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Differential effect of interferon-gamma and interleukin-2 on the induction of IgA and IgM anti-dextran responses.

Supernatants from T-cell lines and T-cell hybridomas can substitute for T cells in the induction of the anti-alpha(1,3) Dextran B1355 plaque-forming cell response in culture. The present study sought to define the lymphokines required for the induction of IgA and IgM anti-alpha (1,3) dextran responses. Recombinant Interferon-gamma (IFN-gamma) supported the induction of low levels of IgA anti-alpha(1,3) dextran plaque-forming cells in splenic B-cell cultures. IgA responses were substantially increased when cultures containing IFN-gamma were supplemented with an interleukin 2 (IL-2)-containing supernatant from the murine T-cell hybridoma BW.Mls, purified murine IL-2, or recombinant human IL-2. In striking contrast, IgM anti-alpha(1,3) dextran plaque-forming cells were not produced in cultures containing IFN-gamma alone or in combination with purified or recombinant IL-2. However, substantial IgM responses could be produced in cultures containing IFN-gamma and BW.Mls supernatant. This data indicates that there may be different lymphokine requirements for the induction of IgA and IgM anti-alpha(1,3) dextran B cells, or alternatively, such B cells may be in different stages of differentiation and therefore, not respond to the same lymphokines.

Animals↗

The role of calcium ions in DEAE-dextran-induced stimulation of neutrophil migration.

The polycation DEAE-dextran caused a strong enhancement of non-directed migration of rabbit peritoneal neutrophils. The activating effect on migration was completely annulled in the presence of the polyanion poly-D-glutamic acid, indicating that the effect depended on the positive charge of the macromolecule. Chemotaxis activated by the chemotactic peptide fMLP was only slightly affected by the polycation. In contrast with fMLP-activation, stimulation of migration by DEAE-dextran was dependent on the presence of extracellular Ca2+. DEAE-dextran also stimulated migration of electroporated neutrophils. The stimulation was absent when calcium was not present; the increase of migration was strongest at Ca(2+)-concentrations between 100 nM and 1 microM Ca2+. This indicates that the requirement for extracellular Ca2+ in intact cells is a reflection of the intracellular requirement. Several types of calcium blockers gave a moderate inhibition of DEAE-dextran activated migration. Activation of migration by DEAE-dextran of electroporated neutrophils was completely inhibited by calcium channel blockers, at very low concentrations. The results suggest that both Ca(2+)-fluxes across the plasma membrane and Ca2+ from intracellular stores are required for DEAE-activated migration, and that the calcium from the intracellular source is required on a place where the extracellular Ca2+ has no, or limited, admittance.

Animals↗

A simple method of measuring aqueous humor flow with intravitreal fluoresceinated dextrans.

Fluoresceinated dextran was injected into the vitreous cavity of rabbits and its concentration was followed in the vitreous and aqueous humors for a period of weeks by slip-lamp fluorometry. The concentrations in both fluids fell in parallel and in an exponential manner where the time constant was determined by the molecular weight of the dextran. The rate of movement of the dextran from the vitreous into the anterior chamber is controlled by passive diffusion and may be assumed to be relatively steady; accordingly, the faster the rate of flow in the aqueous humor the lower will be the concentration of dextran in it. Circadian variations in the aqueous flow were observed, a drop at night time being noted. These were small enough for changes due to systemic influences to be determined; in particular, it was found that general anesthesia with ketamine had no significant effect. The ratio of the concentration in the anterior chambers of the two eyes of one animal normally remains constant with time. Changes in the ratio can be used to determine the influence of unilateral interventions. This was illustrated by the use of adrenergic drops which led to a brief rise in flow rate and then a longer-lasting fall, and by X-ray irradiation with 1000 rads, which caused an immediate fall in flow rate interrupted for a time by a phase of increased flow. An absolute estimate of the aqueous flow can be derived from measurements of the total amount of dextran in the eye and its concentration in the anterior chamber. A definite value has not yet been established owing to ambiguities in determining the former quantity.

Anesthesia, General↗

Dextran-70 versus albumin as plasma expanders in cirrhotic patients with tense ascites treated with total paracentesis. Results of a randomized study.

