Resolution and characterisation of multiple isoforms of bovine kappa-casein by 2-DE following a reversible cysteine-tagging enrichment strategy

Visualisation of multiple isoforms of kappa-casein on 2-D gels is restricted by the abundant alpha- and beta-caseins that not only limit gel loading but also migrate to similar regions as the more acidic kappa-casein isoforms. To overcome this problem, we took advantage of the absence of cysteine residues in alpha(S1)- and beta-casein by devising an affinity enrichment procedure based on reversible biotinylation of cysteine residues. Affinity capture of cysteine-containing proteins on avidin allowed the removal of the vast majority of alpha(S1)- and beta-casein, and on subsequent 2-D gel analysis 16 gel spots were identified as kappa-casein by PMF. Further analysis of the C-terminal tryptic peptide along with structural predictions based on mobility on the 2-D gel allowed us to assign identities to each spot in terms of genetic variant (A or B), phosphorylation status (1, 2 or 3) and glycosylation status (from 0 to 6). Eight isoforms of the A and B variants with the same PTMs were observed. When the casein fraction of milk from a single cow, homozygous for the B variant of kappa-casein, was used as the starting material, 17 isoforms from 13 gel spots were characterised. Analysis of isoforms of low abundance proved challenging due to the low amount of material that could be extracted from the gels as well as the lability of the PTMs during MS analysis. However, we were able to identify a previously unrecognised site, T-166, that could be phosphorylated or glycosylated. Despite many decades of analysis of milk proteins, the reasons for this high level of heterogeneity are still not clear.

Implementation of particle-scale rotation in the 3-d Lattice Solid Model

In this study, 3-D Lattice Solid Model (LSMearth or LSM) was extended by introducing particle-scale rotation. In the new model, for each 3-D particle, we introduce six degrees of freedom: Three for translational motion, and three for orientation. Six kinds of relative motions are permitted between two neighboring particles, and six interactions are transferred, i.e., radial, two shearing forces, twisting and two bending torques. By using quaternion algebra, relative rotation between two particles is decomposed into two sequence-independent rotations such that all interactions due to the relative motions between interactive rigid bodies can be uniquely decided. After incorporating this mechanism and introducing bond breaking under torsion and bending into the LSM, several tests on 2-D and 3-D rock failure under uni-axial compression are carried out. Compared with the simulations without the single particle rotational mechanism, the new simulation results match more closely experimental results of rock fracture and hence, are encouraging. Since more parameters are introduced, an approach for choosing the new parameters is presented.

Depth propagation and surface construction in 3-D vision

In stereo vision, regions with ambiguous or unspecified disparity can acquire perceived depth from unambiguous regions. This has been called stereo capture, depth interpolation or surface completion. We studied some striking induced depth effects suggesting that depth interpolation and surface completion are distinct stages of visual processing. An inducing texture (2-D Gaussian noise) had sinusoidal modulation of disparity, creating a smooth horizontal corrugation. The central region of this surface was replaced by various test patterns whose perceived corrugation was measured. When the test image was horizontal 1-D noise, shown to one eye or to both eyes without disparity, it appeared corrugated in much the same way as the disparity-modulated (DM) flanking regions. But when the test image was 2-D noise, or vertical 1-D noise, little or no depth was induced. This suggests that horizontal orientation was a key factor. For a horizontal sine-wave luminance grating, strong depth was induced, but for a square-wave grating, depth was induced only when its edges were aligned with the peaks and troughs of the DM flanking surface. These and related results suggest that disparity (or local depth) propagates along horizontal 1-D features, and then a 3-D surface is constructed from the depth samples acquired. The shape of the constructed surface can be different from the inducer, and so surface construction appears to operate on the results of a more local depth propagation process.

