Redefining the overtraining syndrome as the unexplained underperformance syndrome.
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Humans↗
Biomedical subjects
M Lehmann
Publications and source records attributed to M Lehmann.
Expression of myosin heavy chain isoforms in skeletal muscle of patients with peripheral arterial occlusive disease.
PURPOSE: Peripheral arterial occlusive diseases (PAODs) not only compromise blood flow but lead to a series of subsequent metabolic and structural changes in the relevant muscles. Changes in myofibrillar proteins (eg, of myosin heavy chain [MHC] isoforms), one of the determinants of muscle structure as well as of muscular function, have not been reported in patients with PAOD and were therefore the aim of this study. METHODS: Thirteen consecutive patients with PAOD were examined (clinical stage according to Fontaine II, three patients; III, three patients, and IV, seven patients) and compared with five age-matched control patients who had been in traffic accidents. A calf muscle sample (gastrocnemius muscle) in the ischemic region was taken for MHC isoform analysis by sodium dodecyl sulfate polyacrylamide gel electrophoresis and silver stain, and the relative content of MHC isoforms was measured. RESULTS: Compared with the control patients, there was no significant change of MHC isoforms in patients with PAOD II. In patients with PAOD III, MHC IIb decreased significantly (P <.05) although MHC IIa remained unchanged; in patients with PAOD IV, both MHC IIa and IIb decreased significantly (P <.05). Accordingly, there was a progressive increase of the relative amount of MHC I with more critical ischemia in PAOD. CONCLUSION: In patients with PAOD, the content of MHC II decreased with a higher grade of ischemia. That seems to be consistent with an increased resistance to ischemia for myosin isoforms in the order of I more than in IIa more than IIb. Whether the decrease of MHC II in patients with PAOD is related to atrophy of muscle fibers or to muscle-fiber transition must be investigated further.
Aged↗
Training of junior rowers before world championships. Effects on performance, mood state and selected hormonal and metabolic responses.
BACKGROUND: Few data have been published on training of competitive athletes and about metabolic, hormonal and psychological reactions to overreaching (transient over-training) and tapering in successful athletes. METHODS: Training was recorded and effects on mood state and metabolic and hormonal responses were examined in 10 rowers and spares of the coxed eight during preparation for the World Championships 1995. Mood state was determined using the Recovery-Stress-Questionnaire for Athletes. Resting morning blood parameters as well as performance were measured every week over a period of five weeks. RESULTS: Very high training loads of approximately 3.2 hours per day were sustained for 18 days. Maximum performance (Pmax) and maximum lactate (Lamax) were decreased during high-load training phases (overreaching), Pmax, Lamax and endurance increased after the tapering period. There were decreases in gonadal and hypothalamic steroid hormones (fsh, 1h, prolactin, testosterone) during overreaching and increases in these hormones in tapering. Both performance and hormonal indices of training load were reflected by deterioration of recovery in the Recovery-Stress-Questionnaire for Athletes. CONCLUSIONS: Clear signs of overreaching were found after 18 days of intense training of about 3 h.d(-1) in these highly-trained athletes, i.e. decreases in performance, gonadal and hypothalamic steroid hormones and deterioration of recovery in the psychological questionnaire. After tapering values returned to baseline values before the World Championship. The findings indicate that overreaching is an integral part of successful training regimens and can be analyzed by a multi-factorial approach involving biological and psychometric data.
Adolescent↗
Intragraft overexpression of interleukin-4 is neither sufficient nor essential for tolerance induction to cardiac allografts in a high-responder strain combination.
BACKGROUND: Recently we have demonstrated that the nondepleting anti-CD4 monoclonal antibody (mAb) RIB5/2 induces long-term acceptance of kidney and heart allografts in all rat strain combinations tested. Cytokine gene expression studies by reverse transcriptase-polymerase chain reaction revealed a reversed intragraft interleukin (IL)-4/interferon-gamma ratio. Whether IL-4 mediated immune deviation contributes to transplantation tolerance is not clear so far. METHODS: To learn more about the functional relevance of the relative IL-4 up-regulation, IL-4 was overexpressed in rat heart allografts by using ex vivo adenoviral gene transfer. The efficiency of gene transfer was analyzed by reporter gene assays as well by reverse transcriptase-polymerase chain reaction analysis of IL-4 mRNA expression. RESULTS: The intragraft overexpression of IL-4 did not prolong the allograft survival compared with controls. Moreover, neutralization of IL-4 by OX81 mAb did not prevent tolerance induction by RIB5/2 treatment. CONCLUSIONS: Anti-CD4 mAb-induced tolerance is associated with an intragraft type1/type2 shift, however, the up-regulation of IL-4 alone is neither sufficient nor essential to induce tolerance to cardiac allografts in a high-responder strain combination.
