scholarly journals Presoaking dried blood spot with water improves efficiency for small-molecule analysis

BioTechniques ◽  
2019 ◽  
Vol 67 (5) ◽  
pp. 219-228
Author(s):  
You-Rim Lee ◽  
Jiyeong Lee ◽  
Hee-Gyoo Kang

The current method of extracting small molecules from dried blood spots (DBSs) and liquid blood is similar. However, owing to their different physical characteristics, a modification of the extraction process for DBS is required. We propose a modified method involving presoaking in water that results in better extraction efficiency for small-molecule analysis than the conventional protein precipitation method. Using blood and DBSs from eight subjects, the similarities, recovery rates and extraction efficiencies of both methods were compared. Quantitative analysis showed that seven and six out of ten conditions for the modified method group exhibited almost 100% recovery and extraction efficiency rates, respectively, compared with the conventional method group. Taken together, the results suggest that a presoaking step is needed for efficient DBS analysis.

2017 ◽  
Vol 9 (13) ◽  
pp. 2014-2020 ◽  
Author(s):  
Ziyu Rao ◽  
Fanglan Geng ◽  
Yiqi Zhou ◽  
Dong Cao ◽  
Yuehui Kang

N-doped graphene quantum dots (N-GQDs) were sythesized by a facile method and applied as a MALDI matrix for small-molecule analysis.


2021 ◽  
Vol 12 ◽  
Author(s):  
Flavia C. G. Reis ◽  
Jonas H. Costa ◽  
Leandro Honorato ◽  
Leonardo Nimrichter ◽  
Taícia P. Fill ◽  
...  

The small molecule (molecular mass <900 Daltons) composition of extracellular vesicles (EVs) produced by the pathogenic fungus Cryptococcus gattii is unknown, which limits the understanding of the functions of cryptococcal EVs. In this study, we analyzed the composition of small molecules in samples obtained from solid cultures of C. gattii by a combination of chromatographic and spectrometric approaches, and untargeted metabolomics. This analysis revealed previously unknown components of EVs, including small peptides with known biological functions in other models. The peptides found in C. gattii EVs had their chemical structure validated by chemical approaches and comparison with authentic standards, and their functions tested in a Galleria mellonella model of cryptococcal infection. One of the vesicular peptides (isoleucine-proline-isoleucine, Ile-Pro-Ile) improved the survival of G. mellonella lethally infected with C. gattii or C. neoformans. These results indicate that small molecules exported in EVs are biologically active in Cryptococcus. Our study is the first to characterize a fungal EV molecule inducing protection, pointing to an immunological potential of extracellular peptides produced by C. gattii.


2021 ◽  
Author(s):  
Flavia C. G. Reis ◽  
Jonas H. Costa ◽  
Leandro Honorato ◽  
Leonardo Nimrichter ◽  
Taícia P. Fill ◽  
...  

AbstractThe small molecule (molecular mass < 900 Daltons) composition of extracellular vesicles (EVs) produced by the pathogenic fungus Cryptococcus gattii is unknown, which limits the understanding of the functions of cryptococcal EVs. In this study, we analyzed the composition of small molecules in samples obtained from solid cultures of C. gattii by a combination of chromatographic and spectrometric approaches, and untargeted metabolomics. This analysis revealed previously unknown components of EVs, including small peptides with known biological functions in other models. The peptides found in C. gattii EVs had their chemical structure validated by chemical approaches and comparison with authentic standards, and their functions tested in a Galleria mellonella model of cryptococcal infection. One of the vesicular peptides (isoleucine-proline-isoleucine, Ile-Pro-Ile) improved the survival of G. mellonella lethally infected with C. gattii or C. neoformans. These results indicate that small molecules exported in EVs are biologically active in Cryptococcus. Our study is the first to characterize a fungal EV molecule inducing protection, pointing to an immunological potential of extracellular peptides produced by C. gattii.


2020 ◽  
Vol 22 (Supplement_2) ◽  
pp. ii200-ii200
Author(s):  
Stephen Skirboll ◽  
Natasha Lucki ◽  
Genaro Villa ◽  
Naja Vergani ◽  
Michael Bollong ◽  
...  

Abstract INTRODUCTION Glioblastoma multiforme (GBM) is the most aggressive form of primary brain cancer. A subpopulation of multipotent cells termed GBM cancer stem cells (CSCs) play a critical role in tumor initiation and maintenance, drug resistance, and recurrence following surgery. New therapeutic strategies for the treatment of GBM have recently focused on targeting CSCs. Here we have used an unbiased large-scale screening approach to identify drug-like small molecules that induce apoptosis in GBM CSCs in a cell type-selective manner. METHODS A luciferase-based survival assay of patient-derived GBM CSC lines was established to perform a large-scale screen of ∼one million drug-like small molecules with the goal of identifying novel compounds that are selectively toxic to chemoresistant GBM CSCs. Compounds found to kill GBM CSC lines as compared to control cell types were further characterized. A caspase activation assay was used to evaluate the mechanism of induced cell death. A xenograft animal model using patient-derived GBM CSCs was employed to test the leading candidate for suppression of in vivo tumor formation. RESULTS We identified a small molecule, termed RIPGBM, from the cell-based chemical screen that induces apoptosis in primary patient-derived GBM CSC cultures. The cell type-dependent selectivity of RIPGBM appears to arise at least in part from redox-dependent formation of a proapoptotic derivative, termed cRIPGBM, in GBM CSCs. cRIPGBM induces caspase 1-dependent apoptosis by binding to receptor-interacting protein kinase 2 (RIPK2) and acting as a molecular switch, which reduces the formation of a prosurvival RIPK2/TAK1 complex and increases the formation of a proapoptotic RIPK2/caspase 1 complex. In an intracranial GBM xenograft mouse model, RIPGBM was found to significantly suppress tumor formation. CONCLUSIONS Our chemical genetics-based approach has identified a small molecule drug candidate and a potential drug target that selectively targets cancer stem cells and provides an approach for the treatment of GBMs.


