sepharose and Disease-Models--Animal

This review presents a summary of various types of scaffold biomaterials used alone or together with therapeutic drugs and cells to regenerate spinal cord injury (SCI). The inhibitory environment and loss of axonal connections after SCI give rise to critical obstacles to regeneration of lost tissues and neuronal functions. Biomaterial scaffolds can provide a bridge to connect lost tissues, an adhesion site for implanted or host cells, and sustained release of therapeutic drugs in the injured spinal cord. In addition, they not only provide a structural platform, but can play active roles by inhibiting apoptosis of cells, inflammation and scar formation, and inducing neurogenesis, axonal growth and angiogenesis. Many synthetic and natural biomaterial scaffolds have been extensively investigated and tested in vitro and in animal SCI models for these purposes. We summarized the literature on the biomaterials commonly used for spinal cord regeneration in terms of historical backgrounds and current approaches.

Topics: Alginates; Animals; Apoptosis; Axons; Biocompatible Materials; Chitosan; Collagen; Disease Models, Animal; Drug Delivery Systems; Fibrin; Humans; Hyaluronic Acid; Inflammation; Lactic Acid; Materials Testing; Peptides; Polyesters; Polymers; Sepharose; Spinal Cord; Spinal Cord Injuries; Spinal Cord Regeneration; Stem Cells; Tissue Engineering; Tissue Scaffolds

Porphyran is known to inhibit immune cell function. Previously, porphyran was shown to prevent lipopolysaccharide-induced sepsis in mice. However, studies on the inhibitory effects of porphyran during colitis are currently lacking. In this study, we evaluated the effects of Pyropia yezoensis-derived porphyran on dextran sodium sulfate (DSS)-induced acute and chronic colitis. The oral or intraperitoneal administration of porphyran inhibited the progression of DSS-induced colitis in mice, with the former also preventing immune cell infiltration in the colon. The levels of intracellular interferon-γ and interleukin-17 in T cells decreased when porphyran was administered orally. Porphyran inhibited T cell activation by suppressing dendritic cells (DCs) and macrophages. Porphyran prevented pathogen-associated molecular pattern and damage-associated molecular pattern-dependent DC and macrophage activation. Finally, porphyran attenuated chronic colitis caused via the long-term administration of DSS. These findings indicate that the oral administration of porphyran can inhibit DSS-induced colitis by suppressing DC and macrophage activation.

Topics: Animals; Colitis; Colon; Dendritic Cells; Dextran Sulfate; Disease Models, Animal; Mice; Mice, Inbred C57BL; Rhodophyta; Sepharose

Antigen-presenting cells express pattern recognition receptors (PRRs), which sense pathogen-associated molecular patterns from microorganisms and lead to the induction of inflammatory responses. C-type lectin receptors (CLRs), the representative PRRs, bind to microbial polysaccharides, among which Dectin-2 and Mincle recognize mannose-containing polysaccharides. Because influenza virus (IFV) hemagglutinin (HA) is rich in mannose polysaccharides, Dectin-2 or Mincle may contribute to the recognition of HA. In this study, we addressed the possible involvement of Dectin-2 and Mincle in the viral recognition and the initiation of cytokine production. Interleukin (IL)-12p40 and IL-6 production by bone marrow-derived dendritic cells (BM-DCs) upon stimulation with HA was significantly reduced in Dectin-2 knockout (KO) mice compared to wild-type (WT) mice whereas there was no difference between WT mice and Mincle KO mice. BM-DCs that were treated with Syk inhibitor resulted in a significant reduction of cytokine production upon stimulation with HA. The treatment of BM-DCs with methyl-α-D-mannopyranoside (ManP) also led to a significant reduction in cytokine production by BM-DCs that were stimulated with HA, except for the A/H1N1pdm09 subtype. IL-12p40 and IL-6 synthesis by BM-DCs was completely diminished upon stimulation with HA treated with concanavalin A (ConA)-bound sepharose beads. Finally, GFP expression was detected in reporter cells that were transfected with the Dectin-2 gene, but not with the Mincle gene, when stimulated with HA derived from the A/H3N2 subtype. These data suggested that Dectin-2 may be a key molecule as the sensor for IFV to initiate the immune response and regulate the pathogenesis of IFV infection.

Topics: Animals; Antigen-Presenting Cells; Bone Marrow Cells; Concanavalin A; Cytokines; Disease Models, Animal; Green Fluorescent Proteins; Hemagglutinin Glycoproteins, Influenza Virus; Humans; Immune System; Influenza, Human; Interleukin-12 Subunit p40; Interleukin-6; Lectins, C-Type; Ligands; Membrane Proteins; Mice; Mice, Inbred C57BL; Mice, Knockout; NFATC Transcription Factors; Sepharose; Syk Kinase

Porphyran and its derivatives possess a variety of biological activities, such as ameliorations of oxidative stress, inflammation, hyperlipemia, and immune deficiencies. In this study, we evaluated the potential efficacy of porphyran-derived oligosaccharides from Porphyra yezoensis (PYOs) in alleviating nonalcoholic fatty liver disease (NAFLD) and preliminarily clarified the underlying mechanism. NAFLD was induced by a high-fat diet for six months in C57BL/6J mice, followed by treatment with PYOs (100 or 300 mg/kg/d) for another six weeks. We found that PYOs reduced hepatic oxidative stress in mice with NAFLD, which plays a critical role in the occurrence and development of NAFLD. In addition, PYOs could markedly decrease lipid accumulation in liver by activating the IRS-1/AKT/GSK-3β signaling pathway and the AMPK signaling pathway in mice with NAFLD. PYOs also apparently relieved the hepatic fibrosis induced by oxidative stress via downregulation of TGF-β and its related proteins, so that liver injury was markedly alleviated. Furthermore, PYOs treatment relieved cecal microbiota dysbiosis (such as increasing the relative abundance of Akkermansia, while decreasing the Helicobacter abundance), which could alleviate oxidative stress, inflammation, and lipid metabolism, and protect the liver to a certain degree. In summary, PYOs treatment remarkably improved NAFLD via a specific molecular mechanism and reshaped the cecal microbiota.

