Lipidomic Profile of Rhodotorula toruloides by GC/MS and Antioxidant Capacity of the Oil by DPPH and TLC-Plate Methods

This work was undertaken to evaluate the antioxidant capacity of Rhodotorula toruloides lipid extract in TLC plate, using the (DPPH) (1,1-diphenyl-2-picril-  hydrazine) method as an innovative way to visualise lipid groups that comprise this activity. Similarly, carotenoids and crude oil were analysed for  antioxidant capacity by the DPPH and β-carotene/linoleic acid methods. The lipidomic profile extract analysis was performed by GC/MS and HPLC/DAD.  The sample preparation for the GC/MS analysis was made by ultrasound-assisted transesterification. Free compounds were silylated with BSTFA (N,O-Bis  (trimethylsilyl) trifluoracetamide) + 1% TMCS (Trimethylchlorosilane). The analysis of the lipid extract showed that in the saponifiable fraction saturated  fatty acids (SFA) and monounsaturated fatty acids (MUFA) were present; and in the unsaponifiable fraction were steroids and carotenoids. The antioxidant  capacity was expressed as IC50 reaching 6.4 mg/L that means relative efficiency. The oil profile, using TLC, shows the chemical groups:  carotenoids, acylglycerols, free fatty acids and steroids. Similarly, the GC/ MS analysis shows the fatty acids and steroids. The HPLC analysis describes the  carotenoids profile, highlighting b-carotene as the majority and the presence of β-carotene-5,8-epoxide, zeaxanthin and b-cryptoxanthin, characterising  the lipidomic study of this yeast

Sodium citrate and potassium phosphate as alternative adsorption buffers in hydrophobic and aromatic thiophilic chromatographic purification of plasmid DNA from neutralized lysate

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)The number of studies on gene therapy using plasmid vectors (pDNA) has increased in recent years. As a result, the demand for preparations of pDNA in compliance with recommendations of regulatory agencies (EMEA, FDA) has also increased. Plasmid DNA is often obtained through fermentation of transformed Escherichia coli and purification by a series of unit operations, including chromatography. Hydrophobic interaction chromatography (HIC) and thiophilic aromatic chromatography (TAC), both using ammonium sulfate buffers, are commonly employed with success. This work was aimed at studying the feasibility of utilizing alternative salts in the purification of pDNA from neutralized lysate with phenyl-agarose (HIC) and mercaptopyrimidine-agarose (TAC) adsorbents. Their selectivity toward sc pDNA was evaluated through adsorption studies using 1.5 mol/L sodium citrate and 2.0 mol/L potassium phosphate as adsorption buffers. Chromatography with mercaptopyrimidine-agarose adsorbent and 1.5 mol/L sodium citrate was able to recover 91.1% of the pDNA with over 99.0% removal of gDNA and endotoxin. This represents a potential alternative for the primary recovery of sc pDNA. However, the most promising result was obtained using 2.0 mol/L potassium phosphate buffer and a mercaptopyrimidine-agarose column. In a single chromatographic step, this latter buffer/adsorbent system recovered 68.5% of the pDNA with 98.8% purity in accordance with the recommendations of regulatory agencies with regard to RNA and endotoxin impurity. (C) 2013 Elsevier B.V. All rights reserved.9196774Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Plant Proteinase Inhibitor BbCI Modulates Lung Inflammatory Responses and Mechanic and Remodeling Alterations Induced by Elastase in Mice

Background. Proteinases play a key role in emphysema. Bauhinia bauhinioides cruzipain inhibitor (BbCI) is a serine-cysteine proteinase inhibitor. We evaluated BbCI treatment in elastase-induced pulmonary alterations. Methods. C57BL/6 mice received intratracheal elastase (ELA group) or saline (SAL group). One group of mice was treated with BbCI (days 1, 15, and 21 after elastase instillation, ELABC group). Controls received saline and BbCI (SALBC group). After 28 days, we evaluated respiratory mechanics, exhaled nitric oxide, and bronchoalveolar lavage fluid. In lung tissue we measured airspace enlargement, quantified neutrophils, TNFα-, MMP-9-, MMP-12-, TIMP-1-, iNOS-, and eNOS-positive cells, 8-iso-PGF2α, collagen, and elastic fibers in alveolar septa and airways. MUC-5-positive cells were quantified only in airways. Results. BbCI reduced elastase-induced changes in pulmonary mechanics, airspace enlargement and elastase-induced increases in total cells, and neutrophils in BALF. BbCI reduced macrophages and neutrophils positive cells in alveolar septa and neutrophils and TNFα-positive cells in airways. BbCI attenuated elastic and collagen fibers, MMP-9- and MMP-12-positive cells, and isoprostane and iNOS-positive cells in alveolar septa and airways. BbCI reduced MUC5ac-positive cells in airways. Conclusions. BbCI improved lung mechanics and reduced lung inflammation and airspace enlargement and increased oxidative stress levels induced by elastase. BbCI may have therapeutic potential in chronic obstructive pulmonary disease

Technological innovation and multinational expansion: A two-way link?

technology, trade, foreign direct investment, multinational company, F23, L12, O33,