Is trehalose special for preserving dry biomaterials?

Simple sugars, especially disaccharides, stabilize biomaterials of various composition during air-drying or freeze-drying. We and others have provided evidence that direct interaction, an interaction that we believe is essential for the stabilization, between the sugar and polar groups in, for example, proteins and phospholipids occurs in the dry state. Some researchers, however, have suggested that the ability of the sugar to form a glass is the only requirement for stabilization. More recently, we have shown that both glass formation and direct interaction of the sugar and headgroup are often required for stabilization. In the present study, we present a state diagram for trehalose glass and suggest that the efficacy of this sugar for stabilization may be related to its higher glass transition temperatures at all water contents. We also show that trehalose and trehalose:liposome preparations form trehalose dihydrate as well as trehalose glass when rehydrated with water vapor. Formation of the dihydrate sequesters water, which might otherwise participate in lowering the glass transition temperature to below ambient. Because samples remain in the glassy state at ambient temperatures, viscosity is high and fusion between liposomes is prevented.     

Sucrose and fructose have qualitatively different flavors to rats

Previous studies suggest that rats might be able to discriminate between sucrose and fructose, but no previous study has examined this possibility in much detail. Rats were conditioned to avoid either sucrose or fructose by injecting them with lithium chloride when they drank these substances. Control rats were given the same injections but were not exposed to either sugar during training. After training, the rats were given a choice of fructose vs. sucrose. Data from control rats provided information about the relative taste intensity of the sugars. If the sugars possess only a single gustatory quality, control rats should prefer the sweeter sugar; under this assumption, sucrose appears to be two-four times sweeter than fructose. The two sugars share a common taste because rats trained to avoid sucrose avoided fructose when the fructose concentration was much greater than the sucrose concentration. Nevertheless, the two sugars are discriminable because, when the apparent sweetness of the sugars was matched, rats showed a greater aversion to the sugar they were trained to avoid. Aversions to sucrose and fructose also generalized to maltodextrins, but sucrose may have a somewhat greater maltodextrin flavor than does fructose. It is proposed that the biological function of maltodextrin taste is to allow animals to sense the ratio of glucose to fructose in foods.     

Characterisation of the glass transition of an amorphous drug using modulated DSC

PURPOSE: The use of modulated differential scanning calorimetry (MDSC) as a novel means of characterising the glass transition of amorphous drugs has been investigated, using the protease inhibitor saquinavir as a model compound. In particular, the effects of measuring variables (temperature cycling, scanning period, heating mode) have been examined. METHODS: Saquinavir samples of known moisture content were examined using a TA Instruments 2920 MDSC at a heating rate of 2 degrees C/min and an amplitude of +/-0.159 degrees C with a period of 30 seconds. These conditions were used to examine the effects of cycling between - 50 degrees C and 150 degrees C. A range of periods between 20 and 50 seconds were then studied. Isothermal measurements were carried out between 85 degrees C and 120 degrees C using an amplitude of +/-0.159 degrees C with a period of 30 seconds. RESULTS: MDSC showed the glass transition of saquinavir (0.98 +/- 0.05%w/w moisture content) in isolation from the relaxation endotherm to give an apparent glass transition temperature of 107.0 degrees C +/- 0.4 degrees C. Subsequent temperature cycling gave reproducible glass transition temperatures of approximately 105 degrees C for both cooling and heating cycles. The enthalpic relaxation peak observed in the initial heating cycle had an additional contribution from a Tg "shift" effect brought about by the difference in response to the glass transition of the total and reversing heat flow signals. Isothermal studies yield a glass transition at 105.9 degrees C +/- 0.1 degrees C. CONCLUSIONS: MDSC has been shown to be capable of separating the glass transition of saquinavir from the relaxation endotherm, thereby facilitating measurement of this parameter without the need for temperature cycling. However, the Tg "shift" effect and the number of modulations through the transition should be taken into account to avoid drawing erroneous conclusions from the experimental data. MDSC has been shown to be an effective method of characterising the glass transition of an amorphous drug, allowing the separate characterisation of the Tg and endothermic relaxation in the first heating cycle.     