Topics: Animals; Cecum; Disease Models, Animal; Dysbiosis; Gastrointestinal Microbiome; Lipid Metabolism; Male; Mice; Mice, Inbred C57BL; Non-alcoholic Fatty Liver Disease; Oligosaccharides; Oxidative Stress; Sepharose; Signal Transduction

Articular cartilage exhibits reduced self-healing following degeneration. This research evaluated the effects of hydrogels derived from various polysaccharides-gellan gum (GG), alginate, and agarose-on cartilage regeneration compared with that of hyaluronic acid (HA), which is commonly used in cartilage tissue engineering. Chondrocytes were isolated from the articular cartilage of New Zealand White (NZW) rabbits and stimulated with IL-1β followed by incubation with polysaccharides. The expressions of NF-κB and Cox-2 were decreased and those of IκBα, Sox-9, aggrecan, and type II collagen were increased in HA, GG, and Alginate groups. Osteochondral defects in NZW rabbits were treated with intra-articular polysaccharide injections; all except alginate resulted in tissue regeneration. Significant improvements were observed in cartilage regeneration in the GG and agarose groups. These results show that GG and agarose improve cartilage regeneration by suppressing inflammatory mediators and inducing cartilage formation and autophagy-related gene expression, indicating their potential for cartilage tissue engineering.

Topics: Alginates; Animals; Autophagy; Biomarkers; Cartilage, Articular; Cell Survival; Cells, Cultured; Chondrocytes; Chondrogenesis; Cross-Linking Reagents; Disease Models, Animal; Gene Expression Regulation; Hyaluronic Acid; Hydrogels; Inflammation; Male; Osteoarthritis; Polysaccharides; Polysaccharides, Bacterial; Rabbits; Regeneration; Rheology; RNA, Messenger; Sepharose; Spectroscopy, Fourier Transform Infrared; X-Ray Diffraction

Nicotinamide adenine dinucleotide (NAD) kinases are essential and ubiquitous enzymes involved in the tight regulation of NAD/nicotinamide adenine dinucleotide phosphate (NADP) levels in many metabolic pathways. Consequently, they represent promising therapeutic targets in cancer and antibacterial treatments. We previously reported diadenosine derivatives as NAD kinase inhibitors with bactericidal activities on

Topics: Adenine; Adenosine; Amino Acid Sequence; Animals; Anti-Bacterial Agents; Bacterial Proteins; Disease Models, Animal; Enzyme Inhibitors; Mice; Models, Molecular; NADP; Phosphotransferases (Alcohol Group Acceptor); Protein Conformation; Sepharose; Staphylococcus aureus

Rats are an early preclinical model for cartilage tissue engineering, and a practical species for investigating the effects of aging. However, rats may be a poor aging model for mesenchymal stem cells (MSCs) based on laboratory reports of a severe decline in chondrogenesis beyond young adulthood. Such testing has not been conducted with MSCs seeded in a scaffold, which can improve the propensity of MSCs to undergo chondrogenesis. Therefore, the objective of this study was to evaluate chondrogenesis of middle-aged rat MSCs encapsulated in agarose.. MSCs from 14- to 15-month-old rats were expanded, seeded into agarose, and cultured in chondrogenic medium with or without 5% serum for 15 days. Samples were evaluated for cell viability and cartilaginous extracellular matrix (ECM) accumulation. Experiments were repeated using MSCs from 6-week-old rats.. During expansion, middle-aged rat MSCs demonstrated a diminishing proliferation rate that was improved ~2-fold in part by transient exposure to chondrogenic medium. In agarose culture in defined medium, middle-aged rat MSCs accumulated ECM to a much greater extent than negative controls. Serum supplementation improved cell survival ~2-fold, and increased ECM accumulation ~3-fold. Histological analysis indicated that defined medium supported chondrogenesis in a subset of cells, while serum-supplementation increased the frequency of chondrogenic cells. In contrast, young rat MSCs experienced robust chondrogenesis in defined medium that was not improved with serum-supplementation.. These data demonstrate a previously-unreported propensity of middle-aged rat MSCs to undergo chondrogenesis, and the potential of serum to enhance chondrogenesis of aging MSCs.

Topics: Animals; Cartilage; Cell Differentiation; Cell Survival; Cellular Senescence; Chondrocytes; Chondrogenesis; Culture Media; Disease Models, Animal; Extracellular Matrix; Mesenchymal Stem Cells; Rats; Sepharose; Serum; Tissue Engineering

Topics: 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine; Animals; Apoptosis; Behavior, Animal; Brain; Disease Models, Animal; Dopamine; Dopaminergic Neurons; Humans; Male; Mice; Mice, Inbred C57BL; MPTP Poisoning; Neuroprotective Agents; Phosphatidylinositol 3-Kinases; Porphyra; Proto-Oncogene Proteins c-akt; Proto-Oncogene Proteins c-bcl-2; Sepharose; Signal Transduction

The present study aimed to investigate the technical feasibility of percutaneous endoscopic mini-hemilaminectomy via a uniportal approach, and to evaluate the possibility of decompression and endoscopic examination of the thoracic and lumbar spinal canals in small dogs during such procedures. Fresh canine cadavers of mixed-breed dogs (n=7) were used in this study. Following injection of a barium and agarose mixture (BA-gel) to stimulate intervertebral disc herniation, percutaneous endoscopic mini-hemilaminectomy was performed using a lateral approach to the thoracic and lumbar vertebrae. BA-gel was removed to decompress the spinal cord using an elevator and rongeurs after mini-hemilaminectomy. Pre and post-operative computed tomography (CT) scans were obtained to evaluate surgical outcomes. Intra-operative complications, incision length, and procedure time were recorded. All procedures were completed with clear visualization of the spinal cord and floor of the spinal canal. The mean total operating time was 58.00 ± 18.06 min. Lengths of incision were under 1 cm in all dogs. Intra-operative complications included iatrogenic nerve root injuries caused by the micro-rongeur in two dogs. CT imaging revealed that removal of BA-gel resulted in sufficient spinal cord decompression. Our findings indicated that percutaneous endoscopic thoracolumbar mini-hemilaminectomy is feasible for spinal cord decompression and allows for adequate observation of the spinal canal. Thus, this technique may be an alternative surgical option for treatment of thoracolumbar disk disease in dogs.

Topics: Animals; Barium Sulfate; Cadaver; Disease Models, Animal; Dog Diseases; Dogs; Endoscopy; Feasibility Studies; Intervertebral Disc Displacement; Laminectomy; Neuroendoscopy; Sepharose; Tomography, X-Ray Computed

In neurodegenerative diseases activation of immune cells is thought to play a major role. Microglia are the main immune cells of the central nervous system. When encountering disease related stimuli microglia adopt an activated phenotype that typically includes a rounded morphology. The exact role of microglia or other potentially infiltrating myeloid cells in different brain diseases is not fully understood. In this chapter we present techniques in zebrafish to induce degeneration of neurons, to activate the microglia, and to study activation phenotypes by immunohistochemistry and in vivo by fluorescence microscopic imaging.