To investigate whether albumin can be substituted by less expensive plasma expanders in cirrhotic patients with tense ascites treated with total paracentesis, 88 patients (16 with renal failure) submitted to this therapeutic procedure were randomly assigned to receive IV albumin (43 patients) or dextran-70. Both substances were given at a dose of 8 g/L of ascitic fluid removed. Patients were discharged from the hospital with diuretics, and cases developing tense ascites during follow-up were treated according to their initial schedule. Total paracentesis was effective in eliminating the ascites in all but two cases in each group. Neither paracentesis plus IV albumin infusion nor paracentesis plus IV dextran-70 infusion was associated with significant changes in renal and hepatic function or serum electrolytes. The incidence of renal impairment (one case in each group), hyponatremia (three and four cases, respectively), and other complications (hepatic encephalopathy, gastrointestinal hemorrhage, bacterial infections) after paracentesis, and the clinical course of the disease as estimated by the probability of readmission to hospital during follow-up, causes of readmission, probability of survival, and causes of death were similar in the two groups of patients. The effect of paracentesis on effective intravascular volume was indirectly assessed by measuring plasma renin activity and aldosterone concentration before and 2 and 6 days after treatment, the patients being without diuretics. In patients treated with albumin, no significant changes in renin and aldosterone were observed during the entire period of observation. In contrast, both parameters increased significantly on the 6th day of treatment in patients receiving dextran-70. A significant increase in plasma renin activity and aldosterone concentration (30% over baseline values) was observed in 51% of patients treated with dextran-70 and in only 15% of those treated with albumin (x2 = 10.4; P = 0.0012). These results indicate that although dextran-70 is less efficacious than albumin in protecting cirrhotic patients treated with total paracentesis from the decrease in effective intravascular volume, it appears to be capable of preventing the renal and electrolyte complications induced by this therapeutic procedure.

Adult↗

Low molecular weight dextran in experimental pancreatitis: effects on pancreatic microcirculation.

UNLABELLED: Although low molecular weight (LMW) dextran has been said to decrease the lethality of experimental acute pancreatitis (AP) by reversing stasis in the pancreatic microcirculation, the actual mechanism(s) of action is unknown. This investigation was designed to measure the effects of low molecular weight dextran on pancreatic capillary flow (QCAP) and arteriovenous shunt flow (QAVS), and on pancreatic oxygen consumption (O2CP) following bile-trypsin-induced AP in dogs. Total pancreatic blood flow (QT) was measured with an electromagnetic flow probe on the superior pancreaticoduodenal artery (SPDA). QAVS was measured by liver trapping of 99mTc-albumin microspheres after SPDA injection. QCAP was calculated as QT minus QAVS. Seventeen dogs were treated with lactated Ringer's (LR) solution at 6.5 ml/kg/hr; 10 dogs were treated with LMW dextran 10% in normal saline at 1.5 ml/kg/hr plus LR at 5.0 ml/kg/hr. Mean arterial and central venous pressures remained constant throughout the 4-hr experiment. In the dogs receiving LR only, QT decreased from 42.7 to 24.4 ml/min (P less than 0.001); QAVS remained constant at 1.35 +/- 0.04 ml/min. During the first 30 min O2CP decreased from 1.17 to 0.76 ml O2/min (P less than 0.05) and remained constant thereafter. LMW dextran treatment altered none of these hemodynamic or metabolic parameters significantly. CONCLUSIONS: bile trypsin AP in the dog causes significant decreases in QT and QCAP without altering QAVS. The decrease in O2CP in association with a constant QAVS suggests a metabolic block to oxygen uptake at the cellular level. Continuous infusion of LMW dextran at a dose of 1.5 ml/kg/hr in the dog does not reverse these abnormalities.

Acute Disease↗

Dextran as a modulator of immune and coagulation activities in trauma patients.

