TNFSF/TNFRSF cytokine gene expression in sickle cell anemia: Up-regulated TNF-like cytokine 1A (TL1A) and its decoy receptor (DcR3) in peripheral blood mononuclear cells and plasma.

Background: Sickle cell anemia (SCA), a disorder with an important inflammatory component, where vasoocclusion is major contributor to the disease pathophysiology. Pro-inflammatory cytokines play an important regulatory role in the process of inflammation. We investigated the expression TL1A/DR3/DcR3 cytokine signaling pathway in peripheral blood mononuclear cells (PBMC) and their corresponding plasma levels in SCA subjects who presented with acute painful episodes.

Fetal hemoglobin in sickle cell anemia: genetic studies of the Arab-Indian haplotype.

Sickle cell anemia is common in the Middle East and India where the HbS gene is sometimes associated with the Arab-Indian (AI) β-globin gene (HBB) cluster haplotype. In this haplotype of sickle cell anemia, fetal hemoglobin (HbF) levels are 3-4 fold higher than those found in patients with HbS haplotypes of African origin. Little is known about the genetic elements that modulate HbF in AI haplotype patients.

Monocytes from sickle cell disease patients induce differential pulmonary endothelial gene expression via activation of NF-κB signaling pathway.

Monocyte-endothelial interactions play an important role in inflammatory diseases and may modulate vasculopathy in sickle cell disease, a disorder with an important inflammatory component. We co-incubated normal and sickle monocytes, lymphocytes and TNF-α with pulmonary microvascular and arterial endothelial cells and compared the expression of genes coding for adhesion molecules and cytokines that might contribute to sickle vasoocclusion. Monocyte-endothelial cell co-incubation resulted in up-regulation of L-selectin, E-selectin, VCAM-1, ICAM-1, MCP-1, MMP-1, TNF-α, IL-6 and IL-1β and down-regulation of eNOS.

Effect of sodium butyrate on lung vascular TNFSF15 (TL1A) expression: differential expression patterns in pulmonary artery and microvascular endothelial cells.

Vascular endothelial growth inhibitor TNFSF15 (TL1A), a ligand for TNFRSF25 (DR3) and decoy receptor TNFRSF6B (DcR3), is expressed in human pulmonary arterial (HPAEC) and lung microvascular (HMVEC) endothelial cells where it might modulate inflammation and sickle vasculopathy. Pulmonary disease, endothelial abnormalities and inflammation are prominent features of sickle cell disease (SCD). Butyrate has opposing effects on endogenous TNFSF15 expression in pulmonary endothelium, acting as an inhibitor in HPAEC and an inducer in HMVEC.

Differential gene expression in pulmonary artery endothelial cells exposed to sickle cell plasma.

Clinical variability in sickle cell disease (SCD) suggests a role for extra-erythrocytic factors in the pathogenesis of vasoocclusion. We hypothesized that endothelial cell (EC) dysfunction, one possible modifier of disease variability, results from induction of phenotypic changes by circulating factors. Accordingly, we analyzed gene expression in cultured human pulmonary artery ECs (HPAEC) exposed to plasma from 1) sickle acute chest syndrome (ACS) patients, 2) SCD patients at steady state, 3) normal volunteers, and 4) serum-free media, using whole genome microarrays (U133A-B GeneChip, Affymetrix).

Gene expression profiling during erythroid differentiation of K562 cells.

We studied the temporal changes in gene expression in K562 cells at intervals from 2 to 48 h following induction using differential display polymerase chain reaction and gene expression arrays. More than 110 cDNA fragments representing 86 unique mRNAs were either up- or downregulated during erythroid differentiation. Sixty-one of the differentially expressed cDNA fragments had more than 95% homology to known GenBank sequences; 21 represented cDNA sequences with only dbEST or high-throughput gene-screening database matches.

Induction of globin synthesis in K562 cells is associated with differential expression of transcription factor genes.

Globin gene switching may be mediated by proteins expressed during different stages of development. Their identification may clarify the mechanisms of the conversion from fetal to adult globin production and lead to new approaches to reversing or retarding the gamma- to beta-globin gene switch. To explore this hypothesis, K562 erythroleukemia cells were induced to differentiate with 1.

Double heterozygosity for the codon beta 39 C-to-T nonsense mutation and a triplicate alpha-globin gene locus can cause "dominantly" inherited beta-thalassemia intermedia.

Background: A beta-thalassemia intermedia phenotype can be caused by multiple genotypes.

Methods: We studied a family where the mother was hematologically normal and both father and daughter had beta-thalassemia intermedia.

Results: Both affected individuals were heterozygous for a codon 39 CAG-to-TAG mutation.

Hematological effects of atypical and Cameroon beta-globin gene haplotypes in adult sickle cell anemia.

To examine the effects of unusual or atypical beta-globin gene cluster haplotypes on the hematological features and Hb F levels of sickle cell anemia, we studied African Americans who had an atypical or Cameroon haplotype chromosome in association with a typical haplotype. We identified over 20 atypical haplotypes. The distribution of 5' sub-haplotypes of the atypical chromosomes mirrored the distribution of common haplotypes in African Americans with sickle cell anemia.

Transcriptional upregulation of gamma-globin by phenylbutyrate and analogous aromatic fatty acids.

