Tenogenic induction of equine mesenchymal stem cells by means of growth factors and low-level laser technology

Tendons regenerate poorly due to a dense extracellular matrix and low cellularity. Cellular therapies aim to improve tendon repair using mesenchymal stem cells and tenocytes; however, a current limitation is the low proliferative potential of tenocytes in cases of severe trauma. The purpose of this study was to develop a method useful in veterinary medicine to improve the differentiation of Peripheral Blood equine mesenchymal stem cells (PB-MSCs) into tenocytes. PB-MSCs were used to study the effects of the addition of some growth factors (GFs) as TGFβ3 (transforming growth factor), EGF2 (Epidermal growth factor), bFGF2 (Fibroblast growth factor) and IGF-1 (insulin-like growth factor) in presence or without Low Level Laser Technology (LLLT) on the mRNA expression levels of genes important in the tenogenic induction as Early Growth Response Protein-1 (EGR1), Tenascin (TNC) and Decorin (DCN). The singular addition of GFs did not show any influence on the mRNA expression of tenogenic genes whereas the specific combinations that arrested cell proliferation in favour of differentiation were the following: bFGF2 + TGFβ3 and bFGF2 + TGFβ3 + LLLT. Indeed, the supplement of bFGF2 and TGFβ3 significantly upregulated the expression of Early Growth Response Protein-1 and Decorin, while the use of LLLT induced a significant increase of Tenascin C levels. In conclusion, the present study might furnish significant suggestions for developing an efficient approach for tenocyte induction since the external administration of bFGF2 and TGFβ3, along with LLLT, influences the differentiation of PB-MSCs towards the tenogenic fate.     

Increased abundance of patatins,lipoxygenase and miraculins in a thaxtomin A-habituated potato Russet Burbank somaclone with enhanced resistance to common scab

Potato common scab caused by the actinobacterium Streptomyces scabiei is characterized by the formation of corky lesions on tubers that reduce their marketability. Management of common scab is very complex and often ineffective under various environmental conditions. Using potato varieties that are more resistant to common scab remains one of the most efficient strategies to control this disease. However, very little is known about the factors associated with resistance to common scab. Somaclone RB9 regenerated from thaxtomin A-habituated potato Russet Burbank calli produced tubers more resistant to common scab than the original variety. Comparison of the RB9 tuber proteome with that of Russet Burbank using label-free quantitative proteomic analysis revealed changes in the accumulation of defence-related proteins from the patatin and lipoxygenase (LOX) families, which are involved in the metabolism of lipids, and of two miraculins of the Kunitz-type protease inhibitors family. The implication of LOX during common scab infection was studied using synchronized minitubers developed from leaf-bud cuttings. S. scabiei infection stimulated the accumulation of LOX in both Russet Burbank and RB9 minitubers, but this accumulation was intensified in RB9 minitubers. Infection also increased LOX activity in Russet Burbank and RB9 minitubers. However, LOX activity measured in noninfected RB9 minitubers was similar to that of infected Russet Burbank minitubers, indicating endogenous activation of LOX activity in RB9 minitubers. We discuss how increased LOX abundance and activity in the somaclone RB9 may contribute to improving tuber defence against common scab.     

The Exploitation of Nematode-Responsive Plant Genes in Novel Nematode Control Methods

The molecular interactions between plants and sedentary nematodes are undergoing intense study, not only for reasons of fundamental research but also for the potential benefits to agriculture. The present technology allows the transformation of an increasing number of crop plants, providing new ways to introduce resistance against plant-parasitic nematodes. The ability of sedentary nematodes to induce specialized feeding sites in plant roots is one of the most fascinating aspects of this host–parasite interaction. Molecular approaches have been initiated to identify and characterize plant genes altered in expression after infection by sedentary nematodes. The results obtained indicate that many genes indeed become up-regulated upon nematode infection. Surprisingly, several so-called constitutive promoters that are normally used to achieve high expression in plant cells are completely ‘silenced’ in the feeding sites within days after nematode infection. Generally, there are two options available for the genetic engineering of nematode resistance: the synthesis of anti-nematode proteins or the localized production of a cytotoxic protein that interferes with the development of feeding cells. Nematode-induced promoters are very useful for the production by plants of sufficiently high levels of anti-nematode proteins at feeding sites. Alternatively, interfering with feeding-cell development is somewhat similar to the hypersensitive response evoked by nematodes in a naturally resistant plant. Here, destruction of specific plant cells can be achieved by the localized expression of a cytotoxin such as barnase, a potent ribonuclease. This approach, however, calls for a highly specific ‘non-leaky’ promoter, which is active only in the feeding cells. Another possibility is to use a two-component system, where the leakiness of the promoter in other tissues is counterbalanced by the constitutive expression of a neutralizing gene.     

