Biomedical subjects

A Aström

Publications and source records attributed to A Aström.

At least 19 recordsLinked to original sources

PubMed1999

Sound exposure as a measure against driver drowsiness.

Twelve lorry drivers participated in a field study, including in total 110 tests of a waking sound system. The system was based on a time and frequency varied sound, generated at intervals between 1 and 15 min for periods of 3-7 s. The waking effect of the sound was analysed through subjective ratings. Exposure to the sounds was correlated to improvements of the immediate as well as the long-term self-reported changes in wakefulness. The results of the study show consistently positive assessments concerning the applied waking system, both regarding the system as a measure for raised wakefulness, increased road safety and the interest in having the system installed in the vehicle. The waking sounds were in no case felt to be annoying. Analyses of the wakefulness and other factors and the interaction related to fatigue and driving are included in the investigation.

Accidents, Traffic↗

PubMed1998

Keratinocyte-specific retinoid regulation of human cellular retinoic acid binding protein-II (hCRABPII) gene promoter requires an evolutionarily conserved DR1 retinoic acid-responsive element.

Transcription of the hCRABPII gene is retinoid inducible in human skin keratinocytes (KC) but, surprisingly, not in cultured cells. The promoter for the gene harbors three putative nuclear receptor binding sites: DR5, upstream of the transcription start site; DR1 (DR1d), distal to the site; and DR1 (DR1p), a proximal variant. DR1d, but not DR1p, is conserved between human and mouse. Although DR5 has been found to be a retinoid receptor target in COS-1 cells, the function of DR1 remains unknown. We examined the functions of these DR in retinoid regulation of the hCRABPII promoter in human KC. In reporter gene assays, no significant retinoid response was observed in the promoter in cultured KC; however, overexpression of retinoid receptor heterodimers RARgamma x RXRalpha restored the response. Gel supershift assays showed that endogenous RARgamma x RXRalpha levels are much lower in cultured KC than in skin in vivo. Ligand-binding assays showed that cultured KC contain only one-third of the level of retinoic acid receptor (RAR) and one-eighth of the level of retinoid X receptor found in KC in skin. Deletion of the DR1d or DR5 sites reduced retinoid-induced promoter activity by 63% and 27%, respectively. Isolated DR1d and DR5 sites, but not DR1p, efficiently bound RARgamma-RXRalpha and conferred RAR-selective retinoid responsiveness on a heterologous promoter. These data indicate that: (i) the previously reported lack of retinoid regulation of endogenous hCRABPII gene transcription in cultured KC is likely due to insufficient levels of RARgamma x RXRalpha, but not their cofactors; (ii) the conserved DR1d site is the major functional target in RARgamma x RXRalpha regulation of hCRABPII in KC; (iii) the DR1p site is nonfunctional due to its lack of affinity for RARgamma x RXRalpha, although its half-sites share high sequence homology with the consensus retinoid receptor-binding half-site.

Base Sequence↗

PubMed1998

Laboratory studies of a sound system that maintains wakefulness.

Two studies investigated the effects of a waking sound that enhances wakefulness. Study I investigated the effect of the sound level and Study 2 the effect of time and frequency variability of the sound. The recordings of EEG and subjective ratings were analysed to study the effect upon wakefulness. The waking effect increased when sound varied in duration and frequency. A number of specific conditions necessary for the waking effect are described. The exposure should be based on high frequency sounds and several tones chosen to produce disharmony. The exposure should be loud enough to be heard over the masking background noise. The duration and tonal quality should be variable from one presentation to another.

Accidents, Traffic↗

PubMed1997

Load pattern and pressure pain threshold in the upper trapezius muscle and psychosocial factors in medical secretaries with and without shoulder/neck disorders.

