Humoral and cell-mediated autoimmune reactions to human acidic ribosomal P2 protein in individuals sensitized to Aspergillus fumigatus P2 protein.

A panel of cDNAs encoding allergenic proteins was isolated from an Aspergillus fumigatus cDNA library displayed on the surface of filamentous phage. Solid phase-immobilized serum immunoglobulin E (IgE) from A. fumigatus-allergic individuals was used to enrich phage displaying IgE-binding molecules. One of the cDNAs encoded a 11.1-kD protein that was identified as acidic ribosomal phosphoprotein type 2 (P2 protein). The allergen, formally termed rAsp f 8, shares >62% sequence identity and >84% sequence homology to corresponding eukaryotic P2 proteins, including human P2 protein. The sequences encoding human and fungal P2 protein were subcloned, expressed in Escherichia coli as His6-tagged fusion proteins, and purified by Ni2+-chelate affinity chromatography. Both recombinant P2 proteins were recognized by IgE antibodies from allergic individuals sensitized to the A. fumigatus P2 protein and elicited strong type 1-specific skin reactions in these individuals. Moreover, human and fungal P2 proteins induced proliferative responses in peripheral blood mononuclear cells of A. fumigatus- allergic subjects sensitized to the fungal P2 protein. These data provide strong evidence for in vitro and in vivo humoral and cell-mediated autoreactivity to human P2 protein in patients suffering from chronic A. fumigatus allergy.

Dissection of immunoglobulin E and T lymphocyte reactivity of isoforms of the major birch pollen allergen Bet v 1: potential use of hypoallergenic isoforms for immunotherapy.

We dissected the T cell activation potency and the immunoglobulin (Ig) E-binding properties (allergenicity) of nine isoforms of Bet v 1 (Bet v 1a-Bet v 1l), the major birch pollen allergen. Immunoblot experiments showed that Bet v 1 isoforms differ in their ability to bind IgE from birch pollen-allergic patients. All patients tested displayed similar IgE-binding patterns toward each particular isoform. Based on these experiments, we grouped Bet v 1 isoforms in three classes: molecules with high IgE-binding activity (isoforms a, e, and j), intermediate IgE-binding (isoforms b, c, and f), and low/no IgE-binding activity (isoforms d, g, and 1). Bet v 1a, a recombinant isoform selected from a cDNA expression library using IgE immunoscreening exhibited the highest IgE-binding activity. Isoforms a, b, d, e, and 1 were chosen as representatives from the three classes for experimentation. The potency of each isoallergen to activate T lymphocytes from birch pollen-allergic patients was assayed using peripheral blood mononuclear cells, allergen-specific T cell lines, and peptide-mapped allergen-specific T cell clones. Among the patients, some displayed a broad range of T cell-recognition patterns for Bet v 1 isoforms whereas others seemed to be restricted to particular isoforms. In spite of this variability, the highest scores for T cell proliferative responses were observed with isoform d (low IgE binder), followed by b, 1, e, and a. In vivo (skin prick) tests showed that the potency of isoforms d and 1 to induce typical urticarial type 1 reactions in Bet v 1-allergic individuals was significantly lower than for isoforms a, b, and e. Taken together, our results indicate that hypoallergenic Bet v 1 isoforms are potent activators of allergen-specific T lymphocytes, and Bet v 1 isoforms with high in vitro IgE-binding activity and in vivo allergenicity can display low T cell antigenicity. Based on these findings, we propose a novel approach for immunotherapy of type I allergies: a treatment with high doses of hypoallergenic isoforms or recombinant variants of atopic allergens. We proceed on the assumption that this measure would modulate the quality of the T helper cell response to allergens in vivo. The therapy form would additionally implicate a reduced risk of anaphylactic side effects.

Identification of profilin as a novel pollen allergen; IgE autoreactivity in sensitized individuals.

A complementary DNA encoding a pollen allergen from white birch (Betula verrucosa) that was isolated from a pollen complementary DNA library with serum immunoglobulin E from a birch pollen-allergic individual revealed significant sequence homology to profilins. The recombinant protein showed high affinity to poly-L-proline. Immunoglobulin E antibodies from allergic individuals bound to natural and recombinant birch profilin and also to human profilin. In addition, birch and human profilin induced histamine release from blood basophils of profilin-allergic individuals, but not of individuals sensitized to other plant allergens. The structural similarity of conserved proteins might therefore be responsible for maintaining immunoglobulin E antibody titers in type I allergy.

