scholarly journals Physical exercise increases urinary excretion of lipoxin A4 and related compounds

2003 ◽  
Vol 94 (6) ◽  
pp. 2237-2240 ◽  
Author(s):  
Sebastiano Gangemi ◽  
Graziella Luciotti ◽  
Etrusca D'Urbano ◽  
Agostino Mallamace ◽  
Domenico Santoro ◽  
...  
Keyword(s):  
Urinary Excretion ◽  
High Performance ◽  
Reversed Phase ◽  
Strenuous Exercise ◽  
Lipoxin A4 ◽  
Safeguard Mechanism ◽  
First Time ◽  
Related Compounds

Lipoxins (LX) are lipoxygenase-derived eicosanoids with potent anti-inflammatory activities and vascular bed-dependent vasodilatory actions. LX can be formed in vitro and in vivo in a number of conditions, and we have reported that immunoreactive LXA4 (iLXA4) is physiologically excreted with human urine. Using a recently developed LX extraction method coupled to an ELISA, we examined whether iLXA4 excretion was modified by strenuous exercise, which is known to trigger potential LX-forming events. Maximal exertion significantly increased iLXA4 urinary excretion in nine healthy volunteers (0.061 ± 0.023 vs. 0.113 ± 0.057 ng/mg creatinine; P = 0.028). iLXA4 levels returned to baseline after 6 h and increased, although at a smaller extent, after 24 h. A significant correlation ( r = 0.988) was denoted between iLXA4 ELISA measurements and reversed-phase high-performance liquid chromatography quantitation of a previously described urinary tetraene, confirming its LXA4-related nature. These findings show for the first time that an increase in excretion of LXA4-related compounds can be observed in response to strenuous exercise. This may be the reflection of an enhanced LX biosynthesis, which may represent a safeguard mechanism that keeps the inflammatory reaction triggered by physical stress under control.

scholarly journals Synthesis of Human Bone Morphogenetic Protein-2 (hBMP-2) in E. coli Periplasmic Space: Its Characterization and Preclinical Testing

Cells ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 3525
Author(s):  
João E. Oliveira ◽  
Miriam F. Suzuki ◽  
Renata Damiani ◽  
Eliana R. Lima ◽  
Kleicy C. Amaral ◽  
...  
Keyword(s):  
High Performance ◽  
Osmotic Shock ◽  
Cytoplasmic Inclusion ◽  
Reversed Phase ◽  
C2c12 Cells ◽  
Size Exclusion ◽  
Bone Morphogenetic Protein 2 ◽  
E Coli

Human BMP-2, a homodimeric protein that belongs to the TGF- β family, is a recognized osteoinductor due to its capacity of inducing bone regeneration and ectopic bone formation. The administration of its recombinant form is an alternative to autologous bone grafting. A variety of E. coli-derived hBMP-2 has been synthesized through refolding of cytoplasmic inclusion bodies. The present work reports the synthesis, purification, and characterization of periplasmic hBMP-2, obtained directly in its correctly folded and authentic form, i.e., without the initial methionine typical of the cytoplasmic product that can induce undesired immunoreactivity. A bacterial expression vector was constructed including the DsbA signal peptide and the cDNA of hBMP-2. The periplasmic fluid was extracted by osmotic shock and analyzed via SDS-PAGE, Western blotting, and reversed-phase high-performance liquid chromatography (RP-HPLC). The purification was carried out by heparin affinity chromatography, followed by high-performance size-exclusion chromatography (HPSEC). HPSEC was used for qualitative and quantitative analysis of the final product, which showed >95% purity. The classical in vitro bioassay based on the induction of alkaline phosphatase activity in myoblastic murine C2C12 cells and the in vivo bioassay consisting of treating calvarial critical-size defects in rats confirmed its bioactivity, which matched the analogous literature data for hBMP-2.

