Distribution by Scientific Domains
Distribution within Medical Sciences

Kinds of Dextran

  • iron dextran

  • Terms modified by Dextran

  • dextran amine
  • dextran sodium
  • dextran sulphate
  • dextran sulphate sodium

  • Selected Abstracts


    JOURNAL OF RENAL CARE, Issue 2 2009
    Smeeta Sinha
    SUMMARY Background: Low-molecular-weight iron dextran (CosmoFer®) is the only form of parenteral iron that can be administered as a total dose infusion (TDI) in the United Kingdom (UK). This study aimed to evaluate the safety and efficacy of TDI CosmoFer in comparison to intravenous iron sucrose infusion (Venofer®) in patients with chronic kidney disease (CKD). Methods and Results: A retrospective study of outpatients with CKD undergoing intravenous TDI CosmoFer or Venofer infusion was conducted at Salford Royal Hospital and Sunderland Royal Hospital. A total of 979 doses of CosmoFer and 504 doses of Venofer were administered. There were three minor adverse events in patients receiving CosmoFer compared with one minor event in a Venofer treated patient. There were no anaphylactoid-type reactions in either group. Serum haemoglobin, ferritin and transferrin saturation (TSAT) improved significantly 4,6 months postinfusion in both treatment groups. Conclusion: TDI CosmoFer is an efficacious method of replenishing iron stores in CKD patients in an outpatient setting. Furthermore, TDI CosmoFer is safe and not associated with an increase in adverse events compared to Venofer. [source]

    CE analysis of the acidic organelles of a single cell

    ELECTROPHORESIS, Issue 14 2007
    Yun Chen
    Abstract The properties of organelles within a cell have been shown to be highly heterogeneous. Until now, it has been unclear just how much of this heterogeneity is endemic to the organelle subpopulations themselves and how much is actually due to stochastic cellular noise. An attractive approach for investigating the origins of heterogeneity among the organelles of a single cell is CE with LIF detection (CE-LIF). As a proof of principle, in this report we optimize and use a single cell CE-LIF method to investigate the properties of endocytic (acidic) organelles. Our results show that the properties of individual acidic organelles containing Alexa Fluor® 488 Dextran suggest that there are two groups of CCRF-CEM cells: a group with a high dextran content per cell, and a group with a low dextran content per cell. Furthermore, the individual organelle measurements of the single cells allow us to compare in each group the distributions of doxorubicin content per acidic organelle and electrophoretic mobilities of these organelles. [source]

    Binding of rat brain hexokinase to recombinant yeast mitochondria

    FEBS JOURNAL, Issue 10 2000
    Identification of necessary physico-chemical determinants
    The association of rat brain hexokinase with heterologous recombinant yeast mitochondria harboring human porin (Yh) is comparable to that with rat liver mitochondria in terms of cation requirements, cooperativity in binding, and the effect of amphipathic compounds. Mg2+, which is required for hexokinase binding to all mitochondria, can be replaced by other cations. The efficiency of hexokinases, however, depends on the valence of hydrophilic cations, or the partition of hydrophobic cations in the membrane, implying that these act by reducing a prohibitive negative surface charge density on the outer membrane rather than fulfilling a specific structural requirement. Macromolecular crowding (using dextran) has dual effects. Dextran added in excess increases hexokinase binding to yeast mitochondria, according to the porin molecule they harbor. This effect, significant with wild-type yeast mitochondria, is only marginal with Yh as well as rat mitochondria. On the other hand, an increase in the number of hexokinase binding sites on mitochondria is also observed. This increase, moderate in wild-type organelles, is more pronounced with Yh. Finally, dextran, which has no effect on the modulation of hexokinase binding by cations, abolishes the inhibitory effect of amphipathic compounds. Thus, while hexokinase binding to mitochondria is predetermined by the porin molecule, the organization of the latter in the membrane plays a critical role as well, indicative that porin must associate with other mitochondrial components to form competent binding sites on the outer membrane. [source]

    Biodegradable Nanogels Prepared by Self-Assembly of Poly(L- lactide)-Grafted Dextran: Entrapment and Release of Proteins

    Koji Nagahama
    Abstract We showed previously that poly(L- lactide)-grafted dextran could form biodegradable nanogels in water. In this paper, various properties of Dex -g- PLLA nanogels were compared with Dex-Chol (dextran-cholesterol conjugate) nanogels to investigate the effects of hydrophobic units. Dex -g- PLLA nanogels exhibited significantly lower CAC and higher colloidal stability, indicating a strong tendency to form nanogels. We prepared lysozyme-loaded Dex -g- PLLA nanogels, and they exhibited a sustained release of lysozyme for 1 week without denaturation in PBS at 37,°C. The Dex -g- PLLA nanogels therefore have great potential as a delivery vehicle for therapeutic protein. [source]

    Gold Nanoparticles Functionalized by a Dextran-Based pH- and Temperature-Sensitive Polymer

    Weipeng Lv
    Abstract A dextran-based dual-sensitive polymer is employed to endow gold nanoparticles with stability and pH- and temperature-sensitivity. The dual-sensitive polymer is prepared by RAFT polymerization of N -isopropylacrylamide from trithiocarbonate groups linked to dextran and succinoylation of dextran after polymerization. The functionalized nanoparticles show excellent stability under various conditions and can be stored in powder-form. UV and DLS measurements confirm that the temperature-induced optical changes and aggregation behaviors of the particles are strongly dependent on pH. [source]

    SIGIRR/TIR-8 is an inhibitor of toll-like receptor signaling in primary human cells and regulates inflammation in models of rheumatoid arthritis

