (ET)-water was constructed, and isotropic solutions were chosen for further optimization. The physicochemical properties of isotropic solutions were evaluated, and the phase structures of liquid crystalline gels formed by isotropic solutions in excess water were confirmed by crossed polarized light microscopy (CPLM) and small-angle X-ray scattering (SAXS). In vitro drug release studies were conducted by using a dialysis membrane diffusion method. The optimal in situ cubic liquid crystal (ISV₂) (PT/ET/water, 64:16:20, w/w/w) loaded with 6 mg/g of SMH showed a suitable pH, showed to be injectable, and formed a cubic liquid crystalline gel in situ with minimum water absorption within the shortest time. The optimal ISV₂ was able to sustain the drug release for 6 days. An in situ hexagonal liquid crystal (ISH₂) system was prepared by addition of 5% vitamin E acetate (VitEA) into PT in the optimal ISV₂ system to improve the sustained release of SMH. This ISH₂ (PT/VitEA/ET/water, 60.8:3.2:16:20, w/w/w/w) was an injectable isotropic solution with a suitable pH range. The developed ISH₂ was found to be able to sustain the drug release for more than 10 days and was suitable for IA injection for the treatment of rheumatoid arthritis (RA).KEY WORDS: in situ cubic liquid crystal, in situ hexagonal liquid crystal, phytantriol, sinomenine hydrochloride, sustained drug release     

Correlation of relaxation time with water content in muscle and brain tissues

Proton T₁ of water in muscle and brain tissues was studied as a function of water content at 25°C and 5, 30, and 100 MHz. Muscle samples were bathed in modified Krebs solutions that have the same ionic strength but different osmolarity in order to change the water content. At all three frequencies their 1/T₁ extrapolated to the same value of 0.6 s^?1 for zero dry weight, indicating that T₁ for the majority of water molecules in normal muscle and brain is frequency-independent. Dehydrated muscle, homogenized brain tissue suspended in modified Krebs solution and dehydrated brain tissue provide samples with wide ranges of water content. Their T₁ values at all three frequencies can be well accounted for by a simple equation derived from the two-state model.     

Polysaccharide Produced by the Genus Pullularia: I. Production of Polysaccharide by Growing Cells

A dextranlike polysaccharide was found to be produced on substrates of sucrose, maltose, glucose, and fructose by growing cells of various strains of the genus Pullularia. The polysaccharide, obtainable in amounts as large as 2 to 3 g per 100 ml of culture medium using various carbohydrates as the carbon source, was soluble in cold water but not in 50% alcohol. The polysaccharide obtained had a α_d = +197.5° (c = 0.2 in water), and its molecular weight, determined by the light-scattering method, was found to be approximately 250,000.     

Calorimetric determination of the conformational transition of kappa carrageenan

This paper deals with measurements of the heat of the helix-coil transition by microcalorimetry for kappa carrageenans; data are given in the absence of gel in various solvents (H₂O, Me₂SO, and formamide) with two counterions (K⁺, Rb⁺). In water, the influence of gel formation on ΔH is pointed out and the influence of the time of ageing is demonstrated. The ΔH values measured in the absence of gel are interpreted in terms of the electrostatic model proposed by Manning; the agreement is quite good if a double helix is formed in water, but a monochain, ordered conformation is suggested for solutions in Me₂SO and formamide.     

Pectic polysaccharide breakdown of cell walls in cucumber roots grown with calcium starvation

Pectic polysaccharides from the roots of cucumber (Cucumis sativus L.) grown in liquid culture medium with or without calcium (1 mm CaCl₂) were studied after extraction successively by hot water and Na hexametaphosphate solution. The Ca²⁺ starvation-treatment caused a striking reduction in content of extracted pectic polysaccharide; from an equivalent weight of cell walls, only 33.1% of the control level was extracted from root cell walls of plants cultured under Ca²⁺ deficiency. The extracted pectic polysaccharides were fractionated into neutral and acidic polymers by a DEAE-Sephadex column. The acidic polymers, which represented more than 76% of the yield, appeared to be a major fraction of extracted pectic polysaccharides. The changes of molecular size and glycosyl residue composition of this fraction were compared for the control and Ca²⁺-deprived samples. The results indicate that Ca²⁺ deficiency caused structural changes which could involve both branching pattern and extent of contiguous galacturonosyl units in the water-solubilized pectic polysaccharides. Ca²⁺ starvation also led to a notable decrease in molecular size of the hexametaphosphate-solubilized polysaccharides and, to a lesser extent, of the water-solubilized fraction as well. In addition, polygalacturonase activity in tissue homogenates increased remarkably with the Ca²⁺ deficiency, whereas β-galactosidase activity did not undergo a change. Thus, it appears that one major effect of Ca²⁺ deprivation was to stimulate polygalacturonase activity, an effect which could be involved in the control of the breakdown of pectic polysaccharides in the cell walls.     

Production and characterization of curdlan by Agrobacterium sp.