Topics: Animals; Animals, Genetically Modified; Apoptosis; Brain; Disease Models, Animal; Gene Expression; Genes, Reporter; Green Fluorescent Proteins; Humans; Immunohistochemistry; Larva; Luminescent Proteins; Microglia; Microscopy, Fluorescence; Neurodegenerative Diseases; Neurons; Phagocytosis; Phase Transition; Red Fluorescent Protein; Sepharose; Zebrafish

Calcific aortic valve disease (CAVD) is the most common valvular disorder. While fluid stresses are presumed to play a role in disease progression, the valvular hemodynamic changes experienced over the course of CAVD remain largely unknown. The study aim was to develop a laboratory protocol for the fabrication of tissue valve models mimicking mild and moderate calcific stenosis, for future use in flow studies.. Different hydroxyapatite (HA)-agarose mixtures were injected into porcine valve leaflets. Micro-computed tomography (micro-CT) was used to quantify HA deposition volume, area fraction and regional distribution, while von Kossa staining was performed to assess tissue mineralization. Particle image velocimetry measurements were carried out in intact and injected valves subjected to in vivo-like hemodynamics to characterize the degree of valvular stenosis in terms of geometric orifice area (GOA) and peak systolic velocity.. The 5% HA-1% agarose solution (solution 1) and the 5% HA-0.5% agarose solution (solution 2) maximized the HA deposition volume. Leaflet injections with solution 1 resulted in a significant 1.9-fold increase in HA area fraction relative to solution 2 injections. While solution 1 injections generated multiple sites of high HA concentration, solution 2 injections produced smaller, discrete spots. Injections of both solution 1 and solution 2 into whole valves generated significant 47% and 32% reductions, respectively, in GOA and 1.8-fold and 1.5-fold increases, respectively, in peak systolic velocity, relative to untreated valves.. Tissue valve models were generated that recapitulated the structure and hemodynamics of mild and moderate valvular calcification. Those models may be used for future investigations of the native valvular hemodynamic alterations that occur during CAVD.

Topics: Animals; Aortic Valve; Aortic Valve Stenosis; Blood Flow Velocity; Calcinosis; Disease Models, Animal; Durapatite; Hemodynamics; Regional Blood Flow; Sepharose; Severity of Illness Index; Swine; X-Ray Microtomography

Arterial thrombosis is an important complication of diabetes and cancer, being an important target for therapeutic intervention. Crataeva tapia bark lectin (CrataBL) has been previously shown to have hypoglycemiant effect and also to induce cancer cell apoptosis. It also showed inhibitory activity against Factor Xa (Kiapp=8.6 μm). In the present study, we evaluated the anti-thrombotic properties of CrataBL in arterial thrombosis model. CrataBL prolongs the activated partial thromboplastin time on human and mouse plasma, and it impairs the heparin-induced potentiation of antithrombin III and heparin-induced platelet activation in the presence of low-dose ADP. It is likely that the dense track of positive charge on CrataBL surface competes with the heparin ability to bind to antithrombin III and to stimulate platelets. In the photochemically induced thrombosis model in mice, in the groups treated with 1.25, 5.0, or 10 mg/kg CrataBL, prior to the thrombus induction, the time of total artery occlusion was prolonged by 33.38%, 65%, and 66.11%, respectively, relative to the time of the control group. In contrast to heparin, the bleeding time in CrataBL-treated mice was no longer than in the control. In conclusion, CrataBL was effective in blocking coagulation and arterial thrombus formation, without increasing bleeding time.

Topics: Animals; Blood Coagulation; Capparaceae; Carotid Arteries; Chromatography, Affinity; Disease Models, Animal; Factor Xa Inhibitors; Humans; Hydrolysis; Mice, Inbred C57BL; Nitric Oxide; Partial Thromboplastin Time; Plant Lectins; Platelet Aggregation; Prothrombin Time; Regional Blood Flow; Sepharose; Substrate Specificity; Thrombosis

This study aimed to evaluate the detectability of stem cells labeled with very small iron oxide particles (VSOP) at 3T with susceptibility weighted (SWI) and T2* weighted imaging as a methodological basis for subsequent examinations in a large animal stroke model (sheep).. We examined ovine mesenchymal stem cells labeled with VSOP in agarose layer phantoms. The experiments were performed in 2 different groups, with quantities of 0-100,000 labeled cells per layer. 15 different SWI- and T2*-weighted sequences and 3 RF coils were used. All measurements were carried out on a clinical 3T MRI. Images of Group A were analyzed by four radiologists blinded for the number of cells, and rated for detectability according to a four-step scale. Images of Group B were subject to a ROI-based analysis of signal intensities. Signal deviations of more than the 0.95 confidence interval in cell containing layers as compared to the mean of the signal intensity of non cell bearing layers were considered significant.. 500 or more labeled cells were judged as confidently visible when examined with a SWI-sequence with 0.15 mm slice thickness. Group B: 500 or more labeled cells showed a significant signal reduction in SWI sequences with a slice thickness of 0.25 mm. Slice thickness and cell number per layer had a significant influence on the amount of detected signal reduction.. 500 VSOP labeled stem cells could be detected with SWI imaging at 3 Tesla using an experimental design suitable for large animal models.

Topics: Animals; Disease Models, Animal; Ferric Compounds; Limit of Detection; Magnetic Resonance Imaging; Mesenchymal Stem Cells; Particle Size; Phantoms, Imaging; Sepharose; Sheep; Staining and Labeling; Stem Cell Transplantation; Stroke

Agarose is hydrolyzed easily to yield oligosaccharides, designated as agaro-oligosaccharides (AGOs). Recently, it has been demonstrated that AGOs induce heme oxygenase-1 (HO-1) expression in macrophages and that they might lead to anti-inflammatory property. Nevertheless, the molecular mechanism of AGO-mediated HO-1 induction remains unknown, as does AGOs' ability to elicit anti-inflammatory activity in vivo. This study was undertaken to uncover the mechanism of AGO-mediated HO-1 induction and to investigate the therapeutic effect of AGOs on intestinal inflammation.. Mice were treated with 2,4,6-trinitrobenzene sulfonic acid (TNBS) to induce colitis. The respective degrees of mucosal injury of mice that had received AGO and control mice were compared. We investigated HO-1 expression using Western blotting, quantitative real-time PCR (qRT-PCR), and immunohistochemistry. The expression of tumor necrosis factor-α (TNF-α) was measured using qRT-PCR and enzyme-linked immunosorbent assay.. AGO administration induced HO-1 expression in colonic mucosa. The induction was observed mainly in F4/80 positive macrophages. Increased colonic damage and myeloperoxidase activity after TNBS treatment were inhibited by AGO administration. TNBS treatment induced TNF-α expression, and AGO administration suppressed induction. However, HO inhibitor canceled AGO-mediated amelioration of colitis. In RAW264 cells, AGOs enhanced HO-1 expression time-dependently and concentration-dependently and suppressed lipopolysaccharide-induced TNF-α expression. Furthermore, agarotetraose-mediated HO-1 induction required NF-E2-related factor 2 function and phosphorylation of c-jun N-terminal kinase.. We infer that AGO administration inhibits TNBS-induced colitis in mice through HO-1 induction in macrophages. Consequently, oral administration of AGOs might be an important therapeutic strategy for inflammatory bowel disease.

