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PubMed2002

Analysis of a dextran-binding domain of the dextranase of Streptococcus mutans.

AIMS: To examine the dextran-binding domain of the dextranase (Dex) of Streptococcus mutans. METHODS AND RESULTS: Deletion mutants of the Dex gene of Strep. mutans were prepared by polymerase chain reaction and expressed in Escherichia coli cells. Binding of the truncated Dexs to dextran was measured with a Sephadex G-150 gel. Although the Dexs which lacked the N-terminal variable region lost enzyme activity, they still retained dextran-binding ability. In addition, further deletion into the conserved region from the N-terminal did not influence the dextran-binding ability. However, the Dex which carried a deletion in the C-terminus still possessed both enzyme activity and dextran-binding ability. Further deletion into the conserved region from the C-terminal resulted in complete disappearance of both enzyme and dextran-binding activities. CONCLUSIONS: Deletion analysis of the Dex gene of Strep. mutans showed that the C-terminal side (about 120 amino acid residues) of the conserved region of the Dex was essential for dextran-binding ability. SIGNIFICANCE AND IMPACT OF THE STUDY: The dextran-binding domain was present in a different area from the catalytic site in the conserved region of the Dex molecule. The amino acid sequence of the dextran-binding domain of the Dex differed from those of glucan-binding regions of other glucan-binding proteins reported.

Amino Acid Sequence↗

PubMed2004

Dextran 40 reduces in vitro platelet aggregation in peripheral arterial disease.

Dextran 40 has been used to prevent post-operative thrombosis. However, its antithrombotic and antiplatelet effects in peripheral arterial disease (PAD) are poorly understood. We studied the in vitro effects of Dextran 40 on platelet function in control subjects and PAD patients using whole blood methods. Platelet function was assessed in 20 control subjects and 20 PAD patients. Spontaneous platelet aggregation (SPA) and agonist-induced platelet aggregation in response to increasing concentrations of Dextran 40 in vitro were measured by whole blood aggregometry. Flow cytometric measurements of platelet P-selectin, GpIIb/IIIa, GpIb and PAC-1 binding were also performed. There was no difference in SPA or ADP-induced aggregation in control patients with Dextran 40 in vitro. However, Dextran 40 inhibited collagen-induced aggregation in control patients (P < 0.05, Friedman test). In PAD patients, SPA and ADP (1 microM)-induced aggregation were significantly reduced by Dextran 40 in vitro (P < 0.001, Friedman test). In PAD patients, collagen-induced platelet aggregation (1 and 5 microg/ml) was significantly reduced by Dextran 40 in vitro (P < 0.01, Friedman test). GpIIb/IIIa, PAC-1 and P-selectin expression were significantly reduced in whole blood samples from PAD patients following incubation with Dextran 40 (P < 0.05, Wilcoxon rank test) but not in samples from control patients. Dextran 40 reduces spontaneous and agonist-induced platelet aggregation as well as the surface expression of markers of platelet activation in PAD patients. This antiplatelet effect may be of benefit to patients undergoing vascular surgical procedures where thrombosis is a significant risk.

Adenosine Diphosphate↗

PubMed1997

Renal imaging with 99Tc(m)-dextran.

Significant uptake and retention of 99Tc(m)-dextran (molecular weight: 81,000) in renal parenchyma was discovered during evaluation of its intravascular use. Renal SPET images confirmed this. This study was designed to evaluate 99Tc(m)-dextran as a renal cortex imaging agent. Stability of parenchymal retention was shown by insignificant outflow at 24 h and by frusemide intervention. Evaluation of the renal parameters of intravenous 99Tc(m)-dextran (n = 71 normal kidneys) and its comparison with 99Tc(m)-DTPA n = 10) and 99Tc(m)-DMSA(III) (n = 23) was undertaken. The early glomerular extraction phase of the renograms of 99Tc(m)-DTPA and 99Tc(m)-dextran appeared identical; parenchymal uptake of 99Tc(m)-dextran continued to increase and reached a near-plateau by 40-60 min. The mean cortex-to-background and cortex-to-liver ratios at 2 h with 99Tc(m)-dextran and 99Tc(m)-DMSA(III) were 14.9 and 9.2, and 16.0 and 8.9, respectively. The target-to-nontarget ratios were similar despite different absolute renal uptake values (12 vs 20% at 2 h) because of faster background clearance of 99Tc(m)-dextran. The mechanism of parenchymal retention of 99Tc(m)-dextran appears to be trapping at the endothelial-epithelial interphase of the glomerulus. Our initial experience suggests 99Tc(m)-dextran is a viable renal parenchyma imaging agent.

