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PubMed1991

Increasing the plasma half-life of trichosanthin by coupling to dextran.

Trichosanthin (TCS) is a plant protein which has a wide spectrum of pharmacological activities. It was demonstrated recently that this compound suppressed the replication of human immunodeficiency virus (HIV-1) in vitro. The mechanism of action is believed to be inhibition of protein synthesis. Trichosanthin is a low molecular weight protein which is expected to be easily filtered and eliminated through the kidney. To minimize renal loss, the molecular size of trichosanthin can be increased by coupling to dextran. The larger complex will not undergo glomerular filtration and therefore renal loss can be prevented. This study investigates the kidney's role in trichosanthin elimination and the beneficial effect afforded by coupling to dextran in prolonging plasma half-life. For this purpose, a radioimmunoassay has been developed to determine the concentration of TCS in plasma and urine. The sensitivity of this assay is in the nanogram range. Trichosanthin was coupled to dextran T40 by a dialdehyde method and successful coupling was confirmed by gel filtration chromatography. The complex retained specific binding to trichosanthin antibodies with decreased affinity which can be partially reversed after incubation with dextranase; an enzyme that digested dextran. The pharmacokinetics of intravenously administered trichosanthin (0.75 mg/kg) was compared between two groups of rats with normal and impaired renal function (bilateral renal arterial ligation). Rats with ligation showed a decrease in plasma clearance from 4780 +/- 570 to 220 +/- 20 microL/min and an increase in the mean residence time from 9 +/- 1 to 145 +/- 16 min. Despite the several-fold difference in these parameters, recovery of trichosanthin from normal rat urine was only 0.38 +/- 0.05%. This value can be increased by using higher injection doses. The data indicate that the kidney is an important organ for the elimination of trichosanthin. When the dextran-trichosanthin complex was injected into normal rats trichosanthin activity was not detected in the urine. All the pharmacokinetic parameters suggest that the dextran-trichosanthin complex stayed longer in the body and maintained a much higher plasma concentration than trichosanthin.

Animals↗
PubMed1990

Serum antibody and cellular immune response in mice to dextran B512.

Serum antibodies to dextran started to appear 3 days after immunization of C57BL/6 mice. Synthesis of IgM antibodies was followed by IgG3 and IgGA. Other immunoglobulin classes (IgG1, IgG2b, and IgG2a) were very low or absent. The immune response to dextran was also thymus independent with regard to IgG3 and IgA synthesis as demonstrated by the use of nu/nu mice. CBA and C57BL/6 mice were high responders to dextran with regard to IgM synthesis. C57BL/6 mice produced high levels of IgG3 and IgA antibodies, whereas CBA, A/J, and A.TL only synthesized IgM antibodies. A/J and A.TL strains were most frequently low responders with regard to IgM synthesis and CBA/N mice were completely nonresponders with regard to all immunoglobulin classes. The ability to produce anti-dextran antibodies increased with age in high responder strains. This was most pronounced for IgG3 and IgA antibodies, which reached adult levels 3 months after birth. The affinity of anti-dextran antibodies was high and homogeneous in antisera from C57BL/6 mice. Preimmune matural antibodies and antibodies from immunized low responder strains had a low and variable affinity for dextran.

Aging↗
PubMed1990

IL4 is not required for proliferative responses in B cells stimulated by anti-IgD-dextran conjugates even under limiting conditions of cell-cell interaction.

The experiments in this manuscript confirm and extend our previous finding that IL4 has minimal enhancing activity on B cell activation stimulated by anti-Ig-dextran conjugates. The absence of significant IL4-mediated enhancement is seen also under conditions which are limiting for optimal B cell proliferation. Thus, even when B cells are cultured at low cell densities where cell to cell contact is minimized and are stimulated with picogram per milliliter concentrations of anti-Ig-dextran, IL4 mediates low levels of enhanced proliferation, if at all. The low level of IL4-induced enhancement does not reflect the anti-Ig-dextran-mediated downregulation of IL4 receptors on B cells, since anti-Ig-dextran stimulates an increase in IL4 receptors similar in magnitude to that stimulated by IL4 by itself. To exclude the possibility that anti-Ig-dextran was stimulating IL4 secretion by B cells and thus masking an effect of added IL4, we added inhibiting concentrations of monoclonal anti-IL4 antibody, with the B cells and found that it was without effect on anti-Ig-dextran-stimulated proliferation. Our results suggest that IL4 may not have a prominent role in influencing B cell growth that is stimulated by multivalent T cell-independent antigens.