Animals↗
Inactivation of Rho signaling pathway promotes CNS axon regeneration.
Regeneration in the CNS is blocked by many different growth inhibitory proteins. To foster regeneration, we have investigated a strategy to block the neuronal response to growth inhibitory signals. Here, we report that injured axons regrow directly on complex inhibitory substrates when Rho GTPase is inactivated. Treatment of PC12 cells with C3 enzyme to inactivate Rho and transfection with dominant negative Rho allowed neurite growth on inhibitory substrates. Primary retinal neurons treated with C3 extended neurites on myelin-associated glycoprotein and myelin substrates. To explore regeneration in vivo, we crushed optic nerves of adult rat. After C3 treatment, numerous cut axons traversed the lesion to regrow in the distal white matter of the optic nerve. These results indicate that targeting signaling mechanisms converging to Rho stimulates axon regeneration on inhibitory CNS substrates.
ADP Ribose Transferases↗
The helix-loop-helix proteins dAP-4 and daughterless bind both in vitro and in vivo to SEBP3 sites required for transcriptional activation of the Drosophila gene Sgs-4.
The expression of Sgs genes in the salivary gland of the third instar larva of Drosophila is a spatially restricted response to signalling by the steroid hormone 20-hydroxyecdysone. For Sgs-4, we have previously demonstrated that its strictly tissue and stage-specific expression is the result of combined action of the ecdysone receptor and secretion enhancer binding proteins (SEBPs). One of these SEBPs, SEBP2, was shown to be the product of the homeotic gene fork head. Together with SEBP3, SEBP2 appears to be responsible for the spatial restriction of the hormone response of Sgs-4. Here, we show that SEBP3 is a heterogeneous binding activity that consists of different helix-loop-helix (HLH) proteins. We cloned the Drosophila homologue of human transcription factor AP-4 (dAP-4) and identified it as one of these HLH proteins. The dAP-4 protein shows great similarity to its human and Caenorhabditis counterparts within the bHLHZip domain, the second leucine zipper dimerization motif, and a third region of unknown function. The expression pattern of dAP-4 indicates that it is a ubiquitously expressed HLH protein in Drosophila. As a second component of SEBP3 we identified the Daughterless (Da) protein, which is also ubiquitously expressed and binds to SEBP3 sites independent of dAP-4. Since both dAP-4 and Da can be detected in situ at transposed Sgs-4 transcriptional control elements in polytene salivary gland chromosomes, we propose that each of the two proteins contributes to the transcriptional control of Sgs-4.
Amino Acid Sequence↗
Distinct tolerance pathways in sensitized allograft recipients after selective blockade of activation signal 1 or signal 2.
BACKGROUND: CD4-targeted therapy or blocking of CD28-B7 T-cell costimulation may produce indefinite cardiac allograft survival in presensitized rats. This study analyzes the immune events associated with tolerance pathways after the blockade of activation signal 1 (CD4 monoclonal antibody [mAb]) or signal 2 (CTLA4Ig). METHODS AND RESULTS: Lewis rats sensitized with Brown Norway skin grafts reject LBNF1 cardiac allografts in <36 hr. Animals were treated with RIB-5/2, a nondepleting CD4 mAb, or with CTLA4Ig + LBNF1 spleen cells. RIB-5/2 monotherapy uniformly produced permanent cardiac graft acceptance, whereas CTLA4Ig produced indefinite graft survival in about 50% of sensitized rats. Spleen cells (100 x 10(6)) from CD4 mAb-treated rats conferred donor-specific tolerance after transfer into new sets of recipients. This tolerant state could be then transferred with regulatory cells in an infectious manner into new cohorts of engrafted rats. In contrast, features of infectious tolerance could be detected in CTLA4Ig-treated hosts after infusion of >300 x 10(6) of splenocytes. CD4 mAb therapy abolished the transcription of both T helper (Th)1 and Th2 cytokines compared with rejecting controls. In contrast, CTLA4Ig treatment resulted in a selective sparing of Th2-type cytokines. Surviving grafts in both groups were largely protected from signs of chronic rejection. CONCLUSIONS: CD4 mAb-induced blockage of activation signal 1 or CTLA4Ig-mediated blockage of costimulatory signal 2 may induce a true transplantation tolerance in sensitized rats, as documented by permanent graft acceptance and attenuation of chronic injury. The infectious pathway operates in a cell dose-dependent manner. Th2-type deviation in the graft itself is not required for tolerance maintenance, and it does not necessarily lead to chronic injury.
