Urine congophilia associated with preeclampsia does not persist 6-months postpartum.

INTRODUCTION: Preeclampsia is a common hypertensive disorder of pregnancy. Several studies have demonstrated that protein aggregates, detected through urine congophilia, is associated with preeclampsia; however, it has yet to be investigated whether urine congophilia remains postpartum in these women. In this study, we aimed to augment prior studies and determine whether urine congophilia is present postpartum. METHODS: Women were recruited from Lyell McEwin Hospital, South Australia. Urine samples were collected during pregnancy and 6-months postpartum from women with non-preeclampsia pregnancies (n = 48) and women with pregnancies complicated by preeclampsia (n = 42). A Congo Red Dot blot test, total protein and creatinine levels from urine, as well as serum Soluble fms-like tyrosine kinase 1 to placental growth factor ratio (sFlt-1:PlGF), were assessed and correlated. RESULTS: Preeclamptic women exhibited increased urine congophilia (P < 0.01), sFlt-1:PlGF ratio (P < 0.0001) and total protein (P < 0.01) during pregnancy; with a positive correlation between urine congophilia and total protein across the entire cohort (P < 0.0001). Although urine congophilia was no longer detected 6-months postpartum in preeclamptic women, total protein remained elevated (P < 0.05). sFlt-1:PlGF ratio during pregnancy was positively correlated with congophilia across the cohort (P = 0.0007). Serum creatinine was also higher in preeclamptic women during pregnancy (P < 0.001). DISCUSSION: These results support that urine congophilia is significantly elevated in pregnancies complicated with preeclampsia and show that it does not continue postpartum, although larger cohort studies are needed to determine its feasibility as a diagnostic marker.

Aetiology of resuscitated out-of-hospital cardiac arrest treated at hospital.

INTRODUCTION: Precipitating aetiology of out-of-hospital cardiac arrest (OHCA), as confirmed by diagnostic testing or autopsy, provides important insights into burden of OHCA and has potential implications for improving OHCA survivorship. This study aimed to describe the aetiology of non-traumatic resuscitated OHCAs treated at hospital within a local health network according to available documentation, and to investigate differences in outcome between aetiologies. METHODS: Observational retrospective cohort study of consecutive OHCA treated at hospital within a local health network between 2011-2016. Cases without sustained ROSC (≥20 minutes), unverified cardiac arrest, and retrievals to external acute care facilities were excluded. A single aetiology was determined from the hospital medical record and available autopsy results. Survival to hospital discharge was compared between adjudicated aetiologies. RESULTS: In the 314 included cases, distribution of precipitating aetiology was 53% cardiac, 18% respiratory, 3% neurological, 6% toxicological, 9% other, and 11% unknown. A presumed cardiac pre-hospital diagnosis was assigned in 235 (84%) cases, 20% of which were incorrect after exclusion of unknown cases. Rates of survival to hospital discharge varied significantly across aetiologies: cardiac 64%, respiratory 21%, neurological 0%, toxicological 58%, other 32% (p < 0.001). A two-fold difference in survival was observed between cardiac and non-cardiac aetiologies (64% versus 29%, excluding unknown, p < 0.001). CONCLUSIONS: Non-cardiac aetiologies represented a substantial burden of resuscitated OHCA treated at hospital within a local health network and were associated with poor outcome. The results confirmed that true aetiology was not evident on initial examination in 1 in 5 cases with a pre-hospital cardiac diagnosis.

[Evaluation of medical insurance consultant reports within the German Statutory Accident Insurance-Methodology and results of a peer review procedure]. / Evaluation der Durchgangsarztberichte in der Deutschen Gesetzlichen Unfallversicherung ­ Methodik und Ergebnisse eines Peer-Review-Verfahrens.

