Characterization of rat vertebrae cortical bone microstructures using confocal Raman microscopy combined to tomography and electron microscopy.

BACKGROUND: The rat vertebrae is a good model to study bone regeneration after implantation of biomaterials used to treat bone loss, a major problem in oral and dental surgery. However, the precise characterization of bone microstructures in the rat vertebrae has not been reported. Therefore, the aim of this study was to achieve the complete analysis of such bone, at different scales, in order to have a clear model of healthy bone for comparison with regenerated bone. METHODS: In order to image the cortical bone of rat caudal vertebra, confocal Raman microscopy was combined with high resolution X-ray micro computed tomography (micro-CT), with scanning electron microscopy (SEM) using backscatter electron imaging and with more conventional histology coloration techniques. SEM and Raman microscopy were done in various regions of the cortical bone corresponding to external, middle and internal areas. The spongy bone was imaged in parallel. Micro-CT was performed on the whole vertebra to monitor the network of haversian canals in the cortical bone. Osteonic canals characteristics, and relative chemical composition were analysed in several regions of interest, in cortical and spongy bone. Five rats were included in this study. RESULTS: On micro-CT images, differences in intensity were observed in the cortical bone, substantiated by SEM. Chemical analysis with Raman spectra confirmed the difference in composition between the different regions of the cortical and spongy bone. PCA and k-mean cluster analysis separated these groups, except for the external and middle cortical bone. Peak intensity ratio confirmed these results with a CO3 to ν2 PO4 ratio significantly different for the internal cortical bone. Grayscale images stack extracted from micro-CT showed that global architecture of cortical bone was characterized by a dense and complex network of haversian osteonic canals, starting from the surface towards the vertebrae center. The mean diameter of the canals was 18.4 µm (SD 8.6 µm) and the mean length was 450 µm (SD 152 µm). Finally, Raman reconstructed images of the lamellar bone showed an enlargement of the lamellar layer width, both in circumferential lamellar bone and around haversian canals. CONCLUSIONS: Micro-CT and confocal Raman microscopy are good tools to complete classical analysis using optical and electron microscopy. The results and measurements presented in a rat model known for its small inter-individual differences provide the main characteristics of a mature bone. This study will allow the community working on this rat vertebrate model to have a set of characteristics, in particular on the structure of the haversian canals.

Assessing transmission of crop diseases by insect vectors in a landscape context.

Theory indicates that landscape composition affects transmission of vector-borne crop diseases, but few empirical studies have investigated how landscape composition affects plant disease epidemiology. Since 2006, Bemisia tabaci (Gennadius) has vectored the cucurbit yellow stunting disorder virus (CYSDV) to cantaloupe and honeydew melons (Cucumis melo L.) in the southwestern United States and northern Mexico, causing significant reductions in yield of fall melons and increased use of insecticides. Here, we show that a landscape-based approach allowing simultaneous assessment of impacts of local (i.e., planting date) and regional (i.e., landscape composition) factors provides valuable insights on how to reduce crop disease risks. Specifically, we found that planting fall melon fields early in the growing season, eliminating plants germinating from seeds produced by spring melons after harvest, and planting fall melon fields away from cotton and spring melon fields may significantly reduce the incidence of CYSDV infection in fall melons. Because the largest scale of significance of the positive association between abundance of cotton and spring melon fields and CYSDV incidence was 1,750 and 3,000 m, respectively, reducing areas of cotton and spring melon fields within these distances from fall melon fields may decrease CYSDV incidence. Our results indicate that landscape-based studies will be fruitful to alleviate limitations imposed on crop production by vector-borne diseases.

Spatial distribution of single-nucleotide polymorphisms related to fungicide resistance and implications for sampling.