Animals↗
PubMed1990

Interaction of dextran sulfate with phospholipid surfaces and liposome aggregation and fusion.

The binding of dextran sulfate to phospholipid liposomes was investigated by microelectrophoresis experiments. The polyanion binds to neutral phospholipid liposomes (DMPC and PE) only in the presence of Ca2+. If positively charged stearylamine is incorporated in the vesicles dextran sulfate is bound without Ca2+. Negatively charged phospholipids as PS do not bind dextran sulfate, even in the presence of millimolar concentrations of Ca2+. The adsorption of dextran sulfate results in an aggregation of vesicles due to a bridging mechanism. In all cases the aggregation is followed by a disaggregation toward higher dextran sulfate concentrations. The disaggregation process starts at polymer concentrations smaller than the concentration of the onset of saturation of the adsorption. By use of the probe dilution method a fusion of small DMPC and DMPC/PE vesicles in the presence of Ca2+ and dextran sulfate was found.

Calcium↗
PubMed1992

Dextran sulfate-dependent fusion of liposomes containing cationic stearylamine.

The incorporation of the positively charged stearylamine into phosphatidylcholine liposomes was studied by measuring electrophoretic mobilities. Up to a molar ratio SA/PC = 0.5 an increase of the positive zeta potential can be observed. Addition of the negatively charged macromolecule dextran sulfate leads to a change of the sign of the surface potential of the PC/SA liposomes indicating binding of the macromolecule to the surface. This process is accompanied by an increase in turbidity, which is dependent on the molecular weight of the dextran sulfate and the SA concentration (measured by turbidimetry). Using the NBD/Rh and Pyr-PC fluorescence assays the fusion of SA containing liposomes was investigated. A strong influence of the SA content and molecular weight of dextran sulfate on the fusion extent was observed. The fusion extent is proportional to the SA content in the PC membrane and the molecular weight of dextran sulfate. PC/SA/PE liposomes exhibit a higher fusion extent after addition of dextran sulfate compared to PC/SA liposomes indicating that PE additionally destabilizes the bilayer. Freeze-fracture electron microscopy reveals that the reaction products are large complexes composed of multilamellar stacks of tightly packed, straight membranes and aggregated vesicles. The tight packing of the membranes in the stacks (and the narrow contact of the aggregated vesicles) indicates a strong adherence of opposite membrane surfaces induced by dextran sulfate.

Amines↗
PubMed1987

Size-dependent blood-brain barrier opening demonstrated with [14C]sucrose and a 200,000-Da [3H]dextran.

The reclosure of the blood-brain barrier following osmotic opening was investigated by determining the product of permeability times surface area for two neutral, water-soluble compounds differing widely in molecular size. [14C]Sucrose (Mr 340 Da, radius 5A) and [3H]dextran (Mr 200,000 Da, radius 100 A) were simultaneously injected i.v., and their regional permeability times surface areas were calculated at 6, 10, 35, and 55 min after the blood-brain barrier was opened by a 30-s infusion of 1.8 m L(+)-arabinose into the right external carotid artery. The control permeability times surface area product was about 10(-5) cm3 s-1 g-1 brain for sucrose and negligible for dextran. It increased to 4 X 10(-4) cm3 s-1 g-1 brain and 10(-4) cm3 s-1 g-1 brain for sucrose and dextran, respectively, at 6 min after opening of the blood-brain barrier. Thereafter, permeability-surface area products for both substances declined. Dextran had significantly lower (P less than 0.05) values than sucrose at all times. The ratios of permeability times surface areas of [14C]sucrose to those to [3H]dextran were consistent with restricted diffusion through pores or slits at 35 and 55 min after blood-brain barrier opening, but at 6 and 10 min these ratios were less than the ratio of their free diffusion coefficients, indicative of bulk fluid flow with solute drag from blood to brain. A previously measured increase in brain water content following opening of the blood-brain barrier together with the present results, suggest the creation of slits approximately 400 A in width after osmotic treatment. Reduction in bulk fluid flow from blood to brain appears to be the major cause for the reduction of permeability times surface areas for both sucrose and dextran as the blood-brain barrier recloses.