BACKGROUND: Accident insurance consultants (D-physicians) are qualified specialists with particular expertise in occupational medicine. Within the medical treatment procedure of the German Statutory Accident Insurance (DGUV), D­physicians must make a report on the medical care after occupational accidents. This nationwide evaluation aimed to systematically measure the quality of documentation of these medical reports. Peer review is a common method to ensure process quality. MATERIAL AND METHODS: For each included D­physician 30 reports of more severe cases from 2017 were randomly selected. The reports were anonymized and randomly assigned to a peer reviewer. Peer reviewers used a web-based checklist with nine rating categories and dichotomous response format (deficiency/no deficiency). To evaluate overall quality each report was rated with an overall grade from 1 (very good) to 6 (insufficient). RESULTS: A total of 30,384 reports were evaluated by 82 peer reviewers. One third of the reports contained no deficiencies. Most deficiencies were found in the category on information about the accident. The mean overall grade for each D­physician was 2.6 and ranged from 1.5 (best) to 4.1 (worst). All evaluated D­physicians were given an individual quality report which described the main findings. CONCLUSION: The first nationwide peer review of the DGUV proved to be a practical and valid quality assurance procedure to evaluate the medical reports of D­physicians. The quality of the reports was in general good. The DGUV plans to repeat the peer review process taking further groups of D­physicians into consideration.

Estatus de selenio en equinos Criollo-Chileno a pastoreo y su respuesta a la suplementación / Selenium metabolic status and response to supplementation in grazing Chilean-Criollo horses

Objetivo. Dos experimentos se realizaron para determinar el estatus de selenio en equinos Criollo-Chileno a pastoreo y, evaluar la respuesta a la suplementación con Na2SeO3. Materiales y métodos. Exp.1A caballos pertenecientes a 10 criaderos del sur de Chile se les determinó la actividad sanguínea de glutatión peroxidasa (GPx, EC.1.11.1.9) (AcGPx). Los animales pertenecieron a 3 grupos, pastoreo de otoño (PO, n=40), pastoreo de otoño y suplementados con avena (PO+A, n=47), y, pastoreo de primavera (PP, n=41). Exp2Se utilizaron equinos con carencia de selenio, distribuidos en tres grupos: G1, n=7, tratado con Na2SeO3 el día 0 (Se= 0.05 mg/kg pv, im); G2, n=8, suplementado con Na2SeO3, los días 0 y 15 y GC, n=8, control. La AcGPx se determinó al día 0, 30 y 120. Resultados. La mediana (Me) de la AcGPx en todos los grupos fue menor al límite adecuado (>130 U/gHb), mayor en PO+A (Me=92, P25%=47, P75%=129 U/gHb) y PP (Me=85, P25%=45, P75%=114 U/gHb) que en PO (Me=35, P25%=20, P75%=85 U/gHb) (p<0.05). En el experimento 2 la AcGPx fue similar y menor al adecuado en todos los grupos. La suplementación con Na2SeO3 aumentó (p<0.05) la AcGPx en G1 (121±52 U/gHb) y G2 (124±69 U/gHb), sin alcanzar valores adecuados. Conclusiones. Los equinos Criollo-Chileno a pastoreo en el sur de Chile presentan carencia de Se, mayor en otoño que primavera y menor al suplementar con avena. Adicionalmente, la administración parenteral de Na2SeO3 mejora el estatus de Se, pero sin revertir el cuadro carencial.

Objective. Two experiments were conducted to determine the selenium status in grazing Chilean-Criollo horses and evaluate their response to a Na2SeO3 supplement. Material and methods. Exp.1The blood activity of glutathione peroxidase (GPx, EC.1.11.1.9) (AcGPx) was determined in horses kept grazing in 10 breeding farms from the south of Chile: PO, n= 40 horses were kept grazing during autumn (PO), 47 horses were kept grazing during autumn and supplemented with oat grain (PO+A), and 23 horses were kept grazing during spring (PP). Exp.223 horses kept on pasture were allotted into three groups: G1, n=7, supplemented with a dose of Na2SeO3 (Se= 0.05mg/kg bw, im) on day 0; G2, n=8, supplemented with similar doses of Na2SeO3, on days 0 and 15; and GC, n=8, control. AcGPx was determined on days 0, 30 and 120. Results. Median (Me) of the AcGPx on all the farms were below the adequate limit (>130 U/g Hb), being higher in PO+A (Me= 92, P25%= 47, P75%= 129 U/g Hb) and PP (Me = 85, P25%= 45, P75%=114 U/g Hb) than PO (Me= 35, P25%= 20, P75%= 85 U/g Hb) (p<0.05). Initial AcGPx was similar and below adequate in the three groups and increased after Na2SeO3 administration (p<0.05) in G1 (121 ± 52 U/g Hb) and G2 (124 ± 69 U/g Hb). Conclusions. A deficiency is observed in Chilean-Criollo horses grazing during autumn and spring in southern Chile, parenteral administration of Na2SeO3 (0.05mg/kg bw) in single or double doses improves the Se status without achieving adequate values.