GermOnline, a cross-species community knowledgebase on germ cell differentiation.

GermOnline provides information and microarray expression data for genes involved in mitosis and meiosis, gamete formation and germ line development across species. The database has been developed, and is being curated and updated, by life scientists in cooperation with bioinformaticists. Information is contributed through an online form using free text, images and the controlled vocabulary developed by the GeneOntology Consortium. Authors provide up to three references in support of their contribution. The database is governed by an international board of scientists to ensure a standardized data format and the highest quality of GermOnline's information content. Release 2.0 provides exclusive access to microarray expression data from Saccharomyces cerevisiae and Rattus norvegicus, as well as curated information on approximately 700 genes from various organisms. The locus report pages include links to external databases that contain relevant annotation, microarray expression and proteome data. Conversely, the Saccharomyces Genome Database (SGD), S.cerevisiae GeneDB and Swiss-Prot link to the budding yeast section of GermOnline from their respective locus pages. GermOnline, a fully operational prototype subject-oriented knowledgebase designed for community annotation and array data visualization, is accessible at http://www.germonline.org. The target audience includes researchers who work on mitotic cell division, meiosis, gametogenesis, germ line development, human reproductive health and comparative genomics.

Molecular dynamics simulation of the rare amino acid LL-dityrosine and a dityrosine-containing peptide: comparison with time-resolved fluorescence.

The fluorescence of the rare amino acid LL-dityrosine, which is found in insoluble biological materials with structural features, was recently shown to decay non-exponentially (Kungl et al. (1992) J. Fluorescence 2, 63-74). Here we investigated the time-resolved fluorescence of a dityrosine-containing peptide (DCP) to study the influence of side chains on the fluorescence decay of the chromophore. The fluorescence decay of DCP was best fitted by three exponential terms including a sub-nanosecond rise term, the values of which are quite similar to the parameters obtained for the decay of free dityrosine. They were found to depend on the pH of the aqueous solution but not on the temperature. Analysis by an exponential series method revealed broad fluorescence lifetime distributions for DCP. Compared to the corresponding analysis of dityrosine transients, similar lifetime centers were found whereas the widths of the distributions were found broader for DCP. Molecular dyamics (MD) simulations of dityrosine at 300 K show that chi 1 and chi 2 side chain conformers (rotamers) of both tyrosine subunits interconvert on a picosecond timescale. The rates of interconversion were shown to depend critically upon the MD technique applied: in vacuo simulations yielded lower interconversion rates compared to stochastic dynamics (SD) and full MD (water explicitly included). However, MD simulations of the dityrosine-containing peptide revealed no interconversions of the chi 1 and chi 2 side chain rotamers of both tyrosine subunits within a 400 ps trajectory. Interconversions could be induced by raising the temperature of the system (DCP plus solvent) to 340 K. Side chain rotamers of dityrosine are not stable on a fluorescence time scale but are stable when a dityrosine-containing peptide is regarded. Nevertheless both molecules yield similar fluorescence decay patterns. We therefore conclude that the rotamer model proposed for the fluorescence decay of tyrosine and tryptophan cannot be applied to the fluorescence decay of dityrosine and peptides containing this chromophore. This should be of future interest when dityrosine is used as an intrinsic sensor to study complex dityrosine-containing macromolecules by fluorescence spectroscopy.

A birch gene family encoding pollen allergens and pathogenesis-related proteins.

Bet v I, the major pollen allergen of birch (Betula verrucosa), shows high sequence homology to a family of pathogenesis-related (PR) proteins that have recently been identified in several other plant species. We have used a pollen Bet v I cDNA clone and anti-Bet v I antibodies as probes to study the expression of Bet v I genes in birch cell suspension cultures under different experimental conditions. Induction of Bet v I-related proteins was detected in immunoblots of cell extracts upon co-cultivation with microbial pathogens. Northern analysis revealed the rapid induction of Bet v I transcripts in the presence of bacteria and fungi, but not by stress treatments (heat shock, metal ions) or by chemical elicitors. RNase protection experiments showed that the pathogen-inducible RNAs did not correspond to the pollen cDNA clone but most likely to the products of transcription of other members of the Bet v I gene family, sharing high sequence homology with the pollen-specific gene within the 5'-half of the coding region. We conclude that the Bet v I gene family of pollen allergens includes a subset of defense-related genes that are transcriptionally activated in the presence of microbial pathogens.