BMC Chemistry ◽  
2021 ◽  
Vol 15 (1) ◽  
Author(s):  
Sanghamitra Khandual ◽  
Edgar Omar Lopez Sanchez ◽  
Hugo Espinosa Andrews ◽  
Jose Daniel Padilla de la Rosa

AbstractPhycocyanin is a blue natural food colorant with multiple health benefits. Here we propose an efficient phycocyanin extraction method from Arthrospira platensis from Mexico. Three extraction methods were applied to optimize the extraction process, using water and buffer as solvents, with three pH values at two agitation times. The highest phycocyanin, 54.65 mg/g, was extracted from dry biomass with water as a solvent using an ultrasonication bar. The optimum condition of extraction was determined to be 1:50 biomass/solvent ratio for dry biomass, with the freeze/thaw method for 20 min repeated twice, and then agitated at 120 rpm for 24 h. The phycocyanin content was 48.88 mg/g biomass, with a purity of 0.47. For scalable phycocyanin productivity, the sonication method is recommended as there is no statistical difference. The phycocyanin stability was best at − 20 °C storage temperature at pH 7 for 35 days. Partial purification with ammonium sulfate was found to be suitable as a fractional precipitation method, first at 0–20% and then 20–65%, to get purity nearly 1. Total protein was found to be 55.52%, and total amino acids after phycocyanin extraction was 33%. The maximum phycocyanin yield using water as a solvent was the most interesting result regardless of the method used for extraction.


Author(s):  
Wei Li ◽  
Cheng Zheng ◽  
Jian Zhao ◽  
Zhengxiang Ning

A novel microwave assisted multi-stage countercurrent extraction (MAMCE) technique was developed for the extraction of dihydromyricetin from Chinese rattan tea, Ampelopsis grossedentata. The technique combined the advantages of microwave heating and dynamic multi-stage countercurrent extraction and achieved marked improvement in extraction efficiency over microwave assisted batch extraction. Analysis of dihydromyricetin concentrations in the solvent and matrix throughout the extraction process showed that by dividing the extraction into multiple stages and exchanging of solvents between stages, steady and substantial concentration gradients were established between the matrix and solvent, thus enabling the achievement of high extraction efficiency. The yield of dihydromyricetin was significantly affected by temperature, pH, solvent/material ratio and extraction time, and optimal extraction conditions were found to be 80-100°C, at acidic pH with a solvent/material ratio of 25-30 to 1 and extraction time of 5-10 min. With the high extraction efficiency and low usage of extraction solvent, MAMCE could prove to be a promising extraction technique which can be applied to the extraction of dihydromyricentin and other bioactive substances from natural materials.


2015 ◽  
Vol 7 (18) ◽  
pp. 7879-7888 ◽  
Author(s):  
Jiafei Wang ◽  
Xiaoya Jiang ◽  
Hang Zhang ◽  
Sha Liu ◽  
Ligai Bai ◽  
...  

A monolith based on an ionic liquid as a porogen was prepared to enhance the column efficiency of small molecule separation in HPLC.


Metals ◽  
2018 ◽  
Vol 8 (9) ◽  
pp. 654 ◽  
Author(s):  
Moussa Toure ◽  
Guilhem Arrachart ◽  
Jean Duhamet ◽  
Stephane Pellet-Rostaing

A study has been carried out on Ta and Nb recovery by a liquid-liquid extraction process using 4-methylacetophenone (4-MAcPh) as the organic phase. The 4-MAcPh was compared to methyl isobutyl ketone (MIBK) with respect to extraction efficiencies (D values) at different concentrations of H2SO4 in the aqueous phase. The results showed a similar extraction of Nb for both solvents. However, for Ta, extraction efficiency is increased by a factor of 1.3 for 4-MAcPh. In addition, the MIBK solubilized completely after 6 mol∙L−1 of H2SO4 against only a loss of 0.14–4% for 4-MAcPh between 6 and 9 mol∙L−1 of H2SO4. The potential of 4-MAcPh has also been studied to selectively recover Ta from a model capacitor waste solution. The results showed a selectivity for Ta in the presence of impurities such as Ag, Fe, Ni and Mn. The 4-MAcPh also presents the advantage of having physicochemical properties adapted to its use in liquid-liquid extraction technologies such as mixer-settlers.


2016 ◽  
Vol 12 ◽  
pp. 125-138 ◽  
Author(s):  
Steven C Zimmerman

This review summarizes part of the author’s research in the area of supramolecular chemistry, beginning with his early life influences and early career efforts in molecular recognition, especially molecular tweezers. Although designed to complex DNA, these hosts proved more applicable to the field of host–guest chemistry. This early experience and interest in intercalation ultimately led to the current efforts to develop small molecule therapeutic agents for myotonic dystrophy using a rational design approach that heavily relies on principles of supramolecular chemistry. How this work was influenced by that of others in the field and the evolution of each area of research is highlighted with selected examples.


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