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PubMed2006

[Research progress on trehalose used in lyophilization of blood cells--review].

Lyophilization is the best method for preservation of blood cells at present. Lyophilized blood cells could be stored at room temperature for long periods of time, while maintaining a high degree of viability. Lyophilized blood cells facilitates transportation and the costs are low. However, the membrane of blood cells is damaged and viability of blood cells is decreased in lyophilization. Trehalose has been shown to protect membrane, proteins and nucleic acids during freezing and desiccation. Now, researchers present a methor for loading blood cells with trehalose. In this paper, damage effect of lyophilization on blood cells, the mechanism of trehalose protection and the experimental studies on trehalose were reviewed.

Blood Preservation↗
PubMed2006

[Trehalose -- a biomembrane protectant applied to lyophilization of human red blood cells: review].

Trehalose, as a nonreducing disaccharide, plays a role in protecting the cytoactivity when the cells is freezing, drying or lyophilization. It has been a biomembrane protectant applied to lyophilization of human blood cells (platelets and erythrocytes), and from which astonishing results have been obtained. Having powerful hydration, distinctive vitrification transform and crystal transform and unique resistance of high temperature and humidification, trehalose is thought of a preferred protectant in the study of cell preservation. In recent years, people concerned trehalose on its protective mechanism, experimental means of transit trehalose to mammal cells and the mechanism of loading in red blood cells. The above aspects were briefly summarized in this article.

Blood Preservation↗
PubMed2006

Content of glycogen and trehalose and activity of alpha-amylase and trehalase in Galleria mellonella larvae infected with entomopathogenic nematodes Steinernema affinis and S. feltiae.

INTRODUCTION: The influence of infection with two species of entomopathogenic nematodes of Steinernematidae family on metabolism of glycogen and trehalose of the host was studied. MATERIAL AND METHODS: Last instar larvae (L7) of Galleria mellonella were experimentally infected with Steinernema affinis and S. feltiae. At 6, 12, 18 and 24 h after infection concentrations of trehalose and glycogen as well as activity of trehalase and alpha-amylase were determined. RESULTS: The content of glycogen was lower in insects infected with S. feltiae than in the controls and animals infected with S. affinis. The content of trehalose was higher in insects from both infected groups than in the controls. Its concentration was slightly higher in larvae infected with S. affinis than in those infected with S. feltiae. The activity of alpha-amylase after infection with S. affinis was low. It was significantly higher in insects infected with S. feltiae. In animals of both infected groups, following a significant reduction at 6 h, the activity of trehalase remained at a similar level, higher than in the controls. In the paper the effects of infection with (i) different species of entomopathogenic nematodes and (ii) the importance of the developmental stage of the insect-host for changes in its metabolism of glycogen and trehalose were discussed.

Animals↗
PubMed1991

Modulation of trehalase activity in two insect cell lines by virus infection and trehalose.

Trehalase (EC 3.2.1.28), an important glycosidase involved in regulating trehalose levels and metabolic energy in insects, was measured in cell lines from fall army worm, Spodoptera frugiperda and salt marsh caterpillar, Estigmene acrea, treated with either glucose or trehalose in the presence or absence of Tipula Iridescent Virus (TIV), a cytoplasmic deoxyribovirus. In medium containing 15-35 mM trehalose, both of these cells increased their trehalase activities by 4.5 to 8x the basal levels from cells in glucose medium. Trehalase activity was rapidly reduced after cells were exposed to TIV. Maximum loss in activity (70-90%), occurring about the same time as peak viral DNA synthesis, was significantly delayed when cells were pre-incubated with 30 mM trehalose. These experiments demonstrate the potential utility of trehalase as a marker for monitoring stresses induced by viral infection and changes in nutrition.

Animals↗
PubMed1981

Biochemical genetics of trehalose metabolism in Saccharomyces cerevisiae.

Different strains of Saccharomyces cerevisiae exhibit alternative patterns of trehalose accumulation during growth on a glucose medium. The active pattern is designated as TAC(+) phenotype and the alternative, low-activity pattern as tac(-) phenotype. The tac(-) phenotype is expressed only during growth, since tac(-) strains actively accumulate trehalose during incubation in a glucose medium lacking a nitrogen source. The tac(-) phenotype appears to be determined by a single, recessive gene tac1. The quantitative expression of the dominant, alternative allele TAC1 is subject to wide variation. A highly active pattern of trehalose accumulation requires TAC1 and an amplification factor, TAM, which consists of one or more dominant gene(s). TAM does not appear to alter significantly the expression of tac1. Highly amplified TAC(+) strains may contain a labile factor not present in a TAC(+) strain which accumulates intermediate levels of trehalose.