Valoración diagnóstica de enzimas hepáticas en perfiles bioquímicos sanguíneos de vacas lecheras / Diagnostic assessment of liver enzymes in blood profiles from dairy cows

Objetivo. El objetivo del estudio fue determinar la asociación, sensibilidad (Se) y especificidad (Es) de las enzimas glutamato deshidrogenasa (GDH, EC: 1.4.1.3), aspartato aminotransferasa (AST, EC: 2.6.1.1) y g-glutamil transpeptidasa (GGT, EC: 2.3.2.2) como indicadores de daño hepático en perfiles bioquímicos sanguíneos de vacas lecheras. Materiales y métodos. Se analizaron los valores de la actividad plasmática de GDH, AST y GGT, obtenidos de 1566 vacas correspondientes a 112 perfiles bioquímicos realizados en rebaños lecheros. Se determinó la asociación entre la actividad de las enzimas y el área bajo la curva (ABC) y la Se y la Es mediante curva ROC considerando la presencia de daño hepático cuando GDH>30 U/L o GGT>39 U/L. Resultados. La AST presentó mayor asociación con la GDH (r=0.55) que la GGT (r=0.44), así como una mayor ABC (p=0.01) al utilizar GDH>30 U/L para indicar daño hepático. El grado de asociación entre AST y GGT fue de r=0.42. La sensibilidad de la AST para detectar daño hepático, utilizando el punto de corte de referencia del laboratorio (>150 U/L), fue inferior que al emplear GGT (>39 U/L). El punto crítico de máxima Se y Es de la curva ROC para daño hepático se obtuvo con AST>110 U/L, valor que incrementó la concordancia diagnóstica con la GDH (Kappa >0.300), pero no con la GGT. Conclusiones. La actividad plasmática de AST en vacas lecheras se asocia mayormente con la de GDH que con la de GGT y su límite de referencia para predecir daño hepático correspondería a >110 U/L.

Objective. The aim of this study was to establish the association, sensibility (Se) and specificity (Sp) of the plasmatic activity of glutamate dehydrogenase (GDH, EC: 1.4.1.3), aspartate aminotransferase (AST, EC: 2.6.1.1) and g-glutamil transpeptidase (GGT, EC: 2.3.2.2) as liver damage markers in blood profiles of dairy cows. Materials and methods. The records of the plasma activity for GDH, AST and GGT obtained from blood profiles taken from 1566 cows from 112 dairy herds. The association between enzymatic activity, area below the curve (ABC), and Se and Sp using a ROC curve were determined, considering liver damage when GDH >30 U/L or GGT >39 U/L. Results. The AST was more likely associated to GDH (r=0.55) than to GGT (r=0.44). AST also had a higher ABC (p=0.01) when using GDH >30 U/L as an indicator of liver damage. The correlation between AST and GGT was of r=0.42. AST Se to detect liver damage with a cut-off level of >150 U/L was lower than the Se of GGT. The critical point for maximum Se and Sp of the ROC curve was obtained with AST>110 U/L and an increase in the diagnostic concordance with GDH (Kappa>0.300), was obtained using this value. Conclusions. Plasma activity of AST is more associated to GDH than to GGT in dairy cows. The cut-off level of plasma activity of AST for liver damage prediction in dairy cows would be >110 U/L.