Animals↗
PubMed1994

Dextran sulfate, a competitive inhibitor for scavenger receptor, prevents the progression of atherosclerosis in Watanabe heritable hyperlipidemic rabbits.

Dextran sulfate competes with binding of modified LDL to the scavenger receptor in macrophages. To elucidate the role of dextran sulfate in the atherosclerotic process, 100 mg of dextran sulfate in drinking water was given to 5 Watanabe heritable hyperlipidemic (WHHL) rabbits for 12 months starting at age 4 months. During the experimental period, there were no significant differences in plasma cholesterol levels between dextran sulfate-treated and untreated rabbits. After 12 months' treatment, accumulation of cholesterol ester in total aorta was significantly suppressed in dextran sulfate-treated rabbits as compared with untreated rabbits (71.4 +/- 22.3 vs. 42.7 +/- 16.5 mg/g dry weight, P < 0.05). Furthermore, lesion area with atherosclerotic plaques in treated rabbits was significantly less than that in untreated rabbits (59.7 +/- 24.5 vs. 30.4 +/- 14.4%, P < 0.05). These results indicate that dextran sulfate might prevent the progression of atherosclerosis by competitively inhibiting the binding of modified LDL to scavenger receptors.

Animals↗
PubMed1984

Effect of Dextran 40 and aprotinin on experimental acute pancreatitis.

This study examines and compares the prophylactic role of aprotinin and Dextran 40 in acute pancreatitis. Experimental acute pancreatitis was induced in 70 male Wistar rats using the closed-duodenal-loop technique. The rats were randomly divided into four groups; sham operation, untreated acute pancreatitis, and therapy with aprotinin or Dextran 40. Samples of blood and urine were collected at the beginning and at the end of the 24-hr period for measurement of amylase and creatinine which allowed calculation of the amylase-creatinine clearance ratio (ACCR). Mortality in the aprotinin group was the same as the untreated rats (20%). Dextran 40 therapy was associated with a lower mortality rate (6.7%). Light microscopic examination confirmed that the histologic changes of acute pancreatitis were less severe in both the aprotinin- and Dextran 40-treated rats. The ACCR was elevated after Dextran 40 therapy, which was due mainly to high urinary amylase levels. These results suggest that Dextran 40 may have a prophylactic role in acute experimental pancreatitis but again emphasizes the high false-positive rate of the ACCR determination.

Acute Disease↗
PubMed1982

Association constants of hybridoma antibodies specific for alpha (1 leads to 6) linked dextran determined by affinity electrophoresis.

Binding constants of monomers of seven BALB/c IgM, four BALB/c IgA, and one C57BL/6 IgA anti-alpha (1 leads to 6) dextran hybridoma antibodies with dextran B512 and with isomaltoheptaose were determined by affinity electrophoresis. Bindings constants to dextran range from 1.52 X 10(5) to 4.43 X 10(5) ml/g for the five IgA monomers and from 1.70 X 10(3) to 6.10 X 10(4) ml/g for the seven IgM monomers. Antibody monomers containing both specific and nonspecific (derived from the myeloma cell that was used to generate the hybridomas) light chains are shown to have association constants with dextran 6 to 30-fold lower than monomers containing only specific light chain, suggesting that the association of specific heavy chain with nonspecific light chain does not result in an anti-dextran combining site. Binding constants with isomaltoheptaose range from 1.45 X 10(4) to 7.01 X 10(4)/M for the IgA proteins and from 6.46 X 10(3) to 7.70 X 10(4)/M for the IgM proteins. The binding constants with dextran and with isomaltoheptaose, and the electrophoretic, immunochemical and idiotypic characteristics of the hybridoma proteins are discussed.

Animals↗
PubMed1986

Immunochemical and related interactions with dextrans reviewed in terms of improved structural information.

