cortisol-succinate--sodium-salt and Disease-Models--Animal

When Zika virus emerged as a public health emergency there were no drugs or vaccines approved for its prevention or treatment. We used a high-throughput screen for Zika virus protease inhibitors to identify several inhibitors of Zika virus infection. We expressed the NS2B-NS3 Zika virus protease and conducted a biochemical screen for small-molecule inhibitors. A quantitative structure-activity relationship model was employed to virtually screen ∼138,000 compounds, which increased the identification of active compounds, while decreasing screening time and resources. Candidate inhibitors were validated in several viral infection assays. Small molecules with favorable clinical profiles, especially the five-lipoxygenase-activating protein inhibitor, MK-591, inhibited the Zika virus protease and infection in neural stem cells. Members of the tetracycline family of antibiotics were more potent inhibitors of Zika virus infection than the protease, suggesting they may have multiple mechanisms of action. The most potent tetracycline, methacycline, reduced the amount of Zika virus present in the brain and the severity of Zika virus-induced motor deficits in an immunocompetent mouse model. As Food and Drug Administration-approved drugs, the tetracyclines could be quickly translated to the clinic. The compounds identified through our screening paradigm have the potential to be used as prophylactics for patients traveling to endemic regions or for the treatment of the neurological complications of Zika virus infection.

Topics: Animals; Antiviral Agents; Artificial Intelligence; Chlorocebus aethiops; Disease Models, Animal; Drug Evaluation, Preclinical; High-Throughput Screening Assays; Immunocompetence; Inhibitory Concentration 50; Methacycline; Mice, Inbred C57BL; Protease Inhibitors; Quantitative Structure-Activity Relationship; Small Molecule Libraries; Vero Cells; Zika Virus; Zika Virus Infection

Improvements in asthma diagnosis and management require deeper understanding of the heterogeneity of the complex airway inflammation. We hypothesise that differences in the two major inflammatory phenotypes of asthma; eosinophilic and neutrophilic asthma, will be reflected in the lung protein expression profile of murine asthma models and can be delineated using proteomics of bronchoalveolar lavage (BAL).. BAL from mice challenged with ovalbumin (OVA/OVA) alone (standard model of asthma, here considered eosinophilic) or OVA in combination with endotoxin (OVA/LPS, model of neutrophilic asthma) was analysed using liquid chromatography coupled to high resolution mass spectrometry, and compared with steroid-treated animals and healthy controls. In addition, conventional inflammatory markers were analysed using multiplexed ELISA (Bio-Plex™ assay). Multivariate statistics was performed on integrative proteomic fingerprints using principal component analysis. Proteomic data were complemented with lung mechanics and BAL cell counts.. Several of the analysed proteins displayed significant differences between the controls and either or both of the two models reflecting eosinophilic and neutrophilic asthma. Most of the proteins found with mass spectrometry analysis displayed a considerable increase in neutrophilic asthma compared with the other groups. Conversely, the larger number of the inflammatory markers analysed with Bio-Plex™ analysis were found to be increased in the eosinophilic model. In addition, major inflammation markers were correlated to peripheral airway closure, while commonly used asthma biomarkers only reflect central inflammation.. Our data suggest that the commercial markers we are currently relying on to diagnose asthma subtypes are not giving us comprehensive or specific enough information. The analysed protein profiles allowed to discriminate the two models and may add useful information for characterization of different asthma phenotypes.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Biomarkers; Bronchial Provocation Tests; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Eosinophils; Female; Hydrocortisone; Inflammation; Inflammation Mediators; Leukocyte Count; Lipopolysaccharides; Mass Spectrometry; Methacholine Chloride; Mice; Mice, Inbred BALB C; Neutrophils; Ovalbumin; Phenotype; Protein Array Analysis; Proteome; Respiratory Mechanics

The present study evaluated the protective action of calcium dobesilate (CDO) in various experimental models of cataract. CDO was studied in hydrocortisone-induced cataract in developing chick embryos and selenite-induced cataract in pups. CDO anti-cataract activity was also evaluated after oral and topical application as eye drops in galactose (30%) fed rats. This study was further extended to evaluate the intraocular penetration of a single dose of 1% CDO (50 microL) in rabbits. CDO exhibited significant protection against cataract in experimental models and considerable penetration after single topical application. Anti-cataract activity may be due to its antioxidant as well as aldose reductase inhibitor properties. Further studies are in progress to evaluate its clinical efficacy in diabetic cataract.