Topics: Animals; Anti-Inflammatory Agents; Blotting, Western; Cell Line; Colitis; Disease Models, Animal; Dose-Response Relationship, Drug; Enzyme-Linked Immunosorbent Assay; Heme Oxygenase-1; Inflammation; Macrophages; Male; Mice; Mice, Inbred C57BL; Oligosaccharides; Real-Time Polymerase Chain Reaction; Sepharose; Time Factors; Trinitrobenzenesulfonic Acid

Object Experimental data about the evolution of intracranial volume and pressure in cases of hydrocephalus are limited due to the lack of available monitoring techniques. In this study, the authors validate intracranial CSF volume measurements within the lateral ventricle, while simultaneously using impedance sensors and pressure transducers in hydrocephalic animals. Methods A volume sensor was fabricated and connected to a catheter that was used as a shunt to withdraw CSF. In vitro bench-top calibration experiments were created to provide data for the animal experiments and to validate the sensors. To validate the measurement technique in a physiological system, hydrocephalus was induced in weanling rats by kaolin injection into the cisterna magna. At 28 days after induction, the sensor was implanted into the lateral ventricles. After sealing the skull using dental cement, an acute CSF drainage/infusion protocol consisting of 4 sequential phases was performed with a pump. Implant location was confirmed via radiography using intraventricular iohexol contrast administration. Results Controlled CSF shunting in vivo with hydrocephalic rats resulted in precise and accurate sensor measurements (r = 0.98). Shunting resulted in a 17.3% maximum measurement error between measured volume and actual volume as assessed by a Bland-Altman plot. A secondary outcome confirmed that both ventricular volume and intracranial pressure decreased during CSF shunting and increased during infusion. Ventricular enlargement consistent with successful hydrocephalus induction was confirmed using imaging, as well as postmortem. These results indicate that volume monitoring is feasible for clinical cases of hydrocephalus. Conclusions This work marks a departure from traditional shunting systems currently used to treat hydrocephalus. The overall clinical application is to provide alternative monitoring and treatment options for patients. Future work includes development and testing of a chronic (long-term) volume monitoring system.

Topics: Animals; Calibration; Catheters; Cerebrospinal Fluid; Cisterna Magna; Dilatation, Pathologic; Disease Models, Animal; Electric Impedance; Equipment Design; Gels; Hydrocephalus; In Vitro Techniques; Injections; Intracranial Pressure; Kaolin; Lateral Ventricles; Monitoring, Physiologic; Rats; Reproducibility of Results; Sepharose

Axonal regeneration depends on many factors, such as the type of injury and repair, age, distance from the cell body and distance of the denervated muscle, loss of surrounding tissue and the type of injured nerve. Experimental models use tubulisation with a silicone tube to research regenerative factors and substances to induce regeneration. Agarose, collagen and DMEM (Dulbecco's modified Eagle's medium) can be used as vehicles. In this study, we compared the ability of these vehicles to induce rat sciatic nerve regeneration with the intent of finding the least active or inert substance. The experiment used 47 female Wistar rats, which were divided into four experimental groups (agarose 4%, agarose 0.4%, collagen, DMEM) and one normal control group. The right sciatic nerve was exposed, and an incision was made that created a 10 mm gap between the distal and proximal stumps. A silicone tube was grafted onto each stump, and the tubes were filled with the respective media. After 70 days, the sciatic nerve was removed. We evaluated the formation of a regeneration cable, nerve fibre growth, and the functional viability of the regenerated fibres.. Comparison among the three vehicles showed that 0.4% agarose gels had almost no effect on provoking the regeneration of peripheral nerves and that 4% agarose gels completely prevented fibre growth. The others substances were associated with profuse nerve fibre growth.. In the appropriate concentration, agarose gel may be an important vehicle for testing factors that induce regeneration without interfering with nerve growth.

Topics: Animals; Axons; Collagen; Disease Models, Animal; Dose-Response Relationship, Drug; Female; Nerve Fibers, Myelinated; Nerve Regeneration; Prostheses and Implants; Rats; Rats, Wistar; Sciatic Nerve; Sciatic Neuropathy; Sepharose; Silicones; Time Factors

Porphyran (POR) from the red alga Porphyra yezoensis is a water soluble dietary fiber. In this study, we investigated the effect of dietary POR on glucose metabolism in KK-Ay mice (a model for type 2 diabetes). Mice were divided into 4 groups and fed a diet containing 5% cellulose (control), POR, POR Arg or POR K. After 3 wk of feeding, plasma insulin levels and the calculated homeostasis model assessment-insulin resistance (HOMA-IR) index were significantly lower in the POR group than in the control group. Compared with the control group, plasma adiponectin levels were significantly increased in the POR, POR Arg and POR K groups. These results suggest that dietary POR should improve glucose metabolism in diabetes via up-regulation of adiponectin levels. In addition, the amount of propionic acid in the cecum of the POR group was significantly higher than in the control group and the profile of bacterial flora was changed by dietary POR. In the cecum of the POR, POR Arg and POR K groups, Bacteroides was significantly increased and Clostridium coccoides was significantly decreased compared with in the control group. The effects of dietary POR on the hindgut environment might contribute to the improvement of glucose metabolism.

Topics: Adiponectin; Animals; Cecum; Cholesterol; Diabetes Mellitus, Type 2; Dietary Fiber; Disease Models, Animal; Fatty Acids, Nonesterified; Glucose; Insulin; Insulin Resistance; Male; Mice; Porphyra; Real-Time Polymerase Chain Reaction; RNA, Bacterial; Sepharose; Triglycerides

This study developed a transplantable platform based on cationic hydrogels to deliver antisense oligodeoxynucleotides (ASOs) targeting the mRNA of TNF-alpha. Cationic agarose (c-agarose) was obtained by conjugating ethylenediamine to agarose via an N,N'-carbonyldiimidazole (CDI)-activation method. ASO-c-agarose system was constructed by mixing ASO in cationic agarose gel of proper concentration and gelation temperature. In vivo assessment of ASO distribution suggested that the system specifically target to spleen, wherein the c-agarose-delivered ASO had a concentration remarkably 50-fold higher than that of the naked ASO. The distribution of c-agarose-delivered ASO was scarcely detectable in liver and kidney. Next, three types of animal models were setup to evaluate the therapeutic efficacies of ASO-Gel, including the adjuvant-induced arthritis (AA), carrageen/lipopolysaccharide (LPS)-induced arthritis (CLA) and collagen-induced arthritis (CIA) models. The effects of ASO-c-agarose in alleviating inflammation and tissue destruction were evidenced in more than 90% of the testing animals, with decrease of main inflammatory cytokines, lightening of joint swelling and tissue damage, as well as increase in their body weights. All these findings suggest that this highly operable devise for the conveyance of antisense nucleotides together with its spleen-targeting property, could become a useful means of antisense-based therapeutics against rheumatoid arthritis and other diseases.