Structurally diverse dextrans from Leuconostoc mesenteroides and related bacteria have been used extensively in fundamental immunochemical studies such as induction and characterization of anti-dextran antibodies, as well as in studies of their interaction with pneumococcal antisera, normal bovine serum, concanavalin A, dextran-binding myeloma immunoglobulins and hybridoma antibodies. The inherent lack of specificity of structural data obtained by POSA and general lack of insight into other limitations of these analyses has often led to inaccurate and superficial interpretations. Proper interpretation of past and future studies necessitates pointing out previous inadequacies of dextran structural data and detailing more recently acquired structural information on the dextrans. Unambiguous terminology has been achieved by a new system of linkage symbols that includes the designation of structural positions, such as (1----3; l)- and (1----3; b) as linear-chain and branch-point positions, respectively. Results of immunological studies are reviewed. Improved interpretations and correlations are made on the basis of structural data from MSA and several other techniques which have become available on the dextrans.

Dextrans↗
PubMed1995

Dextran as protectant against damage caused by sparging for hybridoma cells in a bubble column.

The effect of the addition of dextran as a protective polymer against sparging was examined with hybridoma suspension cells in a bubble column under standardized conditions. The protective effect of high concentrations of high molecular weight dextran showed a correlation with the bulk viscosity of the medium. A distinct protective effect occurs at viscosities greater than 20 x 10(-3) Pa s-1. In contrast, low molecular weight dextrans that cause a minor increase in viscosity, also provide no protection against sparging. There is no strict correlation between surface tension and the protective effect of dextran against sparging. Oxygen transfer is strongly reduced by high concentrations of high molecular weight dextran. Therefore, addition of dextran as protective polymer against sparging for large-scale production processes with animal cells in stirred reactors does not seem feasible.

Animals↗
PubMed1995

Removal of iron dextran by hemodialysis: an in vitro study.

Intravenous iron dextran is frequently prescribed for iron-deficient hemodialysis patients, a practice that has increased during the erythropoietin era. Whether iron dextran is removed by hemodialysis has been a concern, especially for high permeability membranes. The purpose of this in vitro study was to measure iron dextran clearance by nine different hemodialyzers (Fresenius F3, F8, and F80B; Baxter CF25, CA150, CA210, and CT190; Toray BK2.1P; and Hospal Filtral 16) representing six types of membranes (polysulfone, cuprophane, cellulose acetate, cellulose triacetate, polymethylmethacrylate, and polyacrylonitrile) and including membranes considered high efficiency and high flux. Clearances were assessed using a closed-loop, fixed-volume reservoir model. Absolute drug removal also was determined over the 30-minute experiments. Iron dextran clearance did not exceed 25 mL/min, and clearances also were minimal after a single automated reuse with glutaraldehyde sterilant. A maximum of 8% of iron dextran was removed during the experiment. We conclude that iron dextran clearance by the nine hemodialyzers studied was small or too low to be detected in this sensitive in vitro dialysis system and that adjusting dosing schedules is not needed.

Hemodialysis Solutions↗
PubMed1995

Effects of dextran on the self-association of human spectrin.

The self-association of human spectrin is enhanced in the presence of dextran. The equilibrium constant for association of two heterodimers to form a tetramer is increased by an order of magnitude in the presence of 20% dextran. The rate constant for association is also enhanced, while the rate constant for dissociation is almost independent of dextran concentration. The degree of enhancement of association is dependent only on the mass concentration of dextran; for a given mass concentration of dextran the effect is independent of dextran molecular weight. These effects are believed to be due to excluded volume phenomena, and a model is presented that realistically accounts for the effects. These results imply that the association of spectrin within the erythrocyte will be enhanced by the presence of hemoglobin.

Dextrans↗
PubMed1977

Comparative effects of native insulin and insulin-dextran complexes on the metabolism of adipose tissue.