OBJECT: A current hypothesis for the genesis of muscular complaints in the shoulder/neck region postulates that short periods with a completely relaxed muscle are essential to avoid complaints. Another hypothesis is that these disorders are related to psychosocial conditions at work. In order to test these hypotheses, 23 medical secretaries were investigated. METHODS: The load pattern during work in the upper trapezius muscle bilaterally was assessed with electromyographic (EMG) technique and exposure variation analysis (EVA). In addition, pressure pain threshold (PPT) was measured on the trapezius muscle bilaterally and on the sternum. Psychosocial conditions at work were assessed with a questionnaire. RESULTS: The medical secretaries with complaints had significantly fewer episodes with totally or close to totally relaxed muscle compared with the healthy group. The group with complaints tended to have a more monotonous load pattern at low levels (approx. 1%-5% maximum voluntary contraction) while the healthy group had more frequent pauses but also somewhat more frequent short load peaks. The group with complaints showed lower PPT readings compared with the healthy group. However, the whole group had considerably lower PPTs than is usually reported in the literature. Of the 12 questions in the psychosocial questionnaire only one regarding work task satisfaction showed a significant difference between the two groups. CONCLUSION: Support is found for hypothesis that secretaries without complaints have more frequent episodes with totally relaxed muscle. A significant difference is found regarding work task satisfaction.

Adult↗

PubMed1996

Normal levels of the vitamin D receptor and its message in psoriatic skin.

Treatment with vitamin D3 analogs improves psoriasis. The vitamin D receptor (VDR) mediates most, if not all, the effects of vitamin D3. The purpose of this study was to determine the levels of the VDR mRNA and VDR protein in normal and in involved and uninvolved psoriatic skin. Although VDR mRNA was not detected by Northern analysis of human skin samples, it was readily detectable by use of the more sensitive ribonuclease protection assay. The VDR mRNA levels were normal in acute guttate as well as in chronic plaque lesions. There was also no difference in VDR mRNA levels between normal and uninvolved psoriatic skin. The VDR protein was detected by Western analysis using the monoclonal 9A7 gamma anti-VDR antibody and a polyclonal rabbit anti-VDR antibody. For comparison, VDR levels were analyzed in cultures of normal human keratinocytes and the epithelial cell line MCF-7. Studies of the extraction procedures for VDR showed that at least 60% of Escherichia coli-expressed VDR added to the skin biopsy specimens was recovered. The VDR concentration in normal human adult skin was approximately 50 pg/microgram protein, and the concentrations of VDR in involved and uninvolved psoriatic skin were of the same order of magnitude. Using the 9A7 gamma anti-VDR antibody, the VDR (M(r) 53,000) was constantly present in lower amounts than a band of M(r) 80,000 in both skin specimens and keratinocyte cultures. This high-molecular-weight band is most likely a cross-reacting protein not related to VDR, because it was absent when using the polyclonal anti-VDR antibody.

Adult↗

PubMed1995

Retinoic acid antagonizes basal as well as coal tar and glucocorticoid-induced cytochrome P4501A1 expression in human skin.

Cytochrome P4501A1 is known to be expressed in skin and thus has been implicated in the pathogenesis of skin cancer due to certain environmental carcinogens. Retinoic acid (RA) has been used in chemoprevention of certain skin and other epithelial cancers. Therefore, we used Northern and Western analysis to determine the effect of externally applied RA on basal P4501A1 expression. RA reduced basal levels of P4501A1 mRNA and protein by 68 (n = 14, P = 0.005) and 75% (n = 7, P = 0.04) respectively. RA application also reduced basal levels of P4501A2 (another P4501A1 subfamily member) mRNA by 93% (n = 7, P = 0.001) as determined by reverse transcription-polymerase chain reaction. Interestingly, P4501A1 mRNA expression induced by coal tar or glucocorticoid (clobetasol) was reduced 46 (n = 10, P = 0.003) and 69% (n = 5, P < 0.05) respectively by RA co-application. Downregulation of basal P4501A1 expression and antagonism of coal tar mediated P4501A1 induction by RA may be a mechanism involved in chemo-prevention of skin and other epithelial cancer by RA.

Adult↗

PubMed1995

Regulation of cellular retinoic acid binding protein expression in rhino mouse skin by all-trans-retinoic acid.

