Novel isolation and purification method permitting functional cytotoxicity studies of macrophages from milky spots in the greater omentum

1995 ◽  
Vol 184 (2) ◽  
pp. 253-261 ◽  
Author(s):  
Lambert F.G. Krist ◽  
Miranda Kerremans ◽  
Hans Koenen ◽  
Nico Blijleven ◽  
Inge L. Eestermans ◽  
...  
Keyword(s):  
Greater Omentum ◽  
Purification Method ◽  
Isolation And Purification ◽  
Cytotoxicity Studies ◽  
Milky Spots

scholarly journals Green Route for the Isolation and Purification of Hyrdoxytyrosol, Tyrosol, Oleacein and Oleocanthal from Extra Virgin Olive Oil

Molecules ◽  
2020 ◽  
Vol 25 (16) ◽  
pp. 3654
Author(s):  
Antonio Francioso ◽  
Rodolfo Federico ◽  
Anna Maggiore ◽  
Mario Fontana ◽  
Alberto Boffi ◽  
...  
Keyword(s):  
Olive Oil ◽  
High Performance ◽  
Virgin Olive Oil ◽  
Deep Eutectic Solvent ◽  
Extra Virgin Olive Oil ◽  
Hazardous Substances ◽  
Purification Method ◽  
Total Recovery ◽  
Isolation And Purification ◽  
Natural Deep Eutectic Solvent

Extra virgin olive oil (EVOO) phenols represent a significant part of the intake of antioxidants and bioactive compounds in the Mediterranean diet. In particular, hydroxytyrosol (HTyr), tyrosol (Tyr), and the secoiridoids oleacein and oleocanthal play central roles as anti-inflammatory, neuro-protective and anti-cancer agents. These compounds cannot be easily obtained via chemical synthesis, and their isolation and purification from EVOO is cumbersome. Indeed, both processes involve the use of large volumes of organic solvents, hazardous reagents and several chromatographic steps. In this work we propose a novel optimized procedure for the green extraction, isolation and purification of HTyr, Tyr, oleacein and oleocanthal directly from EVOO, by using a Natural Deep Eutectic Solvent (NaDES) as an extracting phase, coupled with preparative high-performance liquid chromatography. This purification method allows the total recovery of the four components as single pure compounds directly from EVOO, in a rapid, economic and ecologically sustainable way, which utilizes biocompatible reagents and strongly limits the use or generation of hazardous substances.

scholarly journals An integrated cell isolation and purification method for rat dorsal root ganglion neurons

2019 ◽  
Vol 47 (7) ◽  
pp. 3253-3260
Author(s):  
Huaishuang Shen ◽  
Minfeng Gan ◽  
Huilin Yang ◽  
Jun Zou
Keyword(s):  
Neuropathic Pain ◽  
Dorsal Root Ganglion ◽  
Primary Culture ◽  
Dorsal Root ◽  
Cell Isolation ◽  
Dorsal Root Ganglion Neurons ◽  
Drg Neurons ◽  
Purification Method ◽  
Isolation And Purification ◽  
Ganglion Neurons

Objective Neurobiology studies are increasingly focused on the dorsal root ganglion (DRG), which plays an important role in neuropathic pain. Existing DRG neuron primary culture methods have considerable limitations, including challenging cell isolation and poor cell yield, which cause difficulty in signaling pathway studies. The present study aimed to establish an integrated primary culture method for DRG neurons. Methods DRGs were obtained from fetal rats by microdissection, and then dissociated with trypsin. The dissociated neurons were treated with 5-fluorouracil to promote growth of neurons from the isolated cells. Then, reverse transcription polymerase chain reaction and immunofluorescence assays were used to identify and purify DRG neurons. Results Isolated DRGs were successfully dissociated and showed robust growth as individual DRG neurons in neurobasal medium. Both mRNA and protein assays confirmed that DRG neurons expressed neurofilament-200 and neuron-specific enolase. Conclusions Highly purified, stable DRG neurons could be easily harvested and grown for extended periods by using this integrated cell isolation and purification method, which may help to elucidate the mechanisms underlying neuropathic pain.

scholarly journals Toxic Indole Diterpenes from Endophyte-Infected Perennial Ryegrass Lolium perenne L.: Isolation and Stability

Toxins ◽  
2019 ◽  
Vol 11 (1) ◽  
pp. 16 ◽  
Author(s):  
Priyanka Reddy ◽  
Myrna A. Deseo ◽  
Vilnis Ezernieks ◽  
Kathryn Guthridge ◽  
German Spangenberg ◽  
...  
Keyword(s):  
Lolium Perenne ◽  
Perennial Ryegrass ◽  
Rapid Development ◽  
Toxicity Testing ◽  
Flash Chromatography ◽  
Preparative Hplc ◽  
Purification Method ◽  
Isolation And Purification ◽  
Lolium Perenne L ◽  
Lolitrem B

The most potent of the indole diterpenes, lolitrem B, is found in perennial ryegrass (Lolium perenne L.) infected with the endophyte Epichloë festucae var. lolii (also termed LpTG-1). Ingestion causes a neurological syndrome in grazing livestock called ryegrass staggers disease. To enable the rapid development of new forage varieties, the toxicity of lolitrem B and its biosynthetic intermediates needs to be established. However, most of these indole diterpenes are not commercially available; thus, isolation of these compounds is paramount. A concentrated endophyte-infected perennial ryegrass seed extract was subjected to silica flash chromatography followed by preparative HPLC and purification by crystallization resulting in lolitrem B and the intermediate compounds lolitrem E, paspaline and terpendole B. The four-step isolation and purification method resulted in a 25% yield of lolitrem B. After isolation, lolitrem B readily degraded to its biosynthetic intermediate, lolitriol. We also found that lolitrem B can readily degrade depending on the solvent and storage conditions. The facile method which takes into consideration the associated instability of lolitrem B, led to the purification of indole diterpenes in quantities sufficient for use as analytical standards for identification in pastures, and/or for toxicity testing in pasture development programs.

