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PubMed1981

Immunochemical specificity of the combining site of murine myeloma protein CAL20 TEPC1035 reactive with dextrans.

The immunochemical specificity of the combining sites of murine myeloma protein CAL20 TEPC1035 was studied by quantitative precipitin and precipitin inhibition assays. Myeloma protein CAL20 TEPC1035 precipitated with only three dextrans, B1355S4, B1498S, and B1501S, with high proportions of alpha(1 leads to 3) linkages, but not with any other dextrans, glycogen, and pullulan. Inhibition tests with various sugars show that the combining site of myeloma protein CAL20 TEPC1035 is most complementary to panose, a trisaccharide DGlc alpha(1 leads to 6)DGlc alpha(1 leads to 4)DGlc. Panose was 3.3 times more potent than a tetrasaccharide DGlc alpha(1 leads to 6)DGlc alpha(1 leads to 4)DGlc alpha(1 leads to 4)DGlc and 8, 23, 42, > 42 times more active than maltose, nigerose, isomaltose, and kojibiose, respectively. These findings were paralleled by their binding properties as determined by affinity electrophoresis. The association constants (Ka) of these three dextrans to myeloma protein CAL20 TEPC1035 ranged from 3.8 X 10(3) ml/g to 5.02 X 10(3) ml/g. The association constant of inhibitor (Kia) of panose was 8.19 X 10(3) M-1. Myeloma protein CAL20 TEPC1035 is an antidextran with specificity different from those of other murine myeloma antidextrans and from human antidextrans reported previously and its combining site size is at least as large as a trisaccharide. The binding constant of methyl alpha-D-glucoside (7.2 X 10(2)) was 73% of that of panose and comparable to that of myeloma protein W3129 (9.4 X 10(2)) with a cavity-type site and 600 times lower (1.6 X 10(0)) for QUPC52 with a groove type site, indicating that the terminal nonreducing residue is held in a cavity. Inhibition data with various alpha(1 leads to 4)-linked oligosaccharides also indicate that the internal portions of these inhibitors may react directly with a portion of the combining site. These findings suggest that myeloma antidextran CAL20 TEPC1035 has a partial cavity-type combining site in which the terminal nonreducing dGlc alpha(1 leads to 6) moiety is held in a cavity with the other two sugars forming a groove. However, oligosaccharides with one or more alternating [leads to 3DGlc alpha(1 leads to 6)DGlc alpha(1 leads to 3)DGlcl leads to] units with and without terminal nonreducing DGlc alpha(1 leads to 6) or DGLc alpha(1 leads to 3) side chains remain to be tested to determine whether structures known to be present in the three dextrans which precipitate CAL20 TEPC1035 may not prove to be more active than panose.

Animals↗
PubMed1998

Cost-effectiveness impact of iron dextran on hemodialysis patients' use of epoetin alfa and blood.

The cost-effectiveness impact of iron dextran administration on the use of epoetin alfa and blood in hemodialysis patients was studied. Subjects were ambulatory hemodialysis patients who had been receiving hemodialysis for at least six months before the start of an iron dextran protocol and who had been given epoetin alfa for at least four of those six months. Clinical data were collected for six months before and six months after the protocol was implemented. Successful treatment was defined as a hematocrit of 33-36%, a transferrin saturation of >10%, a ferritin concentration of >100 ng/mL, and no blood use except for acute blood loss. A total of 33 patients completed the study. Fifty units of blood were used in the first six months and nine units in the second six months. There was significant improvement in mean hematocrit, ferritin, and transferrin saturation values after the protocol began. Average epoetin alfa doses did not change significantly. There was significant improvement in success rates for ferritin and blood use and in the overall success rate. Ten patients met all success criteria in the preprotocol period, versus 27 in the postprotocol period. Monthly cost-effectiveness analysis for the preprotocol and postprotocol periods indicated costs of $1350 and $526, per successful treatment, respectively. The incremental cost-effectiveness of iron dextran was $42 per successful treatment. Iron dextran improved iron indices and reduced the need for blood transfusions but did not reduce the average dose of epoetin alfa. The additional cost of therapy per month seemed justified by the clinical benefits.

Adult↗
PubMed1979

Myocardial blood flow and O2-supply following dextran-haemodilution and methaemoglobinaemia in the dog.