Adolescent↗

PubMed2003

Dextran-related complications in head and neck microsurgery: do the benefits outweigh the risks? A prospective randomized analysis.

Increased experience with free-tissue transfer has minimized flap loss secondary to microvascular thrombosis, yet pharmacologic antithrombotic prophylaxis continues to be used routinely. Currently there is no consensus on the ideal pharmacologic agent, dosing, or efficacy. Low-molecular-weight dextran has been widely used for prophylaxis due to its properties of volume expansion and enhanced microrheology. Significant systemic morbidity (pulmonary morbidity, cardiac morbidity, anaphylaxis) is known to occur with use of low-molecular-weight dextran. The purpose of this study was to evaluate morbidity associated with postoperative low-molecular-weight dextran and aspirin prophylaxis in head and neck microsurgery patients. This study was a randomized prospective analysis of 100 consecutive patients undergoing microvascular reconstruction for head and neck malignancy during a 2-year period. Patients were randomized into one of three postoperative antithrombotic prophylaxis treatment groups: low-molecular-weight dextran 20 cc/hour for 48 hours (n = 35), low-molecular-weight dextran 20 cc/hour for 120 hours (n = 32), or aspirin 325 mg/day for 120 hours (n = 27). Six patients were excluded intraoperatively due to the need for systemic heparin therapy. Treatment groups were compared for age, sex, prior medical problems, duration of anesthesia, and intraoperative fluid intake. Flap outcome and the incidence of local and systemic complications were evaluated in the treatment groups. Patient ages ranged from 12 to 84 years (mean age, 58 years). No significant difference was found among the treatment groups with respect to age, sex, prior medical problems, duration of anesthesia, intraoperative fluid intake, and the distribution of donor and recipient sites. There were no total flap losses and two partial flap losses in this series. Three flaps were reexplored and all were salvaged. The incidence of systemic complications (congestive heart failure, myocardial infarction, pulmonary edema, pleural effusion, and pneumonia) was as follows: low-molecular-weight dextran 120 hours, 51 percent; low-molecular-weight dextran 48 hours, 29 percent; and aspirin, 7 percent. Analysis of these data suggests that the method of prophylaxis had no effect on overall flap survival. However, the incidence of systemic complications was significantly related to the method of prophylaxis, with patients receiving low-molecular-weight dextran 120 hours and 48 hours at a 7.2 and 3.9 times greater relative risk, respectively, of developing a systemic complication compared with patients receiving aspirin. The results of this study have eliminated the routine use of low-molecular-weight dextran prophylaxis at our institution in an effort to reduce morbidity in head and neck microsurgical reconstruction.

Adolescent↗

PubMed1991

Serious complications with dextran-70 despite hapten prophylaxis. Is it best avoided prior to delivery?

Dextran is used clinically for plasma volume expansion, improvement of blood flow and thromboprophylaxis, but has been associated with untoward side effects. Immunoprophylaxis with dextran I (hapten), before the infusion of dextran-70, has reduced the incidence of serious dextran-induced anaphylactoid reactions. We report three cases of severe reactions occurring during anaesthesia in spite of immunoprophylaxis. One patient given dextran-70 before Caesarean section had a mild reaction but gave birth to a child with serious brain damage. One patient with an extremely high titre of dextran-reactive antibodies died from myocardial infarction and another patient recovered without sequelae. From our experience we conclude that dextran-induced anaphylactoid reactions are still a serious problem despite immunoprophylaxis. Dextran-70 should be avoided during pregnancy and should not be given during Caesarean section before delivery of the child. Even in the presence of immunological prophylaxis, vigilant observation of the patient is essential and resuscitation equipment must be available when starting a dextran infusion.