Topics: Administration, Oral; Administration, Topical; Aldehyde Reductase; Animals; Calcium Dobesilate; Cataract; Chick Embryo; Disease Models, Animal; Female; Galactose; Hemostatics; Hydrocortisone; Male; Ophthalmic Solutions; Oxidative Stress; Rabbits; Rats; Rats, Wistar; Sodium Selenite

Glucocorticoids (GC) remain the first choice of treatment in asthma, but GC therapy is not always effective and is associated with side effects. In a porcine study in our laboratory, simultaneous administration of GC and nitric oxide (NO) attenuated the endotoxin-induced inflammatory response and made GC treatment more effective than inhaled NO or steroids alone. In the present study, we aimed to further investigate the interactions between NO and GC treatment in two murine models of asthma. Inflammation was induced by endotoxin, ovalbumin, or a combination of both. With an animal ventilator and a forced oscillation method (FlexiVent), lung mechanics and airway reactivity to methacholine in response to various treatments were assessed. We also describe histology and glucocorticoid receptor (GR) protein expression in response to inhaled NO treatment [40 ppm NO gas or NO donors sodium nitroprusside (SNP) or diethylamine NONOate (DEA/NO)]. SNP and GC provided protection against bronchoconstriction to a similar degree in the model of severe asthma. When GC-treated mice were given SNP, maximum airway reactivity was further reduced. Similar effects were seen after DEA/NO delivery to GC-treated animals. Using 1-H-[1,2,4]-oxadiazolo-[4,3-a]-quinoxalin-1-one (ODQ), a soluble guanylate cyclase inhibitor, we found this effect of NO donors to be mediated through a cGMP-independent mechanism. In the severe model, prolonged NO treatment restored or even increased the nuclear levels of GR. In conclusion, in our murine model of severe asthma GC treatment provided protection to only a limited degree against bronchoconstriction, while concomitant treatment with a NO donor was markedly more potent than the use of either NO or GC alone.

Topics: Administration, Inhalation; Animals; Anti-Asthmatic Agents; Asthma; Bronchial Provocation Tests; Bronchoconstriction; Bronchoconstrictor Agents; Cyclic GMP; Disease Models, Animal; Drug Therapy, Combination; Enzyme Inhibitors; Female; Glucocorticoids; Guanylate Cyclase; Hydrazines; Hydrocortisone; Injections, Intraperitoneal; Lipopolysaccharides; Methacholine Chloride; Mice; Mice, Inbred BALB C; Nitric Oxide; Nitric Oxide Donors; Nitroprusside; Ovalbumin; Oxadiazoles; Pneumonia; Quinoxalines; Receptors, Cytoplasmic and Nuclear; Receptors, Glucocorticoid; Respiration, Artificial; Respiratory Mechanics; Soluble Guanylyl Cyclase

We examined the effects of cyclosporin A (CsA) and glucocorticoid (GC; hydrocortisone sodium succinate) in a mouse model of experimental parkisonism. GC or CsA was administered 30 or 60 min, respectively, prior to intracerebroventricular injection of 6-hydroxydopamine, followed by injection of a similar dose of each drug 3 h later. CsA reduced the extent of depletion of striatal concentrations of dopamine (DA) and dihydroxyphenylacetic acid (DOPAC) associated with dopaminergic neuronal degeneration. GC reduced the extent of homovanillic acid (HVA) depletion in the same region. A combination treatment with CsA and GC did not produce a further enhancement of the recovery of striatal concentrations of monoamines observed with CsA only. Our findings demonstrated the beneficial effects of initial CsA treatment in experimental models of parkinsonism and further support the usefulness of CsA in the treatment of Parkinson's disease.