The adequacy of oxygen-supply to the left ventricular myocardium was compared in anaesthetised dogs following: a) isovolaemic haemodilution with Dextran 70 (to haematocrit = 20 +/- 2%), b) the acute induction of methaemoglobinaemia (to methaemoglobin-concentration = 53 +/- 7%), bu subjecting them to an exchange-transfusion with methaemoglobin-containing erythrocytes in dextran) and c) a control exchange-transfusion with normal homologous erythrocytes in dextran. Myocardial oxygen-supply was assessed by the measurement of myocardial blood flow (using 15 +/- 5 micron microspheres) and of the PO2 in coronary sinus blood. In dextran-haemodiluted dogs, mean myocardial blood flow was 92% greater than in the control group, allowing the maintenance of normal coronary sinus PO2. In the methaemoglobinaemic group, flow was only 15% greater than in the control group; because oxygen-capacity was halved in this group, myocardial oxygen supply was impaired and coronary sinus PO2 fell. Infusions of isoproterenol (0.1 microgram . kg-1 . min-1) produced similar myocardial flow-increments in the haemodiluted and control groups with similar transmural distribution. In the methaemoglobinaemic group, however, the subendocardial flow-increment was reduced, in association with a further reduction in coronary sinus PO2 and an attenuated contractile response to the drug. These findings suggest that the rheological effects of haemodilution play an important role in the maintenance of myocardial oxygen-supply, but in significant methaemoglobinaemia, the oxygen supply of the myocardium may be severely curtailed.

Animals↗
PubMed1982

Comparison of acute cardiovascular effects and oxygen-supply following haemodilution with dextran, stroma-free haemoglobin solution and fluorocarbon suspension.

Changes in haemodynamics and myocardial oxygen-supply were investigated in anaesthetised dogs, when the haematocrit was reduced to 18 to 22% by isovalaemic haemodilution with 8% stroma-free haemoglobin solution, 20% Fluosol-DA and 6% dextran 70. Comparable haemodilution and comparable reduction in whole-blood viscosity was not followed by similar changes in cardiac output: dextran- and Fluosol-diluted dogs showed significantly elevated cardiac output, while haemoglobin-diluted dogs failed to do so. As a result, systemic O2-transport was better maintained with dextran and Fluosol. Myocardial blood flow, estimated by 15 micrometers microspheres, increased in all three haemodiluted groups, but oxygen-supply was not similar. The haemoglobin-diluted dogs showed inadequate O2-supply suggested by a fall of coronary sinus pO2; dextran-diluted dogs exhibited adequate O2-supply suggested by maintained coronary sinus pO2, while the Fluosol-diluted group enjoyed excessive O2-supply, indicated by a markedly elevated pO2 in coronary sinus blood. The observations suggest that similar degrees of haemodilution may not be followed by comparable changes in haemodynamics and O2-supply, depending on O2-unloading characteristics.

Animals↗
PubMed2000

Expression of multilectin receptors and comparative FITC-dextran uptake by human dendritic cells.

Dendritic cells (DC) are potent antigen-presenting cells and understanding their mechanisms of antigen uptake is important for loading DC with antigen for immunotherapy. The multilectin receptors, DEC-205 and macrophage mannose receptor (MMR), are potential antigen-uptake receptors; therefore, we examined their expression and FITC-dextran uptake by various human DC preparations. The RT-PCR analysis detected low levels of DEC-205 mRNA in immature blood DC, Langerhans cells (LC) and immature monocyte-derived DC (Mo-DC). Its mRNA expression increased markedly upon activation, indicating that DEC-205 is an activation-associated molecule. In Mo-DC, the expression of cell-surface DEC-205 increased markedly during maturation. In blood DC, however, the cell-surface expression of DEC-205 did not change during activation, suggesting the presence of a large intracellular pool of DEC-205 or post-transcriptional regulation. Immature Mo-DC expressed abundant MMR, but its expression diminished upon maturation. Blood DC and LC did not express detectable levels of the MMR. FITC-dextran uptake by both immature and activated blood DC was 30- to 70-fold less than that of LC, immature Mo-DC and macrophages. In contrast to immature Mo-DC, the FITC-dextran uptake by LC was not inhibited effectively by mannose, an inhibitor for MMR-mediated FITC-dextran uptake. Thus, unlike Mo-DC, blood DC and LC do not use the MMR for carbohydrate-conjugated antigen uptake and alternative receptors may yet be defined on these DC. Therefore, DEC-205 may have a different specificity as an antigen uptake receptor or contribute to an alternative DC function.

