Inositol- and Galactose-Binding Lectins of Chicken Brain Fractions Enriched with Glial Cells

Galactose- and inositol-binding proteins with lectin activity (GL-GAL and GL-I, respectively) were isolated from membranes enriched with cells of chicken brain fractions. Both lectins are glycoproteins of molecular mass 13.5 and 11.5 kDa, respectively; they show a high affinity to EDTA (GL-I) and EGTA (GL-GAL, GL-I), which indicates an important role of Ca⁺² in molecular organization of these lectins. In brain glial cells of chick embryos, unlike adult chickens, a soluble form of lectins has been revealed; it is easily extracted with 2 mM EDTA and shows sensitivity to L-lactose, D-galactose, and N-acetyl-D-galactosamine. It is suggested that in the course of embryonal and postembryonal development of the chicken brain, a transformation and qualitative changes of the lectin spectrum occur due to a change of function of glial cells.     

Determination of plasma leukotrienes in antigen-induced bronchoconstrictive guinea pigs.

Convenient extraction and radioimmunoassay methods for measurement of leukotrienes C4 and D4 (LTC4 and LTD4) in biological fluids are described. LTC4 or LTD4 in plasma was extracted with acetonitrile, and the extract was washed with dichloromethane then adjusted to pH 3.5 or 6.0, respectively. Each leukotriene was partially purified by using a C18-bonded silica cartridge and quantitated by radioimmunoassay. Amounts of LTC4 and LTD4 in the range of 0.025-1.6 ng could be assayed in plasma. This procedure was employed to examine the increase in plasma LTC4 (0.249 +/- 0.036 ng/ml) and LTD4 (1.399 +/- 0.235 ng/ml) of guinea pigs during intravenous challenge-induced anaphylactic bronchoconstriction, and the suppression of the increase of bronchoconstriction and leukotrienes by the administration of 5-lipoxygenase inhibitors such as E6080 (6-hydroxy-2-(4-sulfamoylbenzyl-amino)- 4,5,7-trimethylbenzothiazole hydrochloride), AA861 (2,3,5-trimethyl-6-(12-hydroxy-5,10-dodecadiynyl)-1,4-benzoquinone ) and phenidone. On the other hand, LTC4 and LTD4 were not detected in plasma after an inhaled challenge, though significant bronchoconstriction was provoked. It was concluded that the present study validates a new technique for quantitating plasma leukotrienes on the basis of pH and a suitable method for evaluating the pharmacological efficacy of 5-lipoxygenase inhibitors.     

A Simple Method for Isolating DNA of High Yield and Quality from Cotton (shape Gossypium hirsutum L.) Leaves

An easy, reproducible and fast procedure to isolate DNA from cotton leaves is described. The addition of 0.5 M glucose in the extraction buffer avoids browning by polyphenolic compounds and improves the quality of DNA for molecular analysis. The DNA yield ranged between 150–400 mg per gram of fresh tissue. The DNA was suitable for digestion by restriction enzymes and amplificatiion by Taq DNA polymerase.