Noninvasive Measurement of Steady and Pulsating Velocity Profiles and Shear Rates in Arteries Using Echo PIV: In Vitro Validation Studies

Although accurate measurement of velocity profiles, multiple velocity vectors, and shear stress in arteries is important, there is still no easy method to obtain such information in vivo. We report on the utility of combining ultrasound contrast imaging with particle image velocimetry (PIV) for noninvasive measurement of velocity vectors. This method (echo PIV) takes advantage of the strong backscatter characteristics of small gas-filled microbubbles (contrast) seeded into the flow. The method was tested in vitro. The steady flow analytical solution and optical PIV measurements (for pulsatile flow) were used for comparison. When compared to the analytical solution, both echo PIV and optical PIV resolved the steady velocity profile well. Error in shear rate as measured by echo PIV (8%) was comparable to the error of optical PIV (6.5%). In pulsatile flow, echo PIV velocity profiles agreed well with optical PIV profiles. Echo PIV followed the general profile of pulsatile shear stress across the artery but underestimated wall shear at certain time points. However, error in shear from echo PIV was an order of magnitude less than error from current shear measurement methods. These studies indicate that echo PIV is a promising technique for noninvasive measurement of velocity profiles and shear stress.     

Evidence for a dose-dependent effect of pulsed magnetic fields on pain processing

Functional magnetic resonance imaging (fMRI) was used to investigate the dose–response relationship (sham, 100, 200, 1000 μT) between a pulsed extremely low frequency magnetic field (ELFMF) and acute thermal pain on the dominant right hand. Forty-seven participants were recruited, and pulsed ELFMF was applied through the MRI gradient system using a novel technique. Regions of interest (ROIs) matching those of previous studies were examined for a potential dose response. Significant correlations between applied field strength and change in BOLD activity were found in the anterior cingulate and the ipsilateral insula, indicating that there might be either a dose response or a threshold effect of the ELFMF.     

Array-based comparative genomic hybridization identifies a high frequency of copy number variations in patients with syndromic overgrowth

Overgrowth syndromes are a heterogeneous group of conditions including endocrine hormone disorders, several genetic syndromes and other disorders with unknown etiopathogenesis. Among genetic causes, chromosomal deletions and duplications such as dup(4)(p16.3), dup(15)(q26qter), del(9)(q22.32q22.33), del(22)(q13) and del(5)(q35) have been identified in patients with overgrowth. Most of them, however, remain undetectable using banding karyotype analysis. In this study, we report on the analysis using a 1-Mb resolution array-based comparative genomic hybridization (CGH) of 93 patients with either a recognizable overgrowth condition (ie, Sotos syndrome or Weaver syndrome) or an unclassified overgrowth syndrome. Five clinically relevant imbalances (three duplications and two deletions) were identified and the pathogenicity of two additional anomalies (one duplication and one deletion) is discussed. Altered segments ranged in size from 0.32 to 18.2 Mb, and no recurrent abnormality was identified. These results show that array-CGH provides a high diagnostic yield in patients with overgrowth syndromes and point to novel chromosomal regions associated with these conditions. Although chromosomal deletions are usually associated with growth retardation, we found that the majority of the imbalances detected in our patients are duplications. Besides their importance for diagnosis and genetic counseling, our results may allow to delineate new contiguous gene syndromes associated with overgrowth, pointing to new genes, the deregulation of which may be responsible for growth defect.     

Characterization of a recombinant canine coronavirus with a distinct receptor-binding (S1) domain

Canine alphacoronaviruses (CCoV) exist in two serotypes, type I and II, both of which can cause severe gastroenteritis. Here, we characterize a canine alphacoronavirus, designated CCoV-A76, first isolated in 1976. Serological studies show that CCoV-A76 is distinct from other CCoVs, such as the prototype CCoV-1-71. Efficient replication of CCoV-A76 is restricted to canine cell lines, in contrast to the prototypical type II strain CCoV-1-71 that more efficiently replicates in feline cells. CCoV-A76 can use canine aminopeptidase N (cAPN) receptor for infection of cells, but was unable to use feline APN (fAPN). In contrast, CCoV-1-71 can utilize both. Genomic analysis shows that CCoV-A76 possesses a distinct spike, which is the result of a recombination between type I and type II CCoV, that occurred between the N- and C-terminal domains (NTD and C-domain) of the S1 subunit. These data suggest that CCoV-A76 represents a recombinant coronavirus form, with distinct host cell tropism.     