Topics: Animals; Arthritis, Experimental; Cations; Disease Models, Animal; Ethylenediamines; Female; Hydrogel, Polyethylene Glycol Dimethacrylate; Mechanical Phenomena; Mice; Microscopy, Electron, Scanning; Oligonucleotides, Antisense; Organ Specificity; Rats; Rats, Sprague-Dawley; Sepharose; Spleen; Tissue Distribution; Tumor Necrosis Factor-alpha

Spiral ganglion neurons often degenerate in the deaf ear, compromising the function of cochlear implants. Cochlear implant function can be improved by good preservation of the spiral ganglion neurons, which are the target of electrical stimulation by the implant. Brain derived neurotrophic factor (BDNF) has previously been shown to enhance spiral ganglion survival in experimentally deafened ears. Providing enhanced levels of BDNF in human ears may be accomplished by one of several different methods. The goal of these experiments was to test a modified design of the cochlear implant electrode that includes a coating of fibroblast cells transduced by a viral vector with a BDNF gene insert. To accomplish this type of ex vivo gene transfer, we transduced guinea pig fibroblasts with an adenovirus with a BDNF gene cassette insert, and determined that these cells secreted BDNF. We then attached BDNF-secreting cells to the cochlear implant electrode via an agarose gel, and implanted the electrode in the scala tympani. We determined that the BDNF expressing electrodes were able to preserve significantly more spiral ganglion neurons in the basal turns of the cochlea after 48 days of implantation when compared to control electrodes. This protective effect decreased in the higher cochlear turns. The data demonstrate the feasibility of combining cochlear implant therapy with ex vivo gene transfer for enhancing spiral ganglion neuron survival.

Topics: Adenoviridae; Animals; Brain-Derived Neurotrophic Factor; Cell Survival; Cells, Cultured; Cochlear Implantation; Cochlear Implants; Deafness; Disease Models, Animal; Ethacrynic Acid; Feasibility Studies; Fibroblasts; Genetic Therapy; Genetic Vectors; Guinea Pigs; Kanamycin; Nerve Degeneration; Neurons; Prosthesis Design; Sepharose; Spiral Ganglion; Transduction, Genetic

We evaluated the functional efficacy of microencapsulated porcine islet xenografts transplanted into nonobese diabetic (NOD) mice. Islets were isolated from the pancreata of CSK miniature swine by manual collagenase digestion and Ficoll purification. Purified porcine islets were immediately encapsulated into microbeads of agarose polystyrene sulfonic acid (Ag-PSSa). They remained morphologically intact by dithizone staining after 7 days in culture. Insulin secretion from encapsulated islets was determined in response to glucose challenge during perifusion. When encapsulated islets were exposed to 200 mg/dL glucose, within 5 minutes, insulin release became 5-fold greater than that at 80 mg/dL. However, a second phase insulin secretion appeared in response to 250 mg/dL glucose challenge. In xenotransplantation, microencapsulated porcine islets (1000 to 1800 MC islets) were transplanted into the peritoneal cavity of diabetic NOD mice (n = 4) without immunosuppression. The survival times after the onset of diabetes were observed after both MC islets transplanted NOD mice and nontransplanted NOD mice (n = 4). MC islets transplant recipients had significantly (P < .05) longer survival (47.5 +/- 18.6; mean +/- SD) than nontransplanted NOD mice (21.0 +/- 9.31), although random blood glucose levels were not normalized.

Topics: Animals; Capsules; Cell Culture Techniques; Cell Survival; Disease Models, Animal; Graft Survival; Islets of Langerhans; Islets of Langerhans Transplantation; Mice; Mice, Inbred NOD; Polystyrenes; Resins, Synthetic; Sepharose; Swine; Swine, Miniature; Transplantation, Heterologous

To study the feasibility to repair the peripheral nerve gap with tissue engineering scaffold complex that is composed of medical biodegradable material agarose hydrogel and nerve growth factor (NGF).. Chitosan tube containing agarose hydrogel and NGF was transplanted to bridge a 10 mm gap of injured sciatic nerve in rat. Chitosan duct without agarose hydrogel and NGF was used as negative control, while autograft nerve as positive control. Sixteen weeks after operation, the regeneration of nerve fiber was observed with morphological and immunohistochemistrical methods.. The number and diameter of regenerating nerve fibers bridged by the scaffold complex of agarose hydrogel and NGF were better than negative control group (P < 0.01) and reached the level of autograft nerve group.. The new type of tissue engineering scaffold complex of agarose hydrogel and NGF may provide a microenvironment, as well as autograft nerve, to promote nerve regeneration. This technique may benefit patients with nerve injury in the future.

Topics: Absorbable Implants; Animals; Biocompatible Materials; Chitosan; Disease Models, Animal; Feasibility Studies; Hydrogel, Polyethylene Glycol Dimethacrylate; Male; Nerve Growth Factors; Nerve Regeneration; Peripheral Nerve Injuries; Peripheral Nerves; Prosthesis Implantation; Rats; Rats, Sprague-Dawley; Sepharose; Stents; Tissue Engineering

The efficacy of agarose in preventing VX2 carcinoma cell leakage was evaluated and the results were compared with two traditional methods. Forty-five rabbits were divided into 3 groups: Group 1, VX2 tumor cells were injected directly into the liver and no special procedure after removal of the needle; Group 2, the puncture site was gently compressed, using an alcoholic cotton gauze, for three minutes; Group 3, 0.2 ml of heated liquid agarose was injected to seal the aperture after injection of VX2 cells. The leakage rates were 80%, 53.3% and 6.6% for group 1, group 2 and group 3, respectively. We consider agarose is a useful material in preventing the leakage in the establishment of VX2 liver tumor models.

