Assessment of cold tolerance at early developmental stages and allelic variation at candidate genes in South American rice germplasm

Rice is susceptible to cold during early developmental stages. Most tolerant cultivars have been developed for other conditions than those occurring in South America’s rice growing areas, or their grain type is not suitable for the local markets. If locally adapted varieties were available, growers could anticipate sowing date, making flowering time coincide with the moment of maximum solar radiation and increasing yields. In this work, 116 rice inbred lines and varieties of diverse origin within the South American gene pool were tested for seedling survival and germination percentage under low temperature in controlled conditions. As a result, lines used as controls responded as expected, whereas lines with similar behavior to controls, intermediate between susceptible and tolerant controls and more extreme than controls were detected at both seedling and germination stages. Allelic variation at candidate genes OsGSTZ1, OsGSTZ2 and OsCDPK13 was analyzed in a subset of ten contrasting lines. Ten out of thirty-four polymorphisms detected in all three genes were associated with cold tolerance in these lines. A functional marker, designed from an aminoacid-changing SNP found in gene OsGSTZ2, was tested in the 116 lines. As a result, a tight association was observed between cold tolerance and marker score. In conclusion, wide variability for cold tolerance at early developmental stages has been detected in rice advanced breeding materials that are adapted to local growing conditions. Also, a functional marker tightly associated with the trait is available for performing marker assisted selection.     

Antiproliferative Effect of Amaranth Proteins and Peptides on HT-29 Human Colon Tumor Cell Line

Antiproliferative effect of Amaranthus mantegazzianus proteins and peptides released after simulated gastrointestinal digestion (DH% 37.8?±?3.8) was investigated on human colon cancer cell line HT-29. Inhibition of proliferation of HT-29 cells was exhibited after a 24 h treatment with different concentrations of amaranth protein isolate (API) and the peptides released after digestion (DGS), presenting IC₅₀ values of 1.35?±?0.12 and 0.30?±?0.07 mg soluble protein/mL, respectively. Lactate dehydrogenase assay indicated that both samples caused the loss of membrane integrity and cell lysis over HT-29 cells, and DAPI fluorescence microscopies evidenced typical apoptotic features. Moreover, Annexin V-FITC flow cytometry showed a significant increase of early apoptotic and late apoptotic/necrotic HT-29 cells compared to untreated ones, and caspase-3 assay confirmed the apoptosis induction with a 43.0?±?10.3 and 65.8?±?12.7% increase of caspase-3 activity produced by a 2 mg/mL treatment of API and DGS, respectively. In conclusion, amaranth peptides successfully released after simulated gastrointestinal digestion would exert a potential antiproliferative activity over HT-29 tumor cells. This effect was linked to the induction of cell necrosis and apoptosis, supporting the idea of using amaranth proteins as a potential food alternative ingredient for functional foods.

     

Alternative and Efficient Extraction Methods for Marine-Derived Compounds

Marine ecosystems cover more than 70% of the globe’s surface. These habitats are occupied by a great diversity of marine organisms that produce highly structural diverse metabolites as a defense mechanism. In the last decades, these metabolites have been extracted and isolated in order to test them in different bioassays and assess their potential to fight human diseases. Since traditional extraction techniques are both solvent- and time-consuming, this review emphasizes alternative extraction techniques, such as supercritical fluid extraction, pressurized solvent extraction, microwave-assisted extraction, ultrasound-assisted extraction, pulsed electric field-assisted extraction, enzyme-assisted extraction, and extraction with switchable solvents and ionic liquids, applied in the search for marine compounds. Only studies published in the 21st century are considered.     

Pharmacokinetics and efficacy of intravenous and extradural tramadol in dogs

Tramadol is a synthetic opioid agonist used extensively in human and, to a lesser extent, veterinary medicine throughout the world. The clinical efficacy and pharmacokinetic profile of intravenous (IV) and extradural (ED) tramadol (2 mg/kg) and its o-desmethyl metabolite were studied in dogs undergoing tibial plateau levelling osteotomy (TPLO). Intra-operative cardiorespiratory variables were monitored and post-operative pain was assessed using the short form of the Glasgow Composite Pain Scale. A rapid (<5 min) and effective production of o-desmethyl tramadol was recorded. The pharmacokinetic profile was similar for tramadol and its metabolite irrespective of the route of administration. ED tramadol provided sufficient intra- and post-operative analgesia without significant clinical side-effects, but the post-operative analgesia was comparable to that following IV administration and the ED route could therefore not be considered a practical alternative to the IV route.     

Biotechnological production of vitamin B2-enriched bread and pasta

Lactic acid bacteria (LAB) were obtained from durum wheat flour samples and screened for roseoflavin-resistant variants to isolate natural riboflavin-overproducing strains. Two riboflavin-overproducing strains of Lactobacillus plantarum isolated as described above were used for the preparation of bread (by means of sourdough fermentation) and pasta (using a prefermentation step) to enhance their vitamin B2 content. Pasta was produced from a monovarietal semolina obtained from the durum wheat cultivar PR22D89 and, for experimental purposes, from a commercial remilled semolina. Several samples were collected during the pasta-making process (dough, extruded, dried, and cooked pasta) and tested for their riboflavin content by a high-performance liquid chromatography method. The applied approaches resulted in a considerable increase of vitamin B2 content (about 2- and 3-fold increases in pasta and bread, respectively), thus representing a convenient and efficient food-grade biotechnological application for the production of vitamin B2-enriched bread and pasta. This methodology may be extended to a wide range of cereal-based foods, feed, and beverages. Additionally, this work exemplifies the production of a functional food by a novel biotechnological exploitation of LAB in pasta-making.