    ARTHRITIS & RHEUMATISM, Issue 8 2010
    Stefan K. Drexler
    Objective Single-immunoglobulin interleukin-1 receptor,related (SIGIRR), which is also known as Toll/interleukin-1 receptor 8 (TIR-8), is a member of the TIR domain,containing family of receptors and was first characterized as an inhibitor of interleukin-1 receptor (IL-1R) and Toll-like receptor (TLR) signaling. In the Dextran sulfate sodium,induced colitis model, SIGIRR,/, mice were shown to have increased inflammation and to be more susceptible to endotoxin challenge. Increasing evidence implicates TLR and IL-1R signaling in the pathology of rheumatoid arthritis (RA). Therefore, the purpose of this study was to investigate the involvement of SIGIRR in regulating inflammation in disease-relevant models. Methods Primary human monocyte-derived macrophages and dendritic cells (DCs) were used to overexpress SIGIRR as well as to knock down endogenously expressed SIGIRR using small interfering RNAs. SIGIRR was also overexpressed in synovial cells derived from RA patients. To investigate the role of SIGIRR in vivo, zymosan-induced arthritis (ZIA) and collagen antibody,induced arthritis (CAIA) were induced in SIGIRR-knockout mice. Results SIGIRR overexpression inhibited TLR-induced cytokine production in macrophages and DCs, while SIGIRR knockdown resulted in increased cytokine production following TLR stimulation. Moreover, SIGIRR overexpression inhibited the spontaneous release of cytokines by human RA synovial cells. The role of SIGIRR as an inhibitor of inflammation was confirmed in vivo, since SIGIRR,/, mice developed a more severe disease in both the ZIA and CAIA models. Conclusion Our study is the first to show the expression pattern and function of SIGIRR in primary human cells. Furthermore, this investigation defines the role of SIGIRR in disease-relevant cell types and demonstrates that SIGIRR is a potential therapeutic target for RA. [source]

    Properties of fucoidan from Cladosiphon okamuranus tokida in gastric mucosal protection

    BIOFACTORS, Issue 4 2000
    Hideyuki Shibata
    Abstract To elucidate the anti-ulcer potential of Cladosiphon fucoidan, anti-peptic activity, bFGF stabilizing activity and inflammatory properties of this and related substances were investigated. Anti-peptic activity was observed with this and other sulfated polysaccharides such as dextran sulfate, carrageenan, and Fucus fucoidan. However, non-sulfated polysaccharides such as mannan and dextran did not exert the anti-peptic activity. The loss of bFGF bioactivity was prevented by all sulfated polysaccharides tested except chondroitin sulfate, at pH 7.4 and at pH 4.0. At pH 2.0, only heparin protected the bFGF activity. The generation of superoxide by macrophages and PMNs was stimulated by dextran sulfate, carrageenan, and Fucus fucoidan, whereas Cladosiphon fucoidan, heparin and chondroitin did not. Dextran sulfate, carrageenan, and Fucus fucoidan also stimulated the secretion of TNF, from macrophages, while Cladosiphon fucoidan did not. Thus, Cladosiphon fucoidan is a sulfated polysaccharide without inflammatory action. These results suggest that Cladosiphon fucoidan is a safe substance with potential for gastric protection. [source]

    3426: Straylight and corneal edema

    Purpose It is known that corneal edema is associated to increased light scatter. It was the purpose of this study to calculate the intensity and angular distribution of scattered light in a series of corneal samples at different hydration, using a sensitive optical technique. Methods Sixteen fresh porcine eyes were obtained from a local abattoir. To isolate the role of corneal stroma the the epithelium was carefully removed with a mechanical brush. The central 8 mm of each cornea was harvested using a Barron's PKP trephine. To establish corneal hydration, corneal buttons were immersed in Dextran (300kDa) solutions, with concentrations ranging from 5 to 20% w/w, for at least 3 hours. The intensity and angular distribution of scattered light was measured for all corneas by means of a purposely-developed camera lens that incorporated excised corneas between its glass elements. This lens was used with a CCD camera to record images projected on a computer screen. Before scatter measurements the thichness of each corneal sample was measured by means of a mechanical pachymeter (Mitutoyo IDC 112T, Japan). Results The mean scatter coefficients for the corneas at normal hydration levels was 0.22 (SD=0.059). This value was effectively double (0.46 ; SD= 0.048) at a moderate increase of stromal thickness by 17% and reached a value of 0.56 (SD=0.079) at a relative increase of corneal thickness of 62%. The angular distribution did not significantly deppend on hydration. Conclusion Scattered light intensity increases with corneal edema. Even small changes in corneal hydration affect significantly the narrow angle light scattering properties of corneal stroma. [source]

    Similarity of permeabilities for Ficoll, pullulan, charge-modified albumin and native albumin across the rat peritoneal membrane