Topics: Animals; Corpus Striatum; Cyclosporine; Disease Models, Animal; Dopamine; Glucocorticoids; Hydrocortisone; Immunosuppressive Agents; Male; Mice; Mice, Inbred ICR; Oxidopamine; Parkinson Disease, Secondary; Sympatholytics

In order to develop an effective screening model for anticataract agents, we examined the age dependence of cataract induction by glucocorticoid in developing chick embryos. Hydrocortisone sodium succinate (0.25 mumol) was administered to chick embryos on day 15 (15-day-old) and cataract formation was examined 48 hr later. Administration earlier than on day 13 or later than on day 15 was a little or ineffective. These results indicate that the formation of glucocorticoid-induced cataract in developing chick embryos depends on developing stages. The embryos treated with hydrocortisone sodium succinate on day 15 decreased GSH amount in the lens, approximately 50% of the control in 48hr. However, the embryos treated at other ages, in which cataract was not induced, showed little or no decrease of GSH. The cataract formation in chick embryos appeared to depend on structure of steroid and was due to biological activities of glucocorticoids. Since cataract is easily produced in a reproducible manner with high incidence by glucocorticoid, our chick embryo model will be a valuable model system for screening anticataract agents.

Topics: Animals; Anti-Inflammatory Agents; Cataract; Chick Embryo; Disease Models, Animal; Drug Evaluation, Preclinical; Glutathione; Hydrocortisone; Incidence; Lens, Crystalline; Reproducibility of Results; Structure-Activity Relationship

Nishigori et al. reported a transient cataract model after administering glucocorticoid to a 15-day-old chick. Biochemically, the mechanism of onset of this cataract was though to be related to damage caused by the formation of oxidative stresses and by a protein-water phase separation. There appear to be no reports on changes in the fine structure. After hydrocortisone succinate sodium was administered to 15-day-old chick embryos, the lenses were removed at 12, 24, 30, 48, 72 and 96 h and put in 4% glutaraldehyde. The specimens were examined by light, transmission and scanning electron microscopy. Twelve to 24 h after administration to chick embryos, lens fibers containing electron-dense cytoplasm began to appear in the bow area of the equator and were still present thereafter. Thirty to 48 h after administration, numerous vacuoles of varying sizes began to appear in the lens in sites corresponding to the opaque region. These vacuoles, ranging from 2 to 8 microns in diameter, were distributed in the intercellular spaces between the lens fibers. The vacuoles had disappeared by 96 h after administration, but during that period, the height of the epithelial cells in the equatorial region and the elongation of the equatorial lens fibers had become irregular. Transient opacity was due to the presence of vacuoles of various sizes, occurring in the intercellular space between the lens fibers around the lens nucleus. Moreover, the effect of glucocorticoid administration was noted in the lens epithelium and the lens fibers in the equatorial region.

Topics: Animals; Cataract; Chick Embryo; Chickens; Disease Models, Animal; Epithelium; Hydrocortisone; Lens Nucleus, Crystalline; Lens, Crystalline; Microscopy, Electron, Scanning; Vacuoles

This study was undertaken to determine the effect of cyclophosphamide, methotrexate, hydrocortisone, and cyclosporin A on a model of granulomatous infiltration in the terminal ileum and draining lymph nodes of the guinea pig. Treatment groups of six animals were used and compared to untreated groups of 12. Epithelioid cell granulomas and primary macrophage granulomas were induced by the inoculation of BCG (Pasteur) and irradiated Mycobacterium leprae respectively into the terminal ileum of the guinea pig. The response to purified protein derivative of tuberculin was reduced in both groups of animals receiving any of these agents. Cyclophosphamide and methotrexate treated animals inoculated with BCG or M leprae showed a significant reduction of granulomatous infiltration at the inoculation site (p less than 0.05 and p less than 0.001 respectively). BCG inoculated animals treated with either hydrocortisone or cyclosporin A showed no reduction in granulomatous infiltration at either the inoculation site or the draining lymph nodes. By contrast M leprae inoculated animals receiving either of these agents showed a significant reduction of granulomatous infiltration at both the inoculation site (p less than 0.001) and in the primary draining lymph node (p less than 0.001). Ziehl Neelsen staining showed an increased proportion of animals with detectable acid fast bacilli (AFB) at the inoculation site in the groups receiving hydrocortisone (50%) and methotrexate (67%) compared to untreated controls (8%). No AFB were observed in any of the animals inoculated with M leprae. In conclusion, this model may be helpful in elucidating the mechanism of T lymphocyte response in Crohn's disease and the variable clinical response seen with the use of immunosuppressive agents in this condition.