Identification of pathogenic Leptospira strains in tissues of a premature foal by use of polymerase chain reaction analysis.

Studies were carried out to determine the cause of death in a prematurely born Thoroughbred foal that died 24 hours after birth. Necropsy revealed gross lesions suggestive of septicemia. A commercial Leptospira polymerase chain reaction (PCR) assay designed to specifically amplify the hemolysis-associated protein 1 (hap1) gene present only in pathogenic Leptospira strains detected the presence of Leptospira DNA in various tissues of the foal. Histologic examination of lung, liver, kidney, and myocardium revealed numerous spirochetes in Warthin-Starry-stained tissue sections. Results of PCR analysis and histologic examination suggested a leptospiral infection in the newborn foal. At the moment of death, the infection coexisted with a streptococcal-associated aspiration bronchopneumonia and postpartum septicemia. These findings indicate that the PCR assay based on the amplification of the hap1 gene represents a useful tool for specific detection of pathogenic leptospira in field samples taken from horses.     

Immunohistochemical characterization of a hepatic neuroendocrine carcinoma in a cat.

A hepatic mass was identified in a 5-year-old, female mixed-breed cat that died spontaneously after a clinical history of progressive emaciation, ptyalism, and persistent coryza. At necropsy, a 7-cm-diameter, yellow-brown, firm, multilobulated tumor was identified in the liver. Microscopically, the mass consisted of neoplastic cells arranged in small, closely packed nests within a thin fibrovascular stroma. These cells were of medium sized and polygonal, with fine argyrophilic cytoplasmic granules. Nuclei were predominantly round with finely stippled chromatin and indistinct nucleoli. Mitotic figures were numerous. Immunohistochemically, most of the neoplastic cells were immunoreactive for chromogranin A, neuron-specific enolase (NSE), and cytokeratin AE1/AE3 and weakly labeled for synaptophysin. The tumor was negative for glial fibrillary acidic protein (GFAP), vimentin, and cytokeratins 5, 6, 8, and 17. Vascular emboli and intrahepatic micrometastasis were also identified with chromogranin A. All these features were consistent with a hepatic neuroendocrine carcinoma and emphasized the importance of using a panel of antibodies to diagnose such rare tumors.     

Influence of complexation between amylose and a flavored model sponge cake on the degree of aroma compound release

Flavoring is used in the food industry to reinforce the aroma profile of baked cereal goods. During the processing of such products, interactions between starch and aroma compounds can occur, and this may have an impact on aroma release and perception. In the present study, 20 aroma compounds were tested to establish whether they formed complexes with amylose. The structure of the complexes was determined by wide-angle X-ray scattering (WAXS). A cocomplexation study proved that several complexing compounds could be present in the same crystalline aggregate. WAXS and differential scanning calorimetry (DSC) experiments were performed in a flavored model sponge cake at different steps of processing and showed that aroma compounds might form complexes with amylose in a sponge cake as they can do in simple system containing only amylose. Some of the aroma compounds trapped in the sponge cake were quantified, and their release behavior was followed by headspace analysis. The V-type structure could partly explain aroma retention in the product and the rate of aroma release.     

Rapid quantitative determination of oleuropein in olive leaves (Olea europaea) using mid-infrared spectroscopy combined with chemometric analyses

Oleuropein, the major active compound in olive leaf, is well known for its benefits for human health. Oleuropein is classically quantified by HPLC, which is time and chemical consuming, laborious and expensive. The aim of this work was to examine the potential of mid-infrared spectroscopy, as a rapid tool, to predict oleuropein content in olive leaf from five Tunisian cultivars (Chemlali, Chetoui, Meski, Sayali and Zarrazi) and one French cultivar (Bouteillan). The reference data of oleuropein content were obtained by the HPLC method. Hundred five samples were analyzed by HPLC and mid-infrared spectroscopy. Samples were randomly divided in a calibration set (73 samples) and in a validation set (32 samples). The spectral data sets were correlated with reference data of oleuropein content by using partial least squares (PLS) regression algorithm. The results showed that the PLS model gave satisfactory model for quantitative prediction of oleuropein content in olive leaf (relative error of prediction = 8.5%). The correlation coefficient was 0.91 and 0.74 for calibration set and validation set, respectively. It can be concluded that mid-infrared spectroscopy constitutes a promising technique for rapid quantification of oleuropein in olive leaf.     

