<Emphasis Type="Italic">Lupinus albus</Emphasis> Conglutin Gamma Modifies the Gene Expressions of Enzymes Involved in Glucose Hepatic Production <Emphasis Type="Italic">In Vivo</Emphasis>

Lupinus albus seeds contain conglutin gamma (Cγ) protein, which exerts a hypoglycemic effect and positively modifies proteins involved in glucose homeostasis. Cγ could potentially be used to manage patients with impaired glucose metabolism, but there remains a need to evaluate its effects on hepatic glucose production. The present study aimed to analyze G6pc, Fbp1, and Pck1 gene expressions in two experimental animal models of impaired glucose metabolism. We also evaluated hepatic and renal tissue integrity following Cγ treatment. To generate an insulin resistance model, male Wistar rats were provided 30% sucrose solution ad libitum for 20 weeks. To generate a type 2 diabetes model (STZ), five-day-old rats were intraperitoneally injected with streptozotocin (150 mg/kg). Each animal model was randomized into three subgroups that received the following oral treatments daily for one week: 0.9% w/v NaCl (vehicle; IR-Ctrl and STZ-Ctrl); metformin 300 mg/kg (IR-Met and STZ-Met); and Cγ 150 mg/kg (IR-Cγ and STZ-Cγ). Biochemical parameters were assessed pre- and post-treatment using colorimetric or enzymatic methods. We also performed histological analysis of hepatic and renal tissue. G6pc, Fbp1, and Pck1 gene expressions were quantified using real-time PCR. No histological changes were observed in any group. Post-treatment G6pc gene expression was decreased in the IR-Cγ and STZ-Cγ groups. Post-treatment Fbp1 and Pck1 gene expressions were reduced in the IR-Cγ group but increased in STZ-Cγ animals. Overall, these findings suggest that Cγ is involved in reducing hepatic glucose production, mainly through G6pc inhibition in impaired glucose metabolism disorders.     

Phytophthora oleae,a new root pathogen of wild olives

Wild olive (Olea europaea subsp. europaea var. sylvestris) is an important component of Mediterranean forests and a key genetic source for olive improvement programmes. Since 2009, a severe decline caused by Phytophthora cryptogea and P. megasperma has been detected in a protected wild olive forest of high ecological value (Dehesa de Abajo, Seville, Spain). In this natural forest, sampling of roots and soil was carried out on 25 wild olives with symptoms in 2014 and 2015. Apart from the already known P. cryptogea A1 and P. megasperma, a third Phytophthora species was consistently isolated from wild olive rootlets with symptoms. These isolates conformed morphologically with the newly described species P. oleae and were confirmed by analysis of their ITS regions and cox1 sequences. Temperature–growth relationships showed a maximum growth at 19.9 °C on carrot agar medium, making it the lowest temperature Phytophthora species infecting wild olive roots. Pathogenicity was confirmed on 1-year-old healthy wild olive seedlings and was similar to the previously known pathogenic phytophthoras. As temperature requirements are quite different, the three Phytophthora species may be active against wild olive roots in different seasons. However, the prevalence of P. oleae infecting wild olives in recent years could be due to its introduction as a new invasive pathogen. The probable invasive nature of P. oleae, together with increasing rain episodes concentrated in short periods frequent in southern Spain, would allow the outbreak of infections in wild olive forests, and also put cultivated olive orchards at risk.     

Methyl jasmonate reduces chilling injury and maintains postharvest quality of mango fruit

Exposure of mango (Mangifera indica cv. Tommy Atkins) fruit to methyl jasmonate (MJ) vapors (10(-)(4) M) for 24 h at 25 degrees C reduced chilling injury during subsequent storage for 21 days at 7 degrees C and after 5 days of shelf life at 20 degrees C. The chilling tolerance induced by MJ was positively correlated with the reduction in the percent ion leakage of mango tissue. The overall quality of MJ-treated fruit was also better than that of control fruit. MJ treatment increased the total soluble solids but did not affect titratable acidity or pH. MJ also did not change the normal climacteric rise in respiration, water loss, and softening rates. The efficacy of MJ to reduce chilling injury and decay of mango could be related to the tolerance induced at low temperature. It was concluded that MJ treatment may prevent chilling injury symptoms of mango without altering the ripening process.     

