RESUMEN
AIM: We aimed to study the feasibility and tolerability of a combination therapy consisting of glutamic acid decarboxylase (GAD-alum), Etanercept and vitamin D in children and adolescents with newly diagnosed with type 1 diabetes (T1D), and evaluate preservation of beta cell function. MATERIAL AND METHODS: Etanercept Diamyd Combination Regimen is an open-labelled multi-centre study pilot trial which enrolled 20 GAD antibodies positive T1D patients (7 girls and 13 boys), aged (mean ±SD): 12.4 ± 2.3 (8.3-16.1) years, with a diabetes duration of 81.4 ± 22.1 days. Baseline fasting C-peptide was 0.24 ± 0.1 (0.10-0.35) nmol/l. The patients received Day 1-450 Vitamin D (Calciferol) 2000 U/d per os, Etanercept sc Day 1-90 0.8 mg/kg once a week and GAD-alum sc injections (20 µg, Diamyd™) Day 30 and 60. They were followed for 30 months. RESULTS: No treatment related serious adverse events were observed. After 6 months 90-min stimulated C-peptide had improved in 8/20 patients and C-peptide area under the curve (AUC) after Mixed Meal Tolerance Test in 5 patients, but declined thereafter, while HbA1c and insulin requirement remained close to baseline. Administration of Etanercept did not reduce tumour necrosis factor (TNF) spontaneous secretion from peripheral blood mononuclear cells, but rather GAD65-induced TNF-α increased. Spontaneous interleukin-17a secretion increased after the administration of Etanercept, and GAD65-induced cytokines and chemokines were also enhanced following 1 month of Etanercept administration. CONCLUSIONS: Combination therapy with parallel treatment with GAD-alum, Etanercept and vitamin D in children and adolescents with type 1 diabetes was feasible and tolerable but had no beneficial effects on the autoimmune process or beta cell function.
Asunto(s)
Diabetes Mellitus Tipo 1 , Adolescente , Anciano , Compuestos de Alumbre , Niño , Etanercept/uso terapéutico , Femenino , Glutamato Descarboxilasa/uso terapéutico , Humanos , Insulina/metabolismo , Leucocitos Mononucleares/metabolismo , Masculino , Proyectos Piloto , Vitamina DRESUMEN
AIM: Double-blind placebo-controlled intervention using glutamic acid decarboxylase (GAD)-alum, vitamin D and Ibuprofen in recent onset Type I diabetes (T1D). METHODS: 64 patients (T1D since <4 months, age 10-17.99, fasting sC-peptide ≥0.12 nmol/l, GADA-positive) were randomized into Day(D) 1-90 400 mg/day Ibuprofen, D1-450 vitamin D 2000 IU/day, D15, 45 sc. 20 µg GAD-alum; as A but placebo instead of Ibuprofen; as B but 40 µg GAD-alum D15, 45; placebo. RESULTS: Treatment was safe and tolerable. No C-peptide preservation was observed. We observed a linear correlation of baseline C-peptide, HbA1c and insulin/per kilogram/24 h with change in C-peptide AUC at 15 months (r = -0.776, p < 0.0001). CONCLUSION: Ibuprofen, vitamin D + GAD-alum did not preserve C-peptide. Treatment efficacy was influenced by baseline clinical and immunological factors and vitamin D concentration. Clinical Trial Registration: NCT01785108 (ClinicalTrials.gov).
RESUMEN
An abnormally high apoptosis has been associated with a number of clinical conditions including embryonal malformations and various pathologies such as neuronal degeneration and diabetes. In this study, boron is reported to inhibit apoptosis in hyperapoptosis conditions as demonstrated in a model of hyperapoptosis. Boron is a metalloid which is present in food in small amounts and is suggested here to inhibit apoptosis by stabilizing the mitochondrial membrane structure, thus preventing matrix remodeling and the release of cytochrome c, an apoptosis-inducer protein from the mitochondrion. The protective effect was assessed by measuring the changes in mitochondrial membrane potential, the levels of cytochrome c and downstream activation of caspase 3, besides phosphatidylserine exposure on the cell surface and DNA damage. The study has implication in clinical conditions characterized by hyperapoptosis as seen in certain embryonal malformations and various pathologies.