Cellular retinoic acid binding proteins (CRABP) are cytoplasmic proteins that bind all-trans-retinoic acid (RA) and other retinoids. The purpose of these studies was to determine the effects of topically applied RA on CRABP expression in rhino mouse skin. CRABP-II mRNA was significantly induced (3- to 4.5-fold) by a single dose of RA at 6 and 16 h after RA treatment, with a return to control levels at 48 h. CRABP-II message was not significantly elevated by 3 or 4 consecutive days of RA treatment, when assessed 24 h after the last treatment. CRABP-I mRNA was undetectable in control and RA-treated skin. We used radiolabelled RA binding combined with non-denaturing PAGE blot autoradiography to distinguish the CRABP subtypes. By this protein assay method, increases in CRABP-II were detected 24 and 48 h after a single application of RA, as well as after 3 and 4 days of RA treatment. RA treatment did not alter CRABP-I expression relative to the vehicle control. These results demonstrate that in mouse skin CRABP-II, but not CRAB-I, is inducible by RA, and is similar to how RA regulates CRABP in human skin.

Animals↗

PubMed1995

Budesonide is metabolized by cytochrome P450 3A (CYP3A) enzymes in human liver.

Budesonide is a synthetic glucocorticosteroid that is commonly used in topical treatment of asthma and rhinitis. The main metabolites formed from budesonide in human liver microsomes have been identified as 16 alpha-hydroxyprednisolone and 6 beta-hydroxy-budesonide. Although it is apparent that the cytochrome P450 (CYP) system is involved, the actual subfamily has not been identified. In attempts to do this, budesonide was incubated with microsomes from ten different human liver samples where various CYP activities had been rank ordered. We found a strong correlation between formation of the two main metabolites and testosterone 6 beta-hydroxylation (correlation 0.98 and 0.95), a marker for CYP3A. When budesonide (10 microM) was incubated with human liver microsomes in the presence of compounds known to interact with different isoforms or subfamilies of CYP, ketoconazole was found to be the strongest inhibitor of budesonide metabolism (IC50: approximately 0.1 microM) followed by troleandomycin (IC50: approximately 1 microM), erythromycin, and cyclosporin, all substances known to interact with CYP3A isoenzymes. Substances known to interact with CYP2C (sulfaphenazole, mephenytoin, and tolbutamide) and with CYP2D6 (bufuralol and quinidine) did not specifically inhibit the metabolism of budesonide. In addition, formation of the budesonide metabolites (16 alpha-hydroxyprednisolone and 6 beta-hydroxybudesonide) was inhibited by antibodies against the CYP3A subfamily, but not by antibodies against the CYP1A subfamily or control immunoglobulin G. We conclude that the formation of 16 alpha-hydroxyprednisolone and 6 beta-hydroxybudesonide from budesonide is catalyzed by isoenzymes within the CYP3A subfamily.

Administration, Topical↗

PubMed1994

Retinoic acid induction of human cellular retinoic acid-binding protein-II gene transcription is mediated by retinoic acid receptor-retinoid X receptor heterodimers bound to one far upstream retinoic acid-responsive element with 5-base pair spacing.

We previously cloned the human cellular retinoic acid-binding protein-II (CRABPII) gene and demonstrated a rapid and transient increase in retinoic acid (RA)-dependent transcription in cultured human skin fibroblasts. To determine whether retinoid receptors could regulate CRABPII gene transcription, cotransfection experiments were performed. When RAR alpha was cotransfected in Cos-1 cells with a reporter construct containing -8.0 kilobases of the upstream region, an 18-fold RA induction was obtained. By deletion analysis, a region essential for RA induction located approximately -5.6 kilobases upstream from the human CRABPII gene start site was identified. Sequencing and mutational analysis identified a direct repeat (GGGTCAttggaAGGACA) with 5-base pair spacing (DR-5) that is critical for RA-mediated induction of human CRABPII gene transcription. This is different from the mouse CRABPII gene in which two RAREs (DR-1 and DR-2) are required for full activation. To determine whether RAR and RXR can bind to the human CRABPII RARE, gel retardation assays were performed. In these assays, in vitro translated RAR alpha and RXR alpha were found to bind efficiently as heterodimers in gel retardation assays; weak binding of RAR alpha homodimers was observed. These data demonstrate that the human CRABPII gene is regulated by a far upstream RARE that most efficiently binds RAR-RXR heterodimers.

Animals↗

PubMed1994

Multiple forms of poly(A) polymerases in human cells.