Topics: Animals; Carcinoma; Disease Models, Animal; Hot Temperature; Injections; Liver Neoplasms, Experimental; Neoplasm Seeding; Neoplasm Transplantation; Rabbits; Sepharose

Topics: Animals; Cystic Fibrosis; Disease Models, Animal; Inflammation; Lung; Mice; Pseudomonas aeruginosa; Sepharose; Spleen

Myasthenia gravis (MG) is a T cell-dependent antibody-mediated autoimmune neuromuscular disease. Antibodies to the nicotinic acetylcholine receptor (AChR) destroy the AChR, thus leading to defective neuromuscular transmission of electrical impulse and to muscle weakness. This unit is a practical guide to the induction and evaluation of experimental autoimmune myasthenia gravis (EAMG) in the mouse, the animal model for MG. Protocols are provided for the extraction and purification of AChR from the electric organs of Torpedo californica, or eel. The purified receptor is used as an immunogen to induce autoimmunity to AChR, thus causing EAMG. The defect in neuromuscular transmission can also be measured quantitatively by electromyography, as described here. In addition, EAMG is frequently characterized by the presence of antibodies to AChR, which are measured by radioimmunoassay and by a marked antibody-mediated reduction in the number of muscle AChRs. AChR extracted from mouse muscle is used in measuring serum antibody levels and for quantifying muscle AChR content.

Topics: Animals; Antibodies; Chromatography, Affinity; Disease Models, Animal; Electromyography; Female; Male; Mice; Mice, Inbred C57BL; Myasthenia Gravis, Autoimmune, Experimental; Neurotoxins; Radioimmunoassay; Receptors, Nicotinic; Sepharose

To purify hsp70 protein and observe its anti-tumor effect on mouse H22 hepatoma.. Cytosolic hsp70 protein of heat treated H22 cells was purified successively by chromatographic procedures, including DEAE 52-cellulose, ConA-sepharose 4B and ADP-argarose chromatography. The molecular weight and the identity of the purified hsp70 protein were confirmed by SDS-PAGE and Western blot. Tumorigenicity of H22 hepatoma was examined in purified hsp70-treated C3H mice.. A sharp stained protein band with a molecular weight of about 70 kD was obtained and shown to be hsp70 as confirmed by Western blot. In C3H mice challenged with H22 hepatoma, tumor developed in 6 of 6 mice while in hsp70 pretreated mice, none of the 6 H22 challenged mice developed tumor. Nevertheless, C57BL/6 mice pretreated with hsp70 or RPMI 1640 all developed tumor upon challenge with EL-4 lymphoma.. The results suggest that the immunoprotective effect of hsp70 protein is due not to hsp70 per se, but rather to the specific antigenic peptides it carries.

Topics: Animals; Antineoplastic Agents; Carcinoma, Hepatocellular; Disease Models, Animal; HSP70 Heat-Shock Proteins; Liver Neoplasms, Experimental; Mice; Mice, Inbred C3H; Mice, Inbred C57BL; Neoplasm Transplantation; Sepharose; Tumor Cells, Cultured; Xenograft Model Antitumor Assays

We tried to induce endolymphatic hydrops in guinea pig cochleas by unilateral, perisaccular deposition of sepharose beads carrying immune complexes. Controls consisted of the deposition of sepharose beads without immune complexes and the contralateral, untreated ear. The effects of the treatment were studied by light microscopy and electrophysiological recordings of the gross cochlear potentials 1, 2, and 6 weeks after treatment. Each condition included six animals. Analysis of variance of the morphometric data concerning the ears treated with deposition of the beads showed a statistically significant difference (P = 0.04) between the degree of hydrops found for the beads with immune complexes and for those without. The difference between the treated ears and the contralateral untreated ears was significant (P = 0.01) for the beads with immune complexes and not significant (P = 0.8) for those without immune complexes while there was no significant effect of post-treatment time interval. Analysis of variance of the electrophysiological data, collected in response to tone bursts at the apex of the cochlea, showed no significant differences between the results for the beads with and without immune complexes. Therefore these results were pooled. One week after treatment the pooled results for the compound action potential showed a small decrease in amplitude, just significant at 2 kHz, but not at 4 and 8 kHz. This decrease disappeared completely after 6 weeks. The pooled results for the negative summating potential (SP) showed a significant increase in magnitude at all frequencies decreasing with post-treatment interval. The cochlear microphonics did not demonstrate any change in amplitude after treatment. The results indicate that deposition of sepharose beads with immune complexes induces endolymphatic hydrops. Also, deposition of the sepharose beads itself may have induced hydrops together with enhancement of the SP. SP enhancement may be related to the development of endolymphatic hydrops rather than to the presence of hydrops as such. This may be based on pressure build-up while hydrops develops.

Topics: Action Potentials; Analysis of Variance; Animals; Antigen-Antibody Complex; Audiometry, Evoked Response; Cochlea; Cochlear Microphonic Potentials; Disease Models, Animal; Drug Carriers; Endolymph; Endolymphatic Hydrops; Female; Guinea Pigs; Immune Complex Diseases; Meniere Disease; Peroxidase; Saccule and Utricle; Sepharose

Bovine enamel and dentin specimens were overlaid with acidogenic Streptococcus mutans suspensions in agarose. In this model, the minimal demineralisation-inhibiting concentrations (MDIC) of hexetidine was determined in the presence of fluoride. A commercially available mouthwash containing 0.1% (2.9 mmol/l) hexetidine was diluted serially and added to the bacterial suspensions together with 0, 5.3, or 26.3 mumol/l fluoride (NaF). After 22 h of incubation at 37 degrees C the bacterial suspensions were removed and assessed for calcium and lactate. The results showed significant inhibitory effects of hexetidine on the demineralisation of the enamel specimens with a MDIC between 15 and 31 mumol/l hexetidine. In the presence of fluoride, approximately fourfold higher concentrations of hexetidine were needed for a significant additional protection of the enamel. No synergistic effect between hexetidine and fluoride was observed. For the demineralisation of the dentin specimens, the MDIC of hexetidine had a value between 31 and 61 mumol/l. At both these concentrations the dentin specimens were relatively less protected in the presence than in the absence of fluoride, and some synergistic effect between hexeditine and fluoride was observed.