Depth propagation and surface construction in 3-D vision

Motion discontinuities can signal object boundaries where few or no other cues, such as luminance, colour, or texture, are available. Hence, motion-defined contours are an ecologically important counterpart to luminance contours. We developed a novel motion-defined Gabor stimulus to investigate the nature of neural operators analysing visual motion fields in order to draw parallels with known luminance operators. Luminance-defined Gabors have been successfully used to discern the spatial-extent and spatial-frequency specificity of possible visual contour detectors. We now extend these studies into the motion domain. We define a stimulus using limited-lifetime moving dots whose velocity is described over 2-D space by a Gabor pattern surrounded by randomly moving dots. Participants were asked to determine whether the orientation of the Gabor pattern (and hence of the motion contours) was vertical or horizontal in a 2AFC task, and the proportion of correct responses was recorded. We found that with practice participants became highly proficient at this task, able in certain cases to reach 90% accuracy with only 12 limited-lifetime dots. However, for both practised and novice participants we found that the ability to detect a single boundary saturates with the size of the Gaussian envelope of the Gabor at approximately 5 deg full-width at half-height. At this optimal size we then varied spatial frequency and found the optimum was at the lowest measured spatial frequency (0.1 cycle deg-1 ) and then steadily decreased with higher spatial frequencies, suggesting that motion contour detectors may be specifically tuned to a single, isolated edge.

The Darboux Problem for a Class of 3-D Weakly Hyperbolic Equations

Недю Попиванов, Цветан Христов - Изследвани са някои тримерни аналози на задачата на Дарбу в равнината. През 1952 М. Протер формулира нови тримерни гранични задачи както за клас слабо хиперболични уравнения, така и за някои хиперболично-елиптични уравнения. За разлика от коректността на двумерната задача на Дарбу, новите задачи са некоректни. За слабо хиперболични уравнения, съдържащи младши членове, ние намираме достатъчни условия както за съществуване и единственост на обобщени решения с изолирана степенна особеност, така и за единственост на квази-регулярни решения на задачата на Протер.

Mechanistic studies on the degradation of gasoline oxygenates by advanced oxidation technologies and the reaction of 4-methyl-1,2,4-triazoline-3,5-dione with tetracyclopropylethylene

Gasoline oxygenates (MTBE, methyl tert-butyl ether; DIPE, di-isopropyl ether; ETBE, ethyl tert-butyl ether; TAME, tert-amyl ether) are added to gasoline to boost octane and enhance combustion. The combination of large scale use, high water solubility and only minor biodegradability has now resulted in a significant gasoline oxygenate contamination occurring in surface, ground, and drinking water systems. Combination of hydroxyl radical formation and the pyrolytic environment generated by ultrasonic irradiation (665 kHz) leads to the rapid degradation of MTBE and other gasoline oxygenates in aqueous media. ^ The presence of oxygen promotes the degradation processes by rapid reaction with carbon centered radicals indicating radical processes involving O 2 are significant pathways. A number of the oxidation products were identified. The formation of products (alcohols, ketones, aldehydes, esters, peroxides, etc) could be rationalized by mechanisms which involve hydrogen abstraction by OH radical and/or pyrolysis to form carboncentered radicals which react with oxygen and follow standard oxidation chain processes. ^ The reactions of N-substituted R-triazolinediones (RTAD; R = CH 3 or phenyl) have attracted considerable interest because they exhibit a number of unusual mechanistic characteristics that are analogous to the reactions of singlet oxygen (1O2) and offer an easy way to provide C-N bond(s) formation. The reactions of triazolinedione with olefins have been widely studied and aziridinium imides are generally accepted to be the reactive intermediates. ^ We observed the rapid formation of an unusual intermediate upon mixing tetracyclopropylethylene with 4-methyl-1,2,4-triazoline-3,5-dione in CDCl 3. Detailed characterization by NMR (proton, 13C, 2-D NMRs) indicates the intermediate is 5,5,6,6-tetracyclopropyl-3-methyl-5,6-dihydro-oxazolo[3,2- b][1,2,4]-triazolium-2-olate. Such products are extremely rare and have not been studied. Upon warming the intermediate is converted to 2 + 2 diazetidine (major) and ene product (minor). ^ To further explore the kinetics and dynamics of the reaction activation energies were obtained using Arrhenius plots. Activation energies for the formation of the intermediate from reactants, and 2+2 adduct from the intermediate were determined as 7.48 kcal moll and 19.8 kcal mol−1 with their pre-exponential values of 2.24 × 105 dm 3 mol−1 sec−1 and 2.75 × 108 sec−1, respectively, meaning net slow reactions because of low pre-exponential values caused by steric hindrance. ^