Spatial distribution of single-nucleotide polymorphisms (SNPs) related to fungicide resistance was studied for Botrytis cinerea populations in vineyards and for B. squamosa populations in onion fields. Heterogeneity in this distribution was characterized by performing geostatistical analyses based on semivariograms and through the fitting of discrete probability distributions. Two SNPs known to be responsible for boscalid resistance (H272R and H272Y), both located on the B subunit of the succinate dehydrogenase gene, and one SNP known to be responsible for dicarboximide resistance (I365S) were chosen for B. cinerea in grape. For B. squamosa in onion, one SNP responsible for dicarboximide resistance (I365S homologous) was chosen. One onion field was sampled in 2009 and another one was sampled in 2010 for B. squamosa, and two vineyards were sampled in 2011 for B. cinerea, for a total of four sampled sites. Cluster sampling was carried on a 10-by-10 grid, each of the 100 nodes being the center of a 10-by-10-m quadrat. In each quadrat, 10 samples were collected and analyzed by restriction fragment length polymorphism polymerase chain reaction (PCR) or allele specific PCR. Mean SNP incidence varied from 16 to 68%, with an overall mean incidence of 43%. In the geostatistical analyses, omnidirectional variograms showed spatial autocorrelation characterized by ranges of 21 to 1 m. Various levels of anisotropy were detected, however, with variograms computed in four directions (at 0°, 45°, 90°, and 135° from the within-row direction used as reference), indicating that spatial autocorrelation was prevalent or characterized by a longer range in one direction. For all eight data sets, the ß-binomial distribution was found to fit the data better than the binomial distribution. This indicates local aggregation of fungicide resistance among sampling units, as supported by estimates of the parameter θ of the ß-binomial distribution of 0.09 to 0.23 (overall median value = 0.20). On the basis of the observed spatial distribution patterns of SNP incidence, sampling curves were computed for different levels of reliability, emphasizing the importance of sample size for the detection of mutation incidence below the risk threshold for control failure.

Human dental pulp stem cells growth and osteodifferentiation on porous resorbable scaffolds.

Short Communication selected from the Oral Presentations of the 56th Congress of the Groupèment International pour la Recherche Scientifique en Stomatologie et Odontologie, Peñafiel (Portugal) May 2012.

[Not Available]. / Fonctionnalisation du Titane par des Peptides Specifiques Bi-Fonctionnels pour l'adhesion Epitheliale.

Short Communication selected from the Oral Presentations of the 56th Congress of the Groupèment International pour la Recherche Scientifique en Stomatologie et Odontologie, Peñafiel (Portugal) May 2012.

Assessment of common scab-inducing pathogen effects on potato underground organs via computed tomography scanning.

Common scab caused by Streptomyces scabies is a major bacterial disease of potato (Solanum tuberosum). Its best known symptom is superficial lesions on the surface of progeny potato tubers, observed at harvesting. In this study, effects of S. scabies on space occupancy by underground organs and on structural complexity of root systems are investigated during growth via computed tomography (CT) scanning. Two groups of potato plants were grown in a greenhouse in middle-sized plastic pots. Using a high-resolution X-ray CT scanner formerly used for medical applications, their underground organs and surrounding medium (sieved and autoclaved homogeneous sand) were submitted to CT scanning 4, 6, and 8 weeks after planting. For one group, sand was inoculated with the common scab-inducing pathogen (S. scabies EF-35) at potting. Space occupancy by underground organs was estimated via curve fitting applied to histograms of CT scan data, while three-dimensional skeletal images were used for fractal analysis. Root systems of diseased plants were found to be less complex than those of healthy plants 4 weeks after planting, and the relative growth rates derived from space occupancy measures were of different sign between the two groups from week 4 to week 8.

Bioremediation of residual fertilizer nitrate: II. Soil redox potential and soluble iron as indicators of soil health during treatment.

The prospect of using wastewater containing high loads of soluble organic matter (OM) for removing residual agricultural chemicals (fertilizer, pesticide, or herbicide) in farm soil, although promising, could have adverse effects on soil agricultural quality as a result of development of redoximorphic features in the soil profile. In this study, the effect of organic carbon supplement for bioremediation of residual fertilizer nitrate on soil properties, redox potential (Eh), pH, and metal ion mobilization was studied using sandy soils packed in columns. The study was included in a general project, described elsewhere (Ugwuegbu et al., 2000), undertaken to evaluate use of controlled water table management (WTM) systems to supply organic carbon for creating a reduced environment conducive to denitrification of residual fertilizer nitrate leaching from the farm to subsurface water. The columns were subjected to subirrigation with water containing soluble organic carbon in the form of glucose. The work was carried out in two experimental setups and the long-term effect of a range of glucose concentrations on the Eh, pH, and soluble levels of Fe and Mn was investigated. From the results obtained, it could be concluded that excessive organic carbon supplement to soil can have adverse effects on soil quality and that Eh and soluble Fe are the two most practical parameters for monitoring soil health during treatment of farm chemicals.