Low Mr dextran has been utilized as a prophylactic therapy in treatment of coagulopathy. There is evidence that monocyte dysfunctions are important contributors to hypercoagulability episodes, as well as to immunoincompetence post-trauma. Dextran is a known monocyte modulator. Consequently, we evaluated the efficacy of dextran infusion in moderating immune dysfunction, monocyte aberrations, and hypercoagulability episodes. Twenty-eight trauma patients were randomly divided into two groups. One group of 15 received dextran at 1 g/kg wt/24 hr for 5 days in addition to standard resuscitation and treatment. The control or nontreated patient group received only standard treatment. Trauma patients in the two groups were retrospectively matched by injury severity score (ISS) to ensure comparability. Blood samples were collected daily for some studies and at 3-day intervals for other assays. In vivo coagulation status was evaluated by assessing the changes in intravascular fibrinopeptide A (FPA). Immune reactivity to the mitogen phytohemagglutinin (PHA) was also evaluated. Both monocyte production of plasminogen activator (PA) and monocyte production of procoagulant activity (PCA) have been shown to correspond to and be augmented by monocyte-T lymphocyte interactions. Consequently, monocyte production of plasminogen activator and procoagulant activity were assessed as measures of monocyte immune activity as well as indicators of monocyte function in controlling the balance between fibrinolysis and coagulation. Only patients with ISS of greater than 25 experienced significant immune, coagulation, or monocyte aberrations. Of those having an injury severity score (ISS) score of 25-35, all of the control and two of the dextran patients had significant perturbations in their immune and monocyte functions.(ABSTRACT TRUNCATED AT 250 WORDS)

Blood Coagulation↗

Components of coagulation and fibrinolysis during thrombosis prophylaxis with a low molecular weight heparin (Enoxaparin) versus Dextran 70 in hip arthroplasty.

In a prospective randomized study heptest, thrombin-antithrombin complexes (TAT), D-dimer, and t-PA:ag were analysed pre- and postoperatively in 206 consecutive patients undergoing hip arthroplasty during thromboprophylaxis with either a LMW heparin (Enoxaparin) or Dextran 70. Deep vein thrombosis (DVT) developed in 6 of 102 (6%) Enoxaparin and in 21 of 104 (20%) Dextran patients diagnosed by bilateral phelobography. In the Enoxaparin group heptest showed a significant increase from the pre- to the postoperative level opposed to a significant decrease in the Dextran group. Postoperative levels of TAT, D-dimer, and t-PA:ag were significantly increased in both groups, however, TAT was significantly higher in patients in the Dextran group than in the Enoxaparin patients. D-dimer was significantly higher in Dextran patients with DVT postoperatively compared with patients without DVT. No differences concerning TAT or t-PA:ag were observed between patients with and without DVT in any of the groups.

Adult↗

Effect on primary haemostasis of prophylactic regimens of low molecular weight heparin, unfractionated heparin, dextran and their combinations. An animal experimental study.

The aim of the study was to determine whether an impairment of the haemostasis could be observed experimentally when thromboprophylactic substances, which act differently on the haemostatic mechanism, were given single or in combination in prophylactic doses. In 36 rabbits we measured the primary haemostatic plug formation time (PHT), rebleedings and total haemostatic plug formation time (THT) after transection of venules and arterioles using an intravital microscope. We combined unfractionated heparin (UH) and low molecular weight heparin (LMWH) in low dose with either dextran 70 or polygeline (placebo volume expander) in a randomized double-dummy set up. In the placebo group (NaCl and polygeline) the median PHT was 55 and 101 seconds for arterioles and venules respectively, which are well-comparable to earlier results from our group. Most prolonged PHT and THT for arterioles were seen for dextran+NaCl, actually less prolongation was seen for UH+dextran. We did not observe any differences, except for a prolongation of THT for venules in rabbits given dextran+NaCl (p less than 0.05). Thus, in thromboprophylactic doses used, there does not seem to be an impaired or additive effect between heparins and dextran 70 in primary haemostasis in rabbits.

Animals↗

Studies on lectins. XLIX. The use of glycosyl derivatives of Dextran T-500 for affinity electrophoresis of lectins.

p-Aminophenyl glycosides and glycosylamines were coupled to periodate oxidized Dextran T-500 either directly or through an epsilon-aminocaproic acid spacer. The new glycosylated derivatives of dextran specifically precipitate lectins having the appropriate carbohydrate specificity, and thus were used in the preparation of affinity gels for affinity electrophoresis of lectins. The apparent strength of interaction of several lectins with carbohydrate residues immobilized in this way was less than with carbohydrates immobilized in O-glycosyl polyacrylamide copolymers. The presence of epsilon-aminocaproic spacer had no effect on the strength of interaction. The advantages of this type of macromolecular derivative of the ligand for affinity electrophoresis and some differences between the glycosylated dextrans and O-glycosyl polyacrylamide copolymers are discussed. Dextrans containing bound p-aminophenyl alpha-D-mannopyranoside and p-aminophenyl alpha-D-glucopyranoside were used to study the binding properties of concanavalin A and the lectin from Lathyrus sativus seeds. For the investigation of interaction of lectins from Ricinus communis and Glycine soja seeds, dextran derivatives containing bound p-aminophenyl alpha- and beta-D-galactopyranosides and alpha- and beta-D-galactopyranosylamines were used.