Cells, Cultured↗
PubMed1995

Trehalose metabolism--new horizons in technological applications.

Trehalose is responsible for the survival of anhydrobiotic organisms when under stress. Trehalose is a unique, non-reducing, extremely stable disaccharide which is able to protect proteins and membranes from damage caused by freezing, high temperatures and dehydration. Yeasts accumulate large amounts of trehalose and constitute excellent models for studying the response of eucaryotic cells to diverse stresses. The regulation of trehalose metabolism is reviewed and new technological applications for preservation of biological materials are discussed.

Animals↗
PubMed1995

Hymenolepis diminuta metacestodes (Cysticeroids) are unable to utilize exogenous trehalose.

Cysticercoids (metacestodes) of Hymenolepis diminuta were incubated in Tyrode's salt solution (pH 7.2) containing 1.25 mM or 40 mM trehalose for 5 or 18 hr, and the amounts of trehalose remaining in the media were determined using high-pressure liquid chromatography. No differences were detected in the amounts of trehalose remaining in control (no cysticercoids) or experimental incubations under any of the experimental conditions. Thus, cysticercoids are apparently unable to utilize trehalose present in the external medium.

Animals↗
PubMed1993

Trehalose metabolism during sporulation in Saccharomyces cerevisiae.

S. cerevisiae has at least two enzymes for trehalose synthesis. In this paper we report some characteristics of the ADPG-dependent trehalose-6-phosphate synthase. As opposed to the UDPG-dependent enzyme, the ADPG-dependent activity did not respond to a glucose signal nor to a heat shock. Mutant strains, deficient in the UDPG-dependent enzyme but possessing the ADPG-dependent activity, did not accumulate trehalose either during growth or during sporulation. A diploid, homozygous for the deficiency in the UDPG-dependent synthase activity, was constructed. During sporulation the ADPG-dependent activity remained constant. However, when mutant spores were subjected to a glucose signal, trehalose was accumulated to the same extent as in control spores.

Adenosine Diphosphate Glucose↗
PubMed1994

Effects of trehalose in canine lung preservation.

BACKGROUND: The effect of trehalose on the preservation of canine lungs was studied with the use of Euro-Collins solution (ECS). METHODS: In group 1, five lungs were perfused and preserved with a modified ECS in which trehalose (35.0 gm/L) was used instead of glucose. In group 2, six lungs were perfused and preserved with a modified ECS in which glucose was replaced by 70.0 gm/L trehalose. In group 3, six lungs were perfused and preserved with ECS. After preservation for 12 hours, left lung transplantation was performed. RESULTS: The PaO2 values in groups 1 and 2 at 130 minutes after reperfusion were 264.9 and 257.5 mm Hg, respectively. These PaO2 values were significantly higher than the corresponding PaO2 value in group 3 (114.8 mm Hg). All the transplanted lungs in groups 1 and 2 had normal structures on histologic examination, whereas five of the group 3 lungs showed severe pulmonary edema. CONCLUSIONS: These findings show that trehalose is effective in the preservation of lungs for 12 hours.

Animals↗
PubMed1995

The correlative evidence suggesting that trehalose stabilizes membrane structure in the yeast Saccharomyces cerevisiae.

The contribution of trehalose and hsp104 to membrane fluidity and the mobility of non-freezing cell water were examined on the basis of whole cell NMR analysis of the yeast Saccharomyces cerevisiae. Membrane fluidity was dependent on the accumulation of trehalose not hsp104 and non-freezing cell water was dependent on the accumulation of hsp104 not trehalose. Thus, the correlative evidence suggesting that trehalose protects yeast cells from temperature extremes by stabilizing the membrane structure was observed in vivo.

Cell Membrane↗
PubMed1996

The role of trehalose in cell stress.