Adult↗

PubMed1984

Immunogenic properties of alpha (1----6) dextran, its protein conjugates, and conjugates of its breakdown products in mice.

Mice were immunized with alpha (1-6) dextran, either as such or coupled to protein carriers, and their anti-dextran response was measured by a solid-phase radioimmunoassay and the Farr assay. Like earlier investigators we found that protein-conjugated dextran was more antigenic than plain dextran. Our novel findings were that (1) a standard dose (30 micrograms of dextran per injection) coupled to strongly antigenic protein (chicken serum albumin (CSA) was three times more antigenic than dextran coupled to weakly antigenic bovine serum albumin (BSA); (2) dextrans of low molecular weight (1000-10,000 daltons) coupled to CSA induced at least ten times stronger secondary responses than did a similarly coupled macromolecular dextran (5-40 million daltons); (3) variation of the CHO/protein ratio from 0.3 to 1 had little effect on the antigenicity of the dextran. Increase of the ratio from one appeared to decrease immunogenicity when BSA was the carrier but not when CSA was the carrier.

Animals↗

PubMed1986

Effect of 100% O2 on passage of uncharged dextrans from blood to lung lymph.

Since charge as well as size may influence the passage of plasma proteins from blood to lung lymph, we used uncharged dextrans as tracers to study the effects of hyperoxic lung injury on the molecular sieving properties of the pulmonary microcirculation in unanesthetized sheep. Polydisperse [3H]dextran was infused intravenously into five sheep before and after the animals breathed 100% O2 until lymph flow increased threefold (66-84 h). Lymph-to-plasma concentration ratios (L/P) were determined for [3H]dextran fractions of graded molecular sizes (1.6-8.4 nm effective radius) from samples obtained during the infusions. Before hyperoxia the blood-lymph barrier was highly restrictive to transport of [3H]dextrans above 5.0 nm in radius; steady-state L/P for these molecules averaged 0.03 or less. After the sheep breathed 100% O2, [3H]dextrans as large as 8.4 nm radius appeared in the lymph. Posthyperoxia, the L/P were significantly increased relative to prehyperoxia base-line values for every [3H]dextran fraction larger than 2.0 nm radius (P less than 0.05). In contrast, neither the L/P for albumin or total protein changed significantly. At autopsy, electron microscopy showed widespread damage to the endothelium of the alveolar capillaries with infrequent gaps between endothelial cells. In two control sheep, inhalation of compressed air for 96 h had no effect on lymph flow or L/P for the [3H]dextrans. We conclude that O2 poisoning reduced the selective sieving of uncharged dextrans across the blood-lymph barrier of the lungs and allowed larger dextrans to enter the lymph. These larger molecules may have leaked from the pulmonary microcirculation via disruptions in the continuity of the endothelial lining.

Animals↗

PubMed1996

Inhibition by dextran of Pseudomonas aeruginosa adherence to epithelial cells.

Adherence of Pseudomonas aeruginosa to cells of the respiratory tract of patients with cystic fibrosis (CF) appears to be a necessary precondition for colonization and infection. To date no effective anti-adhesive strategy has been devised for preventing P. aeruginosa infection in these vulnerable hosts. The purpose of these studies was to evaluate the potential for preventing adhesion of P. aeruginosa to epithelial cells with dextran. Dextran (3,000-70,000 MW) inhibited adhesion of P. aeruginosa to buccal and A549 pulmonary epithelial cells; the 3,000 MW compound, at 10 mM was most inhibitory. Adhesion was inhibited optimally at pH 7.4 and was independent of charge; dextran and dextran sulfate were equally inhibitory. Dextran was most inhibitory if added to the epithelial cells before the P. aeruginosa; adhesion was reversed only minimally by adding dextran after the bacteria were bound. The inhibitory effect appeared to be nonspecific because other neutral polysaccharides (glycogen and mannan) were also inhibitory, dextran blocked attachment of other respiratory tract pathogens (Staphylococcus aureus, Group A streptococcus, and Haemophilus influenzae), and because dextran did not bind specifically to bacteria or to epithelial cells. Dextran is an inexpensive and nontoxic agent and may be useful in patients with CF to prevent colonization and infection with P. aeruginosa.