Binding Sites↗

Effects of dextran on hippocampal brain slice water, extracellular space, calcium kinetics and histology.

Hippocampal brain slices are valuable models for studying brain function but are compromised by several artifacts, including significant water gain and histologic injury, which occur under certain incubation conditions. Addition of colloid to Krebs-Ringer buffer (K-R) has been shown to eliminate water gain but has not achieved widespread acceptance. We confirm prior observations that dextran and PEG lessen the increase in slice mass during incubation in a dose-dependent manner with no water gain occurring at 4% concentrations. However, we also observe that addition of colloid to standard K-R induces severe neuronal pyknosis. Fortunately, the pyknosis can be eliminated by reduction in buffer osmolarity through adjustment of NaCl, producing markedly improved slice histology in dextran buffer, especially in the CA3 and CA4 regions of the hippocampus which are severely injured when incubated submerged in K-R at 37 degrees C. Extracellular space markers are not affected by either colloid. The volume of distribution for 45Ca is much larger in dextran buffers than in K-R and variability of 45Ca kinetics is also reduced. In the presence of dextran, hypoxia induces significant slice water gain, a relatively selective histologic injury and an alteration of tissue Ca2+ kinetics. Use of dextran buffers may eliminate many troubling brain slice artifacts.

Animals↗

Dextran sulfate inhibits the fusion of influenza virus with model membranes, and suppresses influenza virus replication in vivo.

The effect of dextran sulfate and related compounds on the fusion of influenza A virus with model membranes, composed of dioleylphosphatidyl-choline and cholesterol (1:0.5), was investigated by a fusion assay based on de-quenching of fluorescence of octadecyl-rhodamine-HC1 (R18). Dextran sulfate samples of molecular weight of 500,000, 8,000 and 5,000 were found to be potent inhibitors of the virus-liposome fusion process. Polygalacturonic acid also showed anti-fusion activity, but to a lesser extent. Uncharged dextran, positively charged diethylaminoethyldextran, and the monomer glucosamin-1,6-disulfate were ineffective. It was shown that dextran sulfate interacts with the virus. Our results suggest that dextran sulfate binds to and inactivates the viral fusion protein.

Cytopathogenic Effect, Viral↗

Mechanism of inhibitory effect of dextran sulfate and heparin on human T-cell lymphotropic virus type I (HTLV-I)-induced syncytium formation in vitro: role of cell-to-cell contact.

Cell-to-cell contact is usually essential for syncytium formation by HTLV-I-infected cell lines. The present study was undertaken to determine the inhibitory effect of polyanionic compounds, dextran sulfate and heparin, on HTLV-I-induced syncytium formation, as demonstrated by the fusion of HTLV-I-infected cells with target cells. These two compounds almost completely blocked syncytium formation in the early phase of the reaction at a concentration of 125 micrograms/ml, but dextran, as a control, did not inhibit it at concentrations up to 625 micrograms/ml. 50% inhibition of syncytium formation was detected at a concentration of 2 micrograms/ml of dextran sulfate 5000, 3 micrograms/ml of dextran sulfate 8000 and 8 micrograms/ml of heparin. The binding of radiolabeled HTLV-I-infected cells (HCT-1) to the target cells was inhibited by addition of dextran sulfate and heparin, and the inhibitory effects were concentration-dependent. No marked changes were detected in the expression of adhesion molecules on the virus-infected cells and target cells, and in the expression of envelope proteins on the virus-infected cells after exposing them to the polyanionic compounds. These results suggest that the blocking of cell-to-cell contact by polyanionic compounds, probably independent of surface adhesion molecules, is important for their inhibitory effect on HTLV-I-induced syncytium formation.