Ellagic acid, vitamin C, and total phenolic contents and radical scavenging capacity affected by freezing and frozen storage in raspberry fruit

The ellagic acid, total phenolic, and vitamin C contents in four raspberry cultivars (Heritage, Autumn Bliss, Rubi, and Zeva) grown in Spain were detected and quantified by HPLC in fresh, just frozen, and stored fruits at -20 degrees C for a one year period. Ellagic acid [207-244 mg kg(-)(1) of fresh weight (fw)], total phenolic (137-1776 mg kg(-)(1) of fw), and vitamin C (221-312 mg kg(-)(1) of fw) contents in raw material were higher in the late cultivars Zeva and Rubi than in the early cultivars Autumn Bliss and Heritage. The freezing process slightly affected the values of extracted ellagic acid, total phenolic, and vitamin C content. At the end of long-term frozen storage (12 months), no significant change of total phenolic content extracted was observed, but significant decreases of 14-21% in ellagic acid and of 33-55% in vitamin C were quantified. Free radical scavenging capacity measured as antiradical efficiency (AE) depends on the seasonal period of harvest. Late cultivars, Rubi (6.1 x 10(-)(4)) and Zeva (10.17 x 10(-)(4)), showed higher AE than early cultivars, Heritage (4.02 x 10(-)(4)) and Autumn Bliss (4.36 x 10(-)(4)). The freezing process produced a decrease of AE values in the four cultivars ranging between 4 and 26%. During the frozen storage, the AE values reached after the freezing process remained unchanged.     

Partial characterization of polyphenol oxidase activity in raspberry fruits.

A partial characterization of polyphenol oxidase (PPO) activity in raspberry fruits is described. Two early cultivars harvested in May/June (Heritage and Autumm Bliss) and two late cultivars harvested in October-November (Ceva and Rubi) were analyzed for PPO activity. Stable and highly active PPO extracts were obtained using insoluble poly(vinylpyrrolidone) (PVP) and Triton X-100 in sodium phosphate, pH 7.0 buffer. Polyacrylamide gel electrophoresis of raspberry extracts under nondenaturing conditions resolved in one band (R(f)()(1) = 0.25). Raspberry PPO activity has pH optima of 8.0 and 5.5, both with catechol (0.1 M). Maximum activity was with D-catechin (catecholase activity), followed by p-coumaric acid (cresolase activity). Heritage raspberry also showed PPO activity toward 4-methylcatechol. Ceva and Autumm Bliss raspberries showed the higher PPO activity using catechol as substrate.     

Subcellular localization and isoenzyme pattern of peroxidase and polyphenol oxidase in beet root (Beta vulgaris L.)

The two enzymes involved in enzymatic browning reactions, polyphenol oxidase (PPO) and peroxidase (PO), have been partially purified and extracted from different fractions of beet root. PPO is mainly located in the membrane fraction, and it was also found in the soluble fraction. In both cases PPO was in its latent state. However, PO activity was higher in the soluble fraction than in the membrane fraction. Nevertheless, the highest values of specific activity for PO were obtained from the solubilized enzyme from acetone powders. Under native isoelectric focusing (IEF), several PPO isoenzymes were present in the pH range of 4.8-5.8. All of these isoenzymes shared a single band with a similar apparent mass under sodium dodecyl sulfate-polyacrylamide gel electrophoresis. PO was also analyzed by IEF, showing a complex isoenzyme pattern in all fractions. The characteristic basic PO isoenzyme of high pI found in both the soluble fraction and the solubilized enzyme from acetone powders was not detected in the membrane fraction. The kinetic characterization of PPO and PO from all fractions was carried out.     

Effects of rhodium heterogeneous catalyst and isomerization conditions on linoleic acid conjugation of soybean oil

Rhodium heterogeneous catalyst was used to catalyze isomerization of linoleic acid in soybean oil to conjugated linoleic acid (CLA). A central composite rotatable design with five levels of three variables, namely, reaction temperature, stirring speed, and reaction time, was used to determine the maximum CLA yield. The formation of CLA during isomerization was greatly dependent on the reaction temperature and time. The CLA content of soybean oil increased from 0.63 to 202.42 mg/g oil when isomerization was done at 200 degrees C, with a stirring speed of 200 rpm for 49 min. Analysis of triacylglycerol positions showed that linoleic acid at any position in a triacylglyceride could possibly be isomerized to CLA.     

Antifungal activity of strawberry fruit volatile compounds against Colletotrichum acutatum

Eight volatile products characterizing strawberry aroma, which is generated from the oxidative degradation of linoleic and linolenic acids by a lipoxygenase (LOX) pathway, were examined because of their antifungal activity against Colletotrichum acutatum, one of the causal agents of strawberry anthracnose. In this study, the effects of aldehydes, alcohols, and esters on mycelial growth and conidia development were evaluated. (E)-Hex-2-enal was found to be the best inhibitor of mycelial growth [MID (minimum inhibitory doses)=33.65 microL L(-1)] and of spore germination (MID=6.76 microL L(-1)), while hexyl acetate was the least effective of all volatile compounds tested (MID=6441.89 microL L(-1) for mycelial growth and MID=1351.35 microL L(-1) for spore germination). Furthermore, the antifungal activity of (E)-hex-2-enal on susceptibility of strawberry fruits to C. acutatum was also confirmed. The presence of these molecules in jars containing strawberry fruits inoculated with a suspension of spores inhibited the fungus growth and prevented the appearance of symptoms. Moreover, a study of the effects of (E)-hex-2-enal on conidial cells of C. acutatum was evaluated by transmission electron microscopy. This volatile compound altered the structures of the cell wall and plasma membrane, causing disorganization and lysis of organelles and, eventually, cell death.