Water is usually thought to be required for the living state, but many organisms can withstand anhydrobiosis when essentially all of their body water has been removed. The mechanisms for survival to this kind of stress could be similar in microbes, plants and animals. One common feature is the accumulation of sugars by anhydrobiotic organisms. Trehalose, which is one of the most effective saccharides in preventing phase transition events in the lipid bilayer, is accumulated by anhydrobiotic organisms in large amounts. It lowers membrane phase transitions in dry yeast cells, thus preventing imbibitional damages when cells are rehydrated. Yeast cells have a trehalose carrier in the plasma membrane which endows them with the ability to protect both sides of the membrane. Kinetic analysis of the trehalose transport activity in Saccharomyces cerevisiae cells revealed the existence of a multicomponent system with a constitutive low-affinity uptake component and a high-affinity H(+)-trehalose symporter regulated by glucose repression.

Dehydration↗
PubMed1997

The effect of trehalose on human lung fibroblasts stored in euro-collins and low potassium dextran solutions.

BACKGROUND: The research on lung preservation for transplantation is directed to synthesize a solution, in order to prolong the preservation period, thus avoiding the damage to lung tissues caused by ischemia. In recent years some in vivo experimental papers have been published, in which good results were obtained by adding trehalose, a non-reducing disaccharide able to stabilize the cell membrane, to the storage solutions. MATERIALS AND METHODS: In this study we have evaluated the effects on human lung fibroblasts of commonly used storage solutions like Euro-Collins (EC) and Low Potassium Dextran (LPD), in which trehalose has respectively replaced glucose (T-EC) or was added (LPD-T). The cultures were incubated for 8 hours at 10 degrees C in standard and modified solutions. RESULTS: The data relative to EC compared to EC-T showed that the total protein content and the rate of protein synthesis, that is in other words cell viability, are drastically compromised. Concerning LPD-T, compared to standard LPD, a slightly decrease of the total protein content and of the rate of protein synthesis in the cells incubated in the modified solution were observed. The reasons for these unexpected data are related to the hypothesis that during the ischemic period membrane stabilization is neither the main nor the only factor responsible for the cellular damage. Moreover the role of trehalose in preservation solutions, is still not clear and should be further investigated. CONCLUSIONS: With the limits of the study of single cells in vitro, the advantages of trehalose containing solutions proved ineffective and toxic especially as for as EC-T is concerned.

Cell Survival↗
PubMed1980

Stimulation of thymocyte mitogenic protein secretion and of cytostatic activity of mouse peritoneal macrophages by trehalose dimycolate and muramyldipeptide.

Immunomodulators of bacterial origin, such as muramyl-dipeptide (MDP) and trehalose dimycolate, are able to stimulate some important biological functions of macrophages, such as their capacity to secrete a monokine, the thymocyte mitogenic protein (TMP), and to limit the growth of mastocytoma cells in vitro. Adherent cells from the peritoneal cavity of untreated mice do not secrete significant amounts of TMP and are not cytostatic. In contrast, adherent peritoneal cells from mice injected with trehalose dimycolate (emulsified in water) secrete TMP and are strongly cytostatic for P815 cells. Trehalose dimycolate is also active when added in vitro: (a) it enhances the cytostatic action of thioglycollate-elicited macrophages; (b) alone or in sequence with MDP, it induces an appreciable cytostatic activity in resident macrophages; (c) it limits the decline of the strong cytostatic action of trehalose-dimycolate-elicited macrophages occurring during in vitro cultivation. MDP also stimulates the cytostatic activity of the macrophages in vitro. The most interesting effect of MDP is its capacity to induce a strong secretion of TMP after a short contact (1 h) with elicited macrophages.

Acetylmuramyl-Alanyl-Isoglutamine↗
PubMed1998

Trehalose Increases Freeze-Thaw Damage in Liposomes Containing Chloroplast Glycolipids.

Chloroplast thylakoids contain three classes of glycolipids, monogalactosyldiacylglycerol (MGDG), digalactosyldiacylglycerol (DGDG), and sulfoquinovosyldiacylglycerol (SQDG). We have investigated the stability of large unilamellar vesicles made from egg phosphatidylcholine (EPC) and different chloroplast glycolipids during freezing to -18 degreesC, as a function of the presence of three sugars: glucose, sucrose, or trehalose. Contrary to the situation in thylakoids, where cryoprotection increases from glucose < sucrose < trehalose, liposomes containing 50% DGDG showed the opposite behavior. In fact, carboxyfluorescein leakage increased over the control values (freezing in the absence of sugar) in the presence of trehalose. This effect was not seen in vesicles made from pure EPC, or a mixture of EPC and MGDG, or EPC and SQDG. Liposomes made from mixtures of all three glycolipids, however, showed even more leakage in the presence of trehalose than liposomes containing only DGDG and EPC. Copyright 1998 Academic Press.