Antibodies, Monoclonal↗

Beneficial effects of dextran 70 versus Ringer's acetate on pulmonary function, hemodynamics and survival in a porcine endotoxin shock model.

The effects of Dextran 70 with NaCl as against Ringer's acetate on hemodynamics, gas exchange, oxygen transport and survival were evaluated in a porcine model of pulmonary and circulatory insufficiency induced by a continuous i.v. endotoxin infusion over 6 h. Dextran and Ringer's acetate were infused continuously to maintain baseline mean left atrial pressure (MLAP) throughout the endotoxin period. Twelve pigs receiving endotoxin + Ringer's acetate displayed a progressive 45% decline in cardiac output (Qt) and a two peaked increase in pulmonary vascular resistance (PVR) with a late increase of 250%. Venous admixture (Qva/Qt) increased progressively more than 6-fold and extravascular lung water (EVLW) increased by 55%. Mean arterial blood pressure (MAP) fell by 25%, oxygen delivery by 40%, base excess (BE) ranged between - 4.5 and - 9 mmol.1(-1) at the end of the endotoxin period and 4 of 12 animals died. Polymorphonuclear cell count (PMNs) fell rapidly by 90% and was severely decreased throughout the endotoxin period. Contrastingly, the 12 pigs that received endotoxin + Dextran maintained Qt near baseline and PVR was significantly lower in this group. Qva/Qt increased progressively more than 4-fold, but was significantly lower than in the Ringer's group as was the increase in EVLW (23%). MAP only decreased by 10%, oxygen delivery only decreased by 20%. BE ranged between - 1.0 and - 3.0 at the end of the endotoxin period and all animals survived. PMNs fell by 90% at 0.5 h but subsequently tended to return towards baseline. PMNs were significantly increased compared with the Ringer's group. The amount of Ringer's acetate necessary to maintain a stable MLAP averaged 4.6 times the Dextran volume. The superiority of Dextran as compared with Ringer's acetate in this endotoxemic shock model seems to be consequent to better rheological effects combined with pharmacological interactions with granulocytes.

Animals↗

Enhanced stability of beta-galactosidase in parenchymal and nonparenchymal liver cells by conjugation with dextran.

In order to enhance the stability of beta-galactosidase, we conjugated the enzyme with dextran T-10 (Mr approx. 10 000). The conjugate contained 9-10 mol dextran/mol protein (beta-galactosidase, Mr 68 000), and the specific activity retained after conjugation was 90 +/- 4% (n = 3) of the initial activity. Uptake and degradation of native and conjugated beta-galactosidase in isolated hepatocytes and nonparenchymal liver cells was studied. There was a marked increase in stability against degradation in both cell types when beta-galactosidase was conjugated with Dextran. The degradation of dextran-conjugated enzyme was reduced by 35% in hepatocytes and by 43% in nonparenchymal cells, after 80 and 40 min, respectively, as compared with the free enzyme. However, there was insignificant difference between the uptake of native and conjugated enzyme into the liver cells. Upon intravenous infusion into rats, native and conjugated enzyme were cleared from plasma with only a slight difference in the clearance rate. The observed stability of dextran-conjugated beta-galactosidase towards cellular degradation was in accordance with the in vitro experiments. The conjugate showed marked thermal stability at 50 degrees C and enhanced resistance towards proteolysis by the broad specific protease subtilopeptidase A. This demonstrates that dextran conjugation may be used as a means of stabilizing lysosomal enzymes for therapeutic purposes.

Animals↗

Fluorescent dextrans as sensitive anterograde neuroanatomical tracers: applications and pitfalls.

