Engineering and expression of a RhoA peptide against respiratory syncytial virus infection in plants.

MAIN CONCLUSION : A RhoA-derived peptide fused to carrier molecules from plants showed enhanced biological activity of in vitro assays against respiratory syncytial virus compared to the RhoA peptide alone or the synthetic RhoA peptide. A RhoA-derived peptide has been reported for over a decade as a potential inhibitor of respiratory syncytial virus (RSV) infection both in vitro and in vivo and is anticipated to be a promising alternative to monoclonal antibody-based therapy against RSV infection. However, there are several challenges to furthering development of this antiviral peptide, including improvement in the peptide's bioavailability, development of an efficient delivery system and identification of a cost-effective production platform. In this study, we have engineered a RhoA peptide as a genetic fusion to two carrier molecules, either lichenase (LicKM) or the coat protein (CP) of Alfalfa mosaic virus. These constructs were introduced into Nicotiana benthamiana plants using a tobacco mosaic virus-based expression vector and targets purified. The results demonstrated that the RhoA peptide fusion proteins were efficiently expressed in N. benthamiana plants, and that two of the resulting fusion proteins, RhoA-LicKM and RhoA2-FL-d25CP, inhibited RSV growth in vitro by 50 and 80 %, respectively. These data indicate the feasibility of transient expression of this biologically active antiviral RhoA peptide in plants and the advantage of using a carrier molecule to enhance target expression and efficacy.

Pulp and peel of unripe stenospermocarpic mango (Mangifera indica L. cv Ataulfo) as an alternative source of starch, polyphenols and dietary fibre.

As a result of climate change, the production of stenospermocarpic mangoes has increased dramatically. The stenospermocarpic mango, a fruit with reduced size and no seed, is considered to be a by-product that is both underutilised and wasted. Here, we studied the colour, chemical composition, polyphenol content, antioxidant capacity and starch in vitro digestibility of unripe stenospermocarpic mango flours (pulp and peel). The stenospermocarpic mango pulp flour had 11.7 g/100 g of dietary fibre with a balance of soluble and insoluble fractions; additionally, the total starch content of 41 g/100 g in its uncooked flour (resistant starch) can contribute to an increase in the indigestible carbohydrates. The mango peel flour had higher dietary fibre (40.5 g/100 g) and lower total starch content (21 g/100 g) compared with mango pulp flour. The mango pulp flour had higher phenolic compounds content (99.71 mg/g) and antioxidant capacity (248.5 mg/g, DPPH) compared with the peel flour (16.51 mg/g and 92.08 mg/g, DPPH), respectively. The rapidly digestible starch fraction was approximately 50%, with a balance in the content of slowly and resistant starch fractions in the mango pulp flour (approximately 20% per fraction). The flours of the pulp and peel of unripe stenospermocarpic mangoes can be used as alternative ingredients for preparing functional foods with high dietary fibre content and polyphenol compounds with antioxidant capacities.

Physicochemical, microstructural and digestibility analysis of gluten-free spaghetti of whole unripe plantain flour.

Plantain is a climacteric fruit having economic relevance in several tropical regions. Unripe plantain is an alternative source of indigestible carbohydrates (dietary fibre) and undigestible starch fraction. Unripe plantain flour was explored in this work as an alternative ingredient (whole and pulp) in spaghetti formulations. Chemical composition, cooking quality, texture analysis, and microstructure of spaghetti formulations were analyzed. The microstructure results showed that the presence of fiber in the food matrix helped the reduction of the starch granule swelling in the cooking process. Spaghetti made with whole plantain flour exhibited lower rapidly starch fraction, with increased resistant starch fractions. Overall, the whole unripe plantain flour exhibited good potential for gluten-free spaghetti having highest content of fiber and lower starch digestion rates.

Determination of Pinaverium Bromide in Human Plasma by a Sensitive and Robust UPLC-MS-MS Method and Application to a Pharmacokinetic Study in Mexican Subjects.