Milky Spots in the Human Greater Omentum

1989 ◽  
Vol 136 (3) ◽  
pp. 211-216 ◽  
Author(s):  
M. Shimotsuma ◽  
M. Kawata ◽  
A. Hagiwara ◽  
T. Takahashi
Keyword(s):  
Greater Omentum ◽  
Milky Spots

scholarly journals Isolation and purification ofent-pimara-8(14),15-diene from engineeredAspergillus nidulansby accelerated solvent extraction combined with HPLC

2014 ◽  
Vol 6 (4) ◽  
pp. 1227-1234 ◽  
Author(s):  
K. Bromann ◽  
K. Viljanen ◽  
V. M. Moreira ◽  
J. Yli-Kauhaluoma ◽  
L. Ruohonen ◽  
...  
Keyword(s):  
Antioxidant Activity ◽  
Solvent Extraction ◽  
Aspergillus Nidulans ◽  
Accelerated Solvent Extraction ◽  
Purification Method ◽  
Isolation And Purification ◽  
Diterpene Hydrocarbon

This paper describes a purification method for tricyclic diterpene hydrocarbonent-pimara-8(14),15-diene produced inAspergillus nidulansand reports an antioxidant activity for this compound.

scholarly journals Preparative isolation and purification of seven compounds from Hibiscus mutabilis L. leaves by two-step high-speed counter-current chromatography

2015 ◽  
Vol 21 (2) ◽  
pp. 331-341 ◽  
Author(s):  
Zhuoni Hou ◽  
Xianrui Liang ◽  
Feng Su ◽  
Weike Su
Keyword(s):  
Ethyl Acetate ◽  
High Speed ◽  
Solvent System ◽  
Ionization Mass ◽  
Two Phase ◽  
Purification Method ◽  
Isolation And Purification ◽  
Chemical Structures ◽  
13C Nuclear Magnetic Resonance ◽  
Counter Current

Seven compounds from Hibiscus mutabilis L. leaves were first successfully achieved by two-step high-speed counter-current chromatography with two-phase solvent system composed of n-butanol-ethyl acetate-water (1:6:9, v/v/v) and n-hexane-ethyl acetate-methanol-water (3:5:3:5, v/v/v/v/). The critical experimental parameters of first-step separation were optimized with response surface methodology as follows: flow rate was 1.1 mL/min, revolution speed was 800 rpm and temperature was 30?C. Under the optimal conditions, around 5.0 mg of salicylic acid, 13.6 mg of rutin, 5.5 mg of genistein were obtained in 100 mg crude sample. Then, 9.2 mg of potengriffioside A, 4.7 mg of kaempferol 3-O-rutinoside, 3.0 mg of steppogenin and 2.5 mg of emodin were obtained by second-step separation. The purities of the seven compounds determined by UPLC were 96.2%, 93.8%, 95.4%, 94.3%, 98.0%, 94.1% and 90.8%, respectively. Their chemical structures were identified by electron spray ionization mass spectroscopy (ESI-MS) and 1H, 13C nuclear magnetic resonance (NMR). Furthermore, compound steppogenin and genistein were first reported from Hibiscus mutabilis L. The purification method was simple, efficient and evaded tedious separation process.

scholarly journals Antigenic properties of the SARS-CoV-2 nucleoprotein are altered by the RNA admixture

PeerJ ◽  
2022 ◽  
Vol 10 ◽  
pp. e12751
Author(s):  
Denis E. Kolesov ◽  
Maria V. Sinegubova ◽  
Irina V. Safenkova ◽  
Ivan I. Vorobiev ◽  
Nadezhda A. Orlova
Keyword(s):  
Signal To Noise Ratio ◽  
Rnase A ◽  
Soluble Form ◽  
Control Panel ◽  
Signal To Noise ◽  
Purification Method ◽  
Isolation And Purification ◽  
Test Systems ◽  
Antigenic Properties ◽  
Noise Ratio

Determining the presence of antibodies to the SARS-CoV-2 antigens is the best way to identify infected people, regardless of the development of symptoms of COVID-19. The nucleoprotein (NP) of the SARS-CoV-2 is an immunodominant antigen of the virus; anti-NP antibodies are detected in persons previously infected with the virus with the highest titers. Many test systems for detecting antibodies to SARS-CoV-2 contain NP or its fragments as antigen. The sensitivity and specificity of such test systems differ significantly, which can be explained by variations in the antigenic properties of NP caused by differences in the methods of its cultivation, isolation and purification. We investigated this effect for the Escherichia coli-derived SARS-CoV-2 NP, obtained from the cytoplasm in the soluble form. We hypothesized that co-purified nucleic acids that form a strong complex with NP might negatively affect NP’s antigenic properties. Therefore, we have established the NP purification method, which completely eliminates the RNA in the NP preparation. Two stages of RNA removal were used: treatment of the crude lysate of E. coli with RNase A and subsequent selective RNA elution with 2 M NaCl solution. The resulting NP without RNA has a significantly better signal-to-noise ratio when used as an ELISA antigen and tested with a control panel of serum samples with antibodies to SARS-CoV-2; therefore, it is preferable for in vitro diagnostic use. The same increase of the signal-to-noise ratio was detected for the free N-terminal domain of the NP. Complete removal of RNA complexed with NP during purification will significantly improve its antigenic properties, and the absence of RNA in NP preparations should be controlled during the production of this antigen.

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