Topics: Animals; Anti-Infective Agents, Local; Calcium; Cariostatic Agents; Cattle; Culture Media; Dental Enamel; Dentin; Disease Models, Animal; Drug Interactions; Drug Synergism; Fluorides; Fluorides, Topical; Hexetidine; Lactates; Mouthwashes; Sepharose; Sodium Fluoride; Streptococcus mutans; Time Factors; Tooth Demineralization

Autoantibodies from the MRL/lpr mice react with numerous proteins on neuronal cell surfaces. The purpose of this study was to isolate and characterize a population of autoantibodies reactive preferentially or exclusively with nervous system tissue. Using a purified plasma membrane preparation from brain cortex of balb/c mice coupled to diaminopropylamine agarose gel, we affinity-isolated antineuronal antibodies from pooled MRL/lpr immunoglobulins. The isolated immunoglobulins reacted with brain cortex plasma membranes and neuroblastoma cells (but not liver, kidney, or fibroblasts) by Western blot and indirect immunofluorescence with confocal microscopy. By Western blot, the epitopes in the brain cortex were proteins of apparent molecular weights 101, 63, 53, 43, 39, and 33, kd; the epitopes in the neuroblastoma cells were 63, 57, and 53 kd. Lectin column isolation revealed that the 101 and 63 kd epitopes were glycosylated. Indirect immunofluorescence revealed that the antibodies bound to the cell soma more intensely than to the cell processes of viable cultured neuroblastoma cells. The cell surface localization of this binding was confirmed by confocal microscopy. Within the central nervous system the antibodies bound more intensely to primary cultures of isolated neurons from fetal cortex than to hippocampal or neostriatal cells. With these antibodies we can begin studies of their potential pathogenic effects.

Topics: Animals; Antibody Specificity; Antigen-Antibody Reactions; Autoantibodies; Autoimmune Diseases; Blotting, Western; Brain Neoplasms; Cell Membrane; Cerebral Cortex; Chromatography, Affinity; Disease Models, Animal; Female; Fibroblasts; Fluorescent Antibody Technique; Gels; Immunosorbent Techniques; Kidney; Liver; Liver Neoplasms, Experimental; Lupus Erythematosus, Systemic; Membrane Glycoproteins; Mice; Mice, Inbred BALB C; Mice, Mutant Strains; Molecular Weight; Nerve Tissue Proteins; Neuroblastoma; Neurons; Organ Specificity; Sepharose

Adult Sprague-Dawley rats infected intrabronchially with Bordetella pertussis strain 18-323 encased in agarose beads (BP-beads), developed a paroxysmal cough and leucocytosis, both of which peaked at around day 10. When animals were exposed to ether for 2 min after delivery of the beads, there was an enhancement of the number of subsequent coughing episodes. Inclusion of carrageenan in the beads also enhanced coughing. Control rats, given sterile beads or left untreated, showed only a low level of coughing or no coughing, depending upon their source. Rats challenged by the same route with heat-killed B. pertussis in beads, or with live organisms in suspension (without beads) showed no cough induction or leucocytosis. However, intranasal delivery of B. pertussis suspension gave rise to a moderate amount of coughing and leucocytosis. Serum IgG responses to B. pertussis antigens were greatest in rats infected with BP-beads and antibodies against both pertussis toxin and filamentous haemagglutinin were detected. Since the rat is the only conveniently accessible laboratory animal species in which B. pertussis induces an intermittent paroxysmal cough, as in man, it merits further study for determining the mechanisms of pathogenesis and immunity in pertussis.

Topics: Animals; Antibodies, Bacterial; Bordetella pertussis; Carrageenan; Cough; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Immunoglobulin G; Leukocytosis; Male; Microspheres; Rats; Rats, Sprague-Dawley; Sepharose; Virulence; Whooping Cough

In an animal model of hind limb ischemia we documented the levels of endogenous basic fibroblast growth factor (bFGF) in control and ischaemic hind limbs, and evaluated the response to the administration of exogenous recombinant bFGF and heparin. Variations in this model were tested for their ability to alter the development of the collateral circulation. Recovery after acute arterial occlusion was significantly delayed by immediate bilateral mirror-image arterial ligations, when compared with either unilateral arterial ligation or delayed contralateral ligations performed after 2 months. If the major veins were also occluded all limbs developed gangrene, tissue loss and a marked delay in the recovery of blood flow, while none of the animals with unilateral arterial ligations developed gangrene. This indicates that the recovery in blood flow during the acute phase in this model is dependent on collateral vessels from the contralateral iliac artery and that major venous occlusion impedes the development of collateral vessels. Lumbar sympathectomy did not alter the recovery of blood flow after arterial occlusion, suggesting that collateral blood flow is not significantly influenced by autonomic neural supply. Following arterial occlusion there was a ten-fold increase in the levels of endogenous bFGF in all ischaemic muscle groups. Intramuscular implantation of bFGF in heparin-sepharose pellets at the time of arterial ligation markedly enhanced the blood flow for 3 weeks compared with untreated ischaemic limbs. A further increment in blood flow occurred if an additional dose of bFGF was administered 4 weeks after ligation.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Arterial Occlusive Diseases; Collateral Circulation; Disease Models, Animal; Fibroblast Growth Factor 2; Heparin; Hindlimb; Ischemia; Male; Rats; Rats, Wistar; Sepharose

Low levels of high-density lipoproteins (HDLs) may constitute an independent risk factor that may be as important as elevated low-density lipoproteins (LDLs) in coronary artery disease (CAD). Concentrations and distributions of lipids, apolipoprotein (apo) B, and apoA-I in the plasma and lipoprotein subfractions of two groups of swine, one with familial hypercholesterolemia (FHC) and the other with diet-induced hypercholesterolemia (DHC), were examined. Normolipidemic (NL) animals served as controls. All pigs carried the Lpb5 apoB mutation, which is known to influence the formation of atherosclerotic lesions. Mean concentrations of serum total cholesterol in NL, DHC, and FHC were 80.0 +/- 9.3, 774.3 +/- 54.5, and 316.5 +/- 36.1 mg/dL, respectively; HDL cholesterol (HDL-C), 33.5 +/- 1.9, 137.0 +/- 9.9, and 22.3 +/- 2.2 mg/dL; triglycerides, 33.0 +/- 16.3, 40.3 +/- 11.7, and 56.8 +/- 7.2 mg/dL; apoB, 35.7 +/- 3.1, 142.0 +/- 4.8, and 169.3 +/- 13.9 mg/dL; and apoA-I, 62.4 +/- 9.3, 170.9 +/- 6.9, and 42.6 +/- 4.8 mg/dL. The distributions of total cholesterol, apoB, and apoA-I in plasma lipoprotein subfractions were also examined. Compared with NL, FHC had fourfold and 4.7-fold increases in total cholesterol and apoB, respectively, distributed in the lower densities (d < 1.043 g/mL), and low HDL-C and apoA-I levels, resulting in a high total cholesterol/HDL-C ratio (14.4:1) and elevated triglyceride levels. DHC was characterized by 10-fold and fourfold increases in total cholesterol and apoB, respectively, resulting in an LDL particle highly enriched in cholesterol, a fourfold increase of HDL-C, an almost threefold increase in apoA-I, and a normal triglyceride level.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Apolipoprotein A-I; Apolipoproteins B; Cholesterol; Diet; Disease Models, Animal; Electrophoresis, Polyacrylamide Gel; Gels; Hypercholesterolemia; Hyperlipoproteinemia Type II; Immunoelectrophoresis; Lipids; Lipoproteins; Male; Sepharose; Sodium Dodecyl Sulfate; Swine