We have cloned human poly(A) polymerase (PAP) mRNA as cDNA in Escherichia coli. The primary structure of the mRNA was determined and compared to the bovine PAP mRNA sequence. The two sequences were 97% identical at the nucleotide level, which translated into 99% similarity at the amino acid level. Polypeptides representing recombinant PAP were expressed in E. coli, purified, and used as antigens to generate monoclonal antibodies. Western blot analysis using these monoclonal antibodies as probes revealed three PAPs, having estimated molecular masses of 90, 100, and 106 kDa in HeLa cell extracts. Fractionation of HeLa cells showed that the 90-kDa polypeptide was nuclear while the 100- and 106-kDa species were present in both nuclear and cytoplasmic fractions. The 106-kDa PAP was most likely a phosphorylated derivative of the 100-kDa species. PAP activity was recovered in vitro by using purified recombinant human PAP. Subsequent mutational analysis revealed that both the N- and C-terminal regions of PAP were important for activity and suggested that cleavage and polyadenylylation specificity factor (CPSF) interacted with the C-terminal region of PAP. Interestingly, tentative phosphorylation sites have been identified in this region, suggesting that phosphorylation/dephosphorylation may regulate the interaction between the two polyadenylylation factors PAP and CPSF.

Animals↗

PubMed1994

Retinoic acid induces expression of early growth response gene-1 (Egr-1) in human skin in vivo and in cultured skin fibroblasts.

The early growth response gene, Egr-1 (NGFI-A, krox 24, zif 268, TIS 8), is a member of a family of genes with suggested importance in the regulation of cell growth and differentiation. Retinoic acid has been shown to markedly induce Egr-1 gene expression in mouse embryonal carcinoma cells and rat preosteoblastic cells. In this study we demonstrate that treatment of cultured human skin fibroblasts with retinoic acid results in a rapid transient four-fold induction of Egr-1 transcripts, being maximum at 60 min and returning to a basal level by 120 min. However, treatment of cultured human keratinocytes with retinoic acid did not significantly induce Egr-1 transcripts. Expression of Egr-1 message in keratinocytes was observed to be induced by fetal bovine serum and tetra-decanoyl phorbol acetate, whereas calcium, 1,25-dihydroxyvitamin D3, and cyclic adenosine monophosphate caused little or no induction. Topical application of 0.1% retinoic acid cream in vivo resulted in a two- to threefold induction of Egr-1 transcripts following treatment for 24 and 48 h, returning to nearly basal levels by 96 h. Taken together, these data are consistent with the possibility that Egr-1 is a proximal component of an intracellular molecular cascade that may give rise to alterations in cell phenotype in response to retinoic acid.

Blotting, Northern↗

PubMed1994

Retinoic acid induces expression of PA-FABP (psoriasis-associated fatty acid-binding protein) gene in human skin in vivo but not in cultured skin cells.

PA-FABP (psoriasis-associated fatty acid-binding protein) is a new member of a group of low-molecular-weight proteins that are highly up-regulated in psoriatic skin and that share similarity to fatty acid-binding proteins. In this study we demonstrate that PA-FABP transcripts are expressed in human skin in vivo and that topical application of 0.05% retinoic acid (RA) cream results in a rapid induction of PA-FABP transcripts following treatment for 16 hours and persists at increasing levels after 48 and 96 h of RA treatment. The PA-FABP mRNA response to RA was reduced by approximately 50% when patients concurrently were treated with RA and 0.025% clobetasol propionate (CLO) for 48 and 96 h, whereas treatment with CLO alone resulted in PA-FABP transcript levels not significantly different from vehicle-treated skin. When comparing the effects of a well-known irritant, sodium lauryl sulfate (SLS), to those of RA and its vehicle, 0.05% RA cream but not 2% SLS in RA vehicle caused PA-FABP mRNA induction after 16 h. SLS treatment of human skin for 96 h caused a slight increase in PA-FABP transcripts, but markedly less than that observed in response to RA treatment. Incubation of cultured human keratinocytes or skin fibroblasts with RA for up to 48 h did not significantly induce PA-FABP transcripts. Expression of PA-FABP message in keratinocytes was observed to be induced by calcium and fetal calf serum (FCS), while tetra-decanoyl phorbol acetate (TPA) caused little or no induction.(ABSTRACT TRUNCATED AT 250 WORDS)

Base Sequence↗

PubMed1993

Expression, purification, and binding properties of human cellular retinoic acid-binding protein type I and type II.