Kinetics of plasma fructose and glucose when lactose and fructose are used as energy supplements for neonatal calves

Shortly after birth, plasma glucose and fructose concentrations of the neonate decline and thus leave blood sugar below the homeostatic mode. Two trials were conducted to determine the plasma glucose and fructose kinetics in control and supplemented calves for 108 h after birth. In the short-term trial, six Holstein calves were given 40 g of either fructose, lactose, or water (control) orally at 1 and 96 h after birth. Treatments were administered with a colostrum substitute (Life Boost) at 1 h and whole milk at 96 h. Rectal temperatures and changes in plasma glucose and fructose concentrations were monitored at close intervals for 12 h after supplementation. In the long-term trial, 15 Holstein calves were given 40 g of either lactose, fructose, or water (control) at 1 h after birth and at 12-h intervals for 81 h. Plasma glucose and fructose concentrations were determined before and 4 h after each of the seven feedings. Early postpartal feeding of fructose suppressed plasma glucose (approximately 50%), with a reciprocal rise in plasma fructose. Irrespective of treatment, plasma glucose concentrations did not stabilize (approximately 100 mg/dL) until 17 to 24 h after birth. After 24 h, lactose supplements increased concentrations of plasma glucose 4 h after supplementation (169.7 +/- 8.2 mg/dL), compared with those in calves that did not receive the additional lactose. After 24 h, fructose supplements did not affect plasma glucose, but plasma fructose concentrations increased (82.6 +/- 12.4 mg/dL) 4 h after administration. The response to fructose supplements declined by 11.4 mg x dL(-1) x d(-1). Fructose was not detected in the plasma of control or lactose-treated calves after 17 h after birth. Calves that received fructose supplements had rectal temperatures 8 and 10 h after birth that were higher than those of the other calves. The mechanisms of sugar metabolism change quickly following birth. Oral sugar supplements increase the total plasma sugar concentrations of treated calves.     

Taste reception mechanisms in the blowfly: evidence of amiloride-sensitive and insensitive receptor sites

The role of amiloride in the labellar responses to various taste stimuli in the blowfly Protophormia terraenovae was studied with the aim of ascertaining whether amiloride-sensitive cation conductances are present in the chemosensory systems of insects. Results include that: 1) amiloride has no effect on the "salt" cell response to any stimulus; 2) amiloride decreases the "sugar" cell response to fructose, but does not affect that to sucrose; 3) the effects of amiloride on the responses of the "water" cell and the "fifth" cell are less clearly definable, due to the probable superimposition of osmotic mechanisms in the former and the poorly known response modalities of the water. In conclusion, amiloride-sensitive receptor sites seem to exist also in insects. However, unlike most vertebrates investigated, they are principally located in the sugar receptor cell, not on the salt cell.     

Kinetics of Glass Transition and Crystallization in Carbon Nanotube Reinforced Mg-Cu-Gd Bulk Metallic Glass

Mg65Cu25Gd10 bulk metallic glass and its carbon nanotube reinforced composite were prepared. Differential scanning calorimeter （DSC） was used to investigate the kinetics of glass transition and crystallization processes. The influence of CNTs addition to the glass matrix on the glass transition and crystallization kinetics was studied. It is shown that the kinetic effect on glass transition and crystallization are preserved for both the monothetic glass and its glass composite. Adding CNTs in to the glass matrix reduces the influence of the heating rate on the crystallization process. In addition, the CNTs increase the energetic barrier for the glass transition. This results in the decrease of GFA. The mechanism of the GFA decrease was also discussed.     

Improvement of biomass yield and recombinant gene expression in Escherichia coli by using fructose as the primary carbon source