Animals↗
PubMed1987

Accumulation and metabolism of iron-dextran by hepatocytes, Kupffer cells and endothelial cells in the neonatal pig liver.

Treatment of newborn pigs with supplemental iron is a common procedure utilized to prevent neonatal anemia. The aim of this study was to investigate the hepatic distribution and intracellular metabolism of iron-dextran, a widely used colloidal-iron-carbohydrate preparation. Piglets were injected intramuscularly with iron-dextran (50 mg Fe/kg body wt) at 1 d of age. Hepatocytes and sinusoidal cells (Kupffer cells and endothelial cells) were isolated from iron-treated and control (uninjected) piglets at 2, 6 and 11 d of age. The concentrations of iron, copper and zinc in isolated cells were determined by atomic-absorption spectroscopy. In addition, the quantities of ferritin-protein and ferritin-iron were measured by immunoelectrophoresis and ion-exchange chromatography, respectively. At 2 d of age, the concentration (microgram/mg cell protein) of iron was 5-, 62- and 54-fold higher in hepatocytes, Kupffer cells and endothelial cells, respectively, isolated from iron-treated piglets than from control piglets. Hepatocytes, Kupffer cells and endothelial cells accumulated ferritin in response to iron-dextran treatment. Higher concentrations of ferritin-protein and ferritin-iron were present in Kupffer cells and endothelial cells than in hepatocytes at all times after treatment with iron-dextran. The percentage of cellular iron that was associated with ferritin, however, was greater in hepatocytes than in sinusoidal cells. Iron accumulated by all three liver cell types was mobilized to extrahepatic sites. Slight alterations in zinc and copper status of liver cells were evident at 11 d of age as a result of iron treatment.

Animals↗
PubMed1993

Centrifugation of human spermatozoa induces sublethal damage; separation of human spermatozoa from seminal plasma by a dextran swim-up procedure without centrifugation extends their motile lifetime.

While washing of human sperm cells by centrifugation and resuspension is a procedure in widespread use, there have been indications that this procedure per se may be harmful to the cells. The objective of this study was to investigate this question. To this end, a method for the clean separation of motile human spermatozoa from seminal plasma in the absence of centrifugation was developed, using a modified swim-up procedure, in which liquefied semen was mixed with an equal volume of 30 mg/ml dextran in medium, and the mixture overlaid with medium containing 5 mg/ml bovine serum albumin, forming two discreet layers with stable interface. The percentage of motile cells in a given sample was consistently > 80% immediately after recovery. Damage to the cells was assessed by loss of motile cells during incubation up to 96 h post-recovery. Comparison of aliquots of spermatozoa obtained by the dextran swim-up procedure showed that the aliquot subjected to centrifugation had 4 +/- 3% motile cells after 48 h, while the untreated aliquot had 52 +/- 12%. The aliquots showed no difference 1 h post-recovery. Similar results were obtained with spermatozoa that had been centrifuged in seminal plasma and resuspended in fresh plasma, then recovered by dextran swim-up. The delayed onset of motility loss in the centrifuged samples implies that this treatment induces sublethal damage in the cells. Comparison of the standard swim-up and Percoll gradient methods for sperm recovery, both of which involve centrifugation steps, showed decline in motility of the samples similar to that seen with dextran swim-up of centrifuged cells. We conclude that centrifugation per se induces sublethal damage in human spermatozoa, independently of treatment method, and suggest that recovery methods for human spermatozoa which avoid centrifugation might partially alleviate the damage incurred by these cells during cryopreservation.

Adult↗
PubMed1995

Receptor-mediated interaction of the low-density lipoprotein complex with dextran sulfate: potential as a multi-biological coupler.