Bacterial Adhesion↗

PubMed1985

Dextran 70 versus donor plasma as colloid in open-heart surgery under extreme haemodilution.

Dextran 70 and donor plasma were compared as colloid in the priming solution during heart-lung perfusion in open-heart surgery. The patients underwent surgery for coronary artery or aortic valve disease. One group of nine patients had plasma in the priming solution, while ten patients received dextran 70. Prior to perfusion the patients in the dextran group received monovalent hapten to prevent anaphylactic reactions towards dextran. During the perfusion the same level of haemodilution was established in both groups; the haematocrit values fell from about 39 vol. per cent before perfusion to about 19 vol. per cent after 45 min of perfusion. Eighteen hours postoperatively, the haematocrit had increased to about 35 vol. per cent. The mean concentrations of total serum protein in preoperative samples from the plasma and dextran group were 66.0 g/l and 66.3 g/l, respectively, and fell to 42.6 g/l and 28.3 g/l in samples taken after 45 min of perfusion. The serum-protein concentration in the dextran group remained significantly lower than that of the plasma group throughout the postoperative period. A similar pattern was seen for the albumin concentration. Preoperatively the mean values of the colloid osmotic pressure of plasma were 24.2 mmHg and 25.5 mmHg in the plasma and dextran group, respectively. Corresponding figures 45 min after start of perfusion were 13.9 mmHg and 16.9 mmHg, respectively, significantly higher in the dextran group than in the plasma group. The colloid osmotic pressure exerted by dextran was about 9 mmHg during heart-lung perfusion and about 4 mmHg 18 h later.(ABSTRACT TRUNCATED AT 250 WORDS)

Aortic Valve Insufficiency↗

PubMed2000

Improved nasal bioavailability of FITC-dextran (Mw 4300) from mucoadhesive microspheres in rabbits.

The nasal bioavailability of fluorescein isothiocyanate-dextran (FITC-dextran) (Mw = 4300) encapsulated in non-mucoadhesive and mucoadhesive microspheres in New Zealand White rabbits was investigated. FITC-dextran was administered nasally encapsulated in carbopol 934P, chitosan and lactose microspheres and the bioavailability compared to intravenous administration of FITC-dextran solution. Administration of FITC-dextran as carbopol microspheres produced a significantly greater bioavailability (33%) than after administration as chitosan (13%) and non-mucoadhesive rapidly dissolving control lactose microspheres (9%). The FITC-dextran terminal plasma half-lives after carbopol 934P and chitosan microsphere administration were significantly longer than after intravenous administration of FITC-dextran. The FITC-dextran terminal plasma half-life after carbopol 934P microspheres administration was significantly longer than after lactose microsphere administration. This data suggested that the increase in FITC-dextran bioavailability after carbopol 934P microspheres administration was due to increased residence at the absorptive site via mucoadhesion and reduced mucociliary clearance. A change in mucosal permeability could not however be discounted especially for the chitosan microspheres.

Animals↗

PubMed2001

Prospective randomised open-label comparison of danaparoid with dextran 70 in the treatment of heparin-induced thrombocytopaenia with thrombosis: a clinical outcome study.