Time-dependent transition probabilities and the assessment of seasonal effects on within-day variations in chewing behaviour of housed sheep.

State transitions in the chewing behaviour of six half-breed (Île de FrancexTexel) yearling female sheep (Ovis aries L.) were studied by using jaw movements recorded continuously over 5 days at the end of a number of experimental periods from 21 September 1992 to 4 April 1993. The sheep were housed in individual pens. Each of them received the same diet, that is, 250 g/day of concentrate mix (15.5% crude protein (CP), 36.5% neutral detergent fibre (NDF)) fed at 0900 h and natural grass hay (6.7% CP, 69.1% NDF) fed ad libitum at 0915 and 1600 h. Mineral salt blocks and water were continuously available. The main objective was to assess seasonal effects on within-day variations in the chewing behaviour of sheep, at small to large time scales within a day. We therefore focused on two experimental periods characterised by contrasting conditions of daylength and temperature (i.e., 'Period 1': 610 min daylight, mean temperature of 10.9 degrees C, and 'Period 4': 550 min daylight, mean temperature of 7.2 degrees C). In particular, differences between periods in the nycterohemeral pattern of chewing behaviour and the quality of forecasts of chewing states were tested. We submitted our data to a new method of analysis that we developed: the method of time-dependent transition probabilities, and compared the results to those obtained using other methods that were available in the literature.Overall, the sheep spent more time eating in Period 1 than in Period 4. Specifically, a secondary peak in eating activity, which was observed in the early afternoon in Period 1, was absent in Period 4. The nycterohemeral pattern of eating activity showed significant differences between periods, at the main rhythmic component of 24 h and at short components around 2 h. Such differences were not observed for ruminating and idling activities. The quality of forecasts of chewing states decreased from Periods 1 to 4, in terms of accuracy (based on R(2)) and lead of reliable forecasts (i.e., &z.sfnc;forecast-observation&z.sfnc;<0.1). The most (least) accurate forecasts were obtained for the ruminating (eating) state in both periods. We have attributed the differences that we found between periods to daylength instead of temperature because the sheep were mostly within the thermoneutral zone in our study. By comparison, using mean hourly times of eating activity, significant differences between periods were detected for the 24-h rhythmic component and the 4-h component, instead of the 2-h component, probably because of aliasing (i.e., when the sampling time interval used is longer than suited; the minute was found to be a suitable interval length in the calculation of time-dependent transition probabilities). Using the age-dependent model of Rook and Penning, minor differences between periods were detected. On that basis, the method of time-dependent transition probabilities may be brought forward as a complement of value to existing methods of behavioural data analysis.

A Generalized Two-Dimensional Gaussian Model of Disease Foci of Head Blight of Wheat Caused by Gibberella zeae.

ABSTRACT A generalized two-dimensional Gaussian model is proposed to describe disease foci of head blight of wheat in plots (100 to 2,500 m(2)) originating from small areas (1 to 16 m(2)) inoculated with Gibberella zeae-colonized corn kernels. These anisotropic, asymmetrical foci arose from ascospores produced in perithecia. The model is Z = exp[-(AX(2) + BY(2) + CXY + DX + EY + F)], in which Z = the incidence of seed or spikelet infection at point (X,Y) located in the plot, exp = the exponential function, X = the abscissa or spatial coordinate of the point along a given axis (approximately parallel to the average wind vector during the period of spore release in these experiments), Y = the ordinate or spatial coordinate of the point along the axis perpendicular to the X axis (approximately perpendicular to the wind direction in these experiments), A and B = the quadratic coefficients of the second-order polynomial AX(2) + BY(2) + CXY + DX + EY + F, C = the bilinear coefficient, D and E = the linear coefficients, and exp(-F) = the incidence of seed or spikelet infection at the focus peak in which X = 0 and Y = 0. The generalized two-dimensional Gaussian model was tested on data from a circular or isotropic focus, an elliptical or anisotropic focus with two axes of symmetry, and two anisotropic foci with one and zero axis of symmetry. Its goodness-of-fit (r(2) and adjusted r(2)) was compared with the inverse power, modified inverse power, exponential, and classical Gaussian models. Submodels using only the linear terms, only the quadratic terms, or combinations selected from stepwise regression procedures using various probabilities to enter and to stay and a procedure maximizing the adjusted r (2) were also considered. Spatial analysis of the residuals was performed using Geary's c coefficient at the first distance class. For the circular and elliptical foci, our model provided a fit similar to the modified inverse power and exponential models. However, for anisotropic foci with one or zero axis of symmetry arising from ascospores influenced by wind direction, the generalized two-dimensional Gaussian model provided a better fit. For these anisotropic foci, the linear term X but not the quadratic term X(2) was generally retained in the model, indicating an exponential gradient in the direction parallel to the wind. In all models, the quadratic term Y(2) was retained, along with Y in some cases, indicating that the gradient in the direction roughly perpendicular to the wind was Gaussian or Gaussian-exponential in shape. The bilinear term XY provided an indication of the orientation of the focus in relation to the axes of the sampling grid. This model has the versatility and parameters (quadratic, bilinear, and linear) to better describe the anisotropy of foci from wind-dispersed spores.