Abatacept↗
An ISFET-algal (Chlamydomonas) hybrid provides a system for eco-toxicological tests.
A cellular sensoring system was designed in which metabolism-dedicated pH-ISFETs and the unicellular green alga Chlamydomonas reinhardtii as a biological component, were combined. The system permits on-line detection of pH changes caused by the metabolic and photosynthetic activities of the cells. Photosynthetic activity results in a basification of the medium caused by uptake of CO2. In darkness, an acidification of the medium, resulting from the production of CO2 by degradation of starch was observed. Both, acidification and basification, are sensitive indicators for the physiological activity of the alga. Experiments using inhibitors of energy metabolism or photosynthesis illustrate the utility of this system for an on-line monitoring of substances of eco-toxicological importance.
Animals↗
Measurement of biodegradable substances using the salt-tolerant yeast Arxula adeninivorans for a microbial sensor immobilized with poly(carbamoyl)sulfonate (PCS). Part II: Application of the novel biosensor to real samples from coastal and island regions.
A microbial sensor for rapid measurement of the amount of biodegradable substances based on the salt-tolerant yeast Arxula adeninivorans LS3 has been developed especially for coastal and island regions. Our parameter, the so-called sensorBOD, that is available after only a few minutes, agrees with the 5-day value for the biochemical oxygen demand (BOD5) very well. We have employed the Arxula sensor in the short-time estimation and supervision of the BOD of both domestic and industrial wastewater with high salinity. The novel sensor makes it possible to monitor the different types of wastewater rapidly without pretreatment, and it can be used for an active process control of sewage treatment works. Compared to a commercially available sensor, the novel sensor achieves better agreement between sensorBOD and BOD5 measurements with salt containing samples.
Ascomycota↗
Organization of point contacts in neuronal growth cones.
Growth cones from rat dorsal root ganglia plated on laminin contain integrin clusters over the entire growth cone surface, and growth cones make transient adhesions at sites called point contacts. We examined, by immunocytochemistry and confocal microscopy, the composition and distribution of point contacts in neuronal growth cones. Vinculin was concentrated in the central domain of growth cones and at the tips of filopodia. Vinculin was specifically associated with integrin clusters at the membrane-substrate interface and thus marked point contacts. The cytoskeletal proteins paxillin and talin colocalized with beta1 integrin in a subpopulation of clusters restricted to the central domain of the growth cone and to the tips of filopodia. The neuron-specific kinase, FAK+ also distributed with the vinculin-positive clusters. The Rho family proteins RhoA, RhoB, and Cdc42 were present in growth cones, and a few Rho clusters were colocalized with vinculin. Examination of proteins resistant to detergent extraction in PC12 cells confirmed the retention of beta1 integrin, paxillin, talin, and vinculin with the cytoskeleton. Moreover, we detected FAK+ and RhoA in the detergent-resistant cytoskeleton, supporting their distribution to point contacts. Our observations indicate that two types of integrin clusters are present in growth cones: those associated with vinculin at the cell substratum interface, and those not associated with vinculin. Point contacts are mature adhesion sites defined by the presence of both beta1 integrin and vinculin, and they are associated with signaling proteins.
Animals↗
Neutron Laue diffraction studies of coenzyme cob(II)alamin.