    ACTA PHYSIOLOGICA, Issue 4 2009
    D. Asgeirsson
    Abstract Aim:, Compared to neutral globular proteins, neutral polysaccharides, such as dextran, pullulan and Ficoll, appear hyperpermeable across the glomerular filtration barrier. This has been attributed to an increased flexibility and/or asymmetry of polysaccharides. The present study investigates whether polysaccharides are hyperpermeable also across the continuous capillaries in the rat peritoneum. Methods:, In anaesthetized Wistar rats, FITC,Ficoll or FITC,pullulan together with 125I-human serum albumin (RISA) or neutralized 125I-bovine serum albumin (nBSA) were given intravenously, after which peritoneal dialysis (PD) using conventional PD fluid (Gambrosol 1.5%) was performed for 120 min. Concentrations of FITC-polysaccharides and radioactive albumin species in plasma and dialysis fluid were analysed with high-performance size exclusion chromatography and a gamma counter respectively. Transperitoneal clearance values were calculated for polysaccharides in the molecular radius range 36,150 Å, and for RISA and nBSA. Results:, Ficoll and pullulan showed more or less identical permeabilities, compared to RISA and nBSA, across the peritoneal membrane. Although RISA-clearance, 5.50 ± 0.28 (,L min,1; ±SEM), tended to be lower than the clearances of Ficoll36Å (6.55 ± 0.25), pullulan36Å (6.08 ± 0.22) and nBSA (6.56 ± 0.23), the difference was not statistically significant. This is in contrast to the hyperpermeability exhibited by polysaccharides across the glomerular filtration barrier and also contrasts with the charge selectivity of the latter. Conclusion:, The phenomenon of molecular flexibility is more important for a macromolecule's permeability through the glomerular filter than across the continuous peritoneal capillary endothelium. Furthermore, it seems that charge plays a subordinate role in the steady-state transport across the combined peritoneal capillary,interstitial barrier. [source]

    Protective role of pigment epithelium-derived factor (PEDF) in early phase of experimental diabetic retinopathy

    Yumiko Yoshida
    Abstract Background Pigment epithelium-derived factor (PEDF) is the most potent inhibitor of angiogenesis in the mammalian eye, thus suggesting that PEDF may protect against proliferative diabetic retinopathy. However, a role for PEDF in early diabetic retinopathy remains to be elucidated. We investigated here whether and how PEDF could prevent the development of diabetic retinopathy. Methods Streptozotocin-induced diabetic rats were treated with or without intravenous injection of PEDF for 4 weeks. Early neuronal derangements were evaluated by electroretinogram (ERG) and immunofluorescent staining of glial fibrillary acidic protein (GFAP). Expression of PEDF and 8-hydroxydeoxyguanosine (8-OHdG), a marker of oxidative stress, was localized by immunofluorescence. Vascular endothelial growth factor (VEGF) and p22phox expression were evaluated with western blots. Breakdown of blood retinal barrier (BRB) was quantified with fluorescein isothiocynate (FITC)-conjugated dextran. NADPH oxidase activity was measured with lucigenin luminescence. Results Retinal PEDF levels were reduced, and amplitudes of a- and b-wave in the ERG were decreased in diabetic rats, which were in parallel with GFAP overexpression in the Müller cells. Further, retinal 8-OHdG, p22phox and VEGF levels and NADPH oxidase activity were increased, and BRB was broken in diabetic rats. Administration of PEDF ameliorated all of the characteristic changes in early diabetic retinopathy. Conclusions Results suggest that PEDF could prevent neuronal derangements and vascular hyperpermeability in early diabetic retinopathy via inhibition of NADPH oxidase-driven oxidative stress generation. Substitution of PEDF may offer a promising strategy for halting the development of diabetic retinopathy. Copyright © 2009 John Wiley & Sons, Ltd. [source]

    Decreased red blood cell aggregation subsequent to improved glycaemic control in Type 2 diabetes mellitus

    DIABETIC MEDICINE, Issue 4 2003
    B. Chong-Martinez
    Abstract Aims Reports of rheological changes following intensification of metabolic control are limited and not concordant. The present study was designed to test the hypothesis that intensification of management of Type 2 diabetes (T2DM) with diet, exercise and insulin improves haemorheological behaviour by reducing red blood cell (RBC) aggregation. Methods Blood was sampled from 55 subjects before and following 14 ± 3 weeks of intensified management. RBC aggregation was measured in vitro for cells in plasma or in an aggregating 70 kD dextran solution. Plasma viscosity and whole blood viscosity were also measured. Results During treatment, fasting glucose fell 27%, HbA1c fell 21%, and serum triglycerides and total cholesterol fell 28% and 12%, respectively (P < 0.0001 for each). The extent and strength of RBC aggregation in plasma fell by 10,13% (P < 0.002). Similar decreases of RBC aggregation were seen for cells suspended in dextran (P < 0.002). Plasma viscosity decreased by 3% (P < 0.02) and high shear blood viscosity by 6,7% (P < 0.0001). Changes of RBC aggregation in plasma and in dextran were significantly correlated, supporting a cellular rather than a plasmatic origin for these changes. However, there were no significant correlations between RBC aggregation changes and changes of fasting glucose, HbA1c, serum triglycerides, serum cholesterol, or plasma fibrinogen. Conclusions Intensified metabolic control results in a reduction of RBC aggregation that appears to be intrinsic to RBC. Since increased RBC aggregation can impair microcirculatory flow, it is possible that haemorheological factors may contribute to the reduction of microvascular complications resulting from improved metabolic control in T2DM. [source]