The feasibility of substituting glucose with fructose as a carbon source in Escherichia coli fermentations was investigated. Glucose, the most commonly used sugar in bacterial cultivations, is well-known to pose a number of drawbacks; the most important of which is the Crabtree effect, which results in acidogenesis. Fructose, a glucose structural isomer, offers a reasonable alternative for glucose, since its uptake and utilization are more tightly regulated. Comparative fermentation studies indicate that lower acetate excretion and higher biomass yields were attained in fructose-supplemented growth media compared with those of glucose media. More specifically, cells grown in defined media supplemented with fructose do not excrete detectable amounts of acetate, while about 40 mM of acetate was detected extracellularly in similar glucose cultures. A reduction in the initial growth rate of about 20% was observed with fructose, but final cell densities were about 70% higher compared with glucose supplements. Growth in complex LB media supplemented with fructose again resulted in higher biomass yields (up to 40%) and lower acetate excretion (30-40%) than the comparable glucose media. In bioreactor studies using LB media, acetate levels were reduced from 90 to less than 6 mM, while achieving a 25% improvement in biomass yield. When using richer media, cell densities of more than 40 g L-1 dry cell weight were attained in batch cultivation using fructose compared with 30 g L-1 for glucose. These results have immense applicability in the area of recombinant protein processes. Recombinant E. coli, overexpressing beta-galactosidase under the control of the strong pH-inducible promoter, achieved a volumetric recombinant protein yield of 2.2 million U mL-1 (corresponding to approximately 1.5 g L-1) in batch fructose cultures. This represents a 65% recombinant protein yield enhancement when compared to similar glucose cultivations.     

Removal of cytokine inducing substances by polymyxin-B immobilized polystyrene-derivative fibers during in vitro hemoperfusion of 10% human plasma containing Staphylococcus aureus challenge

Dietary wheat gluten has been associated with the risk of diabetes in animal models of human insulin-dependent diabetes mellitus (IDDM). To evaluate the role of wheat gluten as a T cell antigen in human IDDM, we studied the cell-mediated immune response to wheat gluten in patients with IDDM and in control subjects. The cellular response to gluten was measured by the peripheral blood mononuclear cell (PBMC) proliferation test, and the results were expressed as a stimulation index (SI). We observed an enhanced cellular immune response to gluten (SI > or = 3) in seven of 29 patients with newly diagnosed IDDM (24.1%), in six of 39 patients with a longer duration of IDDM (15.4%), and in two of 37 non-diabetic controls (5.4%). Reactivity of T cells to gluten was associated with IDDM at diagnosis (P = 0.03), whereas patients with longer duration of IDDM did not differ from controls (P = 0.16). Responses of T cells to gluten were low in general: the median SI (range) was 2.0 (1-8.6) in patients with newly diagnosed IDDM and 1.5 (1-5.8) in control subjects (P = 0.03). Cellular responsiveness to gluten was not associated with HLA-DQB1 risk alleles for IDDM in patients. Although T cell responses to gluten were slightly increased in newly diagnosed patients the responsiveness was rare, and thus our results do not support a major role of gluten in the pathogenesis of human IDDM.     

Effect of meal dilution on the postprandial glycemic response. Implications for glycemic testing

OBJECTIVE: To investigate the effect of varying the volume of sugar meals on the post-prandial glycemic response (PGR). RESEARCH DESIGN AND METHODS: On six separate occasions, after an overnight fast, blood glucose concentrations were measured in eight healthy subjects (34 +/- 4 years of age, BMI 22.9 +/- 0.9 kg/m2) after the consumption of 25 g glucose, sucrose, or fructose dissolved in either 200 or 600 ml of water. Blood was obtained at fasting and then at times 15, 30, 45, 60, and 90 min after the start of the test meal. RESULTS: PGR was found to be influenced by carbohydrate type (P < 0.001). Mean response areas (min.mmol.l-1) to the three sugars were statistically different (P < 0.05). Glucose had the highest response area (90.0 +/- 8.1), followed by sucrose (61.3 +/- 5.0) and then fructose (14.7 +/- 2.8). Independent of this effect, PGR was also found to be influenced by volume dose (P < 0.01). By tripling meal volume from 200 to 600 ml, PGR areas were significantly increased for all three sugars, glucose (79.3 +/- 10.3 vs. 100.8 +/- 12.0, P = 0.035), sucrose (52.6 +/- 5.5 vs. 70 +/- 7.4, P = 0.0094), and fructose (11.0 +/- 3.8 vs. 18.4 +/- 3.9, P = 0.012). Where the effects of time (P < 0.05) and dose (P < 0.05) were determined to be independent (interaction nonsignificant) for all three sugars, this increase in volume also significantly increased glycemic concentrations at 15 min, for glucose (P = 0.033) and sucrose (P = 0.026), suggesting that changes in gastric emptying time may be a mechanism of action. CONCLUSIONS: Varying the volume of liquid sugar meals alters PGR. Understanding this concept may help to reduce variability both in the glycemic testing of foods and oral glucose tolerance testing.     