AIM: To compare clinical outcomes in a randomised comparison of treatment with danaparoid sodium (a heparinoid), or dextran 70, for heparin-induced thrombocytopaenia (HIT) plus thrombosis. METHODS: Forty-two patients with recent thrombosis and a clinical diagnosis of probable HIT who presented at ten Australian hospitals during a study period of six and one half years were randomly assigned to open-label treatment with intravenous danaparoid or dextran 70, each combined with oral warfarin. Thirty-four patients (83%) had a positive platelet aggregation or 14C-serotonin release test for HIT antibody. Twenty-five received danaparoid as a bolus injection of 2400 anti-Xa units followed by 400 units per hour for 2 h, 300 units per hour for 2 h, and then 200 units per hour for five days. Seventeen received 1000 mL dextran 70 on day one and then 500 mL on days 2-5. Patients were reviewed daily for clinical evidence of thrombus progression or resolution, fresh thrombosis or embolism, bleeding or other complications. The primary trial endpoint was the proportion of thromboembolic events with complete clinical resolution by the time of discharge from hospital. RESULTS: With danaparoid, there was complete clinical recovery from 56% of thromboembolic events compared to 14% after dextran 70 (Odds Ratio 10.53, 95% Confidence Interval 1.6-71.4; p = 0.02). Clinical recovery with danaparoid was complete or partial in 86% of thromboembolic events compared with 53% after dextran 70 (Odds Ratio 4.55, 95% Confidence Interval 1.2-16.7; p = 0.03). Overall clinical effectiveness of danaparoid was rated as high or moderate in 88% of patients compared with 47% for dextran 70 (p = 0.01). One patient given danaparoid died of thrombosis compared with three patients given dextran 70. The platelet count returned to normal after a mean of 6.7 days with danaparoid and 7.3 days with dextran 70. There was no major bleeding with either treatment. CONCLUSION: danaparoid plus warfarin treatment for HIT with thrombosis is effective, safe, and superior to dextran 70 plus warfarin.

Aged↗

PubMed2002

Targeting of interferon to choroidal neovascularization by use of dextran and metal coordination.

PURPOSE: Bioactive proteins such as interferon (IFN) have been reported to be combined with water-soluble polymers, such as dextran, through metal coordination, without need for complicated procedures. In the current study, the targeting and inhibitory effects of IFN combined with dextran on experimental choroidal neovascularization (CNV) were studied in vivo. METHODS: Interferon (IFN)beta was conjugated to dextran, which has metal-chelating, diethylenetriaminepentaacetic acid (DTPA) residues. Based on metal coordination, conjugation of IFNbeta with DTPA-dextran resulted from simply mixing both substances in an aqueous solution containing Zn(2+). The effects of IFNbeta on the proliferation of human umbilical vein endothelial cells (HUVECs) and bovine retinal pigment epithelial cells (BRPECs) were evaluated. To evaluate the activity loss of IFNbeta by conjugation, the effect of the conjugate on HUVECs was compared with that of free IFNbeta. Experimental CNV was induced by subretinal injection of gelatin microspheres containing basic fibroblast growth factor in rabbits. The rabbits with CNV were intravenously treated twice weekly with 7.5 million international units (MIU)/kg per day free IFNbeta (for 4 weeks), with IFNbeta-DTPA-dextran conjugate containing 7.5 (for 2 weeks) or 0.75 (for 4 weeks) MIU/kg per day IFNbeta, or with saline. The effects of these substances were evaluated by fluorescein angiography and histology. To observe the accumulation of conjugate, the doses of IFNbeta in CNV tissues were measured by enzyme-linked immunosorbent assay. RESULTS: IFNbeta inhibited the growth of HUVECs and enhanced the proliferation of BRPECs. The conjugate seemed to preserve approximately 44% of IFNbeta activity. Although both doses of IFNbeta-DTPA-dextran inhibited progression of CNV in rabbits, longer term administration of a lower dose of IFNbeta-DTPA-dextran had a sustained inhibitory effect on progression of CNV (P < 0.05). Histologic studies revealed the inhibitory effect of IFNbeta-DTPA-dextran on progression of CNV. This conjugate prolonged the plasma half-life of IFNbeta and enabled IFNbeta to accumulate in the CNV in rabbits. CONCLUSIONS: In this study, human IFNbeta was successfully used to target CNV, an enhanced antiangiogenic effect was achieved by combining it with dextran, based on metal coordination. This targeted delivery of IFNbeta may have potential as a treatment modality for CNV.