The Level II aggregated forest soil condition database links soil physicochemical and hydraulic properties with long-term observations of forest condition in Europe

Key message

Aggregated, consolidated, and derived soil physicochemical data of 286 ICP Forests Level II plots were completed with soil hydraulic properties for integrated use with forest monitoring data. Database access should be requested at http://icp-forests.net . Metadata associated available at https://metadata-afs.nancy.inra.fr/geonetwork/apps/georchestra/?uuid=153e599e-6624-4e2b-b862-8124386ea9cd&hl=eng

Context

The ICP Forests database is one of the most comprehensive forest ecosystem datasets in Europe and contains the accumulated results of more than two decades of harmonised forest monitoring all over Europe.

Aims

The aim of this paper is to share knowledge on the ICP Forests Level II soil data for broader use among forest scientists.

Methods

After standard analysis, quality checks, aggregation, and calculation of derived variables (e.g. nutrient stocks, base saturation, C:N ratio, and water retention parameters), data have been gathered into a static database (AFSCDB.LII.2.2), which will be updated to new versions as soon as new measurements become available.

Results

The database provides a basis for the combined evaluation of up to 130 unique soil variables of 286 plots with dynamic data on tree growth, ground vegetation, foliar chemistry, crown condition, tree phenology, leaf area index, ozone injury, litterfall, soil solution chemistry, deposition, ambient air quality, and meteorological data assessed on the same plots.

Conclusion

The unprecedented comprehensiveness and level of detail in this newly aggregated database may overcome existing restrictions so far impeding the realisation of large-scale forest ecosystem studies in Europe.

     

Tree-root systems and herbaceous species-characteristics under tree species introduced into grazing lands in subhumid Cameroon

The effect of tree species on the characteristics of the herbaceous stratum, during the first five years of a fallow, was evaluated in the North of Cameroon (average annual temperature 28.2 °C, total annual rainfall 1050 mm). Treatments included a natural grazed herbaceous fallow, a natural ungrazed herbaceous fallow and three planted tree fallows (Acacia polyacantha Willd. ssp. campylacantha (Hochst. ex A. Rich.), Senna siamea Lam. and Eucalyptus camaldulensis Dehnh.), which were protected against grazing. Because tree species influenced light interception in different ways, as well as having different root patterns, they had different effects on the herbaceous stratum in terms of species composition and biomass. The grazed herbaceous fallow maintained the greatest species richness. Protection against grazing or the introduction of tree species associated with the absence of grazing induced both a progressive evolution to a particular species composition. The ungrazed herbaceous fallow consisted mainly of Andropogon gayanus Kunth, which provided the greatest biomass (8 t dry matter ha^–1 at the end of the fallow period). E. camaldulensis provided little shade and the lowest fine root mass in the top layer allowing the growth of A. gayanus and thus a greater herbaceous biomass (3.5 t DM ha^–1) than that found under the other tree species. Under the heavy shade of A. polyacantha, the herbaceous stratum consisted mainly of annual Pennisetum spp. (2.2 t DM ha^–1) and showed the greatest N concentration (1.3%), probably due to N₂ fixation by the tree species. After the fourth year, despite the relatively open tree canopy, S. siamea, which showed the highest fine root mass, had a strong depressive effect on the herbaceous stratum. This revised version was published online in June 2006 with corrections to the Cover Date.     

Origin of French virgin olive oil registered designation of origins predicted by chemometric analysis of synchronous excitation-emission fluorescence spectra

The authentication of virgin olive oil samples requires usually the use of sophisticated and very expensive analytical techniques, so there is a need for fast and inexpensive analytical techniques for use in a quality control methodology. Virgin olive oils present an intense fluorescence spectra. Synchronous excitation-emission fluorescence spectroscopy (SEEFS) was assessed for origin determination of virgin olive oil samples from five French registered designation of origins (RDOs) (Nyons, Vallée des Baux, Aix-en-Provence, Haute-Provence, and Nice). The spectra present bands between 600 and 700 nm in emission due to chlorophylls a and b and pheophytins a and b. The bands between 275 and 400 nm in emission were attributed to alpha-, beta-, and gamma-tocopherols and to phenolic compounds, which characterize the virgin olive oils compared to other edible oils. The chemometric treatment (PLS1) of synchronous excitation-emission fluorescence spectra allows one to determine the origin of the oils from five French RDOs (Baux, Aix, Haute-Provence, Nice, and Nyons). Results were quite satisfactory, despite the similarity between two denominations of origin (Baux and Aix) that are composed by some common cultivars (Aglandau and Salonenque). The interpretation of the regression coefficients shows that RDOs are correlated to chlorophylls, pheophytins, tocopherols, and phenols compounds, which are different for each origin. SEEFS is part of a global analytic methodology that associates spectroscopic and chromatographic techniques. This approach can be used for traceability and vindicates the RDOs.