Molecular cloning of major and minor allergens of Alternaria alternata and Cladosporium herbarum.

The two moulds, Alternaria alternata and Cladosporium herbarum, are recognized as major causes of fungal allergies. Cloning, sequencing and heterologous expression of the allergens of the two moulds is a necessary step in understanding fungal allergy and in the development of new and improved methods of diagnosis and therapy. The seven new mould allergens presented here represent four new allergen proteins: aldehyde dehydrogenase (ALDH), enolase, YCP4 (previously found as a Saccharomyces cerevisiae protein of unknown function), and the acidic ribosomal protein, P2. Three of them (ALDH, YCP4 and P2) were found to be allergens in both fungi, Alternaria and Cladosporium. All allergens found so far are cytoplasmic proteins and are rather well conserved in evolution even when comparing distant species. Most of the allergens have "household" functions (ALDH, enolase). One allergen (P2) is a homolog of a very highly conserved human lupus erythematodes (LE) antigen. None of the fungal allergens is clearly related to other known non-fungal allergens.

The immunological relationship of epitopes on major tree pollen allergens.

The major allergens of birch (Bet v I), alder (Aln g I), hazel (Cor a I) and hornbeam (Car b I) were investigated by means of high-resolution two-dimensional electrophoresis combined with immunoblotting. Eleven sera derived from patients allergic to birch pollen as well as mouse monoclonal antibodies BIP 1 and BIP 4, raised against Bet v I, were used as probes. Human IgE antibodies detected 10 spots in birch (Mr 17 kDa, pI 4.9-5.9); four spots in alder (Mr 18.5 kDa, pI 4.7-5.3); four spots in hazel (Mr 17 kDa, pI 5.0-5.8); and 12 + 7 spots in hornbeam (Mr 16.5 kDa, pI 4.9-6.6 and Mr 18 kDa, pI 5.2-6.7), respectively, representing major allergens. Each patient tested reacted in a similar fashion with the spot cluster(s) of a certain allergen. BIP 1 detected the same spot clusters as patients' IgE. BIP 4 reacted with the 17-, 18.5- and 18-kDa spots of birch, alder and hornbeam, but did not react with the 17-kDa spots of hazel and the 16.5-kDa spots of hornbeam. In inhibition experiments with birch pollen extract as inhibitor, IgE binding to Bet v I, as well as to Aln g I, Cor a I and Car b I was abolished, thus suggesting that IgE binding to major tree pollen allergens is confined to shared epitopes. These findings indicate that it might be sufficient to use only Bet v I for diagnostic procedures as well as for immunotherapy in patients with tree pollen allergy.

Distribution of allergens and allergen-coding mRNAs in various tissues of white birch.

The distribution of allergenic proteins was investigated in various tissues of white birch, Betula verrucosa (pollen, leaves and male inflorescences containing immature pollen). In addition, callus and suspension culture cells were investigated for expression of IgE-binding proteins. Furthermore, RNA was extracted from all these tissues and subjected to in vitro translation in a cell-free wheat germ system. Bet v I, the major birch pollen allergen, could be extracted easily from pollen, and in low amounts from callus and leaves. No Bet v I could be extracted from immature male inflorescences. Minor allergens were expressed in high concentrations in pollen and in low concentrations in immature male inflorescences. No minor allergens could be detected in callus and leaves. In contrast to these observations, RNA from all the tissues as well as from callus could be translated in vitro into Bet v I as well as into minor allergens, in particular birch profilin (Bet v II), an important minor allergen. These data suggest that IgE-binding proteins of B. verrucosa, especially Bet v I, under certain circumstances can readily be synthesized in tissues other than pollen. This concept is corroborated by the recent observation that Bet v I reveals high homology with disease resistance response gene products from other plants, suggesting a similar function of Bet v I for the birch.

The influence of oxygen toxicity on yeast mother cell-specific aging.