We have investigated the receptor-mediated binding and uptake of low-density lipoprotein (LDL) and its insoluble complex with dextran sulfate to determine the contribution of positively charged sites in LDL to receptor-mediated interactions. Rabbit plasma LDL derivatized with FITC retained its sedimentation ability with dextran sulfate, as well as intact LDL, and binding of fluorescent LDL and its complex to liver cells was assayed by flow cytometry. Flow cytometry revealed that the binding of complex LDL with dextran sulfate, as well as that of pure LDL, increased on rat liver parenchymal cells treated with estrogen, which enhanced the expression of LDL receptors, and decreased on Hep G2 and Chang Liver cells treated with a monoclonal antibody against LDL receptors. The binding of pure LDL and complex LDL to hepatocytes was depressed by pretreatment with unlabeled LDL in a similar manner, but not with asialoglycoprotein, a ligand of asialoglycoprotein receptors on liver cells. Furthermore, we carried out a stable primary culture of rat hepatocytes, and then pure LDL and complex LDL were applied to the cultured hepatocytes. Hepatic-differentiated functions such as albumin and bile acid secretion decreased on the uptake of pure LDL and complex LDL in a similar manner. Consequently, comparative studies using pure LDL and complex LDL allowed us to determine that the complex formation with dextran sulfate had no influence on the receptor-mediated binding or uptake of LDL, and that LDL possessed binding domains for LDL receptors and sulfonic carbohydrates, containing positively charged amino acids.

Animals↗
PubMed1975

The effect of dextran on the lysability of ex vivo thrombi.

The lysability was determined of thrombi formed in Chandler tubes before and after infusion of 500 ml dextran 70 to patients undergoing cholecystectomy. 125I-labelled fibrinogen was given the day before operation. Following incubation of the thrombi formed in plasmin, radioactivity remaining in the thrombi and released to the supernatant was determined, as well as fibrinolytic degradation products in the supernatant, using an immunoelectrophoretic method. The dextran infusion was found to increase the radioactivity released from the thrombi to the supernatant from 15.5 plus or minus 7.6% to a maximum of 27.3 plus or minus 8.2% four hours after the infusion (P less than 0.001). A corresponding significant rise of the FDP concentration in the supernatant from 16.8 mug/ml to 44.1 mug/ml was found at the same time. After 24 hours the radioactivity had returned to initial values. The results indicate that dextran infused into patients during surgery increases the lysability of thrombi. It is suggested that this finding at least partly explains the antithrombotic effect of dextran.

Cholecystectomy↗
PubMed1979

Effects of aspirin and dextran on patency of bovine heterografts in the venous system.

The effect of short-term treatment with aspirin or dextran on the patency of bovine heterografts in the venous system was investigated. Animals treated preoperatively and for three days postoperatively with either aspirin (600 mgm BID) or low molecular weight dextran (500 cc/day) had statistically significant increased graft patency as compared to controls during the treatment interval. Significance was lost, however, after cessation of therapy. These results suggest that either aspirin or low molecular weight dextran would be useful adjuvants to venous reconstruction. If used in the venous system, bovine heterografts might be expected to yield better short term patency with adjuvant therapy employing either dextran or aspirin.

Administration, Oral↗
PubMed1992

Inhibitory effect of substituted dextrans on MCF7 human breast cancer cell growth in vitro.

Substituted dextrans can reproduce some of the properties of heparin and can thus be used to alter cellular growth. We studied the effect of heparin (H108), dextran (D), carboxymethylbenzylamide dextran (CMDB) and carboxymethylbenzylamide sulfonate dextran (CMDBS) on the growth of human mammary cells of the MCF7 tumor line. The cells were cultured in minimum Eagle's medium containing 2% fetal calf serum without biopolymer, or with increasing concentrations of H108, D, CMDB or CMDBS. Growth curves were accurately based on cell counting using a Coulter counter. Cell distribution in the various phases of the cycle was analyzed by flow cytometry. Dose-dependent growth inhibitory effects (400-4000 micrograms/ml) were observed. The effect on MCF7 tumor cells was most apparent with CMDBS. The percentage of cells in the S phase decreased with preferential blocking in the G0/G1 phase. Pre-clinical studies can be anticipated as there is an absence of in vivo toxicity.

Aged↗
PubMed2000

Clinical production of pharmaceutical grade Tc-99m dextran 70 for lymphoscintigraphy.

