Biological standardization of pollen allergens from India.

Standardization of allergens are achieved by in vitro and in vivo methods. Some of the allergens from Western countries are standardized using biological potency of the extracts but no attempt has been made till now to standardize any of the pollen extracts from India based on biological units. Therefore, we have attempted to standardize two important pollen allergens Ricinus communis and Holoptelea integrifolia by biological methods. Broadly the methods adopted by Dreborg and Grimmer (1983) was followed. Skin prick tests were carried out with the extracts of R. communis and H. integrifolia on 15 allergic patients in five three fold log dilutions starting with 1:10, in 50% glycerinated buffer. Glycerinated buffer (50%) and histamine dihydrochloride (1 mg/ml) were used as negative and positive controls, respectively. The mean wheal diameter obtained with different concentrations showed a gradual systematic fall with increase in dilution. The mean relative diameter (% of histamine reaction) varied from 124.1 +/- 8.9 to 33.7 +/- 6.1 and 78.9 +/- 5.5 to 21.4 +/- 3.8 with the highest and lowest concentrations of R. communis and H. integrifolia pollen antigens, respectively. The histamine equivalent concentration of antigen 1,000 Biological Units (BU) obtained for crude pollen extracts of R. communis and H. integrifolia was 1:17 and 1:22 respectively.     

Effect of various stabilizing agents on Imperata cylindrica grass pollen allergen extract

BACKGROUND: Allergen extracts are unstable, heat labile or susceptible to proteases. Stability of allergen extracts is important for proper diagnosis and therapy of allergic disorders. OBJECTIVE: The present study was undertaken to determine the preservation and stabilization conditions of Imperata cylindrica (Ic) grass pollen extract. METHODS: The Ic extract was kept with 0.1 mepsilon-aminocaproic acid (EACA), 0.75 m sucrose, 5% glycerol, 0.03% human serum albumin (HSA) or 0.4% phenol for different time periods. The extracts were stored for 3, 6 and 12 months each at 4 degrees C, 4 degrees C with daily exposure to room temperature (RT) for 1 h, and RT. The quality of extracts was analysed by SDS-PAGE, Western blot, ELISA, ELISA inhibition and skin test. RESULTS: Extracts kept with EACA and sucrose retained most of the protein bands followed by glycerol as determined by SDS-PAGE and Western blot during all storage periods and conditions in comparison with standard extracts. The extracts kept with HSA, phenol and without preservative (WP) showed protein degradation below 33 kDa after 3 months storage at all conditions. However, a 67-kDa allergen was stable in these extracts. EACA extract required 75 to 120 ng of protein for 50% inhibition in IgE binding under different conditions, whereas standard extract required 70 ng for the same. ELISA also demonstrated high allergenic reactivity of EACA extract. ID test on allergy patients with EACA extract demonstrated same allergenic potency as that of standard extract. CONCLUSION: EACA is the best preservative/stabilizing agent of Ic pollen extract, followed by sucrose and glycerol. Ic extract kept with phenol, HSA and without preservative showed degradation within 3 months. EACA preserved extract is equally potent as that of standard extract up to 1 year's storage.     

Myopia and plasma cortisol

Estimation of plasma cortisol by fluorometric technique desorbed by Mallingly's was carried out in 56 cases, including 38 cases of myopia (19 cases of simple myopia and 19 cases of degenerative myopia) and 18 normal individuals. Urinary 17-keto steroids/24 hours was also estimated by Zimmermann technique in 12 out of the 56 cases, which include 8 cases of myopia (4 cases of simple myopia and 4 cases of degenerative myopia) and 4 normal individuals as control. Plasma cortisol level is lower in simple and degenerative myopia than in normal subjects, but on statistical analysis the difference was not of much significance. Patients with simple myopia with positive family history showed lower value of plasma cortisol than patients with simple myopia with negative family history, the difference was also statistically insignificant (P-0.1). In degenerative myopia patients, with and without family history, there was very little difference in plasma cortisol level and statistically highly insignificant (P-0.8). Urinary 17 keto steroid/24 hour values are lower in simple and degenerative myopia than in normal subjects.     

Occult spinal dysraphism in children: need for early neurosurgical referral

The natural history of occult spinal dysraphism in children is one of progressive and unpredictable neurological deficit. The modern role of neurosurgery is in the prevention of deterioration rather than correction of established disability. A prospective study was carried out on 40 consecutive patients to analyse the referral pattern of children with this condition. The age, reason for referral, clinical history and source of referral was recorded in each case. The reason for referral varied with age. Of the 40 patients studied, only 12 were found to be without neurological deficit at the time of referral. The incidence of progressive neurological deterioration increased with increasing age. The referral source in different age groups varied, but in all age groups relatively few were referred by the general paediatricians or paediatric neurologists. For optimum surgical results, early referral and treatment is desirable and should become our objective.