Spectroscopic Properties and Judd-Ofelt Theory Analysis of Er^3＋-Doped Heavy Metal Oxyfluoride Silicate Glass

Er^3 -doped heavy metal oxyfluoride silicate glass was fabricated and characterized, and the absorption spectrum and fluorescence spectrum of the glass were studied. The Judd-Ofelt intensity parameters Ωt(t=2, 4, 6), spontaneous transition probability, fluorescence branching ratio and radiative lifetime of each energy levels for Er^3 were calculated by Judd-Ofelt theory, and stimulated emission cross-section of ^4I13/2→^4I15/2 transition was calculated by McCumber theory. The results show that fluorescence full width at half maximum and stimulated emission cross-section of Er^3 -doped heavy metal oxyfluoride silicate glass are broad and large, respectively. Compared with other host glasses, the gain bandwidth property of Er^3 -doped heavy metal oxyfluoride silicate glass is close to those of tellurite and bismuth glasses, and has advantage over those of silicate, phosphate and germante glasses.     

Effect of sugars on headgroup mobility in freeze-dried dipalmitoylphosphatidylcholine bilayers: solid-state 31P NMR and FTIR studies

The effect of the carbohydrates trehalose, glucose, and hydroxyethyl starch (HES) on the motional properties of the phosphate headgroup of freeze-dried dipalmitoylphosphatidylcholine (DPPC) liposomes was studied by means of 31P NMR, Fourier transform infrared spectroscopy (FTIR), and differential scanning calorimetry (DSC). The results show that trehalose, which is a strong glass former (Tg = 115 degreesC), elevates the onset of the lipid headgroup rotations and preserves some rotational mobility of the phosphate headgroups after cooling from the liquid-crystalline state. Glucose (Tg = 30 degreesC), a very effective depressant of the phase transition temperature of freeze-dried DPPC, markedly elevates the initiation of the temperature of headgroup rotations. On the other hand, the monosaccharide does not preserve the headgroup disordering when cooled from the liquid-crystalline state. These effects are consistent with formation of hydrogen bonds between the OH groups of the sugar and the polar headgroups of DPPC. They show, however, that hydrogen bonding is not sufficient for preservation of the dynamic properties of freeze-dried DPPC. HES, although a very good glass former (Tg > 110 degreesC), does not depress the phase transition temperature and affects only slightly the rotational properties of freeze-dried DPPC. This lack of effect of HES is associated with the absence of direct interactions with the lipid phosphates, as evidenced by the FTIR results. These data show that vitrification of the additive is not sufficient to affect the dynamic properties of dried DPPC.     

Disruption of the Candida albicans TPS1 gene encoding trehalose-6-phosphate synthase impairs formation of hyphae and decreases infectivity

The TPS1 gene from Candida albicans, which encodes trehalose-6-phosphate synthase, has been cloned by functional complementation of a tps1 mutant from Saccharomyces cerevisiae. In contrast with the wild-type strain, the double tps1/tps1 disruptant did not accumulate trehalose at stationary phase or after heat shock. Growth of the tps1/tps1 disruptant at 30 degreesC was indistinguishable from that of the wild type. However, at 42 degreesC it did not grow on glucose or fructose but grew normally on galactose or glycerol. At 37 degreesC, the yeast-hypha transition in the mutant in glucose-calf serum medium did not occur. During growth at 42 degreesC, the mutant did not form hyphae in galactose or in glycerol. Some of the growth defects observed may be traced to an unbalanced sugar metabolism that reduces the cellular content of ATP. Mice inoculated with 10(6) CFU of the tps1/tps1 mutant did not show visible symptoms of infection 16 days after inoculation, while those similarly inoculated with wild-type cells were dead 12 days after inoculation.     