Cytokines incorporated into agarose blocks and implanted subcutaneously into mice establish an in vivo gradient which can be used to mimic a local inflammatory process. We have developed a model in which cellular influx into cytokine impregnated blocks parallels the normal cellular reaction to infections or wounds. Agarose blocks containing supernatants from ConA activated rat spleen cells attracted neutrophils within 4 h. These cells were followed by lymphocytes and macrophages in 24 h. Flow cytometry analysis of lymphoid cells on day 1 revealed that 38% were Ig+ (B cell marker), 60% MAC-2,3+ and 20% Thy 1.2+ of which only a small fraction were expressing CD4 on their surface. These numbers changed with time following implantation of the blocks. Initially, isolated adherent cells (macrophages) were resting, with low phagocytic activity. Cells isolated from blocks at later time points were activated, as evidenced by their increased ability to ingest fluoresceinated beads. The secretion patterns of cells trafficking to murine rIL-1 containing agarose blocks were examined. TNF, IL-6 and antibody secreting cells were found. No IL-2 was detected at any time. We believe that this model will be of value in studies of local actions of cytokines.

Topics: Animals; Cell Adhesion; Cells, Cultured; Cytokines; Disease Models, Animal; Female; Humans; Inflammation; Injections, Subcutaneous; Lymphocytes; Macrophages; Mice; Mice, Inbred BALB C; Models, Biological; Phagocytosis; Rats; Recombinant Proteins; Sepharose; Spleen; Wound Infection

A gel consisting of agarose and oxyhemoglobin (OxyHb) was developed so that, when placed in the subarachnoid space, OxyHb would be slowly released, simulating lysis of erythocytes after subarachnoid hemorrhage. The system was used to investigate the importance of reactions mediated by free radicals in the genesis of OxyHb-induced vasospasm in monkeys. Seventeen monkeys were randomly assigned to have subarachnoid placement, on Day 0, of one of the following: 1) agarose gel alone (n = 2); 2) agarose plus OxyHb (n = 3); 3) agarose plus OxyHb plus intrathecal administration of superoxide dismutase and catalase (n = 6); and 4) agarose plus OxyHb plus intrathecal administration of placebo (n = 6). Vasospasm was assessed by comparison of angiograms performed on Day 0 and 7 days after subarachnoid placement of compounds, and by electron microscopy. OxyHb alone caused significant reduction in the diameter of the middle cerebral artery (40 +/- 8%, P less than 0.005, paired t test), which was associated with ultrastructural damage to smooth muscle. Treatment with superoxide dismutase plus catalase or with placebo attenuated vasospasm of the middle cerebral artery, although significant narrowing persisted in both groups (27 +/- 12% and 26 +/- 13%, respectively, P less than 0.05, paired t test). Analysis of variance showed no difference in the degree of vasospasm between groups exposed to subarachnoid placement of OxyHb. Cerebrospinal fluid aspirated from the cisterna magna on Day 7 contained elevated activity of superoxide dismutase in animals that received treatment. Malondialdehyde was undetectable in cerebrospinal fluid after subarachnoid placement of agarose alone, although it was present in similar amounts in all groups that received subarachnoid placement of OxyHb. Since intrathecal superoxide dismutase and catalase failed to protect against OxyHb-induced vasospasm, mechanisms mediated by free radicals may not be important in its genesis. As only one combination of doses of superoxide dismutase and catalase was administered, however, it may be that other dosage schedules might be efficacious.

Topics: Animals; Antioxidants; Catalase; Cerebral Angiography; Delayed-Action Preparations; Disease Models, Animal; Free Radical Scavengers; Free Radicals; Gels; Hemolysis; Injections, Spinal; Ischemic Attack, Transient; Macaca fascicularis; Male; Malondialdehyde; Oxyhemoglobins; Random Allocation; Sepharose; Subarachnoid Hemorrhage; Subarachnoid Space; Superoxide Dismutase

Peripheral blood mononuclear cells (PBMC) from Rhesus monkeys previously immunized with bovine type II collagen to induce arthritis were cultured with the same antigen. Because the native protein is poorly soluble in culture medium a heating step is often used. The antigen in this form induced PBMC proliferation, but epitopes for the induction of antibody production and arthritis were lost. To keep the native protein intact it was coated on affigel beads. With the immobilized antigen specific antibody production could be induced.

Topics: Animals; Antibody Specificity; Arthritis; Autoantibodies; Collagen; Disease Models, Animal; Hot Temperature; Immunization; Lymphocytes; Macaca mulatta; Sepharose

Simultaneous feeding of gliadin and swainsonine, an inhibitor of alpha-D-mannosidases, in rats disturbed enterocytic maturation as shown by a marked loss of activities of alkaline phosphatase and gamma-glutamyltransferase. Morphologically, simultaneous treatment with gliadin and swainsonine caused destruction and decreased density of microvilli, as shown by electron microscopy. Neither gliadin nor swainsonine when given alone had significant effects on enterocytic enzyme activities or enterocytic morphology. Binding of enterocytic glycoproteins to both gliadin-Sepharose and concanavalin A-Sepharose was significantly increased in rats treated with swainsonine. Because swainsonine causes the formation of hybrid-type oligosaccharides with a high binding affinity to mannose-specific lectins, the observed alterations of enterocytic maturation and morphology are presumably caused by the increased binding of gliadin to enterocytic glycoproteins. A possible analogy in the etiology of celiac disease is discussed.

Topics: Alkaline Phosphatase; Alkaloids; Animals; Celiac Disease; Concanavalin A; Disease Models, Animal; Gliadin; Glycoproteins; Intestine, Small; L-Lactate Dehydrogenase; Male; Mannosidases; Microvilli; Plant Proteins; Protein Binding; Rats; Rats, Inbred Strains; Sepharose; Swainsonine

Topics: Adenosine; Adenosine Diphosphate; Adenosine Monophosphate; Adenosine Triphosphate; Animals; Blood Pressure; Coronary Disease; Disease Models, Animal; Hypoxanthines; Hypoxia; In Vitro Techniques; Inosine; Male; Microspheres; Myocardial Contraction; Myocardium; Perfusion; Phosphocreatine; Polysaccharides; Rats; Sepharose