Analysis of Schizosaccharomyces pombe mediator reveals a set of essential subunits conserved between yeast and metazoan cells.

With the identification of eight new polypeptides, we here complete the subunit characterization of the Schizosaccharomyces pombe RNA polymerase II holoenzyme. The complex contains homologs to all 10 essential gene products present in the Saccharomyces cerevisiae Mediator, but lacks clear homologs to any of the 10 S. cerevisiae components encoded by nonessential genes.

Quantitative assay for group M (subtype A-H) and group O HIV-1 RNA detection in plasma.

A quantitative HIV-1 test is described based on a competitive RT-PCR assay combined with a sandwich hybridization as a detection system. The internal RNA standard (IS) was designed specifically to be competitive during the amplification and during the hybridization step. Sample viral load determination was carried out with one RT-PCR in the presence of 10(3) IS copies.

Development of a sensitive multi-well colorimetric assay for active NFkappaB.

The transcription factor nuclear factor kappaB (NFkappaB) is a key factor in the immune response triggered by a wide variety of molecules such as inflammatory cytokines, or some bacterial and viral products. This transcription factor represents a new target for the development of anti-inflammatory molecules, but this type of research is currently hampered by the lack of a convenient and rapid screening assay for NFkappaB activation. Indeed, NFkappaB DNA-binding capacity is traditionally estimated by radioactive gel shift assay.

A model for the complex between the hypoxia-inducible factor-1 (HIF-1) and its consensus DNA sequence.

Hypoxia-inducible factor-1 (HIF-1) is a heterodimeric transcription factor activated by hypoxia. When activated, HIF-1 mediates the differential expression of genes such as erythropoietin and Vascular Endothelial Growth Factor (VEGF) during hypoxia. It is composed of two different subunits, HIF-1alpha and ARNT (Aryl Receptor Nuclear Translocator).

Identification of hypoxia-responsive messengers expressed in human microvascular endothelial cells using differential display RT-PCR.

In order to identify new genes overexpressed in endothelial cells exposed to hypoxia, differential display RT-PCR was performed on total RNA extracted from human microvascular endothelial cells incubated under hypoxia or under normoxic conditions. Northern blot and reverse Northern blot analyses were used to confirm the results. Sequences corresponding to tissue inhibitor of metalloproteinase-1, prostate tumor inducing factor-1, enolase-alpha and prothymosin-alpha were evidenced as overexpressed in hypoxia.

HIF1A gene transcription is dependent on a core promoter sequence encompassing activating and inhibiting sequences located upstream from the transcription initiation site and cis elements located within the 5'UTR.

Hypoxia inducible factor-1 (HIF-1) is a transcription factor composed of two subunits, HIF-1alpha and ARNT, which is activated under hypoxia. HIF-1alpha mRNA is expressed constitutively in a wide variety of cell types, whereas in some others HIF1A gene expression is upregulated by hypoxia. In this report, we show that in endothelial cells (HMEC-1) the HIF-1alpha mRNA expression level is the same in both normoxia and hypoxia.

Follitropin (FSH) deficiency in an infertile male due to FSHbeta gene mutation. A syndrome of normal puberty and virilization but underdeveloped testicles with azoospermia, low FSH but high lutropin and normal serum testosterone concentrations.

We studied a man who sought medical attention at age 28 years because of infertility in both his first and second marriages. His sexual development appeared to have been normal, with normal puberty and virilization, and normal libido and sexual potency. At examination, his testicles were small and soft; otherwise he had a normal physical appearance.

Quantitative determination of CMV DNA using a combination of competitive PCR amplification and sandwich hybridization.

A quantitative PCR method is proposed that combines the use of a competitive internal standard with the sandwich hybridization of the products. The variability of the PCR efficiency was corrected using a specifically designed internal standard, competitive not only for the PCR amplification, but also for the hybridization on capture probes fixed onto microwells. The design of such standard gave a dynamic range extending from 30-1 million copies of target DNA when the internal standard copy number was fixed to 1000 using a simple colorimetric detection.

Pyruvate kinase of Trypanosoma brucei: overexpression, purification, and functional characterization of wild-type and mutated enzyme.

A procedure was developed for overexpression of Trypanosoma brucei pyruvate kinase in Escherichia coli. The enzyme was purified to near-homogeneity from the bacterial lysate by first removing nucleic acids and contaminating proteins by protamine sulfate precipitation and subsequent passage over a phosphocellulose column. The purified protein is essentially indistinguishable in its physicochemical and kinetic properties from the enzyme purified from trypanosomes.