The effect of deleting both catalase genes and of increased oxygen as well as paraquat (a pro-oxidant) on the replicative life span of yeast mother cells has been investigated to test the so-called oxygen theory of aging. This is well established in higher organisms, but has not been extensively tested in the unicellular yeast model system. Life span determinations were performed in ambient air or in a controlled atmosphere (55% oxygen) and an isogenic series of strains deleted for one or both yeast catalases was used and compared with wild type. In the absence of cellular catalase, increased oxygen caused a marked decrease in life span that could be completely reversed by adding 1 mM GSH, a physiological antioxidant, to the yeast growth medium. In a second unrelated strain, the effects were similar although even the wild type showed a decrease in life span when oxygen was increased. The effect could again be compensated by addition of extracellular GSH. Our results show that manipulating the detoxification of reactive oxygen species has a profound effect on yeast aging. These findings are discussed in the light of recent results relating to oxygen toxicity in the aging process of higher organisms.

Ultrasonic transmission in viscoelastic substances.

PURPOSE: To study the propagation of ultrasonic shock waves in viscoelastic agents and the resulting corneal load. SETTING: University Siegen, Institute for Mechanics and Control Engineering, Siegen, Germany. METHODS: The anterior chamber of a manufactured artificial eye was constructed according to anatomic dimensions. Three openings were drilled--for the phaco tip, for the exchange of a viscoelastic agent or water, and for the shock-wave sensor. The sensor was fixed to the area corresponding to the corneal apex. The sensor signal was analyzed using a direct oscilloscope that measured the amplitude reaching the corneal apex. Shock-wave propagation in several viscoelastic agents was compared with that in balanced salt solution. RESULTS: In hydroxypropyl methylcellulose, the shock wave was amplified or influenced slightly. In hyaluronic-acid preparations, acoustic dampening occurred. CONCLUSION: Removal of hyaluronic-acid derivatives prior to phacoemulsification is not necessary.

Does bracing affect bone density in adolescent scoliosis?

STUDY DESIGN: This is a case-control, cross-sectional analysis of bone density. OBJECTIVES: To determine if bracing during growth affects bone density in adolescent idiopathic scoliosis and whether the effect is local or systemic. SUMMARY OF BACKGROUND DATA: Data concerning the effect of bracing on bone mass in adolescents with idiopathic scoliosis are nonexistent. We were concerned that bone mass loss resulting from long-term brace use may be permanent and may predispose to problems with osteoporosis. METHODS: Healthy adolescent females (n = 85) with scoliosis measuring 20-45 degrees and treated either by brace or observation were studied. Dietary calcium, activity level, body mass index, and pubertal status were evaluated. Scoliosis was measured by Cobb angle. Bone mineral density at the hip and spine were measured by dual energy x-ray absorptiometry to differentiate local versus systemic effects of bracing. Lateral scans of the L3 vertebral body were used to minimize the influence of the pedicles, the effect of the scoliosis, and the interference of the ilium. RESULTS: Mean age, height, and weight were similar between braced and observed groups. After adjusting for curve, Cobb angle, body mass index, activity, and diet, two-way analysis of covariance showed L3 and femoral bone mineral density was the same for braced and observed patients, and pubertal status affected spinal bone mineral density but had no effect on femoral bone mineral density. Pubertal status and body mass index accounted for 53% of the variation in spine bone mineral density and was not affected by brace use. Cobb angle, curve pattern, activity, and diet were not associated with bone mineral density. CONCLUSION: Brace treatment does not adversely affect bone mass at the spine and hip in children with idiopathic scoliosis.

Lack of correlation between peroxidase activity and iodothyronine levels in human and murine thyroid tissues.

Since thyroid peroxidase (TPO) is essential in thyroid hormone biosynthesis, we evaluated whether a correlation exists between TPO activity and iodothyronine stores in human thyroid tissues from nodular goiters with scintigraphically "cold" (N = 10) or "hot" (N = 7) nodules, diffuse toxic goiters (N = 6), and in normal rat thyroids (N = 18). T4, T3 and rT3 were significantly correlated (rs greater than 0.84, P less than 0.001) in human thyroid tissues, although no correlation between TPO activity and any of the iodothyronines was found. Both TPO and the iodothyronines in human goitrous thyroid glands showed great variation which may mask an underlying relationship. However, this is unlikely since no correlation was found between the much less variable TPO and T4 or T3 in the normal murine thyroid. Other factors related to iodothyronine synthesis and secretion must be at least as important as TPO in determining thyroidal stores.

Analysis of thyroglobulins 27S and 19S and their hormonal content in human thyroid glands.

