Evaluación preclínica y estudio de estabilidad de extractos a partir del follaje de Momordica charantia Lin / Obtaining and characterizing the Momordica charantia Linn leaf extracts

El objetivo del presente trabajo fue la evaluación preclínica y el estudio de estabilidad de extractos a partir del follaje de Momordica charantia Lin. Se obtuvieron extractos acuoso e hidroalcohólico para los cuales se establecieron las especificaciones de calidad mediante la evaluación de tres lotes y se estudió su estabilidad por el método de vida de estante durante 12 meses. A los extractos se le evaluó el potencial genotóxico mediante ensayos de micronúcleos en médula ósea de ratón y aberraciones cromosómicas en linfocitos de sangre periférica. La actividad hipoglicemiante oral fue evaluada en animales con hiperglicemia temporal inducida por carga de glucosa. Como resultados se establecieron las especificaciones de calidad de los extractos acuoso e hidroalcohólico, los mismos mostraron estabilidad por 6 meses para el extracto acuoso y 12 meses para el extracto hidroalcohólico. No mostraron efecto genotóxico en los ensayos evaluados y mostraron efecto hipoglicemiante oral a la dosis de 450 mg/kg.

The objective of this investigation was the preclinical evaluation and the stability study of the Momordica charantia Linn hydroalcoholic and aqueous leaf extracts. The hydroalcoholic and aqueous extracts were obtained and the quality specifications were determined by evaluating three lots. The stability of the extracts was evaluated for 12 months. The genotoxic potential of the extracts was evaluated by mouse bone marrow micronucleus test and chromosome aberration test. The hypoglycemic effect was determined by oral glucose tolerance test. As results, the quality specifications were established and the aqueous extract was stable for 6 months and the hydroalcoholic extract for 12 months. A genotoxic effect was not observed in both extracts and the hypoglycemic effect was observed at the oral dose of 450 mg/kg of body weight.

Production of native and modified recombinant Der p 1 molecules in tobacco plants.

BACKGROUND: As a complex molecule requiring post-translational processing, it has been difficult to produce the Der p 1 major allergen from the Dermatophagoides pteronyssinus house dust mite in a recombinant form. OBJECTIVE: Here, we tested whether transgenic tobacco plants are suitable to express Der p 1, either as a wild-type molecule or as variants lacking N-glycosylation sites (Gly(-)) and/or cysteine protease activity (Enz(-)). Methods Using Agrobacterium tumefaciens-based transformation, pro Der p 1 molecules bearing mutations within either the N-glycosylation sites (N34Q, N150Q) and/or the cysteine protease-active site (C132V) were expressed in tobacco plants. After purification by ion exchange chromatography, allergens were characterized using immunoblotting, circular dichroism (CD), as well as basophil and T lymphocyte stimulation assays. RESULTS: Four forms of recombinant Der p 1 (i.e. wild-type Gly(+)/Enz(+), as well as Gly(-)/Enz(+), Gly(+)/Enz(-) or Gly(-)/Enz(-) variants) were successfully expressed in tobacco leaves as pro Der p 1 molecules. Spontaneous cleavage of the pro-peptide was observed in tobacco leaf extracts for all forms of recombinant Der p 1 (r Der p 1). CD confirmed that all r Der p 1 molecules, with the exception of the Gly(-)/Enz(-) variant, exhibited secondary structures comparable to the natural protein. A cysteine protease activity was associated only with the Gly(+)/Enz(+) form. All these molecules exhibit a profile similar to natural Der p 1 with respect to IgE immunoreactivity, basophil activation and T cell recognition. CONCLUSION: A tobacco plant expression system allows the production of various forms of mature Der p 1, which could be used for diagnostic or immunotherapeutic purposes.

Tear IgE detected by a new method: Stallerdiag-IgE.

A new method for the tear IgE measurement Stallerdiag-IgE is presented. For this assay, the tears are collected on a strip of filter paper introduced into the conjunctival cul-de-sac. Then the IgE are measured by a simple enzymo-immunological assay. Tear IgE were determined in a normal population and in three groups of patients with keratoconjunctivitis. In the normal population, the tear IgE level is 0.37±0.66 kIU/1. The IgE level observed in the patients with vernal keratoconjunctivitis (19.6±21.5 kIU/1) or chronic allergic keratoconjunctivitis (11.1±16.2 kIU/1) is significantly different (p<0.0005) from the IgE level measured in the patients suffering from non-allergic keratoconjunctivitis (3.4±10.4 kIU/1). By comparison with a non-allergic group, the sensitivity is 80%, the specificity is 89% giving an overall efficiency of 83%.

Precision of the Larsen and the Sharp methods of assessing radiologic change in patients with rheumatoid arthritis.

OBJECTIVE: To compare the sensitivity of Sharp's and Larsen's radiographic scoring methods for detecting change in rheumatoid arthritis (RA) over time. METHODS: Radiographs of the hands and wrists were taken at the beginning and at the end of a 2-year followup period, in 42 patients with active RA. Films were scored blindly using both scoring methods. Patients were under treatment with methotrexate (intramuscular injections). RESULTS: Radiographic evidence of progression or amelioration was detected in 25 patients by Larsen's method and in 35 patients by Sharp's method. The relative sensitivity to change over time was greater for Sharp's method (0.01 less than P less than 0.025). CONCLUSION: Sharp's radiographic scoring method seems to be more sensitive to change over time than is Larsen's method. The clinical importance of the change needs to be definitively established.

Treatment of rheumatoid arthritis with methotrexate: a prospective open longterm study of 191 cases.

One hundred and ninety-one patients with severe rheumatoid arthritis (RA) were included in a prospective open longterm study of the safety, efficacy and maintenance of methotrexate (MTX) treatment. The mean duration of MTX treatment was 19 +/- 13.2 (3-58) months; the mean weekly dose of MTX was 10.2 +/- 0.2 mg. Analysis of the 191 patients in an intent-to-treat manner showed a significant improvement of all the clinical variables and a decrease of erythrocyte sedimentation rate with a steroid sparing effect. The probability of continuing MTX therapy for up to 2 years was 65% and for up to 5 years was 46%. Adverse effects of MTX occurred in 37.1% of the patients, but only 15.7% discontinued MTX permanently.

Radiologic progression during intramuscular methotrexate treatment of rheumatoid arthritis.

The radiographic evolution of joint lesions in 41 patients with severe and longstanding rheumatoid arthritis (RA) (mean duration: 12.9 years) treated with methotrexate (MTX) for more than 24 months (mean 31.2 months) was studied in a double blind manner by 2 rheumatologists using Larsen's score. All the patients were clinically improved. The radiological study of hands and wrists was possible in only 37 of 41 patients for surgical reasons. We found a deterioration of joint lesions in 31 cases (83.8%), a stabilization in 6 cases (16.2%). MTX does not appear to be able to slow down the radiographic evolution of RA. Whether it could prevent the radiologic progression in early RA remains to be elucidated.

Rheumatoid nodulosis: report of two new cases and discussion of diagnostic criteria.

Two patients with typical rheumatoid nodulosis are described and compared with 24 reported cases. Rheumatoid nodulosis is a particular variant of rheumatoid arthritis associated with palindromic rheumatism, subcutaneous rheumatoid nodules, mild or no systemic manifestation and a benign clinical course. Positive rheumatoid factor and radiologic subchondral bone cysts are usual, but their absence should not eliminate the diagnosis of rheumatoid nodulosis, particularly at the onset of the disease.