Synthèse et fonctionnalisation des 4-aminopyrrole-2-carboxylates

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Synthèse et fonctionnalisation des 4-aminopyrrole-2-carboxylates

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Seawater carbonate chemistry and community calcification during Reunion Island coral reef community study, 2007

To assess the contribution of soft-bottoms to the carbon cycle in coral reefs, the net community production (p) was measured in winter at 3 stations on La Saline inner reef flat (Reunion Island). Changes in pH and total alkalinity at different irradiances (I) were assessed using benthic chambers (0.2 m2) during a 1-h incubation. Mean grain size, the silt and clay load and chlorophyll a content of the sediments were analysed in each chamber. Daily community production (P), gross community production (Pg) and community respiration (R) were estimated from p-I curves and daily irradiance variations (PAR, 400-700 nm). Sediment characteristics and chlorophyll a contents did not differ between the three sites, except for the silt and clay fraction at one station. R being higher than Pg (84.88 ± 7.36 and -62.29 ± 3.34 mmolC m-2 d-1 respectively), P value reached 22.59 ± 5.66 mmolC m-2 d-1. The sediments were therefore heterotrophic with a mean Pg/R lower than 1 (0.74 ± 0.05) and appear to be a carbon source. Our data suggested the importance of the degradation process in the functioning of near-reef sediments.

Impacts of high-nitrate freshwater inputs on macrotidal ecosystems. I. Seasonal evolution of nutrient limitation for the diatom-dominated phytoplankton of the Bay of Brest (France)

The chemical factors (inorganic nitrogen, phosphate, silicic acid) that potentially or actually control primary production were determined for the Bay of Brest, France, a macrotidal ecosystem submitted to high-nitrate-loaded freshwater inputs (winter nitrate freshwater concentrations >700 mu M, Si:N molar ratio as low as 0.2, i.e. among the lowest ever published). Intensive data collection and observations were carried out from February 1993 to March 1994 to determine the variations of physical [salinity, temperature, photosynthetically active radiation (PAR), freshwater discharges] and chemical (oxygen and nutrients) parameters and their impacts on the phytoplankton cycle (fluorescence, pigments, primary production). With insufficient PAR the winter stocks of nutrients were almost nonutilized and the nitrate excess was exported to the adjacent ocean, due to rapid tidal exchange. By early April, a diatom-dominated spring bloom developed (chlorophyll a maximum = 7.7 mu g l(-1); primary production maximum = 2.34 g C m(-2) d(-1)) under high initial nutrient concentrations. Silicic acid was rapidly exhausted over the whole water column; it is inferred to be the primary limiting factor responsible for the collapse of the spring bloom by mid-May. Successive phytoplankton developments characterized the period of secondary blooms during summer and fall (successive surface chlorophyll a maxima = 3.5, 1.6, 1.8 and 1.0 mu g l(-1); primary production = 1.24, 1.18 and 0.35 g C m(-2) d(-1)). Those secondary blooms developed under lower nutrient concentrations, mostly originating from nutrient recycling. Until August, Si and P most likely limited primary production, whereas the last stage of the productive period in September seemed to be N limited instead, this being a period of total nitrate depletion in almost the whole water column. Si limitation of spring blooms has become a common feature in coastal ecosystems that receive freshwater inputs with Si:N molar ratios <1. The peculiarity of Si Limitation in the Bay of Brest is its extension through the summer period.

Neuroradiological findings expand the phenotype of OPA1-related mitochondrial dysfunction

International audience

Jacobi quartic curves revisited

This paper provides new results about efficient arithmetic on Jacobi quartic form elliptic curves, y 2 = d x 4 + 2 a x 2 + 1. With recent bandwidth-efficient proposals, the arithmetic on Jacobi quartic curves became solidly faster than that of Weierstrass curves. These proposals use up to 7 coordinates to represent a single point. However, fast scalar multiplication algorithms based on windowing techniques, precompute and store several points which require more space than what it takes with 3 coordinates. Also note that some of these proposals require d = 1 for full speed. Unfortunately, elliptic curves having 2-times-a-prime number of points, cannot be written in Jacobi quartic form if d = 1. Even worse the contemporary formulae may fail to output correct coordinates for some inputs. This paper provides improved speeds using fewer coordinates without causing the above mentioned problems. For instance, our proposed point doubling algorithm takes only 2 multiplications, 5 squarings, and no multiplication with curve constants when d is arbitrary and a = ±1/2.