Topics: Adjuvants, Immunologic; Animals; Cyclophosphamide; Cyclosporins; Disease Models, Animal; Female; Granuloma; Guinea Pigs; Hydrocortisone; Ileal Diseases; Lymph Nodes; Methotrexate; Organ Size

In this study the role of free radicals, lipid peroxidation, and neutrophil infiltration as mediators of ischemia and reperfusion-induced intestinal mucosal damage were investigated. We used a rat experimental model in which a ligated loop of the distal ileum was subjected to ischemia and reperfusion and the ensuing mucosal damage was assessed by means of lysosomal enzyme release and intestinal permeability measurements. We also determined the mucosal content of malondialdehyde, a lipid peroxidation product, and the mucosal activity of myeloperoxidase, a neutrophil granulocyte marker. Ischemia and revascularization alone caused increased mucosal permeability to sodium fluorescein, increased N-acetyl-beta-glucosaminidase release from the mucosa into the lumen, increased malondialdehyde content in the mucosa, and increased myeloperoxidase activity in the mucosa. Intravenous injection of enzymatic antioxidant, superoxide dismutase, together with xanthine oxidase inhibitor, allopurinol, prevented the malondialdehyde accumulation and caused attenuation of all the other effects of ischemia. Intravenous pretreatment of hydrocortisone sodium succinate (Solu-Cortef), a steroid and also a nonenzymatic antioxidant, prevented not only malondialdehyde accumulation but also neutrophil infiltration and mucosal damage. These data support a concept that neutrophil infiltration is an important element in ischemic mucosal damage. In addition, the blocking of this phenomenon may have clinical significance in attempts to modulate the potential damaging effects of the increased neutrophil infiltration associated with small-intestinal ischemia.

Topics: Acetylglucosaminidase; Allopurinol; Animals; Disease Models, Animal; Fluoresceins; Free Radicals; Hydrocortisone; Intestinal Absorption; Intestinal Mucosa; Ischemia; Lipid Peroxidation; Malondialdehyde; Neutrophils; Oxygen; Peroxidase; Rats; Reperfusion Injury; Superoxide Dismutase

The relationship between host immune state and hepatic inflammation and infection pattern of the Duck hepatitis B virus (DHBV) was investigated by experimental transmission of DHBV into 98 Japanese 7-day-old ducklings that had been pretreated with immunoregulatory drugs including cyclophosphamide, OK 432, and a steroid hormone. Immunosuppressive treatment with cyclophosphamide revealed an extension of the viremic period associated with an absence of inflammatory changes in the liver. Although immunostimulating treatment with OK 432 showed a remarkable accumulation of inflammatory cells in the liver, the viremic period was not shortened. Treatment with a steroid used as a immunosuppressant did not suppress the hepatitis; moreover, it increased viral DNA replication and extended the viremic period. This phenomenon of viral replication seemed to be caused by the direct effects of the steroid. Alteration of DHBV infection by modifying the host immune state is quite similar to that of hepatitis B virus (HBV) in humans. In DHBV infection, the host immune state seemed to have a considerable role in determining the infection pattern and degree of hepatitis activity. DHBV may be a helpful model of HBV for studying host-viral interaction and the immunological mechanism of viral hepatitis.

Topics: Adjuvants, Immunologic; Animals; Carrier State; Cyclophosphamide; Disease Models, Animal; DNA-Directed DNA Polymerase; DNA, Viral; Ducks; Hepatitis B; Hepatitis B virus; Hepatitis, Viral, Animal; Hydrocortisone; Immunosuppressive Agents; Picibanil; Viremia; Virus Replication

We sought to develop an experimental animal model in order to study the effects of hypothermia on host defences under conditions which were similar to those used for humans. We required a large animal which could tolerate arterial and venous catheters and serial blood sampling without significantly altering its blood volume and blood pressure. The animal should be intubated and ventilated to control blood gases and fluid and electrolyte balance. Finally the model should have anatomic, metabolic and physiologic similarities to humans. We describe an experimental pig model which appears to fulfill these criteria and provide important information relevant to man.

Topics: Animals; Blood Gas Analysis; Cell Movement; Disease Models, Animal; Epinephrine; Hydrocortisone; Inflammation; Leukocyte Count; Neutrophils; Pleurisy; Research Design; Swine

Topics: Aldosterone; Animals; Cataract; Chick Embryo; Disease Models, Animal; Hydrocortisone