55 research outputs found
Anaerobic work capacity in cycling: the effect of computational method
Purpose To compare the anaerobic work capacity (AnWC, i.e., attributable anaerobic mechanical work) assessed using four
diferent approaches/models applied to time-trial (TT) cycle-ergometry exercise.
Methods Fifteen male cyclists completed a 7Ă4-min submaximal protocol and a 3-min all-out TT (TTAO). Linear relationships between power output (PO) and submaximal metabolic rate were constructed to estimate TT-specifc gross efciency
(GE) and AnWC, using either a measured resting metabolic rate as a Y-intercept (7+ YLIN) or no measured Y-intercept
(7-YLIN). In addition, GE of the last submaximal bout (GELAST) was used to estimate AnWC, and critical power (CP) from
TTAO (CP3ÂŽAO) was used to estimate mechanical work above CP (Wâ, i.e., âAnWCâ).
Results Average PO during TTAO was 5.43±0.30 and CP was 4.48±0.23 Wâkgâ1. The TT-associated GE values were~22.0%
for both 7+ YLIN and 7-YLIN and~21.1% for GELAST (both P<0.001). The AnWC were 269±60, 272±55, 299±61, and
196±52 Jâkgâ1 for the 7+ YLIN, 7-YLIN, GELAST, and CP3ÂŽAO models, respectively (7+ YLIN and 7-YLIN versus GELAST, both
PLIN, 7-YLIN, and GELAST versus CP3ÂŽAO, all PLIN,
7-YLIN, and GELAST, typical errors in AnWC values ranged from 7 to 11 Jâkgâ1, whereas 7+ YLIN, 7-YLIN, and GELAST versus
CP3ÂŽAO revealed typical errors of 55â59 Jâkgâ1.
Conclusion These fndings demonstrate a substantial disagreement in AnWC between CP3ÂŽAO and the other models. The
7+ YLIN and 7-YLIN generated 10% lower AnWC values than the GELAST model, whereas 7+ YLIN and 7-YLIN generated
similar values of AnWC
Dissection of the Role of PfEMP1 and ICAM-1 in the Sensing of Plasmodium falciparum-Infected Erythrocytes by Natural Killer Cells
BACKGROUND: Host innate immunity contributes to malaria clinical outcome by providing protective inflammatory cytokines such as interferon-Îł, and by shaping the adaptive immune response. Plasmodium falciparum (Pf) is the etiologic agent of the most severe forms of human malaria. Natural Killer (NK) cells are lymphocytes of the innate immune system that are the first effectors to produce interferon-Îł in response to Pf. However, the molecular bases of Pf-NK cell recognition events are unknown. Our study focuses on the role of Pf erythrocyte membrane protein 1 (PfEMP1), a major Pf virulence factor. PfEMP1 is expressed on parasitized-erythrocytes and participates to vascular obstruction through the binding to several host receptors. PfEMP1 is also a pivotal target for host antibody response to Pf infection. METHODOLOGY/PRINCIPAL FINDINGS: Using genetically-engineered parasite mutant strains, a human genetic deficiency, and blocking antibodies, we identified two receptor-ligand pairs involved in two uncoupled events occurring during the sensing of Pf infection by NK cells. First, PfEMP1 interaction with one of its host receptor, chondroitin sulfate A, mediates the cytoadhesion of Pf-infected erythrocytes to human NK cell lines, but is not required for primary NK cell activation. Second, intercellular adhesion molecule-1 (ICAM-1), another host receptor for PfEMP1, is mandatory for NK cell interferon-Îł response. In this case, ICAM-1 acts via its engagement with its host ligand, LFA-1, and not with PfEMP1, consistent with the obligatory cross-talk of NK cells with macrophages for their production of interferon-Îł. CONCLUSION/SIGNIFICANCE: PfEMP1-independent but ICAM-1/LFA-1-dependent events occurring during NK cell activation by Pf highlight the fundamental role of cellular cooperation during innate immune response to malaria
Efficient transfer of two large secondary metabolite pathway gene clusters into heterologous hosts by transposition
Horizontal gene transfer by transposition has been widely used for transgenesis in prokaryotes. However, conjugation has been preferred for transfer of large transgenes, despite greater restrictions of host range. We examine the possibility that transposons can be used to deliver large transgenes to heterologous hosts. This possibility is particularly relevant to the expression of large secondary metabolite gene clusters in various heterologous hosts. Recently, we showed that the engineering of large gene clusters like type I polyketide/nonribosomal peptide pathways for heterologous expression is no longer a bottleneck. Here, we apply recombineering to engineer either the epothilone (epo) or myxochromide S (mchS) gene cluster for transpositional delivery and expression in heterologous hosts. The 58-kb epo gene cluster was fully reconstituted from two clones by stitching. Then, the epo promoter was exchanged for a promoter active in the heterologous host, followed by engineering into the MycoMar transposon. A similar process was applied to the mchS gene cluster. The engineered gene clusters were transferred and expressed in the heterologous hosts Myxococcus xanthus and Pseudomonas putida. We achieved the largest transposition yet reported for any system and suggest that delivery by transposon will become the method of choice for delivery of large transgenes, particularly not only for metabolic engineering but also for general transgenesis in prokaryotes and eukaryotes
A comparative analysis of body psychotherapy and dance movement psychotherapy from a European perspective
Brugia malayi microfilariae adhere to human vascular endothelial cells in a C3-dependent manner
Brugia malayi causes the human tropical disease, lymphatic filariasis. Microfilariae (Mf) of this nematode live in the bloodstream and are ingested by a feeding mosquito vector. Interestingly, in a remarkable co-evolutionary adaptation, Mf appearance in the peripheral blood follows a circadian periodicity and reaches a peak when the mosquito is most likely to feed. For the remaining hours, the majority of Mf sequester in the lung capillaries. This circadian phenomenon has been widely reported and is likely to maximise parasite fitness and optimise transmission potential. However, the mechanism of Mf sequestration in the lungs remains largely unresolved. In this study, we demonstrate that B. malayi Mf can, directly adhere to vascular endothelial cells under static conditions and under flow conditions, they can bind at high (but not low) flow rates. High flow rates are more likely to be experienced diurnally. Furthermore, a non-periodic nematode adheres less efficiently to endothelial cells. Strikingly C3, the central component of complement, plays a crucial role in the adherence interaction. These novel results show that microfilariae have the ability to bind to endothelial cells, which may explain their sequestration in the lungs, and this binding is increased in the presence of inflammatory mediators