Angiogenesis Inhibitors↗

PubMedDate not supplied

Development of dextran derivatives with cytotoxic effects in human urinary bladder cancer cell lines.

BACKGROUND: In a previous study we reported on a new approach describing intravesical instillation of charged dextran in patients with superficial bladder carcinoma. The cationic derivative showed a strong tumor-selective accumulation. To develop this approach, the present study investigates the cytotoxic effect of cationic dextran derivatives on two urinary bladder cancer cell lines (J82 and 5637). METHODS: The dextran conjugates were prepared by periodate activation and subsequent coupling by reductive amination. A fluorimetric cytotoxicity assay (FMCA) was used for the cytotoxicity assay. The tumor cells were seeded into 96-well microtiter plates and different cationic dextran derivatives were added and incubated for 72 hours. RESULTS: The results showed that cationic epirubicin-dextran had a clear inhibitory effect on the growth in both cell lines (40-95% growth inhibition). The corresponding values for epirubicin (the reference) was 90-100% inhibition. Interestingly, cationic dextran had, by itself, a growth inhibitory effect. This cytotoxic effect could be strongly enhanced to be almost equal to the reference by changing the cationic sidegroup to aminohexane. Dextran alone showed no effect. CONCLUSION: The finding that cationic dextran by itself can be made cytotoxic, together with its capacity to accumulate in superficial bladder cancer, suggests possibilities for new therapeutic constructs. Cationic dextran with different cationic side-groups and in combination with cytotoxic drugs will be studied further. The cytotoxic mechanism needs to be elucidated.

Antineoplastic Agents↗

PubMed1992

Improved lung preservation with dextran 40 is not mediated by a superoxide radical scavenging mechanism.

Improved lung preservation with a low-potassium dextran-containing solution has been previously demonstrated. In a subsequent study, it was shown that dextran 40 contributes significantly to this improved preservation. In the current in vitro study, human neutrophils suspended in lung preservation solutions (low potassium with dextran and low potassium without dextran) were stimulated to produce superoxide radicals. The presence of dextran in the solution did not significantly alter the amount of superoxide measured in the assay (low potassium with dextran, 4.149 +/- 0.144 nmol/10(6) cells/20 min; low potassium without dextran, 3.896 +/- 0.215; p greater than 0.2). This suggests that dextran 40 did not appreciably scavenge superoxide radicals, nor did it alter the production of superoxide radicals by stimulated neutrophils. Thus the significantly improved lung preservation seen with the use of dextran 40 is probably not mediated by a superoxide radical scavenging process.

Dextrans↗

PubMed1990

Colorimetry and constant-potential coulometry determinations of transferrin-bound iron, total iron-binding capacity, and total iron in serum containing iron-dextran, with use of sodium dithionite and alumina columns.

After the parenteral administration of iron-dextran (imferon), the increased total iron concentrations in serum can be determined by atomic absorption spectroscopy and by colorimetric methods involving sodium dithionite, which reductively dissociates iron from the dextran complex. We report that constant-potential coulometry detects only about 55-70% of dextran-bound iron before dithionite reduction and variable amounts after reaction with the reducing agent. In addition, we have developed a procedure for determining transferrin-bound iron, total iron-binding capacity (TIBC), total iron, and dextran-bound iron with the Kodak Ektachem colorimetric system. In determining total serum iron, the sample is first mixed with sodium dithionite, which rapidly dissociates all dextran-bound iron, but does not remove iron from either transferrin or hemoglobin. After the mixture is applied to an Ektachem slide, transferrin-bound iron is released at pH 4 and is detected together with the iron previously bound to dextran. TIBC is determined by mixing serum with ferric citrate in moderate excess and filtering through a small alumina (Al2O3) column, which binds excess free iron and iron-dextran; the iron in the column eluate represents the TIBC. Transferrin-bound iron is determined by applying diluted serum without added ferric citrate to an alumina column and measuring the iron in the column eluate. Dextran-bound iron is equivalent to the difference between total and transferrin-bound iron. Using this method, we found that transferrin iron-binding sites are saturated in vitro by excess iron-dextran less efficiently than by ferric citrate.