Thyroglobulins (Tg) 27S and 19S and their hormonal content were determined in normal (N = 6), paranodular (N = 4) and hypofunctioning nodular (N = 8) human thyroid tissues. The proportion of Tg 27S and the hormonal content of Tg 19S were significantly decreased in cold nodules. In normal tissues, the molar ratio of T4 in Tg 27S (2.8) was almost twice that found in Tg 19S (1.7), while for T3 the molar ratio in Tg 27S (0.06) was lower than in Tg 19S (0.09). These results indicate that Tg 27S is predominantly formed from well iodinated T4-containing Tg 19S molecules.

Pituitary-thyroid axis in short- and long-term experimental diabetes mellitus.

Short-term experimental diabetes mellitus (DM) produces a significant decrease in serum thyroid hormones, a decreased or normal serum thyroid-stimulating hormone (TSH) and a reduction in hepatic and renal T4-5'-deiodination. However, little is known about the effects of chronic diabetes mellitus on the pituitary-thyroid axis function. We evaluated the changes induced by very short-term (6 days), short-term (15 days) and chronic (6 months) streptozotocin-induced diabetes mellitus in 3-month old female Dutch-Miranda rat serum T4, serum TSH and T4-5'-deiodinase activity in the thyroid and pituitary glands. Serum hormones were determined by specific radioimmunoassays. Iodothyronine-5'-deiodinase activities were assayed in the thyroid and pituitary microsomal fractions using 2 microM T4 as substrate. Mean serum T4 was significantly decreased from 3.3 to 2.0 micrograms/dl 6 days after diabetes mellitus induction, and from 2.2 to 1.5 micrograms/dl after 15 days of DM, with no significant changes in serum TSH, indicating a decreased pituitary TSH responsiveness to the diminished suppression by T4, even though pituitary T4-5'-deiodinase activity was unchanged. Thyroid T4-5'-deiodinase was unchanged after 6 days of diabetes mellitus, but was significantly increased from 20.6 to 37.0 pmol T3/mg protein after 15 days. Six months after diabetes mellitus induction, both serum T4 and thyroid T4-5'-deiodinase returned to normal ranges and serum TSH was unchanged, although pituitary T4-5'-deiodinase was now significantly decreased from 2.7 to 1.7 pmol T3/mg protein. These findings indicate that some kind of adaptation to chronic insulinopenia may occur at the thyroid level, but this does not seem to be true for the pituitary.

Thyroid function in fasting rats: variations in 131I uptake and transient decrease in peroxidase activity.

Serum thyroxine and triiodothyronine, radioiodide thyroid uptake and thyroid peroxidase (TPO) activity were studied over a 2 to 5 day period in fasting rats treated (F+) or not (F-) with TSH. In F- rats, TPO activity was transiently decreased on the 3rd day, whereas in F+ it was always higher than in controls. On the 5th day, the 2 h thyroid uptake of 131I decreased in F-, while the 24 h uptake increased in both F- and F+. Serum T3 and T4 decreased in both fasting groups. Thus, not all effects of fasting on rat thyroid function are reverted by TSH administration, suggesting intrinsic impairment of glandular function.

Nailfold capillaroscopy in diabetes mellitus: morphological abnormalities and relationship with microangiopathy.

The morphology and density of nailfold capillaries were evaluated in patients with Type I and Type II diabetes mellitus and in healthy controls, using videocapillaroscopy. A protocol of well-defined capillary patterns was used. Diabetic patients had more tortuous and enlarged capillaries than controls. Nodular apical elongation was found only in diabetics and was more frequent in patients with a longer history of disease. Type II diabetic patients with chronic clinical complications had a higher frequency of enlarged capillaries and nodular apical elongation. Glycemic control was without influence. Capillary density did not differ among the groups.

Is the infertility in hypothyroidism mainly due to ovarian or pituitary functional changes?