Physiological demand and pacing strategy during the new combined event in elite pentathletes

To evaluate the physiological demands and effects of different pacing strategies on performance during the new combined event (CE) of the modern pentathlon (consisting of three pistol shooting sessions interspersed by three 1-km running legs). Nine elite pentathletes realised five tests: a free-paced CE during an international competition; an incremental running test to determine [Formula: see text] and its related velocity ([Formula: see text]) and three experimental time-trial CE, where the pacing strategy was manipulated (CE(ref), CE(100%), CE(105%)). CE(ref) reproduced the international competition strategy with a 170-m fast running start within the first 2 km. CE(100%) and CE(105%) imposed a constant strategy over km-1 and km-2 with a velocity of 100 and 105% of the mean speed adopted over the same sections during the international competition, respectively. Km-3 was always self-paced. The subjects ran CE(ref) at 99 ± 4% of [Formula: see text] and reached 100 ± 5, 100 ± 7, 99 ± 8% of [Formula: see text] at the end of kilometres 1, 2 and 3, respectively ([Formula: see text]: 72 ± 6 mL O(2) min(-1) kg(-1)), with a peak blood lactate concentration of 13.6 ± 1.5 mmol L(-1). No significant differences in overall performance were found between the pacing conditions (753 ± 30, 770 ± 39, 768 ± 27 s for CE(ref), CE(100%) and CE(105%), respectively, p = 0.63), but all of the shooting performance parameters were only stable in CE(ref). Completion of CE by elite pentathletes elicits a maximal aerobic contribution coupled with a high glycolytic supply. Manipulating the mean running speed over km-1 and km-2 had strong influence on the overall pacing strategy and induced minor differences in shooting performance, but it did not affect overall performance.     

Putative use of a Bacillus subtilis L194 strain for biocontrol of Phoma medicaginis in Medicago truncatula seedlings

An antagonist L194 strain against Phoma medicaginis pathogenic fungi was isolated from Tunisian soil (vicinity of Tunis) and identified as Bacillus subtilis based on biochemical characteristics and partial 16S rDNA sequence. When cells were grown in a minimal medium for 24 h, spore culture supernatant exhibited 2-fold higher antifungal activity than vegetative cells. MALDI-TOF mass spectrometry analysis showed that L194 spores produced mainly iturins, surfactins and fengycins with long-chain fatty acids, and other not yet identified compounds. Both vegetative cells and spores of L194 efficiently reduced germination of P. medicaginis conidia. As revealed by atomic force microscopy, L194 spores modified conidia morphology from a regular to a deflated shape. Data suggest that lipopeptides interacted with the cytoplasmic membrane, causing pore formation. In vivo, L194 spores were highly protective against P. medicaginis by reducing disease symptoms and alleviating growth disturbance of Medicago truncatula seedlings. As a whole, the lipopeptide-producing L194 strain may be successfully used in biocontrol of plant diseases induced by phytopathogenic fungi such as P. medicaginis.     

Remodelling of the SUR–Kir6.2 interface of the KATP channel upon ATP binding revealed by the conformational blocker rhodamine 123

ATP-sensitive K⁺ channels (K_ATP channels) are metabolic sensors formed by association of a K⁺ channel, Kir6, and an ATP-binding cassette (ABC) protein, SUR, which allosterically regulates channel gating in response to nucleotides and pharmaceutical openers and blockers. How nucleotide binding to SUR translates into modulation of Kir6 gating remains largely unknown. To address this issue, we have used a novel conformational K_ATP channel inhibitor, rhodamine 123 (Rho123) which targets the Kir6 subunit in a SUR-dependent manner. Rho123 blocked SUR-less Kir6.2 channels with an affinity of ∼1 μ m , regardless of the presence of nucleotides, but it had no effect on channels formed by the association of Kir6.2 and the N-terminal transmembrane domain TMD0 of SUR. Rho123 blocked SUR + Kir6.2 channels with the same affinity as Kir6.2 but this effect was antagonized by ATP. Protection from Rho123 block by ATP was due to direct binding of ATP to SUR and did not entail hydrolysis because it was not mimicked by AMP, did not require Mg²⁺ and was reduced by mutations in the nucleotide-binding domains of SUR. These results suggest that Rho123 binds at the TMD0–Kir6.2 interface and that binding of ATP to SUR triggers a change in the structure of the contact zone between Kir6.2 and domain TMD0 of SUR that causes masking of the Rho123 site on Kir6.2.