Fe60CoxZr10Mo5W2B23-x(x=1,3,5,7,9)块状金属玻璃的非晶形成能力

The bulk Fe60CoxZr10Mo5W2B23-x (x= 1, 3, 5, 7, 9) amorphous rods with diameters of1.5 mm were successfully prepared by copper mold casting method with the low purity raw materials.The amorphous and crystalline states, and thermal parameters, such as the glass transition temperature (Tg), the initial crystallization temperature (Tx), the supercooled liquid region (ΔTx = TxTg), the reduced glass transition temperature Trg (Tg/Tm, Tm: the onset temperature of melting of the alloy, and Tg/T1, T1 : the finished temperature of melting of the alloy) were investigated by X-ray diffractometry (XRD) and differential scanning calorimetry (DSC) analysis. Glass forming ability of Fe60CoxZr10Mo5W2B23-x (x=1, 3, 5, 7, 9)bulk metallic glasses has been studied. According to the results, the alloy (x=7) with the highest Trg (Tg/T1 =0. 607, Tg/T1 =0.590) value, has the strongest glass forming ability among these alloys because its composition is near eutectic composition.The wide supercooled liquid region over 72 K indicates the high thermal stability for this alloy system.This bulk metallic glass exhibits quite high strength (Hv 1020). The success of production of the Febased bulk metallic glass with industrial materials is of great significance for the future progress of basic research and practical application.     

A mammary epithelial cell line is transiently stimulated towards milk lipid synthesis by lactogenic treatments

The effect of moisture sorption at different relative humidities on the tensile strength and the physical stability of compacts of crystalline and partly amorphous lactose, alone and in binary mixtures with PVP, has been studied. Furthermore, the role of moisture as a plasticizer and its effect on the glass transition temperature, Tg, are related to the compactibiltiy. Samples were conditioned for 2 hr using a climate test chamber at different relative humidities. Moisture sorption was determined, the radial crushing strength for compacts was measured immediately and after storage, and the tensile strength was calculated. The glass transition temperature, Tg, was determined using DSC. The tensile strength of the compacts was found to depend on both the conditioning humidity and the humidity during storage. An increase in humidity to a level at which the glass transition temperature, Tg, fell below the operating temperature, T, resulted in transition from a rigid glassy state to a mobile rubbery state. For compacts of partly amorphous lactose, an increase in the tensile strength was observed during storage of tablets, due to recrystallization of the amorphous regions above Tg. Tablets of mixtures of lactose and PVP exhibit a sharp decrease in tensile strength at humidities above 70% RH, due to the glass-to-rubber transition of PVP.     

Std1, a gene involved in glucose transport in Schizosaccharomyces pombe

A wild-type strain, Sp972 h-, of Schizosaccharomyces pombe was mutagenized with ethylmethanesulfonate (EMS), and 2-deoxyglucose (2-DOG)-resistant mutants were isolated. Out of 300 independent 2-DOG-resistant mutants, 2 failed to grow on glucose and fructose (mutants 3/8 and 3/23); however, their hexokinase activity was normal. They have been characterized as defective in their sugar transport properties, and the mutations have been designated as std1-8 and std1-23 (sugar transport defective). The mutations are allelic and segregate as part of a single gene when the mutants carrying them are crossed to a wild-type strain. We confirmed the transport deficiency of these mutants by [14C]glucose uptake. They also fail to grow on other monosaccharides, such as fructose, mannose, and xylulose, as well as disaccharides, such as sucrose and maltose, unlike the wild-type strain. Lack of growth of the glucose transport-deficient mutants on maltose revealed the extracellular breakdown of maltose in S. pombe, unlike in Saccharomyces cerevisiae. Both of the mutants are unable to grow on low concentrations of glucose (10 to 20 mM), while one of them, 3/23, grows on high concentrations (50 to 100 mM) as if altered in its affinity for glucose. This mutant (3/23) shows a lag period of 12 to 18 h when grown on high concentrations of glucose. The lag disappears when the culture is transferred from the log phase of its growth on high concentrations. These mutants complement phenotypically similar sugar transport mutants (YGS4 and YGS5) reported earlier by Milbradt and Hoefer (Microbiology 140:2617-2623, 1994), and the clone complementing YGS4 and YGS5 was identified as the only glucose transporter in fission yeast having 12 transmembrane domains. These mutants also demonstrate two other defects: lack of induction and repression of shunt pathway enzymes and defective mating.     