    Evaluation of CE methods for global metabolic profiling of urine

    ELECTROPHORESIS, Issue 14 2010
    Rawi Ramautar
    Abstract In this study, the usefulness of noncovalently coated capillaries with layers of charged polymers is investigated to obtain global electrophoretic profiles of urinary metabolites covering a broad range of different compound classes in a highly repeatable way. Capillaries were coated with a bilayer of polybrene (PB) and poly(vinyl sulfonate) (PVS), or with a triple layer of PB, dextran sulfate (DS) and PB. The bilayer and triple layer coatings were evaluated at acidic (pH 2.0) and alkaline (pH 9.0) separation conditions, thereby providing separation conditions for basic and acidic compounds. A representative metabolite mixture and spiked urine samples were used for the evaluation of the four CE methods. Migration time repeatability (RSD<2%) and plate numbers (N, 100,000,400,000) were similar for the test compounds in all CE methods, except for some multivalent ions that may exhibit adsorption to oppositely charged coatings. The analysis of cationic compounds with the PB-DS-PB CE method at low pH (i.e. after the EOF time) provided a larger separation window and number of separated peaks in urine compared to the analysis with the PB-PVS CE method at low pH (i.e. before the EOF time). Approximately, 600 molecular features were detected in rat urine by the PB-DS-PB CE-MS method whereas about 300 features were found with the PB-PVS CE-MS method. This difference can be attributed to reduced comigration of compounds with the PB-DS-PB CE-MS method and a related decrease of ion suppression. With regard to the analysis of anionic compounds by CE-MS, in general analyte responses were significantly lower than that for cationic compounds, most probably due to less efficient ionization and to ion suppression effects caused by the background electrolyte. Hence, further optimization is required for the sensitive CE-MS analysis of anionic compounds in body fluids. It is concluded that the selection of a CE method for profiling of cationic metabolites in urine depends on the purpose of the study. For high-throughput analyses, the PB-PVS CE-MS method is favored whereas the PB-DS-PB CE-MS method provides a more information-rich metabolic profile, but at the cost of prolonged analysis time. [source]

    Chiral ion-exchange capillary electrochromatography of arylglycine amides with dextran sulfate as a pseudostationary phase

    ELECTROPHORESIS, Issue 4-5 2005
    Yi Chen
    Abstract A low-cost tunable chiral ion-exchange capillary electrochromatographic method has been developed for the separation of arylglycine amide racemic mixtures with dextran sulfate (DS) as an anionic and chiral pseudostationary phase and Tris-tartrate as a buffer system. The concentrations of DS and Tris had opposite influences on retention and resolution and could serve as ideal factors to finely tune the running speed and chiral resolution. Tartrate and pH largely impact the separation but pH should be confined within 3.0,5.5, only suitable for coarse tuning, while tartrate was preserved as the key buffering reagent, normally maintained at 40 mmol/L. With a working system composed of 0.1,1.0% DS, 20,60 mmol/L Tris, and 40 mmol/L tartrate at pH 3.50,4.50, the enantioresolution of arylglycine amides was shown to be dependent on their chemical structure: The chiral resolution increased when the hydrogen at the ,-amino group or at the p -position of phenyl ring was replaced by other larger group(s) but the resolution decreased when the group at the o- or m- site on the phenyl ring was enlarged. Further, the electronegative substitute of -Cl had larger resolution increment than methyl or methoxy at the position p- of phenyl ring but much lower increment at position m- . It is possible to well explain the resolution variation phenomenon by considering the group resistance and the variation of hydrogen-bonds formed inside the amino amides and between the solutes and DS. The amido group was shown irreplaceable to have chiral resolution with DS alone as an ionic and chiral pseudostationary phase. [source]

    Integrated Enzymatic Synthesis and Adsorption of Isomaltose in a Multiphase Fluidized Bed Reactor

    M. Ergezinger
    Abstract Dextransucrase catalyzes the formation of dextran, but also of numerous oligosaccharides from sucrose and different acceptors, if appropriate conditions are chosen. A process on a technical scale with immobilized enzyme was established to produce isomaltose, a disaccharide of industrial interest. Isomaltose is also a reactant for dextransucrase and has to be quickly taken out of the reaction solution. This was realized by integrated adsorption of isomaltose on zeolites. In the case of biotransformation the reactor works with a fluidized bed of immobilized enzyme and the in situ separation is realized with a suspension flow of adsorbent. This process was investigated experimentally and theoretically. With a design model consisting of hydrodynamics, kinetics of enzymatic synthesis, and thermodynamics of adsorption, a comparison was made between experimental and calculated data. [source]

    Idiotype-specific CD4+CD25+ T suppressor cells prevent, by limiting antibody diversity, the occurrence of anti-dextran antibodies crossreacting with histone H3

    Christoph Specht
    Abstract CD25+ suppressor T cells regulate the immune response against the type-2 "thymus independent" bacterial polysaccharide antigen ,(1,3)dextran (Dex) in BALB/c mice. These T cells, represented by the clone 178-4 Ts, restrict the Dex-specific IgG antibody repertoire such that the J558 idiotype dominates. Antibodies with other structures in the heavy-chain variable region (VH region), predominantly within the CDR3 domain, occur when the T cell control fails. This increase of antibody diversity caused by a lack of CD25+ Ts cells, e.g. in nude mice, does not result in the appearance of antibodies with enhanced affinity to the antigen Dex, but often leads to a crossreactivity with autologous proteins. Twenty-two out of sixty Dex-specific hybridomas from nude mice, but no hybridomas from euthymic mice, crossreact with a nuclear protein, as tested by ELISA. This nuclear protein was identified as histone H3. Ten of the sixty hybridomas from nude mice were sequenced and show VH sequences that deviate from the original J558 sequence. Three of these ten hybridomas crossreact with the histone H3. Adoptive transfer of CD25+ Ts cells to nude mice leads to a marked increase of antibodies carrying the original J558 idiotype within the IgG pool after immunization with Dex. Our data demonstrate a CD25+ Ts cell-mediated restriction of VH usage, which prevents the appearance of crossreactive autoantibodies. [source]

    Reciprocal connections between olfactory structures and the cortex of the rostral superior temporal sulcus in the Macaca fascicularis monkey

