Common genetic variants of the human steroid 21-hydroxylase gene (CYP21A2) are related to differences in circulating hormone levels.

PURPOSE: Systematic evaluation of the potential relationship between the common genetic variants of CYP21A2 and hormone levels. METHODS: The relationships of CYP21A2 intron 2 polymorphisms and haplotypes with diverse baseline and stimulated blood hormone levels were studied in 106 subjects with non-functioning adrenal incidentaloma (NFAI). The rationale for using NFAI subjects is dual: i) their baseline hormone profiles do not differ from those of healthy subjects and ii) hormone levels after stimulation tests are available. RESULTS: The carriers (N = 27) of a well-defined CYP21A2 haplotype cluster (c5) had significantly elevated levels of cortisol (p = 0.0110), and 17-hydroxyprogesterone (p = 0.0001) after ACTH stimulation, and 11-deoxycortisol after metyrapone administration (p = 0.0017), but the hormone values were in normal ranges. In addition, the carriers (N = 33) of the C allele of the rs6462 polymorphism had a higher baseline aldosterone level (p = 0.0006). The prevalence of these genetic variants of CYP21A2 did not differ between NFAI and healthy subjects. CONCLUSIONS: The common CYP21A2 variants presumably exert the same effect on hormone levels in the healthy and disease-affected populations. Therefore, they may contribute to complex diseases such as some cardiovascular diseases, and may influence the genotype-phenotype correlation in patients with congenital adrenal hyperplasia (CAH) including the individual need for hormone substitution.

Both positive and negative selection pressures contribute to the polymorphism pattern of the duplicated human CYP21A2 gene.

The human steroid 21-hydroxylase gene (CYP21A2) participates in cortisol and aldosterone biosynthesis, and resides together with its paralogous (duplicated) pseudogene in a multiallelic copy number variation (CNV), called RCCX CNV. Concerted evolution caused by non-allelic gene conversion has been described in great ape CYP21 genes, and the same conversion activity is responsible for a serious genetic disorder of CYP21A2, congenital adrenal hyperplasia (CAH). In the current study, 33 CYP21A2 haplotype variants encoding 6 protein variants were determined from a European population. CYP21A2 was shown to be one of the most diverse human genes (HHe=0.949), but the diversity of intron 2 was greater still. Contrary to previous findings, the evolution of intron 2 did not follow concerted evolution, although the remaining part of the gene did. Fixed sites (different fixed alleles of sites in human CYP21 paralogues) significantly accumulated in intron 2, indicating that the excess of fixed sites was connected to the lack of effective non-allelic conversion and concerted evolution. Furthermore, positive selection was presumably focused on intron 2, and possibly associated with the previous genetic features. However, the positive selection detected by several neutrality tests was discerned along the whole gene. In addition, the clear signature of negative selection was observed in the coding sequence. The maintenance of the CYP21 enzyme function is critical, and could lead to negative selection, whereas the presumed gene regulation altering steroid hormone levels via intron 2 might help fast adaptation, which broadly characterizes the genes of human CNVs responding to the environment.

Intraspecific evolution of human RCCX copy number variation traced by haplotypes of the CYP21A2 gene.

The RCCX region is a complex, multiallelic, tandem copy number variation (CNV). Two complete genes, complement component 4 (C4) and steroid 21-hydroxylase (CYP21A2, formerly CYP21B), reside in its variable region. RCCX is prone to nonallelic homologous recombination (NAHR) such as unequal crossover, generating duplications and deletions of RCCX modules, and gene conversion. A series of allele-specific long-range polymerase chain reaction coupled to the whole-gene sequencing of CYP21A2 was developed for molecular haplotyping. By means of the developed techniques, 35 different kinds of CYP21A2 haplotype variant were experimentally determined from 112 unrelated European subjects. The number of the resolved CYP21A2 haplotype variants was increased to 61 by bioinformatic haplotype reconstruction. The CYP21A2 haplotype variants could be assigned to the haplotypic RCCX CNV structures (the copy number of RCCX modules) in most cases. The genealogy network constructed from the CYP21A2 haplotype variants delineated the origin of RCCX structures. The different RCCX structures were located in tight groups. The minority of groups with identical RCCX structure occurred once in the network, implying monophyletic origin, but the majority of groups occurred several times and in different locations, indicating polyphyletic origin. The monophyletic groups were often created by single unequal crossover, whereas recurrent unequal crossover events generated some of the polyphyletic groups. As a result of recurrent NAHR events, more CYP21A2 haplotype variants with different allele patterns belonged to the same RCCX structure. The intraspecific evolution of RCCX CNV described here has provided a reasonable expectation for that of complex, multiallelic, tandem CNVs in humans.

Astronaut's organ doses inferred from measurements in a human phantom outside the international space station.

Space radiation hazards are recognized as a key concern for human space flight. For long-term interplanetary missions, they constitute a potentially limiting factor since current protection limits for low-Earth orbit missions may be approached or even exceeded. In such a situation, an accurate risk assessment requires knowledge of equivalent doses in critical radiosensitive organs rather than only skin doses or ambient doses from area monitoring. To achieve this, the MATROSHKA experiment uses a human phantom torso equipped with dedicated detector systems. We measured for the first time the doses from the diverse components of ionizing space radiation at the surface and at different locations inside the phantom positioned outside the International Space Station, thereby simulating an extravehicular activity of an astronaut. The relationships between the skin and organ absorbed doses obtained in such an exposure show a steep gradient between the doses in the uppermost layer of the skin and the deep organs with a ratio close to 20. This decrease due to the body self-shielding and a concomitant increase of the radiation quality factor by 1.7 highlight the complexities of an adequate dosimetry of space radiation. The depth-dose distributions established by MATROSHKA serve as benchmarks for space radiation models and radiation transport calculations that are needed for mission planning.

Developing multimedia software and virtual reality worlds and their use in rehabilitation and psychology.

The multimedia and virtual reality projects performed at our laboratory during the last ten years can be grouped into the following groups: 1) tutorial and entertainment programs for handicapped children, 2) rehabilitation programs for stroke patients and patients with phobias. We have developed multimedia software for handicapped children with various impairments: partial vision, hearing difficulties, locomotive difficulties, mental retardation, dyslexia etc. In the present paper we show the advantages of using multimedia software to develop mental skills in handicapped people and deal with the special needs of handicapped children. For the rehabilitation of stroke patients we have developed a computer-controlled method, which enables - contrary to methods used internationally - not only the establishment of a diagnosis, but also measurement of therapy effectiveness: 1) it enables us to produce a database of patients, which contains not only their personal data but also test results, their drawings and audio recordings, 2) it is in itself an intensive therapeutic test and contains tutorial programs. We are currently collecting test results. We have also developed some virtual worlds for treating phobias: a virtual balcony and a ten-story building with an external glass elevator as well as an internal glass elevator in the virtual Atrium Hyatt hotel. We have developed a virtual environment for treating claustrophobia too: a closed lift and a room where the walls can move. For specific phobias (fear of travelling) we have modelled the underground railway system in Budapest. For autistic children, we have developed virtual shopping software too. In this paper we present the advantages of virtual reality in the investigation, evaluation and treatment of perception, behaviour and neuropsychological disorders.

Development of a computer controlled cognitive diagnostics and rehabilitation method.

In this paper we introduce a computer controlled method (HELp Neuropsychology = HELEN) which enables - as a difference to methods used internationally--not only the establishment of the diagnosis, but permits measurement of the effectiveness of the therapy. It allows: --To produce a database of the patients that contains not only their personal data but also the results of the tests, their drawings and audio recordings. --It is an intensive therapeutic test, which contains tutorial programs too.