Lymphoscintigraphy is an established means to determine the lymphatic drainage patterns from malignant tissues and edematous extremities. Unfortunately, there are no commercially available dedicated radiopharmaceuticals labeled for use in lymphoscintigraphy studies. The authors report a simple way to extemporaneously compound pharmaceutical-grade Tc-99m dextran 70 to meet this need. Pharmaceutical-quality Tc-99m dextran 70 injection is prepared by a simple rapid method from drug intermediates that are marketed as parenteral drug products. The radiolabeling process yields a product of high radiochemical purity, with good in vitro and in vivo stability. The authors illustrate the use of this product in a patient with melanoma to show the lymphatic drainage pattern before surgery. The method described permits rapid compounding of Tc-99m dextran 70 injection from drug components that are intended for parenteral administration. Tc-99m dextran 70 provides the option of performing lymphoscintigraphy in any clinical nuclear medicine setting.

Aged↗
PubMed1994

Transport of fluorescent dextrans across the rat ileum after cutaneous thermal injury.

OBJECTIVE: To determine the time course and spatial distribution of uptake of macromolecules in the small intestine of rats subjected to cutaneous thermal injury. DESIGN: Prospective, controlled animal study. SUBJECTS: Fifty-five female Sprague-Dawley (CD) rats subjected to scald burn injury covering 20% (small injury; n = 29) and 40% (large injury; n = 6) of the total body surface area between 3 and 72 hrs after injury. Animals subjected to sham injury (n = 20) were used as controls. INTERVENTIONS: The intestine was cannulated near the distal ileum and incised 7 cm upstream. After perfusion with physiologic buffer, this intestinal loop was filled with the same buffer containing fluorescent-labeled dextrans (3 and 70 kilodaltons molecular weight) and ligated 4 cm from the injection point. After a 2-hr incubation period, the tissues were fixed with paraformaldehyde and cryosections were examined by laser confocal microscopy. The mesentery was also observed by laser confocal microscopy during incubation with the permeability probes. The disappearance of fluorescence was studied after washing the dextran probes from the gut lumen. MEASUREMENTS AND MAIN RESULTS: In small injuries, there was a transient uptake of the 3-kilodalton dextran by the epithelium in focal regions of the ileum with the effects seen between 7 and 21 hrs after injury. In large injuries, epithelial staining was visible within 3 hrs, and the marker was seen to translocate both to the lymphatics and the blood vessels of the mesentery. In comparison, the 70-kilodalton dextran was visible within the intercellular spaces. Little or no epithelial staining was seen in sham-injured animals. CONCLUSIONS: These results suggest that a transcellular pathway for the translocation of small macromolecules from the lumen to the mesentery can be activated after burn injury. The novel techniques described here will be useful to examine intestinal transport in various pathologic situations.

Animals↗
PubMed1999

The adult respiratory distress syndrome after dextran infusion as an antithrombotic agent in free TRAM flap breast reconstruction.

Adult respiratory distress syndrome occurred in a patient who had received dextran as a routine antithrombotic agent during and after free TRAM breast reconstruction. Although most patients who receive dextran have no adverse reaction, particularly after hapten inhibition by dextran 1 infusion, the serious nature of this complication in an elective operation calls into question the continuing routine use of dextran in microsurgery.

Anticoagulants↗
PubMed2004

Paenibacillus isolates possess diverse dextran-degrading enzymes.

AIMS: To isolate and identify dextran-degrading organisms from sugar mill and compost samples, and to examine the diversity of the dextranolytic enzymes produced. METHODS AND RESULTS: Fifteen dextranolytic prokaryotes were purified at various temperatures from sugar-mill or compost samples, using indicator plates containing blue dextran. A 16S rRNA gene sequence analysis showed that 12 isolates purified at 40, 50 or 70 degrees C were closely aligned to Paenibacillus spp. The three isolates purified at 60 degrees C had identical 16S rDNA sequences, with highest affinity to Bacillus spp. Liquid culture of the 11 isolates purified at 40 or 50 degrees C produced dextranolytic activity in the spent media with maximal activity at 40 or 45 degrees C under the assay conditions used. Hydrolysis of blue dextran in activity gels showed that the 12 Paenibacillus isolates produced from one to five dextranolytic proteins, ranging from 70 to 120 kDa. Based on 16S rDNA sequence, growth habit in liquid culture and dextranolytic enzyme pattern, the 12 Paenibacillus-like isolates could be differentiated into six distinct groups, one of which was capable of growth at 70 degrees C. CONCLUSIONS: The Bacillales, especially the Paenibacillus, are a valuable environmental repository for dextranolytic enzymes of diverse size and potentially diverse activity. SIGNIFICANCE AND IMPACT OF THE STUDY: Dextranolytic enzymes produced by Paenibacillus spp. are an exploitable resource for those interested in modifying the structure of dextrans.