    A. Mohedano-Moriano
    Abstract Convergence of sensory modalities in the nonhuman primate cerebral cortex is still poorly understood. We present an anatomical tracing study in which polysensory association cortex located at the fundus and upper bank of the rostral superior temporal sulcus presents reciprocal connections with primary olfactory structures. At the same time, projections from this polysensory area reach multiple primary olfactory centres. Retrograde (Fast Blue) and anterograde (biotinylated dextran,amine and 3H-amino acids) tracers were injected into primary olfactory structures and rostral superior temporal sulcus. Retrograde tracers restricted to the anterior olfactory nucleus resulted in labelled neurons in the rostral portion of the upper bank and fundus of superior temporal sulcus. Injections of biotinylated dextran,amine at the fundus and upper bank of the superior temporal sulcus confirmed this projection by labelling axons in the dorsal and lateral portions of the anterior olfactory nucleus, as well as piriform, periamygdaloid and entorhinal cortices. Retrograde tracer injections at the rostral superior temporal sulcus resulted in neuronal labelling in the anterior olfactory nucleus, piriform, periamygdaloid and entorhinal cortices, thus providing confirmation of the reciprocity between primary olfactory structures and the cortex at the rostral superior temporal sulcus. The reciprocal connections between the rostral part of superior temporal sulcus and primary olfactory structures represent a convergence for olfactory and other sensory modalities at the cortex of the rostral temporal lobe. [source]

    Presynaptic source of quantal size variability at GABAergic synapses in rat hippocampal neurons in culture

    Andrea Barberis
    Abstract The variability of quantal size depends on both presynaptic (profile of the neurotransmitter concentration in the cleft) and postsynaptic (number and gating properties of postsynaptic receptors) factors. Here we have examined the possibility that at nonsaturated synapses in cultured hippocampal neurons, changes in both the transmitter concentration peak and its clearance from the synaptic cleft may influence the variability of spontaneous miniature synaptic GABAergic currents (mIPSCs). We found that, in contrast to the slow-off GABAA receptor antagonist bicuculline, fast-off competitive antagonists such as SR-95103 and TPMPA differentially blocked small and large mIPSCs. In the presence of flurazepam, a drug believed to increase the affinity of GABA for GABAAR, small mIPSCs were enhanced more efficiently than large events. Moreover, the addition of dextran, which increases the viscosity of the extracellular fluid, preferentially increased small mIPSCs with respect to large ones. These observations suggest that changes in the concentration peak and the speed of GABA clearance in the cleft may be an important source of synaptic variability. The study of the correlation between peak amplitude and kinetics of mIPSCs allowed determination of the relative contribution of transmitter peak concentration vs. time of GABA clearance. Small synaptic responses were associated with fast onset and decay kinetics while large amplitude currents were asociated with slow kinetics, indicating a crucial role for GABA synaptic clearance in variability of mIPSCs. By using model simulations we were able to estimate the range of variability of both the concentration and the speed of clearance of the GABA transient in the synaptic cleft. [source]

    Odour-evoked [Ca2+] transients in mitral cell dendrites of frog olfactory glomeruli

    Kerry Delaney
    Abstract We measured Ca2+ concentration, [Ca2+], transients in mitral cell distal apical dendritic tufts produced by physiological odour stimulation of the olfactory epithelium and electrical stimulation of the olfactory nerve (ON) using two-photon scanning and conventional wide-field microscopy of Ca2+ -Green-1 dextran in an in vitro frog nose,brain preparation. Weak or strong ON shock-evoked fluorescence transients always had short latency with an onset 0,10 ms after the onset of the bulb local field potential, rapidly increasing to a peak of up to 25% fractional fluorescence change (,F/F) in 10,30 ms, were blocked by 10 µm CNQX, decaying with a time constant of about 1 s. With stronger ON shocks that activated many receptor axons, an additional, delayed, sustained AP5-sensitive component (peak at ,,0.5 s, up to 40% ,F/F maximum) could usually be produced. Odour-evoked [Ca2+] transients sometimes displayed a rapid onset phase that peaked within 50 ms but always had a sustained phase that peaked 0.5,1.5 s after onset, regardless of the strength of the odour or the amplitude of the response. These were considerably larger (up to 150% ,F/F) than those evoked by ON shock. Odour-evoked [Ca2+] transients were also distinguished from ON shock-evoked transients by tufts in different glomeruli responding with different delays (time to onset differed by up to 1.5 s between different tufts for the same odour). Odour-evoked [Ca2+] transients were increased by AMPA-kainate receptor blockade, but substantially blocked by AP5. Electrical stimulation of the lateral olfactory tract (5,6 stimuli at 10 Hz) that evoked granule cell feedback inhibition, blocked 60,100% of the odour-evoked [Ca2+] transient in tufts when delivered within about 0.5 s of the odour. LOT-mediated inhibition was blocked by 10 µm bicuculline. [source]

    Activation of hepatocyte growth factor activator zymogen (pro-HGFA) by human kallikrein 1-related peptidases

    FEBS JOURNAL, Issue 5 2008
    Shoichiro Mukai
    Hepatocyte growth factor activator (HGFA) is a serine protease and a potent activator of prohepatocyte growth factor/scatter factor (pro-HGF/SF), a multifunctional growth factor that is critically involved in tissue morphogenesis, regeneration, and tumor progression. HGFA circulates as a zymogen (pro-HGFA) and is activated in response to tissue injury. Although thrombin is considered to be an activator of pro-HGFA, alternative pro-HGFA activation pathways in tumor microenvironments remain to be identified. In this study, we examined the effects of kallikrein 1-related peptidases (KLKs), a family of extracellular serine proteases, on the activation of pro-HGFA. Among the KLKs examined (KLK2, KLK3, KLK4 and KLK5), we identified KLK4 and KLK5 as novel activators of pro-HGFA. Using N-terminal sequencing, the cleavage site was identified as the normal processing site, Arg407,Ile408. The activation of pro-HGFA by KLK5 required a negatively charged substance such as dextran sulfate, whereas KLK4 could process pro-HGFA without dextran sulfate. KLK5 showed more efficient pro-HGFA processing than KLK4, and was expressed in 50% (13/25) of the tumor cell lines examined. HGFA processed by these KLKs efficiently activated pro-HGF/SF, and led to cellular scattering and invasion in vitro. The activities of both KLK4 and KLK5 were strongly inhibited by HGFA inhibitor type 1, an integral membrane Kunitz-type serine protease inhibitor that inhibits HGFA and other pro-HGF/SF-activating proteases. These data suggest that KLK4 and KLK5 mediate HGFA-induced activation of pro-HGF/SF within tumor tissue, which may thereafter trigger a series of events leading to tumor progression via the MET receptor. [source]