Agrobacterium sp. was studied for the production of curdlan by conventional one-factor-at-a-time technique and response surface methodology. Factors such as initial pH, urea concentration, sucrose concentration having the greatest influence on the curdlan production were identified. By using response surface methodology (RSM), the curdlan production by Agrobacterium sp. was increased significantly by 109%, from 2.4 g/L to 5.02 g/L when the strain was cultivated in the optimal medium developed by RSM as compared to conventional one-factor-at-a-time technique. The curdlan production rate of 0.84 g/(L h) was obtained when Agrobacterium sp. was cultivated in the optimal medium developed by RSM, which was the highest curdlan production rate reported to date. The infrared (IR) and NMR spectra, the thermogram of DSC and pattern of X-ray diffraction for the curdlan of the present study were almost identical to those of the authentic curdlan sample (from Alcaligenes faecalis; Sigma). The purified curdlan was a linear polysaccharide composed of exclusively β-(1,3)-glucosidic linkages with the molecular weight of 160,000 Da by GPC. The crystalline melting point (T_m), glass transition temperature (T_g) and X-ray diffraction of the sample indicated low crystallinity in the structure.     

Conductance studies on the interaction of sucrose with symmetrical tetraalkylammonium halides in formamide

The interaction of sucrose with symmetrical tetraalkylammoniurn halides (R₄NX) in formamide has been studied by employing conductance measurements. Conductance data of these salts in formamide saturated by sucrose at 50.0 ±0.05° are reported in the temperature range 25 to 70°. Plots of —log (specific conductance) against 1/T showed a break at the saturation temperature, where two straight lines intersect one another. Divergence of the pairs of straight lines in these homogeneous, ternary systems was found to be highly influenced by the structure-making or -breaking properties of the electrolytes. The results are interpreted in terms of the structural properties of the electrolytes and the hydrogen-bonding capabilities of the formamide and sucrose molecules. A similarity in the conductance behavior of R₄N⁺ ions in both aqueous and formamide solutions containing sucrose at saturation concentration has been observed. The transitional effect is more pronounced for R₄N⁺ ions in formamide than in aqueous systems, and this is attributed to the less-structured nature and higher dielectric constant of formamide.     

Localization of 3,5,3'-L-triiodothyronine receptor in rat kidney mitochondrial membranes

Binding of triiodothyronine(T₃) to submitochondrial fractions from rat kidney was studied. Both inner and outer mitochondrial membranes were purified by sucrose gradient centrifugation. Both membranes had specific binding sites for T₃. Scatchard analysis of T₃ binding by membranes gave different affinity constants between inner and outer membranes. In studies with gel filtration of soluble T₃ receptors, four main T₃ binding activities in outer membranes and two main T₃ binding activities in inner membranes were isolated. The results indicate that both inner and outer mitochondrial membranes have specific binding sites for T₃ and that each membrane has a specific structure in T₃ receptor.     

Galactomannan from the seeds of chinese honey locust (Gleditsia sinensis Lam.)

By the hot water extraction method, galactomannan was extracted (4.5% yield of the seed mass) from the seeds of Chinese honey locust (Gleditsia sinensis Lam). It had a molecular weight of 1230 kDa, and its solutions had a high viscosity [η] of 1064 ml/g and optical activity [α]_D of +21.4°. The polysaccharide consists of mannose and galactose residues in the molar ratio 2.69: 1. In the galactomannan macromolecule the backbone is formed by 1,4-β-D-mannopyranose residues, 37% of which are substituted by α-D-galactopyranose at C6. By ¹³C-NMR-spectroscopy, fragments of differently galactose-substituted mannobiose units were found to be in the galactomannan being studied: Man-Man, (Gal)Man-Man, and Man-Man(Gal) in the ratio of 0.23: 0.47: 0.30.     

Water proton magnetic resonance studies of normal and sickle erythrocytes Temperature and volume dependence

The temperature and cell volume dependence of the NMR water proton linewidth, spin-lattice, and spin-spin relaxation times have been studied for normal and sickle erythrocytes as well as hemoglobin A and hemoglobin S solutions. Upon deoxygenation, the spin-spin relaxation time (T₂) decreases by a factor of 2 for sickle cells and hemoglobin S solutions but remains relatively constant for normal cells and hemoglobin A solutions. The spin-lattice relaxation time (T₁) shows no significant change upon dexygenation for normal or sickle packed red cells. Studies of the change in the NMR linewidth, T₁ and T₂ as the cell hydration is changed indicate that these parameters only slightly by a 10–20% cell dehydration. This result suggests that the reported 10% cell dehydration observed with sickling is not important in the altered NMR properties. Low temperature studies of the linewidth and T₁ for oxy and deoxy hemoglobin A and hemoglobin S solutions suggest that the “bound” water possesses similar properties for all four species. The low temperature linewidth ranges from about 250 Hz at ?15°C to 500 Hz at ?36°C and analysis of the NMR curves yield hydration values near 0.4 g water/g hemoglobin for all four species. The low temperature T₁ data go through a minimum at ?35°C for measurements at 44.4 MHz and ?50°C for measurements at 17.1 MHz and are similar for oxy and deoxy hemoglobin A and hemoglobin S. These similarities in the low temperature NMR data for oxy and deoxy hemoglobin A and hemoglobin S suggest a hydrophobically driven sickling mechanism. The room temperature and low temperature relaxation time data for normal and sickle cells are interpreted in terms of a three-state model for intracellular water. In the context of this model the relaxation time data imply that type III, or irratationally bound water, is altered during the sickling process.