Phenylbutyrate has been shown recently to induce fetal hemoglobin (HbF) production in patients with sickle cell anemia and beta thalassemia. We have now examined related aromatic fatty acids in order to define the range of active structures and identify plausible mechanisms of action. Structure-function analysis revealed that for effective stimulation of HbF in erythroid precursors: (1) the ideal length for the aliphatic side chain is four carbons; (2) oxygen or sulfur substitutions in the carboxylic chain are allowed, as evidenced by the equal or increased activity of phenoxypropionate, benzylthioglycolate, and benzyloxyacetate compared with phenylbutyrate; and (3) blocking the carboxylate group by conversion to the amide form greatly reduces potency.

Augmentation of gamma-globin gene promoter activity by carboxylic acids and components of the human beta-globin locus control region.

Butyric acid increases fetal hemoglobin synthesis in adult animals and in erythroid cells in culture and induces the gamma-globin gene promoter in transient expression experiments in K562 cells (McDonagh KT, Nienhuis AW, Blood 78:255a, 1992 [abstr, suppl 1]). We compared the effect of butyrate and other short-chain carboxylic acids in transient expression studies with K562 cells using an expression plasmid bearing a luciferase reporter gene driven by the normal human A gamma-globin gene promoter. Butyrate (4 carbons) increased the activity of the human A gamma-globin gene promoter up to 123 times.

Studies on the in vitro and in vivo expression of a dysfunctional alpha-globin gene.

In a black family with members having alpha-thalassemia and hemoglobin H (HbH) disease, a deletion of an AG dinucleotide at the 3' end of exon 1 near the junction with intron 1 was shown previously to produce a dysfunctional alpha-thalassemia gene with a reading frame-shift and a nonsense codon (Safaya S, Rieder RF: J Biol Chem 263:4328-4332, 1988). We have found that the same mutation is responsible for alpha-thalassemia and HbH disease in a second unrelated black family (Bellevue R, Dosik H, Rieder RF: Br J Haematol 41:193-202, 1979). Despite the loss of two nucleotides from the consensus sequence at the 5' splice donor site of intron 1, studies employing an in vitro plasmid-based expression system indicated that the mutant alpha-globin mRNA was spliced normally and expressed in amounts equal to normal alpha-globin mRNA in COS-7 cells.

Effect of beta-globin gene cluster haplotype on the hematological and clinical features of sickle cell anemia.

In 113 black American adults with sickle cell anemia (HbSS), we examined nine polymorphic restriction sites, including the Xmnl site 5' to the G gamma gene, to see whether haplotype is related to the level of HbF and the proportion of G gamma chains or if it influences the hematological and clinical features of the disease. Seventy-five percent of the patients were homozygous or heterozygous for the Benin (no. 19) or Central African Republic (Bantu, no.

Homozygous beta-thalassemia without anemia.

A 37-year-old man of Guyanese origin was found to have homozygous beta-thalassemia without anemia. There were no physical stigmata of thalassemia. The hematocrit value was 41 to 45.

Dysfunctional alpha-globin gene in hemoglobin H disease in blacks. A dinucleotide deletion produces a frameshift and a termination codon.

alpha-Thalassemia trait is common in Black Americans; the (-alpha) haplotype occurs in 30% of that population. However, hemoglobin H disease (genotype:- -/-alpha) is very uncommon due to the rarity of the (- -) haplotype. A subject with HbH-HbG Philadelphia (alpha 2(68)Asn----Lys) synthesized only alpha G and no alpha A.

Dysfunctional alpha-globin genes in hemoglobin H disease in blacks: variation in restriction fragment size permits the detection of the -alpha/-alpha T genotype.

Hemoglobin H (HbH) disease is most often due to deletion of three of the four alpha-globin genes (genotype --/--alpha). In black subjects although the -alpha/chromosome is common, the --/haplotype is very rare and few examples of HbH disease have been detected. We have studied three black siblings with HbH by restriction endonuclease mapping of the alpha-like gene complex (5'-zeta-psi zeta-psi alpha 2-psi alpha 1-alpha 2-alpha 1-3') using zeta- and alpha- specific probes.

Similarity of the nucleotide sequences of rat alpha-lactalbumin and chicken lysozyme genes.

alpha-Lactalbumin (alpha-LA) is a milk protein that interacts with the enzyme galactosyltransferase, modifying its substrate specificity in a way which promotes the transfer of galactose to glucose, resulting in a way which promotes the transfer of galactose to glucose, resulting in a beta-1----4 glycosidic linkage and the synthesis of lactose. Lysozyme, an enzyme which catalyses the hydrolysis of a beta-1----4 glycosidic linkage in polysaccharides, has been shown to be structurally related to alpha-LA and it has been proposed that they have arisen from a common ancestral gene. To compare their evolutionary relationships, we report here the complete nucleotide sequence of the rat alpha-LA gene, including its 5'-flanking sequences, and compare its gene structure with the chicken egg-white lysozyme gene.

Low dose injectable contraceptive norethisterone enanthate 20mg monthly - II. Metabolic side effects.

Metabolic effects of a long-acting low dose injectable contraceptive, norethisterone enanthate 20-mg, monthly injections (Neten-20), was tested in 13 women belonging to the low income groups over a period of 1 year. No change was observed in hemoglobin, hematocrit, glucose tolerance, plasma lipids, iron, calcium, or serum glutamate-oxaloacetate transaminase after treatment. Marginal rise in albumin and fall in some globulin fractions was observed.