Effect of permeabilization on the thermostability of catalase in immobilized yeast cells

Summary Catalase in yeast cells is rendered thermolabile on permeabilization using organic solvents or any process of immobilization that eventually permeabilizes the cells. A stable immobilized catalase preparation can be obtained only if yeast cells are immobilized in an intact or viable form.     

Synergistic effect of two β globin gene cluster mutations leading to the hereditary persistence of fetal hemoglobin (HPFH) phenotype

Co-inheritance of gamma and beta globin gene mutations in a compound heterozygous state is rare but of clinical interest as it provides an important data on understanding the HbF expression. Hematological analysis was carried out (Sysmex KX-21). F-cells were enumerated using flow cytometry. Beta globin gene was analysed by CRDB technique and by DNA sequencing. Gamma globin promoter region was sequenced and expression studies were carried out using real time Taqman assay. We report a family, where two inherited defects of the β globin gene cluster segregate. The proband and her sibling were compound heterozygotes for a novel ^Gγ promoter mutation and the 619 bp deletion a common Indian β thalassemia mutation. Molecular characterization revealed that the father (HbA₂ 5.1%, HbF 5.4%), proband (HbA₂ 3.6%, HbF 31.7%) and her brother (HbA₂ 3.9%, HbF 23.6%) were heterozygous for the 619 bp deletion. The mother (HbA₂ 2.1%, HbF 3.4%) had a normal β globin gene. As both the children showed high HbF levels, the γ globin gene work up was carried out. The ^Gγ-globin gene promoter analysis revealed that the mother and the two children were heterozygous for a 5 bp deletion -ATAAG (-533 to -529) that resides in the GATA binding site. These findings suggest that the 5 bp deletion in the ^Gγ globin promoter has a functional role in silencing the γ-globin gene expression in adults by disrupting GATA-1 binding and the associated repressor complex and results in the up-regulation of gamma globin gene expression. When co-inherited with β -thalassemia trait it leads to a phenotype of HPFH.     

Regulation of sialyltransferase activity in intestinal segments of rats

A differential distribution of sialyltransferase (ST) in different regions of intestine has been shown. Jejunum and ileum homogenates from rats showed almost exclusive presence of alpha-2-3 ST (to Gal in Gal beta-1-4GlcNAc and/or to Gal in Gal beta-1-3GalNAc). In contrast, colon homogenates showed the presence of both alpha-2-3 ST (as above) and alpha-2-6 ST. Incubation of intestinal slices in presence of heat-inactivated horse serum (HHS) showed a time- and temperature-dependent secretion of soluble ST into the medium. Both jejunum and ileum slices showed high rates of secretion of alpha-2-3 ST. Colon slices, though rich in alpha-2-6 ST, secreted only alpha-2-3 ST. Colchicine, an anti-mitotic drug, injected into rats caused about 10-fold increase of the serum ST level. Jejunum slices from colchicine-treated rats showed an increased secretion of alpha-2-6 ST, suggesting that intestine undergoes a change in the expression of normal secretion of alpha-2-3 ST to a secretion of alpha-2-6 ST. The secretion of ST from incubated intestinal slices was inhibited by heparin. Certain protein factors (anti-proteases) in HHS bind to heparin-sepharose column and these protein factors are responsible for causing the secretion of ST into the medium. It has also been found that a supernatant fraction of the colon homogenate activated ST. Gel chromatography on HPLC produced 3-4 protein fractions from the colon cytosol and one of this fraction bearing high molecular weight proteins produced the maximum activation of ST.(ABSTRACT TRUNCATED AT 250 WORDS)