    A Generalized System for Photoresponsive Membrane Rupture in Polymersomes

    Neha P. Kamat
    Abstract Polymersomes are vesicles whose membranes comprise self-assembled block copolymers. It has recently been shown that co-encapsulating conjugated multiporphyrin dyes in a polymersome membrane with ferritin protein in the aqueous lumen confers photolability to the polymersome. In the present study, the photolability is shown to be extendable to vesicles containing dextran, an inert and inexpensive polysaccharide, as the luminal solute. How structural features of the polymersome/porphyrin/dextran composite affect its photoresponse is explored. Increasing dextran molecular weight, decreasing block copolymer molecular weight, and altering fluorophore-membrane interactions results in increasing the photoresponsiveness of the polymersomes. Amphiphilic interactions of the luminal encapsulant with the membrane coupled with localized heat production in the hydrophobic bilayer likely cause differential thermal expansion in the membrane and the subsequent membrane rupture. This study suggests a general approach to impart photoresponsiveness to any biomimetic vesicle system without chemical modification, as well as a simple, bio-inert method for constructing photosensitive carriers for controlled release of encapsulants. [source]

    Insights into the reaction mechanism of glycosyl hydrolase family 49

    FEBS JOURNAL, Issue 22 2004
    Site-directed mutagenesis, substrate preference of isopullulanase
    Aspergillus niger isopullulanase (IPU) is the only pullulan-hydrolase in glycosyl hydrolase (GH) family 49 and does not hydrolyse dextran at all, while all other GH family 49 enzymes are dextran-hydrolysing enzymes. To investigate the common catalytic mechanism of GH family 49 enzymes, nine mutants were prepared to replace residues conserved among GH family 49 (four Trp, three Asp and two Glu). Homology modelling of IPU was also carried out based on the structure of Penicillium minioluteum dextranase, and the result showed that Asp353, Glu356, Asp372, Asp373 and Trp402, whose substitutions resulted in the reduction of activity for both pullulan and panose, were predicted to be located in the negatively numbered subsites. Three Asp-mutated enzymes, D353N, D372N and D373N, lost their activities, indicating that these residues are candidates for the catalytic residues of IPU. The W402F enzyme significantly reduced IPU activity, and the Km value was sixfold higher and the k0 value was 500-fold lower than those for the wild-type enzyme, suggesting that Trp402 is a residue participating in subsite ,1. Trp31 and Glu273, whose substitutions caused a decrease in the activity for pullulan but not for panose, were predicted to be located in the interface between N-terminal and ,-helical domains. The substrate preference of the negatively numbered subsites of IPU resembles that of GH family 49 dextranases. These findings suggest that IPU and the GH family 49 dextranases have a similar catalytic mechanism in their negatively numbered subsites in spite of the difference of their substrate specificities. [source]

    Binding of rat brain hexokinase to recombinant yeast mitochondria

    FEBS JOURNAL, Issue 10 2000
    Identification of necessary physico-chemical determinants
    The association of rat brain hexokinase with heterologous recombinant yeast mitochondria harboring human porin (Yh) is comparable to that with rat liver mitochondria in terms of cation requirements, cooperativity in binding, and the effect of amphipathic compounds. Mg2+, which is required for hexokinase binding to all mitochondria, can be replaced by other cations. The efficiency of hexokinases, however, depends on the valence of hydrophilic cations, or the partition of hydrophobic cations in the membrane, implying that these act by reducing a prohibitive negative surface charge density on the outer membrane rather than fulfilling a specific structural requirement. Macromolecular crowding (using dextran) has dual effects. Dextran added in excess increases hexokinase binding to yeast mitochondria, according to the porin molecule they harbor. This effect, significant with wild-type yeast mitochondria, is only marginal with Yh as well as rat mitochondria. On the other hand, an increase in the number of hexokinase binding sites on mitochondria is also observed. This increase, moderate in wild-type organelles, is more pronounced with Yh. Finally, dextran, which has no effect on the modulation of hexokinase binding by cations, abolishes the inhibitory effect of amphipathic compounds. Thus, while hexokinase binding to mitochondria is predetermined by the porin molecule, the organization of the latter in the membrane plays a critical role as well, indicative that porin must associate with other mitochondrial components to form competent binding sites on the outer membrane. [source]

