Frutapin, a lectin from Artocarpus incisa (breadfruit): cloning, expression and molecular insights

Artocarpus incisa (breadfruit) seeds contain three different lectins (Frutalin, Frutapin and Frutackin) with distinct carbohydrate specificities. The most abundant lectin is Frutalin, an α-D-galactose-specific carbohydrate-binding glycoprotein with antitumour properties and potential for tumour biomarker discovery as already reported. Frutapin (FTP) is the second most abundant, but proved difficult to purify with very low yields and contamination with Frutalin frustrating its characterization. Here, we report for the first time high-level production and isolation of biologically-active recombinant FTP in E. coli BL21, optimizing conditions with the best set yielding >40 mg/L culture of soluble active FTP. The minimal concentration for agglutination of red blood cells was 62.5 µg/mL of FTP, a process effectively inhibited by mannose. Apo-FTP, FTP-mannose and FTP-glucose crystals were obtained and diffracted X-rays to a resolution of 1.58 (P212121), 1.70 (P3121) and 1.60 (P3121) Å, respectively. The best solution showed four monomers per asymmetric unit. Molecular Dynamics simulation suggested FTP displays higher affinity for mannose than glucose. Cell studies revealed FTP was non-cytotoxic to cultured mouse fibroblast 3T3 cells below 0.5 mg/mL and also capable of stimulating cell migration at 50 µg/mL. In conclusion, our optimized expression system allowed high amounts of correctly-folded soluble FTP to be isolated. This recombinant bioactive lectin will now be tested in future studies for therapeutic potential; for example, in wound healing and tissue regeneration

A Brazilian regional basic diet-induced chronic malnutrition drives liver inflammation with higher ApoA-I activity in C57BL6J mice

Malnutrition is still considered endemic in many developing countries. Malnutrition-enteric infections may cause lasting deleterious effects on lipid metabolism, especially in children living in poor settings. The regional basic diet (RBD), produced to mimic the Brazilian northeastern dietary characteristics (rich in carbohydrate and low in protein) has been used in experimental malnutrition models, but few studies have explored the effect of chronic RBD on liver function, a central organ involved in cholesterol metabolism. This study aimed to investigate whether RBD leads to liver inflammatory changes and altered reverse cholesterol metabolism in C57BL6/J mice compared to the control group, receiving a standard chow diet. To evaluate liver inflammation, ionized calcium-binding adapter protein-1 (IBA-1) positive cell counting, interleukin (IL)-1b immunohistochemistry, and tumor necrosis factor (TNF)-a and IL-10 transcription levels were analyzed. In addition, we assessed reverse cholesterol transport by measuring liver apolipoprotein (Apo)E, ApoA-I, and lecithin-cholesterol acyltransferase (LCAT) by RT-PCR. Furthermore, serum alanine aminotransferase (ALT) was measured to assess liver function. RBD markedly impaired body weight gain compared with the control group (Po0.05). Higher hepatic TNF-a (Po0.001) and IL-10 (Po0.01) mRNA levels were found in RBD-challenged mice, although without detectable non-alcoholic fatty liver disease. Marked IBA-1 immunolabeling and increased number of positive-IBA-1 cells (presumably Kupffer cells) were found in the undernourished group. No statistical difference in serum ALT was found. There was also a significant increase in ApoA-I mRNA expression in the undernourished group, but not ApoE and LCAT, compared with the control. Altogether our findings suggested that chronic RBD-induced malnutrition leads to liver inflammation with increased ApoA-I activity

Evaluation of babassu meal in feed for layer hens during the growth phase

Studies were conducted to evaluate the nutritional value and inclusion levels of babassu meal (BM) in the diet of grower layer pullets in substitution to wheat meal. Digestibility, metabolism and growth trials were conducted. Twelve cecectomized roosters were used in the digestibility assay to determine the coefficients of standardized digestibility of amino acids (CSDAA). The metabolism trial was conducted with 30 adult roosters to determine the apparent metabolizable energy corrected for nitrogen (AMEn) of BM. A growth trial was performed to determine replacement levels of wheat midds by BM diet using 360 six-week-old commercial layer pullets. BM was included at the 0, 75 and 150 g/kg of BM, during grower and development rearing phases, respectively. Feed intake, body weight gain, and feed conversion were evaluated. BM AMEn was determined as 1,474 kcal/kg, on as-fed basis. The CSDAA determined for BM were below 88% for all AA. The inclusion of BM in the feed of grower layers (7-18 week) significantly decreased feed intake (p < 0.05), but significantly improved body weight gain and feedconversion ratio (p < 0.05) at 15% inclusion level. Considering the nutritional value and performance results, BM can replace wheat midds in diets of grower layer pullets

Perda de massa de flores de capuchinha após armazenamento Loss of nasturtium flower mass after storage

Avaliou-se a perda de massa pós-colheita de flores de capuchinha, em experimento conduzido no Laboratório de Bioquímica da UFMS, em setembro de 2004. Flores das cores vermelha e amarela foram imersas durante trinta minutos em água, ácido ascórbico a 2% e 4%, cloreto de cálcio a 2% e 5% e sacarose a 5% e 10%, e armazenadas com e sem o uso de filme de PVC em câmara fria a 5ºC±2ºC. Foi analisada a perda de massa fresca, utilizando-se o delineamento inteiramente casualizado em fatorial 2 x 2 x 7, com três repetições (8 flores por bandeja) e as médias comparadas pelo teste de Tukey, a 5% de significância. A longevidade média das flores de capuchinha foi de oito dias para as flores acondicionadas na embalagem de PVC e armazenadas em câmara fria, enquanto as flores não acondicionadas apresentaram-se murchas a partir do segundo dia. Os tratamentos químicos, nas concentrações estudadas, não beneficiaram a conservação pós-colheita das flores da capuchinha.<br>The post-harvest behavior of nasturtium flowers was evaluated using sucrose, ascorbic acid and calcium chloride which were associated or not with polyethylene pack and stored under refrigeration. This work was carried out in a biochemical laboratory, in Dourados, Mato Grosso do Sul State, Brazil. Red and yellow flowers were immersed during thirty minutes in water, ascorbic acid at 2% and 4%, calcium chloride at 2% and 5% and sucrose at 5% and 10%, and then stored with and without package in a refrigerated chamber at 5°C ±2°C. The trial was carried out in a complete randomized experimental design in a 2 x 2 x 7 factorial scheme with three replications (8 flowers/tray). Nasturtium flowers stored with package in refrigerated chamber last for eight days, while flowers kept at environmental conditions withered after the second day. Treatments with ascorbic acid, calcium chloride and sucrose did not benefit post-harvest conservation of nasturtium flowers