Using a recently designed neutron single-crystal diffractometer utilizing a narrow-band Laue concept (LADI), diffraction data were collected from a crystal of the coenzyme cob(II)alamin (B12r), crystallized from a mixture of D2O and perdeuterated acetone. The instrument was placed at the end of a cold neutron guide at the Institute Laue Langevin (ILL, Grenoble, France), and data collection with neutrons of 1.8-8.0 A wavelength to a crystallographic resolution of 1.43 A was complete after about 36 h. This compares favourably with a previous experiment utilizing the same crystal specimen, where more than four weeks were required to collect monochromatic diffraction data to about 1 A resolution. Using the Laue data, the structure was solved by molecular replacement with the known X-ray crystal structure. Difference density maps revealed the atomic positions (including deuterium atoms) of seven ordered solvent water molecules and two (partially disordered) acetone molecules. These density maps were compared with corresponding maps computed with monochromatic neutron-diffraction data collected to 1. 0 A resolution using the same crystal specimen, as well as to maps derived from high-resolution (0.90 A) synchrotron X-ray data. In spite of the better definition of atomic positions in the two high-resolution maps, the 1.43 A LADI maps show considerable power for the determination of the location of hydrogen and deuterium positions.
Coenzymes↗
Prevention of autoimmune but not allogeneic destruction of grafted islets by different therapeutic strategies.
Grafting autoimmune-diabetic recipients with allogeneic islets, graft rejection and disease recurrence as major problems of reaching indefinite survival and tolerance induction have to be solved. Anti-CD25 and anti-CD4 monoclonal antibodies were successfully used after allogeneic islet transplantation in experimentally diabetic rats. A temporary anti-CD25 therapy also prevented disease recurrence in autoimmune-diabetic BB rats, while this was not yet reported for an anti-CD4 treatment. In autoimmune-diabetic NOD mice disease recurrence can be successfully treated using an anti-CD4 monoclonal antibody. We, therefore, compared the efficacy of a short-term anti-CD25 and anti-CD4 treatment regarding the prevention of allograft rejection and disease recurrence in autoimmune-diabetic BB/OK rats. Both monoclonal antibodies were combined with low doses of Cyclosporin A. Untreated BB/OK rats relapsed into hyperglycaemia within 3 weeks independent of the islet donor, LEW.1A, LEW.1BB/OK or BB/OK rats. However, after grafting MHC-identical allogeneic (LEW.1BB/OK) or syngeneic (BB/OK) islets we observed about 30% spontaneous acceptance. Both the anti-CD25 and anti-CD4 therapy significantly prolonged the survival of allogeneic grafted islets. After MHC-identical allogeneic and syngeneic islet transplantation the temporary immunotherapy increased the proportion of permanent acceptors to 63% and 75%, respectively. The efficacy of both treatment strategies in prolonging allograft survival and prevention of disease recurrence was identical. In summary, anti-CD25 as well as anti-CD4 therapy prevented autoimmune but not allogeneic islet destruction in autoimmune-diabetic BB/OK rats. In conclusion, targeting different immune cells by monoclonal antibodies with different specificities can lead to very similar results with respect to an interruption of allograft rejection and autoimmune reaction.
Animals↗
Expression of interferon regulatory factors and indoleamine 2,3-dioxygenase in Chlamydia trachomatis-infected synovial fibroblasts.
Synovial fibroblasts probably represent host cells for Chlamydia trachomatis during initial intra-articular infection in reactive arthritis. In vitro synovial cells produce interferon-beta (IFN-beta) in response to chlamydial infection. IFN-beta expression can be activated by interferon regulatory factor-1 (IRF-1) and interferon-stimulated gene factor 3gamma (ISGF3gamma). In this study, we demonstrate that infection of synovial fibroblasts with C. trachomatis serotype D induced the expression of IRF-1 mRNA as shown by reverse transcription-PCR. Tumor necrosis factor-alpha (TNF-alpha) stimulation enhanced IRF-1 mRNA levels in infected cells and was required to detect IRF-1 protein by immunoblotting. The level of constitutively expressed IRF-2 was not significantly affected after infection. C. trachomatis was found to cause an up-regulation of ISGF3gamma protein in synovial cells. Induction of the tryptophan-catabolizing enzyme indoleamine 2,3-dioxygenase (IDO) is an important mechanism of the host cell response to control intracellular infection by chlamydiae. It has been described that IRF-1 can induce IDO gene expression. Infection of synovial fibroblasts alone in the absence of exogenous cytokine induced the expression of IDO mRNA which was enhanced by TNF-alpha treatment. The stimulation of IRF-1, ISGF3gamma, and IDO expression was most effective when viable chlamydiae were used as inoculum. Neutralization of IFN-beta in the culture medium of infected cells diminished but did not abrogate expression of IRF-1, ISGF3gamma, and IDO. The increased production of IRF-1 and ISGF3gamma in C. trachomatis-infected synovial fibroblasts may contribute to induction of IFN-beta and IDO.