A Multilaboratory Comparison of Calibration Accuracy and the Performance of External References in Analytical Ultracentrifugation
Analytical ultracentrifugation (AUC) is a first principles based method to determine absolute sedimentation coefficients and buoyant molar masses of macromolecules and their complexes, reporting on their size and shape in free solution. The purpose of this multi-laboratory study was to establish the precision and accuracy of basic data dimensions in AUC and validate previously proposed calibration techniques. Three kits of AUC cell assemblies containing radial and temperature calibration tools and a bovine serum albumin (BSA) reference sample were shared among 67 laboratories, generating 129 comprehensive data sets. These allowed for an assessment of many parameters of instrument performance, including accuracy of the reported scan time after the start of centrifugation, the accuracy of the temperature calibration, and the accuracy of the radial magnification. The range of sedimentation coefficients obtained for BSA monomer in different instruments and using different optical systems was from 3.655 S to 4.949 S, with a mean and standard deviation of (4.304 ± 0.188) S (4.4%). After the combined application of correction factors derived from the external calibration references for elapsed time, scan velocity, temperature, and radial magnification, the range of s-values was reduced 7-fold with a mean of 4.325 S and a 6-fold reduced standard deviation of ± 0.030 S (0.7%). In addition, the large data set provided an opportunity to determine the instrument-to-instrument variation of the absolute radial positions reported in the scan files, the precision of photometric or refractometric signal magnitudes, and the precision of the calculated apparent molar mass of BSA monomer and the fraction of BSA dimers. These results highlight the necessity and effectiveness of independent calibration of basic AUC data dimensions for reliable quantitative studies
A multilaboratory comparison of calibration accuracy and the performance of external references in analytical ultracentrifugation.
Analytical ultracentrifugation (AUC) is a first principles based method to determine absolute sedimentation coefficients and buoyant molar masses of macromolecules and their complexes, reporting on their size and shape in free solution. The purpose of this multi-laboratory study was to establish the precision and accuracy of basic data dimensions in AUC and validate previously proposed calibration techniques. Three kits of AUC cell assemblies containing radial and temperature calibration tools and a bovine serum albumin (BSA) reference sample were shared among 67 laboratories, generating 129 comprehensive data sets. These allowed for an assessment of many parameters of instrument performance, including accuracy of the reported scan time after the start of centrifugation, the accuracy of the temperature calibration, and the accuracy of the radial magnification. The range of sedimentation coefficients obtained for BSA monomer in different instruments and using different optical systems was from 3.655 S to 4.949 S, with a mean and standard deviation of (4.304 ± 0.188) S (4.4%). After the combined application of correction factors derived from the external calibration references for elapsed time, scan velocity, temperature, and radial magnification, the range of s-values was reduced 7-fold with a mean of 4.325 S and a 6-fold reduced standard deviation of ± 0.030 S (0.7%). In addition, the large data set provided an opportunity to determine the instrument-to-instrument variation of the absolute radial positions reported in the scan files, the precision of photometric or refractometric signal magnitudes, and the precision of the calculated apparent molar mass of BSA monomer and the fraction of BSA dimers. These results highlight the necessity and effectiveness of independent calibration of basic AUC data dimensions for reliable quantitative studies
Heart Rate Does Not Accurately Predict Metabolic Intensity During Variable-Intensity Roller Skiing or Cycling
Purpose: To critically appraise the utility of heart rate (HR) and power output (PO) to predict metabolic rate (MR) and oxygen consumption (
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) during variable-intensity roller skiing and cycling. Methods: National-level cyclists (nâ=â8) and cross-country skiers (nâ=â9) completed a preliminary session to determine
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max
, and a variable-intensity protocol with 3 high-intensity stages at 90%
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for 3 minutes interspersed with 3 moderate-intensity stages at 70%
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max
for 6 minutes. Cardiorespiratory measures were recorded throughout. Linear HRâMR,
HR
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, POâMR, and
PO
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regressions were computed from the preliminary session, individually, for all athletes and used to predict MR and
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from both HR and PO, separately, during the variable-intensity protocol. Mean differences with 95% limits of agreement (LOA) between measured and predicted MR and
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were calculated. Results: MR and
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estimated from HR displayed a mean bias close to zero but wide LOA. HR overestimated MR and
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during moderate intensity but underestimated MR and
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during high intensity, for both roller skiing and cycling. MR and
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estimated from PO were more consistent across the experimental trial, displaying a mean bias farther from zero but with tighter LOA. Conclusions: This study has demonstrated that HR has limited utility to predict metabolic intensity during variable-intensity roller skiing and cycling because of wide LOA. On the other hand, metabolic intensity predicted from PO had tighter LOA, suggesting better consistency. PO might provide a better prediction of metabolic intensity compared with HR, particularly when longer-duration steps are performed during preliminary testing
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