Hemoglobinuria posparto en vacas de tres rebaños lecheros de la región del Bío-Bío, Chile / Postpartum haemoglobinuria in dairy cows of the Bio-Bio region, Chile

La hemoglobinuria posparto es una enfermedad metabólica esporádica que presentan las vacas lecheras, manifestándose dentro de las 6 semanas posparto asociada al rápido incremento de la producción de leche al inicio de la lactancia. Se caracteriza por una anemia aguda regenerativa por hemólisis intravascular con hemoglobinuria. Entre los factores que predisponen a su presentación están la ingesta de una dieta carente en fósforo, cobre o selenio, o bien concomitante a la presencia de agentes hemolíticos en plantas crucíferas como el nabo o colza. El trabajo describe aspectos relacionados a la presentación de brotes de la enfermedad en tres rebaños lecheros de la región del Bío-bío, Chile, con hipofosfatemia asociada a la incorporación de derivados de la remolacha azucarera en la dieta.

Characterization of a Yersinia enterocolitica biotype 1A strain harbouring an ail gene.

AIMS: The chromosomal ail gene (attachment and invasion locus) is commonly used as target gene for the detection of pathogenic Y. enterocolitica strains in food testing. The ail PCR does not detect strains of biotype 1A (BT1A), which are regarded as non-pathogenic because BT1A strains lack the virulence plasmid and chromosomally encoded virulence genes. In some recent reports, however, BT1A strains were discovered that harboured the ail gene. We isolated an ail-positive strain and characterized this strain with phenotypic and genotypic methods to study its possible relation to pathogenic Y. enterocolitica strains. METHODS AND RESULTS: The ail region of the BT1A strain was sequenced and compared with the corresponding region of nonpathogenic BT1A strains and pathogenic strains. Pulsed field gel electrophoresis (PFGE) analysis was applied revealing no similarity of the PFGE pattern of this strain to the patterns of pathogenic strains. Virulence-gene-based PCR analyses showed the strain to be positive for ystB, but negative for virulence genes ystA, virF and yadA. Whole-cell MALDI-TOF MS combined with a shrinkage discriminant analysis approach was applied and clearly classified the ail-positive biotype 1A strain within the cluster of BT1A strains. CONCLUSIONS: PCR detection of ail sequences in food matrices should be followed by the isolation of the responsible strain and its characterization using phenotypic or genotypic methods. SIGNIFICANCE AND IMPACT OF THE STUDY: The ail gene may be present in Y. enterocolitica BT1A strains, which are commonly considered as nonpathogenic. Efficient methods such as PCR typing of other virulence genes or rapid MALDI-TOF MS-based bacterial profiling allow a more comprehensive assessment of the pathogenicity potential of Yersinia strains.

Distribution and genetic variability among Campylobacter spp. isolates from different animal species and humans in Switzerland.

In Switzerland, a national database with 1028 Campylobacter isolates from poultry, pigs, cats, dogs, cattle, humans, zoo animals and water has been created. The database contains the genetic fingerprint and background information of each Campylobacter isolate. Dominant species could be identified in the different sources with a majority of Campylobacter jejuni in poultry (73%), humans (79%), cattle (95%), zoo animals (40%) and water (100%), of Campylobacter coli in pigs (72%), and of Campylobacter upsaliensis/helveticus in cats and dogs (55%). The comparison of three genotyping methods, amplified fragment length polymorphism (AFLP), pulsed field gel electrophoresis and restriction fragment length polymorphism, revealed that AFLP allows discrimination between the different Campylobacter species and is the most appropriate method to distinguish specific strains within the same species. Genotyping analysis demonstrated that the Campylobacter population is heterogeneous among the different sources and that no dominant clone is spread in the country. Genotyping and the resulting database are useful tools to trace back future Campylobacter infections.

Effect of acute hypoxia on maximal oxygen uptake and maximal performance during leg and upper-body exercise in Nordic combined skiers.