Antibodies↗
Detection of Chlamydia trachomatis DNA in cervical samples with regard to infection by human papillomavirus.
OBJECTIVE: The correlation between human papillomavirus (HPV) and Chlamydia, trachomatis infections was evaluated in 144 patients with normal cytology or with atypical squamous cells of undetermined significance (ASCUS). METHODS: Cervical samples were analysed using polymerase chain reaction (PCR) and non-radioactive Southern blot analysis. Specificity and sensitivity of two C. trachomatis PCR systems: major outer membrane protein (MOMP)-PCR and plasmid-PCR were determined. Southern blot hybridization of the PCR amplicons was done using 5' and 3' biotinylated oligonucleotide probes. RESULTS: All cervical samples were tested by the plasmid-PCR due to a 10 times higher sensitivity compared to the MOMP-PCR. To determine the specificity of our C. trachomatis primer sets different bacteria and viruses which can cause urogenital infections were analysed. Comparison of the probes revealed an increased sensitivity of the 5' and 3' double-biotinylated probe vs. the 5' biotinylated probe. The infection rate of C. trachomatis in cervical samples of HPV-positive patients was 10.3% (three out of 29) vs. 1.7% (two out of 115; P< or =0.05) in HPV-negative patients. In patients HPV-X (unsequenced HPV-types) positive the rate was 14.3% (one out of seven) vs. 2.9% (four out of 137: P = 0.2) in HPV-X negative patients. In high risk (HR) HPV-positive cervical samples the infection rate was 9.1% (two out of 22) vs. 2.5% (three out of 122; P = 0.14) in HR HPV-negative samples. Chlamydia trachomatis frequency of patients with cytological changes (ASCUS) was 27.3% (three out of 11) vs. 1.5% (two out of 1 33) in patients with normal cytology (P = 0.003). The highest prevalence rate of C. trachomatis-positive cervical samples (50%; one out of two) was found in HR HPV-positive patients with cytological changes (ASCUS) vs. 5% (one out of 20) in HR HPV-positive patients with normal cytology (P = 0.17). Patients negative for HPV and positive for ASCIIS have a C. trachomatis rate of 22.2% (two out of nine) vs. HPV-negative patients with normal cytology (none out of 106; P = 0.006) and vs. HR HPV-negative patients with normal cytology (0.9%; one out of 113; P = 0.014). CONCLUSIONS: There appears to be a correlation between cervical HPV and cervical C. trachomatis infections. The prevalence rate of C. trachomatis was significantly higher in patients with abnormal cytology (ASCUS) vs. normal cytology.
Blotting, Southern↗
Measurement of biodegradable substances using the salt-tolerant yeast Arxula adeninivorans for a microbial sensor immobilized with poly(carbamoyl) sulfonate (PCS) Part I: Construction and characterization of the microbial sensor.
A microbial biosensor based on the yeast Arxula adeninivorans LS3 has been developed for measurement of biodegradable substances. Arxula is immobilized in the hydrogel poly(carbamoyl) sulfonate (PCS). The immobilized yeast membrane is placed in front of an oxygen electrode with -600 mV versus Ag/AgCl. Arxula is salt tolerant; it can give a stable signal up to 2.5 M NaCl in sample (120 mM in measuring cell). The sensor's measurements are highly correlated to BOD5 measurements. It has a very high stability which can last for 40 day without any decrease in signal. The linear range of the sensor is up to a corresponding BOD value of 550 mg/l.
Biodegradation, Environmental↗
Biogenesis of alpha6beta4 integrin in a human colonic adenocarcinoma cell line involvement of calnexin.