We have examined five conjugated 10,000 mol. wt. dextrans as potential anterograde tract tracers: Lucifer Yellow, Texas Red, fluorescein, Cascade Blue and tetramethylrhodamine. Pressure injections were made into the brain, dorsal root ganglia or footpads of adult rats. The retrograde tracer Fluoro-Gold was injected alone or mixed with the dextrans before injection. Three-14 days after injection, animals were perfused and sections cut with a freezing microtome. Texas Red-, fluorescein- and tetramethylrhodamine-conjugated dextrans produced intense labeling of neuronal cell bodies, axons and dendritic processes at the injection site and were transported by neurons predominantly in an anterograde direction to yield terminal and preterminal labeling. Relative to the fluorescein and tetramethylrhodamine conjugates, the quality and intensity of the anterograde labeling produced by Texas Red was variable. Results with Lucifer Yellow and Cascade Blue conjugates were negative. Optimal results were produced by slow-pressure injections via glass micropipettes. In comparison with Fluoro-Gold, retrograde transport by the dextran conjugates was present, but limited in its extent. Injections of the tetramethylrhodamine conjugate into dorsal root ganglia produced anterograde labeling of afferent fibers in visceral organs and injections into the nucleus ambiguous labeled motor fibers in the esophagus. Double/triple labeling was observed in the brain and spinal cord following multiple injections of fluorescein, tetramethylrhodamine and Fluoro-Gold. Also, Fluoro-Gold could be mixed with one of the dextrans in order to produce specific retrograde and anterograde labeling from the same injection site. The conjugates were compatible with fluorescent immunocytochemical procedures, but proved unsuitable for peripheral injections.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Effects of storage and homogenization methods on the hepatic recovery of dextrans determined by size-exclusion chromatography.

The effects of storage and homogenization methods on the analytical recovery of dextran macromolecules from rat livers were investigated using a high-performance size-exclusion chromatographic (HPSEC) method. Livers were collected from rats dosed with fluorescein-labeled dextrans with molecular weights of 150 or 70 kD. Subsequently, the livers were subjected to different methods to study the effects of the following parameters on the hepatic recovery of dextrans: storage method (freezing the livers before homogenization or freezing the homogenates); contents of the homogenization buffer (addition of 1% Triton X-100); and sample type (HPSEC analysis of the whole homogenate or the supernatant after centrifugation). It is shown that in the absence of Triton in the homogenization buffer, the hepatic recovery of dextrans is substantially affected by all the factors studied. However, in the presence of 1% Triton in the buffer, the hepatic recoveries were maximal and independent of the storage method or sample type. These studies suggest that for optimal recovery of dextran macromolecules from the liver, a sample preparation method capable of disrupting the subcellular membranes should be used.

Analysis of Variance↗

The use of dextran post free tissue transfer.

Dextran has been used in microsurgery to reduce the risk of free tissue transfer loss. A number of regimens which vary considerably in dosage and timing have been published in the literature. Using a postal questionnaire, a survey was conducted to delineate the current practise of UK plastic surgeons. Data were received from 161 plastic surgeons in 51 units (response rate of 61%). Forty-five percent of microsurgeons routinely use dextran post-operatively whilst 29% use alternative thromboprophylaxis. The indications, post-operative regimes and duration of administration of dextran vary significantly amongst surgeons and units. The reported success rates of free tissue transfer and digital replants were 97 and 85.1%, respectively, and was not significantly affected by the use of dextran. We conclude that there is considerable variation amongst UK plastic surgeons regarding thromboprophylaxis post microsurgery. Our data suggest that the use of dextrans does not affect free tissue transfer success rates.

Anticoagulants↗

Release characteristics of salmon calcitonin from dextran hydrogels for colon-specific delivery.

Biodegradable dextran hydrogels were synthesized by crosslinking dextran (T-70) with epichlorohydrin (ECH) for the in vitro colon-specific delivery of salmon calcitonin (sCT). Crosslinking reaction was performed in 2.8 M NaOH solution both in the presence and absence of ethanol at 10 and 23 degrees C. Biodegradation kinetics of dextran hydrogels were studied and, in the presence of 0.7 IU ml-1 dextranase, dextran discs lost 71.0% and 56.5% of their dry weight within 80 h at pH 5.5 and 7.0, respectively. sCT was derivatized with the fluorescamine (FSM) at borate buffer (pH 9.0) and the quantitative determinations were performed using spectrofluorimetric method (lambdaex: 390 nm, lambdaem: 475 nm). In vitro release studies for the hydrogels prepared in the presence of ethanol were carried out in simulated gastrointestinal fluids. Results indicated that 84.9% of the loaded-sCT was released for 17 h and dextran hydrogel prepared in the presence of ethanol may be a good delivery device for the colon-specific delivery of other peptide-type drugs as well as sCT.

Animals↗