Head Blight Gradients Caused by Gibberella zeae from Area Sources of Inoculum in Wheat Field Plots.

ABSTRACT The spread of Fusarium head blight of wheat from a small area inoculum source was examined in wheat plots (100, 625, or 2,500 m(2)) inoculated in the center with Gibberella zeae-colonized corn kernels or macro-conidia sprayed on heads at anthesis. With the first inoculation method, disease foci were produced from ascospores released from perithecia formed on inoculated kernels. With the second inoculation method, disease foci were produced by macroconidia directly applied to the heads. Some plots were misted during anthesis. Plots were divided into grids, and disease incidence on spikelets and seeds was assessed at the grid intersections. Isopath contour maps were constructed using an interpolation procedure based on a weighted least squares method. Disease gradients were constructed from the isopath contours in the direction parallel to average nightly wind vectors using an exponential model. This study was conducted over a 3-year period at two sites: one in Quebec and one in Ontario. Both inoculation methods resulted in a discrete, primary focus of head blight in each plot, with one or two smaller secondary foci in some plots. The highest incidence of disease on spikelets or seed was commonly displaced somewhat from the inoculum source, usually downwind. The gradient slopes of seed and spikelet infection ranged from -0.10 to -0.43 m(1) in plots with ascospore inoculum and from -0.48 to -0.79 m(1) in plots inoculated with macroconidia. Seed infection declined to 10% of the maximum within 5 to 22 m from the focal center in asco-spore-inoculated plots, and within 5 m in a macroconidia-inoculated plot. Gradients were usually steeper upwind compared with downwind of the inoculum source. In misted plots, incidence of disease was higher and more diffuse than in nonirrigated plots. Based on gradients and dispersal patterns, disease foci in plots inoculated with G. zeae-colonized corn kernels probably arose from airborne ascospores rather than from splash-borne macroconidia and were the result of infection events that occurred over a short period of time. Comparison of conidial- and ascospore-derived disease gradients indicated a lack of secondary infection, confirming that Fusarium head blight is primarily a monocyclic disease.

Among-environment heteroscedasticity and genetic autocorrelation: implications for the study of phenotypic plasticity.

The impact of among-environment heteroscedasticity and genetic autocorrelation on the analysis of phenotypic plasticity is examined. Among-environment heteroscedasticity occurs when genotypic variances differ among environments. Genetic autocorrelation arises whenever the responses of a genotype to different environments are more or less similar than expected for observations randomly associated. In a multivariate analysis-of-variance model, three transformations of genotypic profiles (reaction norms), which apply to the residuals of the model while preserving the mean responses within environments, are derived. The transformations remove either among-environment heteroscedasticity, genetic autocorrelation or both. When both nuisances are not removed, statistical tests are corrected in a modified univariate approach using the sample covariance matrix of the genotypic profiles. Methods are illustrated on a Chlamydomonas reinhardtii data set. When heteroscedasticity was removed, the variance component associated with the genotype-by-environment interaction increased proportionally to the genotype variance component. As a result, the genetic correlation rg was altered. Genetic autocorrelation was responsible for statistical significance of the genotype-by-environment interaction and genotype main effects on raw data. When autocorrelation was removed, the ranking of genotypes according to their stability index dramatically changed. Evolutionary implications of our methods and results are discussed.

Nycterohemeral eating and ruminating patterns in heifers fed grass or corn silage: analysis by finite Fourier transform.