We examined the effect of normobaric hypoxia (3200 m) on maximal oxygen uptake (VO2max) and maximal power output (Pmax) during leg and upper-body exercise to identify functional and structural correlates of the variability in the decrement of VO2max (DeltaVO2max) and of maximal power output (DeltaPmax). Seven well trained male Nordic combined skiers performed incremental exercise tests to exhaustion on a cycle ergometer (leg exercise) and on a custom built doublepoling ergometer for cross-country skiing (upper-body exercise). Tests were carried out in normoxia (560 m) and normobaric hypoxia (3200 m); biopsies were taken from m. deltoideus. DeltaVO2max was not significantly different between leg (-9.1+/-4.9%) and upper-body exercise (-7.9+/-5.8%). By contrast, Pmax was significantly more reduced during leg exercise (-17.3+/-3.3%) than during upper-body exercise (-9.6+/-6.4%, p<0.05). Correlation analysis did not reveal any significant relationship between leg and upper-body exercise neither for DeltaVO2max nor for DeltaPmax. Furthermore, no relationship was observed between individual DeltaVO2max and DeltaPmax. Analysis of structural data of m. deltoideus revealed a significant correlation between capillary density and DeltaPmax (R=-0.80, p=0.03), as well as between volume density of mitochondria and DeltaPmax (R=-0.75, p=0.05). In conclusion, it seems that VO2max and Pmax are differently affected by hypoxia. The ability to tolerate hypoxia is a characteristic of the individual depending in part on the exercise mode. We present evidence that athletes with a high capillarity and a high muscular oxidative capacity are more sensitive to hypoxia.

Phenon cluster analysis as a method to investigate epidemiological relatedness between sources of Campylobacter jejuni.

AIMS: To develop a method for assessing the relative epidemiological significance of possible infection sources for human campylobacteriosis. METHODS AND RESULTS: Using fluorescent amplified fragment length polymorphism (AFLP), 243 apparently epidemiologically unrelated Campylobacter jejuni isolates were genotyped (77 human, 46 cattle, 49 pet and 71 poultry isolates). In total 136 different phena were identified, of which 48 were clusters grouping at least two isolates. Isolates from different sources were frequently clustered together, underlining the high degree of source mixing and the lack of host specificity of C. jejuni. The phena were classified into different phenon types according to the sources of the isolates they contained. The occurrence of these phenon types was analysed using an area-proportional Euler diagram to describe epidemiological relatedness among C. jejuni isolates. Group separation statistics revealed that 43% of analysed human isolates expressed maximum similarity to other human isolates, 9% to cattle isolates, 21% to pet isolates and 27% to poultry isolates; these results were in accordance with the pattern observed in the phenon cluster analysis. CONCLUSIONS: Based on the grouping of strains into molecular similarity clusters, ecological patterns between sources can be investigated. SIGNIFICANCE AND IMPACT OF THE STUDY: This approach is a new methodological contribution to establish the relative epidemiological significance of concurrent infection sources.

Detection of Mycobacterium avium subspecies paratuberculosis in Swiss dairy cattle by real-time PCR and culture: a comparison of the two assays.

AIMS: To compare the two different diagnostic assays for the detection of Mycobacterium avium ssp. paratuberculosis, the aetiological agent of paratuberculosis. METHODS AND RESULTS: Faecal samples were derived from 310 cows, representing 13 commercial dairy herds in various locations in Switzerland with expected increased risk because of a past history of disease. Detection assays for M. avium ssp. paratuberculosis were culture (gold standard) and a newly designed real-time PCR. Real-time PCR identified 31 of 310 animals as positive within this risk population whereas culture identified 20 positive animals. The specificity of real-time PCR was confirmed by DNA sequencing of the PCR product. Depending on the test used, the paratuberculosis prevalence in our tested risk population ranged from 6.5 to 10%. CONCLUSIONS: Real-time PCR and culture data were in good agreement, and real-time PCR generates data in a short time in contrast to culture. SIGNIFICANCE AND IMPACT OF THE STUDY: We consider real-time PCR as a suitable alternative method to culture for the detection of M. avium ssp. paratuberculosis in a national surveillance programme.

Campylobacter spp. in dogs and cats in Switzerland: risk factor analysis and molecular characterization with AFLP.

A 1-year cross-sectional study was carried out to determine the prevalence, risk factors for carriage, and genetic diversity of Campylobacter spp. in healthy dogs and cats in Switzerland. Veterinary practitioners collected samples from 1268 animals (all ages) presented for vaccination. The prevalence of Campylobacter spp. in 634 dogs and 596 cats that were eligible for the study was 41.2% (confidence interval 95%: 37.3-45.1%) and 41.9% (CI 95%: 37.9-46%), respectively. Risk factors identified for carriage of Campylobacter jejuni were found to be different from risk factors for C. upsaliensis/C. helveticus. Young animals (< or =3 years) had significantly higher odds of carrying C. upsaliensis/C. helveticus than older animals (OR 1.8-3.3), whereas for C. jejuni carriage, the age was not a risk factor. Amplified fragment length polymorphism (AFLP) genotyping revealed heterogeneity among C. jejuni strains and was found to clearly separate C. helveticus from C. upsaliensis. It was shown that cats more often carry C. helveticus with an estimated prevalence of 28.2%, whereas dogs mainly are carrying C. upsaliensis.