    B,Z DNA Transition Triggered by a Cationic Comb-Type Copolymer

    Naohiko Shimada
    Abstract The conformational transition from right-handed B,DNA to left-handed Z,DNA,the B,Z transition,has received increased attention recently because of its potential roles in biological systems and its applicability to bionanotechnology. Though the B,Z transition of poly(dG,dC),·,poly(dG,dC) is inducible under high salt concentration conditions (over 4,M NaCl) or by addition of multivalent cations, such as hexaamminecobalt(III), no cationic polymer were known to induce the transition. In this study, it is shown by circular dichroism and UV spectroscopy that the cationic comb-type copolymer, poly(L -lysine)- graft -dextran, but not poly(L -lysine) homopolymer or a basic peptide, induces the B,Z transition of poly(dG,dC),·,poly(dG,dC). At a cationic amino group concentration of 10,4,M the copolymer stabilizes Z,DNA. The transition pathway from the B to the Z form is different to that observed previously. We speculate that the cationic backbone of the copolymer, which reduces electrostatic repulsion among DNA phosphate groups, and the hydrophilic dextran chains, which reduce activity of water, cooperate to induce the B,Z transition. The copolymer specifically modified the micro-environment around DNA molecules to induce Z,DNA formation through stable and spontaneous inter-polyelectrolyte complex formation. [source]

    Fabrication of Microbeads with a Controllable Hollow Interior and Porous Wall Using a Capillary Fluidic Device

    Sung-Wook Choi
    Abstract Poly(D,L -lactide-co-glycolide) (PLGA) microbeads with a hollow interior and porous wall are prepared using a simple fluidic device fabricated with PVC tubes, glass capillaries, and a needle. Using the fluidic device with three flow channels, uniform water-in-oil-in-water (W-O-W) emulsions with a single inner water droplet can be achieved with controllable dimensions by varying the flow rate of each phase. The resultant W-O-W emulsions evolve into PLGA microbeads with a hollow interior and porous wall after the organic solvent in the middle oil phase evaporates. Two approaches are employed for developing a porous structure in the wall: emulsion templating and fast solvent evaporation. For emulsion templating, a homogenized, water-in-oil (W/O) emulsion is introduced as the middle phase instead of the pure oil phase. Low-molecular-weight fluorescein isothiocyanate (FITC) and high-molecular-weight fluorescein isothiocyanate,dextran conjugate (FITC,DEX) is added to the inner water phase to elucidate both the pore size and their interconnectivity in the wall of the microbeads. From optical fluorescence microscopy and scanning electron microscopy images, it is confirmed that the emulsion-templated microbeads (W-W/O-W) have larger and better interconnected pores than the W-O-W microbeads. These microstructured microbeads can potentially be employed for cell encapsulation and tissue engineering, as well as protection of active agents. [source]

    Self-Rupturing and Hollow Microcapsules Prepared from Bio-polyelectrolyte - Coated Microgels,

    G. De, Geest
    Abstract This paper reports on microcapsules obtained by layer-by-layer deposition of bio-polyelectrolyte multilayers at the surface of biodegradable dextran microgels. The behavior of the layer-by-layer coating upon degradation of the microgel core strongly depends on the bio-polyelectrolytes used. Two types of microcapsules, "self-rupturing" microcapsules and "hollow" microcapsules, are presented. Self-rupturing microcapsules are obtained when the swelling pressure of the degrading microgel core is strong enough to rupture the surrounding bio-polyelectrolyte membrane. Self-rupturing microcapsules could be of interest as a pulsed drug delivery system. Hollow microcapsules are obtained after applying multiple layers of bio-polyelectrolyte that can withstand the swelling pressure of the degrading microgel core. Biomacromolecules (such as albumin and dextran) spontaneously accumulate in the hollow microcapsules prepared from dex-HEMA microgels, which could be of interest for drug-encapsulation purposes. [source]

    Aldehyde-Amine Chemistry Enables Modulated Biosealants with Tissue-Specific Adhesion

    ADVANCED MATERIALS, Issue 32-33 2009
    Natalie Artzi
    The interfacial regions between PEG: dextran-based adhesive sealant and excised rat heart, lung, liver, and duodenum tissues exhibit three distinct domains; target tissue (red and blue), bulk material (green), and an adhesive regime interposed between the two. The variation in adhesive regime morphology when applied to different tissues provides a rational approach for the engineering of application-specific surgical sealants. [source]

    Cord blood mesenchymal stem cells propel human dendritic cells to an intermediate maturation state and boost interleukin-12 production by mature dendritic cells

    IMMUNOLOGY, Issue 4 2009
    Lieke C. J. Van Den Berk
    Summary Pathogen-derived entities force the tissue-resident dendritic cells (DCs) towards a mature state, followed by migration to the draining lymph node to present antigens to T cells. Bone marrow mesenchymal stem cells (MSCs) modulate the differentiation, maturation and function of DCs. In umbilical cord blood an immature MSC population was identified. Remarkably, these immature stem cells modulated DCs in a different way. Marker expression was unchanged during the differentiation of monocytes towards immature DCs (iDCs) when cocultured with cord blood MSC [unrestricted somatic stem cells (USSCs)]. The maturation to mature DCs (mDCs) was enhanced when DCs were co-cultured with USSC, as evidenced by the up-regulation of costimulatory molecules. Endocytosis of dextran by iDCs was hampered in the presence of USSCs, which is indicative for the maturation of iDCs. Despite this maturation, the migration of iDCs cocultured with USSCs appeared to be identical to iDCs cultured alone. However, USSCs increased the migration of mDCs towards CCL21 and boosted interleukin-12 production. So, USSCs mature iDCs, thereby redirecting the antigen-uptake phenotype towards a mature phenotype. Furthermore, DC maturation by lipopolysaccharide (LPS) or USSCs reflects two distinct pathways because migration was unaffected when iDCs were matured by coculture with USSCs, while it was strongly enhanced in the presence of LPS. DCs are able to discriminate the different MSC subtypes, resulting in diverse differentiation programmes. [source]

    DEC-205lo Langerinlo neonatal Langerhans' cells preferentially utilize a wortmannin-sensitive, fluid-phase pathway to internalize exogenous antigen