Insulin-dextran complexes have been prepared and their biological activities compared to that of native insulin. Complexes referred to as T-70 had a molecular weight of about 450 000, T-2000 being more than 2 million. Insulin-dextran T-70 was stable and no release of free insulin from the complex was observed upon its incubation with adipose tissue. Insulin-dextran T-70 mimicked the effects of native insulin upon adipose tissue metabolism. It also lowered blood sugar. These effects necessitated, on an insulin molar basis, concentrations of the complex that were 10-times greater than those needed for native insulin. Maximal concentrations of insulin complex T-70 or native insulin elicited similar quantitative effects. This suggested that when the concentration of insulin-dextran T-70 was high enough, the complex occupied a sufficient number of receptor sites to produce maximal stimulation of the tissue. In contrast, insulin-dextran T-2000 was barely effective, indicating that, probably due to its size, it was unable to reach receptor sites. The size, stability and metabolic effects of insulin-dextran T-70 observed in this study give additional support to the concept that insulin action is probably mediated via a series of events initiated at the level of the plasma membrane of adipocytes.

Adipose Tissue↗
PubMed1994

Dextran 70 administration after trauma-hemorrhagic shock does not impair cellular immune functions.

PURPOSE: Although the effects of the colloid dextran 70 on induction of anaphylactoid reactions or reticuloendothelial phagocytosis have been examined previously, its effects on specific cell-mediated immunity after trauma-hemorrhage shock remain unknown. METHODS: Nonheparinized C3H/HeN mice underwent a laparotomy, were bled, and then maintained at a blood pressure of 35 mm Hg for 60 minutes. Then they were resuscitated with either 4 x the shed blood volume as lactated Ringer's solution (LRS) or 2 x LRS + 1 x dextran 70. Control mice underwent all operative protocols but were neither hemorrhaged, nor resuscitated. At 2 or 24 hours posthemorrhage, serum, splenocytes (SPL), and peritoneal macrophages (pM phi, splenic Mo (sM phi) were obtained. Bioassays were used to determine interleukin-2 (IL-2), IL-3, IL-6, and SPL proliferation. RESULTS: Trauma-hemorrhage markedly depressed lymphokine release, splenocyte proliferation, and IL-6 release at 2 hours after the insult. The combination of LRS + dextran did not restore lymphocyte functions, but also did not further suppress them. The release of IL-6 by pM phi and sM phi at 2 and 24 hours after dextran infusion and serum IL-6 remained at the same level as in LRS-treated animals. CONCLUSIONS: The combination of LRS and colloid dextran 70 does not adversely affect ex vivo cell-mediated immune functions during the first 24 hours after its administration after trauma-hemorrhage. Thus, from the immunological standpoint, dextran is a safe resuscitation adjunct.

Analysis of Variance↗
PubMed1996

Functionalisation of Si/SiO2 and glass surfaces with ultrathin dextran films and deposition of lipid bilayers.

Dextran with molecular weight of 500 kDalton was covalently coupled to glass and Si/SiO2-surfaces by epoxy functionalisation or by photo reactive functionalisation of the solid surface. With the described methods we can control the deposited mass density between 0.3 and 4.8 ng/m2 corresponding to mean film thicknesses of a dry dextran film between 2 A and 30 A. We studied the structural properties (thickness, density) of ultrathin dextran layers coupled to silicon wafers of glass surfaces in humid atmosphere and under water by ellipsometry and reflection interference contrast microscopy (RICM) and developed a new method to measure the interfacial forces in ultrathin films. We demonstrate, that these hydrophilic polymer films from soft cushions which can be reversibly swollen both under water and in humidified air. In water the films swell up to thicknesses of about 600 to 800 A as measured by reflection interference contrast microscopy (RICM) and ellipsometry. The interaction of the polymer films with Si/SiO2-surfaces in contact with humidified air was studied by ellipsometry. We measured the thickness of the polymer layers as a function of the relative humidity of the atmosphere surrounding the sample in a hydration chamber. Depending on the humidity of the surrounding air and induced by the hydration the film thickness changes by about a factor of 10. Coupling of N-hydroxysuccinimide to dextran enables the functionalisation of the dextran cushions with a broad range of different specific binding molecules for various detection tasks. The possibility of soft hydrated dextran films as a cushion for the deposition of self healing lipid bilayers is shown.

Biocompatible Materials↗
PubMed2006

Glycine intravenous donor preconditioning is superior to glycine supplementation to low-potassium dextran flush preservation and improves graft function in a large animal lung transplantation model after 24 hours of cold ischemia.