Average daily and within-day nycterohemeral patterns of eating and ruminating behavior were determined in six Holstein-Friesian heifers (average BW = 427 kg) given ad libitum access to either corn or grass silage in a two-period crossover design. Rhythm components (number of cycles/24 h) were characterized by finite Fourier transform of the 24-h mastication activities as measured during 4 d by continuous jaw movement recordings. Average daily voluntary intake of corn silage was 8.2% greater (P = .05) than that for grass silage and was associated (P < .05) with fewer meals and shorter daily, unitary eating and ruminating times, and smaller number of rumination boli. Analysis of variance of the daily mean of hourly activities and Rhythm Components 1 to 12 indicated effects of (P < .05) silage type (S), animal (A), period (P), and a significant interaction (S x A x P) for each mastication activity. The finite Fourier transform was reparameterized to express the amplitude (as periodograms) and phase of each rhythm component. Rhythm Components 1, 3, and 4 contributed primarily to explaining the total dispersion of the 24-h series of time spent eating and ruminating, for both silage types and individual heifers. Relative importance of Rhythm Component 1 of time spent eating, indicative of a main circadian pattern, was related positively to pedigree value for milk production (P = .01) and negatively to milk protein concentration (P = .09).(ABSTRACT TRUNCATED AT 250 WORDS)

Maternal behaviour in the Wistar rat under atypical Zeitgeber.

In previous experiments, neonatal rats subjected to atypical Zeitgeber (light and temperature) produce adult animals that present a different reactivity in some test situations. On the other hand, a great body of evidence indicates that physiological and psychological processes, including the maturation of the circadian system, are regulated by maternal behaviour. In order to investigate in which manner the atypical Zeitgeber, mentioned above, influences maternal behaviour, mothers with their young were observed systematically. More precisely, the question was: is the changed reactivity due to the fact that the young experience a situation of classical early privation (e.g., a decrease in social or other sensory stimulation or in nutrition) or is it due to a situation in which only the temporal pattern of the maternal behaviour is modified and thus disturbs the proper development of the circadian system of the young. Results tend to show that the only difference between the experimental and the control group is a modification in the circadian rhythm of several behavioural items. Only one activity (licking of the young) shows a significant difference in the overall mean value but in favour of the experimental group. We would suggest that the modified temporal pattern of the maternal care could be a poor timegiver for the young, in such a way that the development of their circadian rhythms is disturbed. The unusual reactivity in adulthood could depend on this abnormal development of the circadian system and not on a care privation.

[Quantitative analysis of the rythm of feeding and merycismal activities]. / Analyse quantitative du rythme des activités alimentaire et mérycique.

The nycterohemeral eating and ruminating patients were studied in 6 heifers given ad libitum access to grass or corn silage in a 2 period cross-over design. Rhythm components (no of cycles/24 h) were characterized by the finite Fourier transform of hourly means of the 24 h mastication activities. Rhythm components 1, 3 and 4 contributed considerably to explaining the total dispersion of 24 h eating and ruminating mastication series.

Quantitative analysis of nycterohemeral eating and ruminating patterns in heifers with different voluntary intakes and effects of monensin.

The nycterohemeral pattern of eating and ruminating behavior was examined in six heifers given ad libitum access to a corn silage-based diet with or without 100 mg monensin.hd-1.d-1 in a two-period crossover design. Rhythm components (no. of cycles/24 h) were characterized by the finite Fourier transform of 24-h mastication activities series measured over 7 d. Analysis of variance of the daily mean of hourly activities and rhythm components 1 to 12 showed significant effects of heifer (H), monensin treatment (T), period (P) and T x P x H interactions. A reparameterization of the finite Fourier transform yielded the amplitude and the phase for each rhythm component and allowed the plot of periodograms and phase diagrams, respectively. Rhythm components 1, 2 and 3 contributed primarily in explaining the total dispersion of 24-h eating and ruminating mastication series. The major effect of monensin was to increase the phase at rhythm component 1, delaying by 1 h in the onset of rumination after the morning feeding. Heifer effects were significant and appeared related to variations in daily voluntary intake (VI) of individual heifers. Heifer 5, with the largest VI, had two main eating periods and the highest rate of eating. Heifer 4, with the smallest VI, partitioned mastication activity throughout the day into four main eating and ruminating periods. Differences among animals in their nycterohemeral chewing behavior patterns may be related to differences in forestomach structure associated with a larger VI capacity by animals having mastication rhythm components 1 and 3.