Anneal and Concentration Effect on PL Properties of Sol-Gel Derived Eu3+ Doped SiO2 Glass

Eu3 doped SiO2 nano-crystalline glasses were prepared by sol-gel method. The broad peak of XRD pattern indicates an amorphous SiO2 matrix. The affection of anneal time and anneal temperatures on photoluminescence (PL) properties of SiO2 glass under different Eu3 doping concentration were studied systematically. It is found that the optimized anneal time is about 3 h. The excitation spectra of 2% Eu3 doped SiO2 glass powder were measured under various anneal temperatures, and the optimized anneal temperature is observed around 700 ℃. The fluorescence-quenching effect can be observed in the emission spectra when the annealing temperature exceeds 700 ℃. The emission spectra of different molar ratio dopants were measured at an annealed temperature of 500 ℃, and the concentration-quenching phenomenon has also been observed in SiO2 glass powder when the molar ratio of Eu3 ion exceeds 3%. The result shows that the PL intensity approaches its maximum when the molar ratio of Eu3 ions in the sample is about 3%. In addition, a comparatively stronger emission spectrum at wavelength of 703 nm which is corresponding to the energy transition 5D0→7F4 of Eu ions is also obtained.     

Calorimetric studies of the interaction between DNA and poly-L-lysine

The transition enthalpy deltaH of the helix-random coil transition of the DNA-polylysine complex was measured as a function of the peptide:nucleotide ratio by the help of an adiabatic scanning differential calorimeter. Furthermore the transition enthalpy of a complex with a specific peptide:nucleotide ratio was determined as a function of the cation concentration of the solution. Finally the reaction enthalpy of the interaction of polylysine with native and denatured DNA was measured with the help of a LKB batch calorimeter. From the results of the calorimetric measurements one can conclude that the transition enthalpy of the DNA-polylysine complexes is linearly dependent on the nucleotide: peptide ratio. The extrapolated value for the 1:1 complex is 14.4 kcal per mole base pairs.     

Differential effects of sucrose, fructose, glucose, and lactose on crying in 1- to 3-day-old human infants: Qualitative and quantitative considerations.

Assessed the potency of different sugars as calming agents in human infants. Five 0.1-ml aliquots of 0.51M sucrose, fructose, glucose, or lactose were presented to 1- to 3-day-old infants who were crying spontaneously. Sucrose and fructose were equally effective calming agents, glucose slightly less so. Lactose, the milk sugar, was not at all effective and did not reduce crying any more than did water. In fact, some babies cried more when given lactose. A 2nd experiment established quantitative, dose-response functions for sucrose as a calming agent: 0.17M, 0.42M, and 0.51M sucrose reduced crying equally effectively. Moreover, crying reduction was not differentially affected by the volume of ingested sucrose, because 0.2 ml of 0.34M sucrose was as effective as 0.6 or 1.0 ml of 0.34M sucrose. Results suggest that sucrose calms in a stepwise manner with a flat suprathreshold function and that the calming basis of milk must be sought in components other than its sugar. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Mannitol 1-phosphate mediates an inhibitory effect of mannitol on the activity and the translocation of glucokinase in isolated rat hepatocytes

When tested in the presence of an inhibitor of sorbitol dehydrogenase, both mannitol and sorbitol caused a progressive inhibition of the detritiation of [2-3H]glucose in isolated rat hepatocytes. The purpose of the present work was to investigate the possibility that this effect was mediated by the regulatory protein of glucokinase. When added to hepatocytes, mannitol decreased the apparent affinity of glucokinase for glucose and increased the concentration of fructose required to stimulate detritiation, without affecting the concentration of fructose 1-phosphate. Its effect could be attributed to the formation of mannitol 1-phosphate, a potent agonist of the regulatory protein, which, similarly to fructose 6-phosphate, reinforces its inhibitory action. Formation of mannitol 1-phosphate in hepatocytes was dependent on the presence of mannitol and was stimulated by compounds that increase the concentration of glucose 6-phosphate. Liver extracts catalysed the conversion of mannitol to mannitol 1-phosphate about 7 times more rapidly in the presence of glucose 6-phosphate than of ATP. The glucose 6-phosphate-dependent formation was entirely accounted for by a microsomal enzyme, glucose-6-phosphatase and was not due to a loss of latency of this enzyme. In hepatocytes in primary culture, mannitol decreased the detritiation rate and counteracted the effect of fructose to stimulate glucokinase translocation. Taken together, these results strongly support a central role played by the regulatory protein in the control of glucokinase activity and translocation in the liver, as well as a feedback control exerted by fructose 6-phosphate on this enzyme.