OBJECTIVES: The potential role of glycine in combination with standard lung preservation with low-potassium dextran solution in lung ischemia-reperfusion injury has not been investigated in a preclinical porcine transplant model. METHODS: In a control group (n = 6), donor lungs were flushed with 1 liter of low-potassium dextran solution. In a second group (LPD-glyc, n = 6), low-potassium dextran solution was supplemented with 3.75 g of glycine. In a third group (IV-glyc, n = 6), donor preconditioning was performed by intravenous administration of 3.75 g glycine 1 hour before low-potassium dextran preservation. Grafts were stored in low-potassium dextran at 4 degrees C for 24 hours. Posttransplant graft function was assessed throughout a 7-hour observation period. RESULTS: In the control group, 2 recipients died of right-sided heart failure caused by severe ischemia-reperfusion injury. All animals of the glycine groups survived the entire observation period. Pulmonary vascular resistance remained significantly (P < .01) lower in both glycine groups when compared with controls. At the end of the observation period pulmonary vascular resistance in the control group was higher (P < .01) compared with the glycine groups (1310 +/- 319 dyn x sec x cm(-5) vs 879 +/- 127 dyn x sec x cm(-5) [LPD-glyc] vs 663 +/- 191 dyn x sec x cm(-5) [IV-glyc]). Changes of lung tissue water content were lower in the IV-glyc group compared with the LPD-control (P < .01) and LPD-glyc lungs (P < .05). Oxygenation (PO2/FiO2) was higher in the IV-glyc group compared with the LPD-glyc and control lungs (445 +/- 110 mm Hg vs 388 +/- 124 mm Hg [P < .01] vs 341 +/- 224 mm Hg [P < .001], respectively). DISCUSSION: Modification of low-potassium dextran solution with glycine or donor preconditioning ameliorates ischemia-reperfusion injury in lung transplantation. This intriguing approach merits further evaluation with respect to the mechanisms involved and may improve results in clinical lung preservation.

Animals↗
PubMed2003

Low potassium dextran lung preservation solution reduces reactive oxygen species production.

BACKGROUND: Low potassium dextran lung preservation solution has reduced primary graft failure in animal and human studies. Though the mechanism of reducing primary graft failure is unknown, low potassium dextran differs most significantly from solutions such as Euro-Collins (EC) and University of Wisconsin in its potassium concentration. The aim of this study was to investigate the impact that potassium concentration in lung preservation solutions had on pulmonary arterial smooth muscle cell depolarization and production of reactive oxygen species. METHODS: Using isolated pulmonary artery smooth muscle cells from Sprague-Dawley rats, the patch-clamp technique was used to measure resting cellular membrane potential and whole cell potassium current. Measurements were recorded at base line and after exposure to low potassium dextran, EC, and University of Wisconsin solutions. Pulmonary arteries from rats were isolated from the main pulmonary artery to the fourth segmental branch. Arteries were placed into vials containing low potassium dextran, EC, low potassium EC, Celsior, and University of Wisconsin solutions with reactive oxygen species measured by lucigenin-enhanced chemiluminescence. RESULTS: Pulmonary artery smooth muscle cell membrane potentials had a significant depolarization when placed in the University of Wisconsin or EC solutions, with changes probably related to inhibition of voltage-gated potassium channels. Low potassium dextran solution did not alter the membrane potential. Production of reactive oxygen species as measured by chemiluminescence was significantly higher when pulmonary arteries were exposed to University of Wisconsin or EC solutions (51,289 +/- 5,615 and 35,702 +/- 4353 counts/0.1 minute, respectively) compared with low potassium dextran, Celsior, and low potassium EC (12,537 +/- 3623, 13,717 +/- 3,844 and 15,187 +/- 3,792 counts/0.1 minute, respectively). CONCLUSIONS: Preservation solutions with high potassium concentration are clearly able to depolarize the pulmonary artery smooth muscle cells and increase pulmonary artery reactive oxygen species production. Low potassium preservations solutions may limit reactive oxygen species production and thus reduce the incidence of primary graft failure in lung transplantation.

Adenosine↗