A high-throughput ultra-performance liquid chromatography coupled to tandem mass spectrometry (LC-ESI-MS-MS) method was developed for the determination of pinaverium bromide in human plasma. Protein precipitation with acetonitrile was used to extract pinaverium and itraconazole (as internal standard) from 500 µL plasma samples. The chromatographic separation was achieved with an Acquity UPLC BEH C18 column (1.7 µm, 2.1 × 100 mm) using a mixture of acetonitrile-5 mM ammonium formate (80:20, v/v) as mobile phase. Isocratic elution at 0.3 mL/min was used. Detection was performed by positive ion electrospray tandem mass spectrometry on a XEVO TQ-S by multiple reaction monitoring mode. The mass transitions monitorized were as follows: m/z 511.2 → 230 for pinaverium bromide, and m/z 705.29 → 392.18 for the itraconazole. The method was validated over a concentration range of 12-12,000 pg/mL. The chromatographic method runtime is 2.5 min and was applied to characterize the pharmacokinetics of pinaverium bromide after the oral administration of 100 mg to healthy Mexican subjects.

Absence of a significant pharmacokinetic interaction between atorvastatin and fenofibrate: a randomized, crossover, study of a fixed-dose formulation in healthy Mexican subjects.

Several clinical trials have substantiated the efficacy of the co-administration of statins like atorvastatin (ATO) and fibrates. Without information currently available about the interaction between the two drugs, a pharmacokinetic study was conducted to investigate the effect when both drugs were co-administered. The purpose of this study was to investigate the pharmacokinetic profile of tablets containing ATO 20 mg, or the combination of ATO 20 mg with fenofibrate (FNO) 160 mg administered to healthy Mexican volunteers. This was a randomized, two-period, two-sequence, crossover study; 36 eligible subjects aged between 20-50 years were included. Blood samples were collected up to 96 h after dosing, and pharmacokinetic parameters were obtained by non-compartmental analysis. Adverse events were evaluated based on subject interviews and physical examinations. Area under the concentration-time curve (AUC) and maximum plasma drug concentration (Cmax) were measured for ATO as the reference and ATO and FNO as the test product for bioequivalence design. The estimation computed (90% confidence intervals) for ATO and FNO combination versus ATO for Cmax, AUC0-t and AUC0-∞, were 102,09, 125,95, and 120,97%, respectively. These results suggest that ATO and FNO have no relevant clinical-pharmacokinetic drug interaction.

Pharmacokinetic non-interaction analysis in a fixed-dose formulation in combination of atorvastatin and ezetimibe.

Recent clinical research has shown that atorvastatin (ATO) in combination with cholesterol absorption inhibitor ezetimibe (EZE) significantly reduces LDL-C level in patients with hypercholesterolemia, showing a superior lipid-lowering efficacy compared to statin alone. With no information currently available on the interaction between the two drugs, a pharmacokinetic study was conducted to investigate the influence of EZE on ATO and conversely when the two drugs were coadministered. The purpose of this study was to investigate the presence of differences in the pharmacokinetic profiles of capsules containing ATO 80 mg, EZE 10 mg or the combination of both 80/10 mg administered to healthy Mexican volunteers. This was a randomized, three-period, six-sequences crossover study. 36 eligible subjects aged between 20 to 50 years were included. Blood samples were collected up to 96 h after dosing, and pharmacokinetic parameters were obtained by non-compartmental analysis. Adverse events were evaluated based on subject interviews and physical examinations. Area under the concentration-time curve (AUC) and maximum plasma drug concentration (Cmax) were measured for each drug alone or together and tested for bioequivalence-based hypothesis. The estimation computed (90% confidence intervals) for AUC and Cmax, were 96.04% (85.88-107.42%) and 97.04% (82.36-114.35%), respectively for ATO-EZE combination versus ATO alone, while 84.42% (77.19-92.32%) and 95.60% (82.43-110.88%), respectively, for ATO-EZE combination versus EZE alone were estimated. These results suggest that ATO and EZE have no relevant pharmacokinetic drug-drug interaction.