Journal Article↗
PubMed2005

Identification and structural characterisation of novel trehalose dinocardiomycolates from n-alkane-grown Rhodococcus opacus 1CP.

Rhodococcus opacus 1CP, a potent degrader of (chloro-) aromatic compounds was found to utilise C10-C16 n-alkanes as sole carbon sources. Highest conversion rates were observed with n-tetradecane and n-hexadecane, whereas the utilisation of n-dodecane and n-decane was considerably slower. Thin-layer chromatography of organic extracts of n-alkane-grown 1CP cultures indicated the growth-associated formation of a glycolipid which was characterised as a trehalose dimycolate by 1H-NMR spectroscopy and mass spectrometry. Total chain lengths between 48 and 54 carbons classify the fatty acid residues as nocardiomycolic acids. The presence of two double bonds in each mycolic acid is another feature that distinguishes the corresponding trehalose dinocardiomycolates from trehalose dicorynomycolates reported for Rhodococcus erythropolis DSM43215 and Rhodococcus ruber IEGM231. R. opacus 1CP was not found, even under nitrogen limitation, to produce anionic trehalose tetraesters which have previously been reported for R. erythropolis DSM43215.

Alkanes↗
PubMed1984

Fructose 2,6-bisphosphate, a potent regulator of carbohydrate metabolism, inhibits trehalose phosphorylase from protist Euglena gracilis.

Partially purified trehalose phosphorylase (EC 2.4.1.64) from Euglena gracilis SM-ZK was inhibited by fructose 2,6-bisphosphate in both synthetic and degradative directions. Ki value for trehalose phosphorolysis was 1.2 microM and that for trehalose synthesis was 0.5 microM. Functions of fructose 2,6-bisphosphate in Euglena, particularly in the regulative mechanism of the two reserve carbohydrates, paramylon and trehalose, are discussed.

Adenine Nucleotides↗
PubMed1987

Dissemination of beads coated with trehalose 6,6'-dimycolate: a possible role for coagulation in the dissemination process.

When spread as a monolayer on the surface of hydrophobic beads and injected into mice, the mycobacterial glycolipid, trehalose 6,6'-dimycolate, reproduces the biologic effects traditionally associated with virulent mycobacteria, including acute inflammation, granuloma formation, and immune adjuvancy. Repeated intraperitoneal injection of glycolipid-coated beads into young C57Bl/6 mice elicits a granulomatous peritonitis, with concomitant dissemination of beads from the peritoneum to distant organs. Glycolipid-coated beads which disseminate from the peritoneum to other sites elicit neither acute inflammation nor granulomata. The coagulation system may be involved in the dissemination of glycolipid-coated beads as evidenced by the following: fibrinogen is a necessary cofactor of the trehalose dimycolate monolayer; diffuse peritoneal and pulmonary hemorrhage accompanies bead dissemination; peritoneal exudate collected shortly after intraperitoneal injection of glycolipid-coated beads is enriched in coagulant activity; coagulability of blood from trehalose dimycolate-treated animals is reduced; and anticoagulation inhibits the inflammatory response to glycolipid-coated beads. In this report, the dissemination of trehalose dimycolate-coated beads is characterized, and a role for the coagulation system in this process is proposed.

Animals↗
PubMed1990

The effect of sucrose and trehalose on viability of one- and two-cell rabbit embryos.

The effect of sucrose and trehalose on the viability of one- and two-cell rabbit embryos was investigated. A significant decrease in the viability of one- and two-cell embryos exposed for 30 min. at 20 degrees C was observed. At 38 degrees C none of the two-cell embryos in a sucrose solution survived after 30 min exposure, while approximately 50% of the embryos survived in a trehalose solution. The cleveage rate in culture of two-cell embryos exposed both to 2.0 M or 1.45 M trehalose was significantly lower in comparison with the control group. However the survival rate after transfer of two-cell embryos exposed to 1.45 M trehalose solution at 20 degrees C remained the same as that of the control group.

Journal Article↗