Cotransfected human chondrocytes: over-expression of IGF-I and SOX9 enhances the synthesis of cartilage matrix components collagen-II and glycosaminoglycans

Damage to cartilage causes a loss of type II collagen (Col-II) and glycosaminoglycans (GAG). To restore the original cartilage architecture, cell factors that stimulate Col-II and GAG production are needed. Insulin-like growth factor I (IGF-I) and transcription factor SOX9are essential for the synthesis of cartilage matrix, chondrocyte proliferation, and phenotype maintenance. We evaluated the combined effect of IGF-I and SOX9 transgene expression on Col-II and GAG production by cultured human articular chondrocytes. Transient transfection and cotransfection were performed using two mammalian expression plasmids (pCMV-SPORT6), one for each transgene. At day 9 post-transfection, the chondrocytes that were over-expressing IGF-I/SOX9 showed 2-fold increased mRNA expression of the Col-II gene, as well as a 57% increase in Col-II protein, whereas type I collagen expression (Col-I) was decreased by 59.3% compared with controls. The production of GAG by these cells increased significantly compared with the controls at day 9 (3.3- vs 1.8-times, an increase of almost 83%). Thus, IGF-I/SOX9 cotransfected chondrocytes may be useful for cell-based articular cartilage therapies.

Technological properties of precooked flour containing coffee powder and rice by thermoplastic extrusion

Although Brazil is a country of tradition in both the production and consumption of coffee, the most of the coffee is consumed as a beverage, which reduces greatly the competitiveness on international market, for reducing the chances of supplying the product under other forms of consumption. Owing to that, the aim of this study was developing a precooked mixed flour containing coffee powder and rice for use in coffee flavored products. Mixtures of rice and coffee in the proportions of 900:100, 850:150 and 800:200 g, respectively, were processed in a single screw extruder (Brabender DS-20, Duisburg, German) and the effect of the extrusion process on the variables moisture content (16%, 18% and 20%) and temperature in the third extruding zone (140 °C, 160 °C and 180 °C) was studied. The results for expansion index ranged from 2.91 to 11.11 mm in diameter; the water absorption index from 4.59 to 6.33 g gel/g sample and the water solubility index varied from 4.05% to 8.57%. These results showed that, despite coffee powder influenced the variables studied, the expanded product after milling resulted in a extruded mixture with good absorption and water solubility indices, which favors the use of the precooked mixed flour for human consumption.

Characterization of pre-gelatinized rice and bean flour

The objective of this study was to develop a pre-gelatinized flour using a mixture of broken rice and split beans by thermoplastic extrusion, and to evaluate the physicochemical, nutritional, and technological quality of the final product. The extrusion parameters were maintained using three heating zones with temperatures of 30 ºC, 40 ºC, and 70 ºC; screw speed of 177 rpm; feed rate of 257 g/m, and circular matrix of 3.85 mm. The following characterization analyses were performed: physicochemical, nutritional, water absorption index (WAI), water solubility index (WSI), and paste viscosity. The pre-gelatinized rice and bean flour had an intermediate value of WAI, 7.51 g/g, and high WSI value, 24.61%. Regarding proteins, it was verified an average content of 12.9% in the final product. The amino acid contents found in the pre-gelatinized flour indicate that the mixture has the essential amino acids. It was also found that the pre-gelatinized flour supplies more than 60% of the essential amino acids recommended for children aged one to three years old. The gelatinized flour composed of broken rice and split beans is an alternative to the use of these by-products of the manufacture process of rice and beans to obtain a product with viable technological characteristics and high nutritional value.