The objective of the present study was to examine whether hypothyroidism affects the reproductive system of adult female rats by evaluating ovarian morphology, uterus weight and the changes in serum and pituitary concentrations of prolactin and gonadotropins. Three-month-old female rats were divided into three groups: control (N = 10), hypothyroid (N = 10), treated with 0.05% 6-propyl-2-thiouracil (PTU) in drinking water for 60 days, and T4-treated group (N = 10), receiving daily sc injections of L-thyroxine (0.8 microg/100 g body weight) during the last 10 days of the experiment. At the end of 50 days of hypothyroidism no hypothyroid animal showed a regular cycle, while 71% of controls as well as the T4-treated rats showed regular cycles. Corpora lutea, growing follicles and mature Graafian follicles were found in all ovaries studied. The corpora lutea were smaller in both the hypothyroid and T4-replaced rats. Graafian follicles were found in 72% of controls and only in 34% of hypothyroid and 43% of T4-treated animals. Serum LH, FSH, progesterone and estradiol concentrations did not differ among the three groups. Serum prolactin concentration and the pituitary content of the three hormones studied were higher in the hypothyroid animals compared to control. T4 treatment restored serum prolactin concentration to the level found in controls, but only partially normalized the pituitary content of gonadotropins and prolactin. In conclusion, the morphological changes caused by hypothyroidism can be a consequence of higher prolactin production that can block the secretion and action of gonadotropins, being the main cause of the changes observed.

Biological and immunological importance of Bet v 1 isoforms.

In 2D-PAGE analysis of Bet v 1, the major birch pollen allergen, up to 12 isoforms can be demonstrated that differ in their isoelectric points from about pH 4.9 to pH 5.9. The molecular weights of these isoforms seem to be rather similar, but minor variations can also be seen. Preliminary experiments with birch leaves seem to indicate that in aging leaves some isoforms can be found that do not occur in pollen. In birch cells cultured in vitro, Bet v 1 isoforms can be induced by bacterial infection that do not occur in pollen (Swoboda et al. (1995), Pant, Cell and Environment 18, 865-874). In a recent paper (Swoboda et al (1995)., J. Biol. Chem. 270, 2607-2613) we show that in natural Bet v 1 from pollen the isoforms are due to different protein sequences. The derived protein sequences of 10 different isoforms (corresponding to 13 different cDNAs) were determined and confirmed by plasma desorption mass spectrometry of purified natural Bet v 1 after trypsin and endoproteinase Glu-C digestion. These experiments also showed that pollen Bet v 1 isoforms were reactive to patients' sera to different degrees and that common post-synthetic modifications (besides N-terminal methionine cleavage) did not occur on Bet v 1. Recombinant isoforms were produced in E. coli, purified and tested with selected patients allergic to birch pollen (Ferreira et al., J. Exp. Med., in the press). The pattern of IgE binding to Bet v 1 isoforms widely differs. Also, T-cell clones from individual patients in some cases are specific to peptides occurring only in certain isoforms. It was of particular interest that three of the naturally occurring pollen Bet v 1 isoforms do not or hardly bind IgE of untreated patients allergic to Bet v 1. However, a comparison of IgE reactivity in patients before and after conventional immunotherapy with natural pollen extract clearly showed that this form of immunotherapy induced IgE to the isoforms that had been unreactive in untreated patients. One of these, Bet v 1d, showed a particularly strong potency towards T-cell stimulation. The isoform(s) that do not bind IgE in untreated patients but still show T-cell reactivity could be potentially utilized for a new form of immunotherapy that avoids the risk of anaphylaxis.

Nailfold capillaroscopy in hypothyroidism and hyperthyroidism: blood flow velocity during rest and postocclusive reactive hyperemia.

Direct intravital microscopic examinations of nailfold capillaries were made in three groups of subjects: 15 healthy volunteers (C) and 11 patients, six with hypothyroidism (h) and five with hyperthyroidism (H). The groups h and H were examined twice, before the onset of treatment and when they returned to euthyroidism. Capillary blood flow velocity (CBFV) was measured during rest and after release of 60-second arterial occlusion. To assess autoregulatory capacity the authors determined peak CBFV postocclusion and time to reach it in single capillaries. In patients with hypothyroidism, before the onset of the treatment, the mean resting and the mean peak CBFV were significantly lower (resting CBFV-group C: 0.93+/-0.11 mm/s (mean+/-SE); group h: 0.33+/-0.09 mm/s; and mean peak CBFV-group C: 1.49+/-0.14 mm/s; group h: 0.79+/-0.19 mm/s). The time to reach mean peak CBFV postocclusion was significantly prolonged (group C: 8.9+/-0.65 s and group h: 19.2+/-2.0 s) compared with the group of healthy volunteers. When these patients achieved euthyroidism, all the studied parameters returned to control levels. In patients with hyperthyroidism only minor changes in CBFV could be detected. In patients with hypothyroidism, the skin microvascular autoregulatory mechanisms are disturbed. The impairments of the reactive hyperemia response could be correlated with the control of the disease (thyroid state).