Aluminum Oxide↗

PubMed1987

Intestinal vascular obstruction shock in the rat. Effect of 2 dextran solutions on blood and plasma volumes, cardioinhibitory activity in blood and pulmonary platelet trapping.

Pathophysiologic mechanisms involved in the dextran effect on mortality in intestinal shock in rats were studied, using a standardized model for intestinal vascular obstruction. Both dextran 70 and dextran 40 (respective mean molecular weights 70 X 10(3) and 40 X 10(3) d) given to shocked rats reduced but did not prevent the cardioinhibitory action of intestinal venous plasma in vitro. Both dextrans prevented platelet trapping in the lungs. Reduction in blood and plasma volumes was found in shocked rats given saline or dextran 40, but not following dextran 70. These differences were found after 120 min, but not after 240 min, when all shocked groups had lower blood and plasma volumes than in non-shocked controls. The results indicate that several mechanisms influence the effect of dextran 70 on mortality of rats in intestinal shock. Previously observed difference in mortality rates after infusion of dextran 70 and dextran 40 could not be explained by mechanisms studied in this series of experiments.

Animals↗

PubMed1989

Dextran efflux from McCarey-Kaufman-stored corneas as measured by nuclear magnetic resonance.

Dextran, a high molecular weight polymer of glucose, is used as an osmotic agent in McCarey-Kaufman (MK) (corneal storage) medium. It has been proposed that Dextran may cause deleterious effects on living cells either by high molecular weight toxicity or by the induction of an immunological reaction against transplanted tissues. Prolonged retention of Dextran in transplanted corneas, therefore, may produce deleterious side effects. The efflux of Dextran from MK-stored human corneas was monitored by transferring corneas to 2 ml of fresh Krebs Ringer bicarbonate solution (BSS) containing 5.5 mM 1-13C-labeled glucose at approximately 1 h intervals for 1-24 h. Chemical shifts of 98.7, 74.5, 72.5, 71.2, 70.6, and 66.5 ppm were found for the naturally abundant 13C NMR spectrum of 40,000 molecular weight Dextran. Peak heights for the 70.6 ppm Dextran chemical shift were measured and plotted against total wash time to determine the slope for Dextran efflux from each cornea. The results of this study suggest that all of the Dextran absorbed into human corneas stored for up to 2 weeks in MK medium is eluted in about 19 h and that the rate of Dextran efflux is unaffected by donor age, time of storage in MK, or corneal viability.

Adult↗

PubMed1981

Regulation of clones responding to dextran B1355S. II. Response of T-dependent and T-independent precursors.

The existence of precursors for TI and TD alpha 1 leads to 3 dextran antigens in BALB/c mice was demonstrated. A T-dependent dextran antigen was prepared by coupling dextran B1355S to hemocyanin and subsequent digestion with dextranase. The PFC response of BALB/c mice primed with hemocyanin to dextran-hemocyanin was found to be 8 times higher than in unprimed animals. The splenic focus assay was adapted for the analysis of precursors responding to T-dependent and T-independent dextran antigens. Pretreatment of recipients with anti-thymocyte serum abolished the response in fragment cultures to dextran-hemocyanin but did not affect the response to dextran B1355S. The frequencies of precursors in the adult BALB/c mouse responding to dextran and dextran-hemocyanin were determined by limiting dilution analysis. The frequency of T-dependent precursors was found to be almost 3 times greater than the frequency of T-independent precursors.

Animals↗