Mosaics of gene variations in the Interleukin-10 gene promoter affect interleukin-10 production depending on the stimulation used.

Interleukin-10 (IL-10), a cytokine involved in many aspects of the immune response shows interindividual variations in their expression. However, genetic variations of the 5'-flanking region of the IL-10 gene (PIL-10) are poorly characterised with respect to different stimuli. New extended haplo- and genotypes are identified present at differing frequencies in three geographically separated populations. Their influence on IL-10 expression have been assessed in vitro after stimulation of leukocytes with lipopolysaccharide (LPS), dibutyryl-cAMP or following immortalisation with Epstein-Barr virus (lymphoblastoid cell line (LCL)). Interindividual differences of IL-10 production were found to be related to single-nucleotide polymorphisms (SNP) haplotype -6752/-6208 in LCLs (P<0.02), and for haplotypes comprising SNPs -6752/-6208/-3538 after LPS stimulation (P<0.03). Carriers of the IL10.G microsatellite with 22, 24 or 26 dinucleotide repeats linked with the -1087G SNP, exhibited the highest levels of IL-10 expression. Contrasting IL-10 secretion patterns were found for IL10.R microsatellite alleles characterised by 15 dinucleotide repeats: after LPS stimulation this allele was associated with high IL-10 production (P<0.007), but with low IL-10 levels in LCLs (P< 0.038). Thus, the effects of mosaics of genetic elements in the PIL-10 on the capacity of leukocytes to produce IL-10 depend on the agent inducing IL-10 expression.

Simultaneous analysis of interleukin-10 gene microsatellites and single-nucleotide polymorphisms in parallel with tumour necrosis factor and interferon-gamma short tandem repeats by fluorescence-based polymerase chain reaction.

Different cytokine genotypes exist in the population, for example, as a result of selective pressure of infectious diseases. It may be that specific cytokine genotypes that are beneficial by creating a 'proinflammatory' phenotype predispose to severe inflammatory disease with worse clinical outcome. There is individual variation in the production of certain cytokines in relation to their genotypes. IL-10, IFN-gamma and TNF-alpha are key components in the regulation of immune responses and the balance of their expression levels is predictive in certain diseases. To describe cytokine genotypes, a one-tube PCR reaction was developed to analyse simultaneously DNA sequence variations of cytokine genes IL-10, IFN-gamma, and TNF. This multiplex PCR approach was used to provide genotypic data for two geographically independent donor groups from Germany and Gabon. Significant differences were obtained for the majority of sequence variations comparing both populations. However, the SNPs within the 5'-flanking region of the IL-10 gene at position -1087 and -6208 are comparable in their genic and genotypic behaviour. Comparing allelic and genotypic disequilibrium between pairs of loci revealed different association patterns for both populations according to the geographical polymorphism. This assay may improve immunogenetic studies in disease, characterized by disbalanced IL-10, IFN-gamma and TNF-alpha expression.

A new allelic variation within the 5'-flanking region of the interleukin-10 gene.

At -2471 bp from the transcriptional start site of the interleukin-10 (IL-10) gene, the inverted repeat TG/CA was previously identified and designated IL-10.IR. In an analysis of samples from 200 Germans (Caucasian) and 286 Gabonese (Central African), three different alleles, IL-10.IR6, IL-10.IR7 and IL-10.IR8, were identified. In the Caucasians, IL-10.IR6 and IL-10.IR8 were found only once, in each case with IL-10.IR7 as the corresponding second allele.

Structural characterisation of the distal 5' flanking region of the human interleukin-10 gene.

Interleukin-10 (IL-10) is an important immunoregulatory cytokine. The recent characterisation of the proximal 5' flanking region of IL-10 led to the identification of the promoter region. Two polymorphic dinucleotide repeats and 10 single nucleotide polymorphisms (SNPs) have been identified and suggested to be useful genetic markers in several diseases. We have sequenced a further 5275 bp from -9296 to -4021 of the distal part of the 5' flanking region of the human IL-10 gene from the cosmid clone pWE15-4/11. Our sequence analysis reveals a high density of Alu-repeats within the IL-10 gene locus, including three novel, related structures which we term Alu-IL10 (A-C). Using three overlapping PCR products spanning 5110 bp of this distal part of the IL-10 gene the following single base pair substitutions were identified: at -8571 C/T, -8531 G/A, -6752 A/T, -6208 G/C, -5402 C/G. In addition a heterozygous three base pair deletion at -7400 was observed. The SNPs at -8571 C/T and -8531 G/A are contained within an Alu-repeat. These data should further the understanding of how the IL-10 gene is controlled in man and how its function may vary between individuals.

Effects of goal structures and performance contingencies on the math performance of adolescents with learning disabilities.

This study assessed the effects of assigned versus self-selected goals and contingent versus noncontingent gameplay conditions, delivered during computer-assisted math computation drill-and-practice sessions. Participants were 20 high school pupils with learning disabilities who had active Individualized Education Plan (IEP) goals that targeted the improvement of computational skills. Students were assigned randomly to goal conditions and, within goal conditions, to contingency groups. Math computation performance was measured pre-, mid-, and posttreatment. Analyses of variance indicated that students who selected their goals performed better than pupils with assigned goals. No differences existed between the contingency groups. Implications for practice are discussed.

[Interaction studies with fluconazole, a new triazole antifungal drug]. / Interaktionsstudien mit Fluconazol, einem neuen Triazolantimykotikum.

Fluconazole is a new systemic antifungal agent from the class of triazoles. Its efficacy was documented in clinical studies of patients with candidosis and cryptococcosis. Due to the influence on human cytochrome-P-450-dependent enzymes, special attention should be paid, in the development of new antifungal agents, to the inhibition of the metabolization of other simultaneously given substances and to the inhibition of steroid hormone synthesis in the gonads and adrenal glands. The interaction studies carried out with frequently administered drugs such as oral contraceptives, cimetidine, warfarin, tolbutamide, and cyclosporin (transplantation patients) allow the concomitant intake of these substances with fluconazole. Concomitant anti-coagulant treatment requires a precautionary monitoring of prothrombin time; it may be necessary in individual cases to reduce the anticoagulant dosage. The intake of oral hypoglycemics of the tolbutamide type requires monitoring of blood glucose, although blood glucose lowering effects were not found. One interaction study carried out with phenazone showed the low metabolization tendency of fluconazole. Approximately 80% of the administered fluconazole dose appeared in the urine unchanged. Studies in male and female subjects with multiple doses of fluconazole (4 weeks) did not show any significant effects on the endocrinology and steroid synthesis in man.

Chromosome investigations of workers exposed to cadmium in the manufacturing of cadmium stabilizers and pigments.

Chromosome analyses were undertaken on 14 workers exposed to cadmium and the results were compared with a corresponding control group. All probands have been employed for many years in cadmium pigment and stabilizer production plants (between 6 and 25 years). The results of the clinical investigations were, for the most part, unspecific, although at the same time some parameters were above the normal (transaminase, beta 2-microglobulin deposit in the urine, as well as cadmium in the blood and urine). The chromosome analyses were undertaken on the lymphocytes of the peripheral blood with the aid of the so-called "micromethod." Metaphases, 150 per person, were analyzed. The statistical evaluation was made according to the Fisher-Yates test. The chromosome aberration rate of the group investigated was not higher than that of the control group, inclusive and exclusive of gaps.