Genetic diversity and antibiotic resistance patterns in a campylobacter population isolated from poultry farms in Switzerland.

The diversity and genetic interrelation of Campylobacter jejuni and C. coli isolated from Swiss poultry were assessed by three independent typing methods. Samples were derived prior to slaughter from 100 randomly selected flocks (five birds per flock) raised on three different farm types. The observed flock prevalence was 54% in total, with 50% for conventional and 69% for free-range farms. Birds held on farms with a confined roaming area had the lowest prevalence of 37%. Campylobacter isolates were characterized by amplified fragment length polymorphism (AFLP), restriction fragment length polymorphism of flaA PCR fragments (flaA-RFLP), and disk diffusion testing for eight antimicrobial agents that are commonly used in veterinary or human medicine in Switzerland. Analysis of the genotypic results indicates that the Campylobacter population in Swiss poultry is genetically highly diverse. Nevertheless, occasionally, isolates with identical or nearly identical characteristics were isolated from different farms or farm types in different locations. Genetic typing by AFLP and flaA-RFLP was found to be complementary. The majority of isolates (67%) were susceptible to all tested antibiotics; however, single, double, and triple resistances were observed in 7%, 23%, and 2% of the strains, respectively. There was no correlation between genotype and antibiotic resistance. Surprisingly, sulfonamide resistance was frequently found together with streptomycin resistance. Our findings illustrate the results of common genetic exchange in the studied bacterial population.

[Incidence of zoonoses in petting zoos and evaluation of hygiene measures to prevent the transmission to humans]. / Vorkommen von Zoonosen in Streichelzoos und Beurteilung der Hygienemassnahmen für die Ubertragung auf den Menschen.

In summer 2003, a study was performed in thirty Swiss petting zoos with the objective to determine the prevalence of zoonotic agents, and to describe hygiene measures implemented to reduce the risk of human infection. Fecal samples from different animal species were collected from the floor of pens to determine the prevalence of Salmonella spp., Campylobacter spp., verocytotoxin producing E. coli/ VTEC and Francisella tularensis. A questionnaire on hygiene measures, number of animals per species, housing system, care procedures and feeding was administered to every petting zoo to estimate exposure of visitors to zoonotic microorganisms. In total, 423 fecal samples were examined. Of these samples, 41 were positive for Campylobacter spp., which were mainly isolates from pigs and poultry (35% positive samples from each species). In pigs, 50% of the positive samples (6 samples) were typed as C. jejuni. The others were typed as C. coli (3) and C lan' (3), respectively. Five poultry isolates were typed as C. jejuni, and two as C. coli. Two samples were positive for Salmonella spp. Salmonella typhimurium was isolated from a goat, the other isolate could not be identified by serotyping. Neither Francisella tularensis nor verocytotoxin producing E. coli/ VTEC were found. The low prevalence of zoonotic microorganisms in Swiss petting zoos could be attributed to the cleanness of enclosures and animals, low stocking rates and good animal care. However, there is room for improvement concerning visitors' information on hygiene and hand washing. Furthermore, a strict separation between picnic - areas and animals should be enforced.

Regulatory gene expression in skeletal muscle of highly endurance-trained humans.