Bacillus↗
PubMed1990

A double-blind trial of Dextran-haemodilution vs. placebo in claudicants.

Haemodilution is often recommended for peripheral arterial disease, yet little data is available to support its clinical efficacy. This study was designed to prove or disprove the effectiveness of Dextran-haemodilution in intermittent claudication. Twenty claudicants with long, well-collateralized arterial occlusions were randomized into groups 1 and 2. Group 1 received isovolemic haemodilution with Dextran 40 (500 ml per session) during 3 weeks, which was followed by a wash-out period, followed by placebo treatments for 3 weeks. In group 2 this sequence was reversed. Pain-free and maximal walking distances were measured by standardized treadmill tests along with plethysmographic blood flow, Doppler pressures, haematocrit, blood and plasma viscosity as well as fibrinogen. Walking distances increased significantly by about 50% during haemodilution in both groups. This was paralleled by a fall in haematocrit and blood viscosity. All other variables remained constant. During placebo treatments there were no significant changes of any variable. The treatment was tolerated without complications. Thus Dextran-haemodilution seems safe and effective in selected peripheral occlusive arterial disease (POAD) patients. Potential responders might be identifiable before the start of therapy by angiographic investigations. The clinical effectiveness of Dextran 40 is comparable to that of hydroxyethyl starch 200 as reported in the literature.

Adult↗
PubMed1979

V and C gene contribution in creating anti-alpha-1,3 dextran antibodies in mice. II. Characteristics of Ig-lb to e prototype responses.

The light (L) and heavy (H) chain and the idiotypic (Id) composition of the antibody (Ab) plaque-forming cells (PFC) and serum Ab specific for alpha-1,3 dextran have been characterized in murine strains exhibiting the CH-Ig-lb to e allotypes and in their F1 hybrids with Ig-la1, BALB/c prototypes. The Ab response of the Ig-lb to e mice to the alpha-1,3 dextran was low in the kappa (kappa) L chain class with only a minor, sporadic Ab in the lambda (lambda) L chain class discernible after prolonged immunization. Two of a total of sixty-eight C57Bl/6 Ig-lb mice, in a total of 318 Ig-lb to e mice tested, exhibited, in late responses, a significantly elevated Ab in the lambda L class at both serum and PFC levels, equalling at the PFC level the total non-specific lambda PFC values. An Id analysis showed this lambda Ab and the kappa Ab to lack the Id relatedness to the three BALB/c alpha-1,3 dextran-binding myeloma proteins (MP) Ab prototypes, J-558, 104 E, and UPC-102, exhibited by the lambda Id+ Ab of the Ig-la1 BALB/c prototypes and their F1 hybrids with the Ig-lb to e prototypes. Furthermore, affinity differences could be detected by alpha-1,3 nigerodextrans, PFC inhibition analysis, between the late C57Bl/6 anti-alpha-1,3 dextran lambda Id--Ab PFC and the lambda Id+ Ab PFC of the BALB/c and their F1 progeny.

Animals↗
PubMed1982

Cyclosporin A inhibits thymus-dependent but not thymus-independent immune responses induced by dextran B512.

We have compared previous studies of the immune response to dextran (Dx) B512 in thymus-deficient nu/nu and thymectomized, lethally irradiated, and bone marrow-reconstituted mice with those obtained with cyclosporin A (CyA) as a T-cell-inhibiting drug. Our data show that only immune responses to TD forms of Dx B512 are susceptible to suppression by CyA, whereas anti-alpha 1-6 and anti-DNP antibodies induced by high molecular weight dextran and DNP-native Dx, respectively, were not inhibited. Similar results were obtained when polyclonal responses were studied. The polyclonal response induced by another dextran preparation, the polyanion dextran sulphate (DxS), was inhibited by CyA to the same extent as polyclonal T-cell activation by T-cell mitogens.

Animals↗