Human cellular retinoic acid-binding protein (CRABP) type I and type II were expressed in Escherichia coli from cloned cDNAs. Expressed proteins were purified by gel filtration and ion-exchange chromatography, resulting in highly pure proteins. The yield after gel filtration was approximately 50 mg/liter bacterial culture. In binding studies the equilibrium dissociation constant, Kd, of retinoic acid (RA) for E. coli-derived CRABP-I and CRABP-II was 6.8 and 39 nM, respectively. The Kd of the synthetic retinoid analog CD 367 was 2.2 nM for CRABP-I and 3.0 nM for CRABP-II. RA competed with the binding of CD 367 to CRABP-I and CRABP-II with IC50 values of 20.0 and 90.0 nM, respectively. Retinoid analogs competed with the binding of CD 367 to CRABP-I and CRABP-II in the following order: (p-[(E)-2-(5,6,7,8-tetrahydro-5,5,8,8- tetramethyl-2-naphtyl)-1-propenyl]-benzoic acid (TTNPB) > 4-oxo-RA > 4-OH-RA > 13-cis-RA = 9-cis-RA. m-carboxy-TTNPB and CD 271 were found not to compete with the binding of CD 367 to CRABP-I or CRABP-II even at 500-fold molar excess. These data demonstrate that E. coli-derived CRABP-I has a higher affinity for RA than CRABP-II and that retinoic acid metabolites have a lower affinity for these proteins. The observed difference in affinity for RA supports the idea that CRABP-I, which is constitutively expressed, and CRABP-II, which is induced by RA, have different functions in the cell. In addition, 9-cis-RA, a natural ligand for the retinoid X receptors, is not a physiological ligand for either CRABP-I or CRABP-II.

Binding, Competitive↗

PubMed1992

Structure of the human cellular retinoic acid-binding protein II gene. Early transcriptional regulation by retinoic acid.

The gene for human cellular retinoic acid-binding protein II (CRABP-II) has been cloned. It was isolated from a human placenta genomic library and is contained within one bacteriophage clone. The gene spans 6 kilobases and consists of 4 exons. One major transcription initiation site was mapped to an A residue 137 nucleotides upstream of the ATG initiation codon. The upstream region contains a TATA box and potential AP2, Sp1, and Krox-24 binding sites, as well as a direct repeat with homology to a retinoic acid-responsive element. The CRABP-II mRNA was rapidly induced within 2-6 h in cultured human skin fibroblasts by retinoic acid, reaching a plateau after 6 h of treatment. The rapid increase of CRABP-II message was mainly due to an increased rate of transcription as determined by nuclear run-on experiments. Increased transcription could be detected as early as 1 h after addition of retinoic acid, peaked at 2 h, and returned to basal levels within 6 h. On-going protein synthesis was required for this transient increase of transcription, since the induction was blocked by cycloheximide. These data suggest that the human CRABP-II gene is transcriptionally regulated by a newly synthesized regulatory protein.

Amino Acid Sequence↗

PubMed1992

Properties of a HeLa cell 3' exonuclease specific for degrading poly(A) tails of mammalian mRNA.

A HeLa cell 3'-exonuclease with properties of a mammalian mRNA poly(A) tail-removing enzyme has been characterized. The exonuclease shows high specificity for the poly(A) tail, and it is single strand-specific and requires a 3'-hydroxyl group for its activity. During degradation 5'-AMP is liberated as a product, and a 3'-OH group is left on the last adenosine residue of the remaining poly(A) tail. The activity is inhibited by 5'-AMP and can be competed by poly(A)-containing mRNA or poly(A). Based on these findings we propose a reaction pathway for poly(A) tail removal catalyzed by the HeLa cell poly(A) tail-specific 3' exonuclease.

Base Sequence↗

PubMed1992

Assignment of the human CRABP-II gene to chromosome 1q21 by nonisotopic in situ hybridization.