Comparison between microwell and bead supports for the detection of human cytomegalovirus amplicons by sandwich hybridization.

In this study, we compared the efficiency of capture DNA probes covalently bound onto magnetic beads or microplates for their hybridization with target human cytomegalovirus (HCMV) DNA amplicons. Polystyrene supports were first aminated by wet chemistry to allow covalent grafting of the capture probes. The level of amines grafted was three times higher on beads than on microwells.

Hormones, body composition and cardiovascular risk.

Some 20 compartments of the body may be measured by CT and organ areas determined in 28 CT scans. Advantages of CT are described. While there have been extensive studies of hormones in pre- and postnatal growth, apart from evidence from disease, the role of hormones in adults has been less known.

Changes in body composition and adipose tissue distribution after treatment of women with Cushing's syndrome.

Longitudinal studies on changes in body composition and adipose tissue (AT) distribution in response to altered cortisol exposure have not yet been undertaken. Therefore, we determined body composition in seven women aged 34 +/- 9 years (mean +/- SD) with Cushing's disease/syndrome before and 8 +/- 2 (SD) months after surgical treatment for pituitary (n = 5) or cortical adrenal (n = 2) adenomas. The treatment resulted in a reduction of plasma and urinary cortisol by 78% and 97% (P < .

Cloning and sequence analysis of the gene encoding pyruvate kinase in Trypanoplasma borelli.

The gene coding for pyruvate kinase in Trypanoplasma borelli has been cloned and characterized. A single gene copy was found with an open reading frame for a polypeptide of 496 amino acids and a molecular mass of 54337. The deduced amino acid sequence has a calculated net charge of -3.

Pyruvate kinase of Leishmania mexicana mexicana. Cloning and analysis of the gene, overexpression in Escherichia coli and characterization of the enzyme.

Leishmania mexicana mexicana contains two tandemly arranged genes for pyruvate kinase (PYK). The 5' located gene codes for a polypeptide with a molecular mass of 54,370. The calculated net charge and isoelectric point of the polypeptide are -6 and 6.

Adrenocortical carcinoma--diagnostic and therapeutical implications.

Objective: To evaluate the results of treatment of a consecutive series of patients with adrenocortical carcinoma who presented during the six year period 1985 to 1991.

Design: Open study.

Setting: Departments of Surgery, Pathology, Endocrinology, and Clinical Chemistry, Sahlgren Hospital, Göteborg, Sweden.

Molecular cloning and analysis of two tandemly linked genes for pyruvate kinase of Trypanosoma brucei.

In Trypanosoma brucei (stock 427) genes encoding the glycolytic enzyme pyruvate kinase are present on two homologous chromosomes. We have cloned and characterized one of the alleles. Two large, tandemly arranged open reading frames were found, each coding for a pyruvate kinase polypeptide of 498 amino acids.

The genes encoding fructose bisphosphate aldolase in Trypanosoma brucei are interspersed with unrelated genes.

The fructose bisphosphate aldolase genes of Trypanosoma brucei are interspersed with unrelated genes whose transcript levels show no developmental modulation. Transcription appears approximately constant across the entire locus, suggesting that aldolase mRNA abundance is regulated post-transcriptionally.

Detection of specific antigens for ten serogroups of Clostridium difficile.

We previously described a serogrouping technique for Clostridium difficile based on slide agglutination with rabbit antisera raised against formol-treated cells. It allows the differentiation of ten serogroups, namely A, B, C, D, F, G, H, I, K and X. Each serogroup displays a specific profile with several distinctive bands by polyacrylamide gel electrophoresis (PAGE).

Long-term results in 64 patients operated upon for pheochromocytoma.

During the years 1956-1982, 64 pheochromocytoma patients were operated upon without mortality. Twenty-eight patients had sustained hypertension and 29 paroxysmal hypertension only. In two patients high blood pressure was not related to pheochromocytoma and five subjects were normotensive.

Epidemiology and long-term survival in acromegaly. A study of 166 cases diagnosed between 1955 and 1984.

In 1985, a follow-up investigation was performed of all patients with acromegaly (n = 166, 89 women and 77 men) seen over a 30-year period (1955-1984) at our Endocrine Unit, referral centre for the western region of Sweden (population 1.5 million). At the end of 1984, the prevalence of the disease was 6.