AIM AND BACKGROUND: Changes in regulatory and structural gene expression provide the molecular basis for the adaptation of human skeletal muscle to endurance exercise. HYPOTHESIS: The steady-state levels of multiple mRNAs mainly involved in regulatory functions differ between highly endurance-trained and untrained subjects in a muscle heavily recruited during the exercise. METHODS: Biopsies from musculus vastus lateralis of seven untrained (UT) subjects [maximal oxygen consumption (VO2max) = 39 mL kg-1 min-1] and seven trained (T) professional cyclists (VO2max = 72 mL kg-1 min-1) were analysed for the contents of 597 different mRNAs using commercially available cDNA arrays (Clontech no. 7740-1). Intra-individual expression profiles were compared by least-square linear regression analysis. Differences in gene expression between the two groups were tested for statistical significance using L1 regression analysis combined with the sign test on all permutations of scatter plots of log raw values from UT vs. T subjects. RESULTS: Transcripts for 144 of 597 genes were sufficiently abundant to be analysed quantitatively. The expression profiles of the T group had a better intragroup correlation (R2) than those of the UT group (0.78 vs. 0.65, P < 0.05). An intergroup (T vs. UT) correlation of expression profiles gave an R2 of 0.71. Statistical analysis at a false discovery rate of 5% identified differential expression of nine cell-regulatory genes between T and UT. The mRNA levels of eight genes, including two DNA repair enzymes, transcription factors, signal transducers, a glycolytic enzyme and a factor involved in steroid hormone metabolism were increased in T vs. UT. Conversely, the mRNA of the tumour suppressor APC was downregulated with endurance training. Selective reverse-transcriptase polymerase chain reaction experiments confirmed the signal estimates from the array analysis. CONCLUSIONS: The repetitive impact of the complex exercise stimuli in professional cyclists attenuated the interindividual differences in regulatory gene expression in skeletal muscle. Long-term nuclear reprogramming of regulatory gene expression seems to be characteristic of human musculus vastus lateralis in a highly endurance-trained steady state.

The chick and human collagen alpha1(XII) gene promoter--activity of highly conserved regions around the first exon and in the first intron.

A single gene encodes collagen XII, an extracellular matrix protein with three large fibronectin-related subunits connected via a short collagen triple helix. Since collagen XII is a component of a specific subset of collagen fibrils in tissues bearing high tensile stress, we are interested to know how its restricted expression is regulated. To this aim, we have isolated the region around the first exon of both the chick and human collagen alpha1(XII) gene. The upstream sequences of the two genes share common features but are not related. Strong similarity starts about 100 bp 5' of the first exon and ends 100 bp into the first intron. In addition, two large conserved regions (56-63% similarity) were found in the first intron. A single major and two clusters of minor transcription start sites were identified in both the chick and human gene. To test for promoter activity, conserved fragments from the chick gene were cloned into reporter plasmids for transient transfection of fibroblasts. A 70-bp stretch containing a conserved nuclear factor-1 binding sequence just upstream of the first transcription start site was found to work as a basal promoter. An adjacent, but nonoverlapping short segment including the more downstream start sites and a conserved TATTAA sequence exhibited independent promoter activity. GC-rich sequences just 5' and 3' of the minimal promoter fragments were required for full activity. In contrast, inclusion of more upstream sequences (up to 2.4 kb) had no effect. The two conserved regions in the first intron showed no promoter activity on their own but modulated activity when linked to autologous or heterologous promoters. Specifically, one of these intronic regions might contain enhancer element(s) that respond to mechanical stress acting on the fibroblasts. We conclude that the collagen XII gene is driven by a basal promoter with two halves that can act independently; conserved control regions are located around the first exon and in the first intron.

Expression of fos and jun genes in human skeletal muscle after exercise.

It is believed that the induction of the fos and jun gene family of transcription factors might be at the origin of genetic events leading to the differential regulation of muscle-specific genes. We have investigated the effect of a 30-min running bout in untrained subjects on the expression of the mRNAs of all members of the fos and jun gene families, including c-fos, fosB, fosBdel, fra-1, and fra-2 as well as c-jun, junB, and junD. While the fos family members were transiently upregulated 10- to 20-fold (an exception being fra-2) the induction of the jun family members was up to 3-fold only. The induction of c-fos could also be demonstrated at the protein level. Both c-fos and c-jun mRNAs were coinduced in muscle fiber nuclei. The induction was not restricted to a particular fiber type, as expected from established muscle fiber recruitment schemes, but followed a "patchy" pattern confined to certain regions of the muscle. The signals leading to the expression of these immediate early genes are therefore unclear.