The heterodimer alpha6beta4 is a major integrin and the main laminin receptor in epithelia. The alpha6 integrin subunit is proteolytically cleaved, probably by furin, and glycosylated during its biosynthesis. In the present work, we have investigated the kinetics of the assembly process of alpha6beta4 heterodimers in the colonic adenocarcinoma cell line HT29-D4. We demonstrate that the association of alpha6 and beta4 precursors occurs within the ER, while the endoproteolytic cleavage of pro-alpha6 occurs later, probably in the trans-Golgi network. When pro-alpha6 was blocked within the ER by treatment with brefeldin A, its maturation processing was completely prevented without any consequence on its association with beta4 subunit. Low temperature (20 degrees C) also blocked pro-alpha6 maturation, like brefeldin A, but in addition impaired the integrin assembly. Calnexin, an ER resident protein chaperone, was found to be associated with both the alpha6 and beta4 subunit precursors. Our data suggest that calnexin might be responsible for the prolonged retention of pro-alpha6 within the ER compartment and for the defect of integrin subunit association observed at low temperature.
Adenocarcinoma↗
Structure and regulation of the salivary gland secretion protein gene Sgs-1 of Drosophila melanogaster.
The Drosophila melanogaster gene Sgs-1 belongs to the secretion protein genes, which are coordinately expressed in salivary glands of third instar larvae. Earlier analysis had implied that Sgs-1 is located at the 25B2-3 puff. We cloned Sgs-1 from a YAC covering 25B2-3. Despite using a variety of vectors and Escherichia coli strains, subcloning from the YAC led to deletions within the Sgs-1 coding region. Analysis of clonable and unclonable sequences revealed that Sgs-1 mainly consists of 48-bp tandem repeats encoding a threonine-rich protein. The Sgs-1 inserts from single lambda clones are heterogeneous in length, indicating that repeats are eliminated. By analyzing the expression of Sgs-1/lacZ fusions in transgenic flies, cis-regulatory elements of Sgs-1 were mapped to lie within 1 kb upstream of the transcriptional start site. Band shift assays revealed binding sites for the transcription factor fork head (FKH) and the factor secretion enhancer binding protein 3 (SEBP3) at positions that are functionally relevant. FKH and SEBP3 have been shown previously to be involved in the regulation of Sgs-3 and Sgs-4. Comparison of the levels of steady state RNA and of the transcription rates for Sgs-1 and Sgs-1/lacZ reporter genes indicates that Sgs-1 RNA is 100-fold more stable than Sgs-1/lacZ RNA. This has implications for the model of how Sgs transcripts accumulate in late third instar larvae.
Amino Acid Sequence↗
Biophysical characterization of fungal phytases (myo-inositol hexakisphosphate phosphohydrolases): molecular size, glycosylation pattern, and engineering of proteolytic resistance.
Phytases (myo-inositol hexakisphosphate phosphohydrolases) are found naturally in plants and microorganisms, particularly fungi. Interest in these enzymes has been stimulated by the fact that phytase supplements increase the availability of phosphorus in pig and poultry feed and thereby reduce environmental pollution due to excess phosphate excretion in areas where there is intensive livestock production. The wild-type phytases from six different fungi, Aspergillus niger, Aspergillus terreus, Aspergillus fumigatus, Emericella nidulans, Myceliophthora thermophila, and Talaromyces thermophilus, were overexpressed in either filamentous fungi or yeasts and purified, and their biophysical properties were compared with those of a phytase from Escherichia coli. All of the phytases examined are monomeric proteins. While E. coli phytase is a nonglycosylated enzyme, the glycosylation patterns of the fungal phytases proved to be highly variable, differing for individual phytases, for a given phytase produced in different expression systems, and for individual batches of a given phytase produced in a particular expression system. Whereas the extents of glycosylation were moderate when the fungal phytases were expressed in filamentous fungi, they were excessive when the phytases were expressed in yeasts. However, the different extents of glycosylation had no effect on the specific activity, the thermostability, or the refolding properties of individual phytases. When expressed in A. niger, several fungal phytases were susceptible to limited proteolysis by proteases present in the culture supernatant. N-terminal sequencing of the fragments revealed that cleavage invariably occurred at exposed loops on the surface of the molecule. Site-directed mutagenesis of A. fumigatus and E. nidulans phytases at the cleavage sites yielded mutants that were considerably more resistant to proteolytic attack. Therefore, engineering of exposed surface loops may be a strategy for improving phytase stability during feed processing and in the digestive tract.
6-Phytase↗