Change in colour appearance of small defocused lights

Small long wavelength lights (≤ 1’ arc) change colour appearance with positive defocus, appearing yellow or white. I investigated influences of longitudinal chromatic aberration and monochromatic aberrations on colour appearance of small narrow band lights. Seven cyclopleged participants viewed a small light (1’ arc diameter, λmax range 510 - 628 nm) centred within a 4.6’ black annulus and surrounded by a uniform white field under photopic light levels. An optical trombone varied focus. Participants were required to vary the focus by moving the optical trombone in either positive or negative direction and report when they noticed a change in appearance of the defocused narrow band light. Longitudinal chromatic aberration was controlled using a Powell achromatizing lens and its doublet and triplet components that neutralized, doubled and reversed the eye’s chromatic aberration, respectively. Changes in colour appearance for a 628 nm light occurred without any lens at +0.5 ± 0.2D defocus and with the doublet at +0.6 ± 0.2 D. The achromatizing lens did not affect appearance and the phenomenon was evident with the triplet for negative defocus (-0.5 ± 0.3 D). Adaptive optics correction of astigmatism and higher order monochromatic aberration did not affect magnitude significantly. Colour changes occurred despite a range of participant L/M cone ratios. Direction of change in colour appearance was reversed for short compared to long wavelengths. We conclude that longitudinal chromatic aberrations, but not monochromatic aberrations, are involved in changing appearance of small lights with defocus. Additional neuronal mechanisms that may contribute to the colour changes are considered.

On the use of different spatial reference frames for movement control

Previous work has shown that amplitude and direction are two independently controlled parameters of aimed arm movements, and performance, therefore, suffers when they must be decomposed into Cartesian coordinates. We now compare decomposition into different coordinate systems. Subjects pointed at visual targets in 2-D with a cursor, using a two-axis joystick or two single-axis joysticks. In the latter case, joystick axes were aligned with the subjects’ body axes, were rotated by –45°, or were oblique (i.e., one axis was in an egocentric frame and the other was rotated by –45°). Cursor direction always corresponded to joystick direction. We found that compared with the two-axis joystick, responses with single-axis joysticks were slower and less accurate when the axes were oriented egocentrically; the deficit was even more pronounced when the axes were rotated and was most pronounced when they were oblique. This confirms that decomposition of motor commands is computationally demanding and documents that this demand is lowest for egocentric, higher for rotated, and highest for oblique coordinates. We conclude that most current vehicles use computationally demanding man–machine interfaces.

Porcine bone marrow stromal cell differentiation on heparin-adsorbed poly (e-caprolactone)-tricalcium phosphate-collagen scaffolds

We evaluate the potential of heparin as a substrate component for the fabrication of bone tissue engineering constructs using poly(e- caprolactone)–tricalcium phosphate–collagen type I (PCL–TCP–Col) three-dimensional (3-D) scaffolds. First we explored the ability of porcine bone marrow precursor cells (MPCs) to differentiate down both the adipogenic and osteogenic pathways within 2-D culture systems, with positive results confirmed by Oil-Red-O and Alizarin Red staining, respectively. Secondly, we examined the influence of heparin on the interaction and behaviour of MPCs when seeded onto PCL–TCP–Col 3-D scaffolds, followed by their induction into the osteogenic lineage. Our 3-D findings suggest that cell metabolism and proliferation increased between days 1 and 14, with deposition of extracellular matrix also observed up to 28 days. However, no noticeable difference could be detected in the extent of osteogenesis for PCL–TCP–Col scaffolds groups with the addition of heparin compared to identical control scaffolds without the addition of heparin.