Two highly conserved forms of cellular retinoic acid binding protein (CRABP-I and CRABP-II) have been described, and one, CRABP-II, is highly expressed in human skin. We have utilized a 10-kb fragment containing the human CRABP-II (hCRABP-II) gene (isolated from a human genomic library) to localize hCRABP-II to human chromosome 1 band q21 by fluorescence in situ hybridization. Localization to 1q was confirmed by hybridization of a hCRABP-II cDNA clone against a human-mouse hybrid cell line containing a t(1;6)(q21;q13)) translocation chromosome. The hCRABP-II gene is therefore localized to a band known to contain several other genes that are expressed in the context of epidermal differentiation, including profilaggrin, loricrin, involucrin, and calcyclin.

Animals↗

PubMed1992

Retinoic acid receptors and binding proteins in human skin.

Nuclear retinoic acid receptors (RAR) are likely to mediate many of the pleiotypic cutaneous actions of retinoids by acting as ligand-dependent enhancer factors. The presence of nuclear RAR in skin was confirmed by identification of a 45-kDa nuclear RA binding activity by fast protein liquid chromatography (FPLC). Analysis of RNA extracted from skin specimens demonstrated expression of RAR-alpha and RAR-gamma transcripts, as well as expression of the homologous low-affinity receptor, RXR-alpha. Both isoforms of RAR-gamma RAR-gamma 1 and RAR-gamma 2 were detectable, with RAR-gamma 1 being the more strongly expressed. FPLC analysis also demonstrated a 15-kDa peak of specific RA binding activity, consistent with the presence of cellular retinoic acid binding protein (CRABP). Of the two known forms of CRABP, CRABP-II was much more strongly expressed than CRABP-I at the level of steady-state mRNA. CRABP-II was also expressed in keratinocytes and fibroblasts in vitro. CRABP-II was up-regulated by agents that induce keratinocyte differentiation, and inhibited by prolonged exposure to high concentrations of RA. In contrast, CRABP-II was consistently induced by RA in dermal, but not in lung fibroblasts. CRABP-I was expressed at low to undetectable levels under all these conditions. The presence of tissue-specific and differentiation-related regulation of CRABP-II suggests that it may be an important regulator of RA action in human skin.

Carrier Proteins↗

PubMed1992

Differential regulation of retinoic acid receptors and binding proteins in human skin.

Many of the pleiotropic effects of retinoids are likely to be mediated by nuclear retinoic acid receptors (RAR) acting as ligand-dependent enhancer factors. However, in previous studies we have been unable to document altered RAR expression at the RNA level in response to retinoic acid (RA) treatment or in psoriatic lesions, conditions characterized by marked alterations in keratinocyte proliferation and differentiation, which are either caused by or responsive to RA. In an attempt to identify other potential regulators of RA responsiveness, we have used RNA blot hybridization to study the expression of the cellular retinoic acid binding proteins (CRABP) CRABP-I and CRABP-II, the RAR-gamma isoforms RAR-gamma 1 and RAR-gamma 2, and the low-affinity RAR homologue RXR in normal, RA-treated, and psoriatic human epidermis. CRABP-II is selectively and markedly induced by RA in adult human skin (J Biol Chem 266:17662-17666, 1991). However, in submerged, serum-free keratinocyte cultures, CRABP-II mRNA could not be induced by RA. Comparisons of intact human skin, submerged keratinocyte cultures, and human skin equivalent cultures indicated that induction of CRABP-II by RA requires epidermal stratification, dermal-epidermal interactions, or both. CRABP-II transcripts were also expressed in heat-separated human dermis at levels similar to those found in epidermal keratome biopsies, whereas CRABP-I transcripts were undetectable in dermal RNA. CRABP-II transcripts were markedly elevated in psoriatic lesions, as they were in RA-treated skin. In contrast, CRABP-I mRNA was undetectable and not increased in psoriatic lesions. Expression of RAR-gamma isoforms and RXR was not detectably altered in either psoriatic lesions or in RA-treated skin. Thus, altered expression of CRABP-II appears more likely to regulate the cutaneous actions of RA than does altered expression of CRABP-I, RXR, or RAR-gamma isoforms. From these and other results, a model for regulation of RA action involving sequestration of RA by CRABP-II is proposed.

Base Sequence↗