    IMMUNOLOGY, Issue 4 2003
    Bernadette M. Bellette
    Summary Antigen treatment of neonatal epidermis results in antigen-specific immune suppression. Compared with adult counterparts, neonatal Langerhans' cells (LC) demonstrate an impaired ability to transport antigen to the lymph node (LN). As it is possible that neonatal LC have a reduced ability to endocytose antigen, we evaluated the acquisition of endocytic function, the expression of uptake receptors and the internalization of soluble and small particulate antigens in neonatal, juvenile and adult mice. Although LC from 4-day-old mice were weakly positive for the mannose-type receptor, Langerin, they were capable of internalizing fluorescein isothiocyanate (FITC)-dextran, but to a lesser extent than LC from 6-week-old mice. However, when ratio data were calculated to account for variations in fluorescence intensity at 4°, it was demonstrated that neonatal LC continued to internalize antigen over a longer period of time than adult mice and, as the ratios were much higher, that neonatal cells were also relatively more efficient in antigen uptake. When receptors for mannan and mannose were competitively blocked, LC from neonatal mice, but not adult mice, could still efficiently internalize FITC,dextran. Consequently, the uptake of FITC,dextran, in part, occurred via alternative receptors or a receptor-independent fluid-phase pathway. A feasible pathway is macropinocytosis, as LC from 4-day-old mice demonstrated a reduction in FITC,dextran internalization by the macropinocytosis inhibitor, wortmannin. Evidence of a functional macropinocytosis pathway in neonatal LC was further supported by internalization of the soluble tracer Lucifer Yellow (LY). We conclude that neonatal LC preferentially utilize a wortmannin-sensitive, fluid-phase pathway, rather than receptor-mediated endocytosis, to internalize antigen. As neonatal LC are capable of sampling their environment without inducing immunity, this may serve to avoid inappropriate immune responses during the neonatal period. [source]

    Bifidobacterium lactis inhibits NF-,B in intestinal epithelial cells and prevents acute colitis and colitis-associated colon cancer in mice,,

    Seung Won Kim MS
    Abstract Background: The aim of this study was to investigate the antiinflammatory effects of Bifidobacterium lactis on intestinal epithelial cells (IECs) and on experimental acute murine colitis and its tumor prevention effects on colitis-associated cancer (CAC) in mice. Methods: Human HT-29 cells were stimulated with IL-1,, lipopolysaccharides, or tumor necrosis factor-, with and without B. lactis, and the effects of B. lactis on nuclear factor kappa B (NF-,B) signaling in IEC were examined. For in vivo study, dextran sulfate sodium (DSS)-treated mice were fed with and without B. lactis. Finally, we induced colonic tumors in mice by azoxymethane (AOM) and DSS and evaluated the effects of B. lactis on tumor growth. Results: B. lactis significantly suppressed NF-,B activation, including NF-,B-binding activity and NF-,B-dependent reporter gene expression in a dose-dependent manner, and suppressed I,B-, degradation, which correlated with the downregulation of NF-,B-dependent gene products. Moreover, B. lactis suppressed the development of acute colitis in mice. Compared with the DSS group, the severity of DSS-induced colitis as assessed by disease activity index, colon length, and histological score was reduced in the B. lactis -treated group. In the CAC model, the mean number and size of tumors in the B. lactis -treated group were significantly lower than those in the AOM group. Conclusions: Our data demonstrate that B. lactis inhibits NF-,B and NF-,B-regulated genes in IEC and prevents acute colitis and CAC in mice. These results suggest that B. lactis could be a potential preventive agent for CAC as well as a therapeutic agent for inflammatory bowel disease. (Inflamm Bowel Dis 2010) [source]

    Dipeptidyl peptidase expression during experimental colitis in mice

    Roger Yazbeck PhD
    Abstract Background: We have previously demonstrated that inhibition of dipeptidyl peptidase (DP) activity partially attenuates dextran sulfate sodium (DSS) colitis in mice. The aim of this study was to further investigate the mechanisms of this protection. Materials and Methods: Wildtype (WT) and DPIV,/, mice consumed 2% DSS in drinking water for 6 days to induce colitis. Mice were treated with saline or the DP inhibitors Ile-Pyrr-(2-CN)*TFA or Ile-Thia. DP mRNA and enzyme levels were measured in the colon. Glucagon-like peptide (GLP)-2 and GLP-1 concentrations were determined by radioimmunoassay, regulatory T-cells (Tregs) by fluorescence activated cell sorting (FACS) on FOXp3+T cells in blood, and neutrophil infiltration assessed by myeloperoxidase (MPO) assay. Results: DP8 and DP2 mRNA levels were increased (P < 0.05) in WT+saline mice compared to untreated WT mice with colitis. Cytoplasmic DP enzyme activity was increased (P < 0.05) in DPIV,/, mice at day 6 of DSS, while DP2 activity was increased (P < 0.05) in WT mice with colitis. GLP-1 (63%) and GLP-2 (50%) concentrations increased in WT+Ile-Pyrr-(2-CN)*TFA mice compared to day-0 controls. MPO activity was lower in WT+Ile-Thia and WT+Ile-Pyrr-(2-CN)*TFA treated mice compared to WT+saline (P < 0.001) at day 6 colitis. Conclusions: DP expression and activity are differentially regulated during DSS colitis, suggesting a pathophysiological role for these enzymes in human inflammatory bowel disease (IBD). DP inhibitors impaired neutrophil recruitment and maintenance of the Treg population during DSS-colitis, providing further preclinical evidence for the potential therapeutic use of these inhibitors in IBD. Finally, DPIV appears to play a critical role in mediating the protective effect of DP inhibitors. Inflamm Bowel Dis 2010 [source]