Asunto(s)
Apoptosis , Boro/farmacología , Membranas Mitocondriales/metabolismo , Neuronas/citología , Caspasa 3/metabolismo , Línea Celular Tumoral , Supervivencia Celular , Citocromos c/metabolismo , Daño del ADN , Humanos , Etiquetado Corte-Fin in Situ , Potencial de la Membrana Mitocondrial , Enfermedades Neurodegenerativas/metabolismo , Nitritos/química , Fosfatidilserinas/químicaRESUMEN
Chemical mediators of inflammation (CMI) are important in host defense against infection. The reduced capacity of host to induce the secretion of these mediators following infection is one of the factors in host susceptibility to infection. Boron, which has been suggested for its role in infection, is reported in this study to increase lymphocyte proliferation and the secretion of CMI by the lipopolysaccharide (LPS)-stimulated peritoneal macrophages in BALB/c mice. Boron was administered to mice orally as borax at different doses for 10 consecutive days, followed by the stimulation of animals with ovalbumin and isolation of splenocytes for proliferation assay. The lymphocyte subsets were determined by flow cytometry in spleen cell suspension. The mediators of inflammation, TNF-α, IL-6, IL-1ß and nitric oxide (NO), were measured in culture supernatant of LPS-primed macrophages isolated from borax treated mice. TNF and ILs were measured by ELISA. NO was determined by Griess test. The expression of inducible nitric oxide synthase (iNOS) in macrophages was studied by confocal microscopy. Results showed a significant increase in T and B cell populations, as indicated by an increase in CD4 and CD19, but not CD8, cells. Boron further stimulated the secretion of TNF-α, IL-6, IL-1ß, NO and the expression of iNOS by the LPS-primed macrophages. The effect was dose dependent and most significant at a dose level of 4.6 mg/kg b. wt. Taken together, the study concludes that boron at physiological concentration induces lymphocyte proliferation and increases the synthesis and secretion of pro-inflammatory mediators by the LPS-primed macrophages, more specifically the M1 macrophages, possibly acting through Toll-like receptor. The study implicates boron as a regulator of the immune and inflammatory reactions and macrophage polarization, thus playing an important role in augmenting host defense against infection, with possible role in cancer and other diseases.
Asunto(s)
Boratos/farmacología , Boro/farmacología , Proliferación Celular/efectos de los fármacos , Factores Inmunológicos/farmacología , Lipopolisacáridos/inmunología , Linfocitos/efectos de los fármacos , Macrófagos/efectos de los fármacos , Animales , Boratos/administración & dosificación , Boro/administración & dosificación , Células Cultivadas , Factores Inmunológicos/administración & dosificación , Mediadores de Inflamación/inmunología , Interleucina-1beta/inmunología , Interleucina-6/inmunología , Linfocitos/citología , Linfocitos/inmunología , Macrófagos/citología , Macrófagos/inmunología , Masculino , Ratones , Ratones Endogámicos BALB C , Óxido Nítrico/inmunología , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
BACKGROUND: Dried flower bud of Syzygium aromaticum (clove) is rich in eugenol, an antioxidant and antiinflammatory compound that can protect liver against injury. Clove, besides eugenol, also contains other pharmacologically active phytochemicals such as ß-sitosterol and ascorbic acid. This study reports the effect of eugenol-rich fraction (ERF) of clove on liver cirrhosis induced by thioacetamide. METHODS: Cirrhosis of the liver, which predisposes to hepatocellular carcinoma, was induced by administering thioacetamide (0.03%) in drinking water for 16 weeks. Cirrhotic animals were divided into two groups; the treated group was administered ERF for 9 weeks, one week after discontinuation of thioacetamide, while the other group received normal saline for a similar duration of time. RESULTS: The treatment with ERF, as determined by histopathology and through a battery of biochemical markers of hepatic injury, oxidative stress and drug metabolizing enzymes, significantly ameliorated the signs of liver cirrhosis. It lowered the elevated levels of alkaline phosphatase, γ-glutamyl transferase and other biochemical changes in liver cirrhosis. Histopathology of the liver corroborated the effect of ERF with biochemical findings. ERF treatment further inhibited cell proliferation, as demonstrated by reduced [(3)H]-thymidine uptake. CONCLUSIONS: Data provide evidence supporting the protective action of ERF on liver cirrhosis. The study assumes significance because cirrhosis predisposes the liver to cancer, which is characterized by abnormal cell proliferation. ERF in this study is reported to inhibit hepatic cell proliferation and at the same time decrease oxidative stress, which might be the mechanism of protection against liver cirrhosis.
RESUMEN
Boron is reported in this study to stabilize the structure of heme proteins exposed to peroxides. The oxidized heme protein (15 microM) was treated with H(2)O(2) (10mM) in 1M glycine-NaOH buffer (pH 9.2) at 25 degrees C in absence/presence of boron, and characterized by visible absorption spectroscopy, gel exclusion chromatography, native PAGE, HPLC and DLS. Spectral analysis of exposed heme proteins revealed a decrease in absorbance in the Soret region, which was stabilized by boron. The native PAGE analysis of exposed heme proteins showed high molecular weight products; the band intensity was lesser in presence of boron. Further, elution profile of the exposed heme proteins on Sephadex G-200 column and HPLC revealed more than one peak (aggregate formation) when compared to the respective untreated proteins. DLS, which measures the hydrodynamic radius (R(H)), was used to ascertain whether the peaks correspond to monomer, dimer or aggregate forms. The R(H) of boron pretreated heme proteins was close to R(H) of the respective heme protein. Non-heme protein RNase did not show any change when exposed to peroxide. Taken together, results conclude that boron stabilizes the structure of heme proteins, which might be due to specific sites on heme proteins that can bind to borate ions.