Xenopus laevis oocytes, eggs and tadpoles contain immunoactive insulin

1994 ◽  
Vol 141 (1) ◽  
pp. 123-129 ◽  
Author(s):  
F de Pablo ◽  
R Dashner ◽  
A R Shuldiner ◽  
J Roth
Keyword(s):  
Xenopus Laevis ◽  
High Performance ◽  
Immunoblot Analysis ◽  
Reversed Phase ◽  
Xenopus Laevis Oocytes ◽  
Maternal Origin ◽  
Insulin Mrna ◽  
Low Levels

Abstract Insulin is a multifunctional polypeptide hormone that regulates metabolic processes and promotes mitogenesis and differentiation in vitro in the cells and tissues of several species. Its role in vivo during embryogenesis is still poorly understood. We have previously found insulin mRNA in mature Xenopus laevis oocytes and in embryos during neurulation (before organogenesis of the pancreas takes place). We have now measured insulin immunoactivity in mature oocytes, unfertilized eggs and day-2 tadpoles. Using reversed phase high performance liquid chromatography, we found low levels of insulin in extracts of oocytes (stage VI). Both Xenopus insulin I and II were detected in unfertilized eggs. The day-2 tadpoles (stages 31–33) also contained immunoactive insulin, and in swimming tadpoles (stage 46) a few clusters of cells containing insulin immunoactivity could be identified by indirect immunofluorescence. Immunoblot analysis was relatively insensitive, detecting insulin only in the adult Xenopus pancreas. In summary, insulin (from maternal origin and embryonic expression) appears to be present early enough in Xenopus laevis to influence developmental processes such as neurulation. Journal of Endocrinology (1994) 141, 123–129

Pyclen Based Ln(III) Complexes as Highly Luminescent Bioprobes for in Vitro and in Vivo 1P and 2P Bioimaging Applications

2020 ◽  
Author(s):  
Nadège Hamon ◽  
Amandine Roux ◽  
Maryline Beyler ◽  
Jean-Christophe Mulatier ◽  
Chantal Andraud ◽  
...  
Keyword(s):  
High Performance ◽  
Breast Cancer Cells ◽  
Positive Impact ◽  
Quantum Yields ◽  
Human Breast ◽  
Zebrafish Model ◽  
High Brightness ◽  
First Time

Two pyclen based lanthanide chelators, <b>L<sup>4b</sup> </b>and <b>L<sup>4c</sup>, </b>bearing two specific picolinate 2P antennas (tailor-made for each targeted metal) and one acetate arm arranged in a dissymmetrical manner, have been synthesized to form, with the already described ligand <b>L<sup>4a</sup>,</b> a complete family of lanthanide luminescent bioprobes: [Eu<b>L<sup>4a</sup></b>], [Sm<b>L<sup>4a</sup></b>], [Yb<b>L<sup>4b</sup></b>], [Tb<b>L<sup>4c</sup></b>] and [Dy<b>L<sup>4c</sup></b>]. Additionally, symmetrically arranged regioisomer <b>L<sup>4a’</sup> </b>was also synthesized as well as its [Eu<b>L<sup>4a’</sup></b>] complex to highlight the astonishing positive impact of the dissymmetrical <i>N</i>-distribution of the functional chelating arm. The investigation clearly shows the high performance of each bioprobe, which, depending on the complexed lanthanide, could be used in various applications. Each presents high brightness, quantum yields and lifetimes. Staining of the complexes into living human breast cancer cells was observed. In addition, <i>in vivo</i> 2P-microscopy was performed for the first time on a living Zebrafish model with [Eu<b>L<sup>4a</sup></b>]. No apparent toxicity was detected on the growth of the zebrafish and images of high quality were obtained.

Auto-oxidation of 5,6,7 ,8-tetrahydroneopterin

Pteridines ◽  
1998 ◽  
Vol 9 (1) ◽  
pp. 26-28
Author(s):  
Toshiyuki Arai ◽  
Hiroko Mori ◽  
Hisanari Ishii ◽  
Toshinori Suzuki ◽  
Shuji Kojima ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Free Radicals ◽  
High Performance ◽  
Oxygen Free Radicals ◽  
Phosphate Buffer Solution ◽  
Reversed Phase ◽  
Buffer Solution ◽  
First Time

Summary Auto-oxidation of 5,6,7,8-tetrahydroneopterin (NPH4 ) in phosphate buffer solution (PBS) was examined using reversed-phase high-performance liquid chromatography. In the chromatograms obtained every 30 min after NPH4 was dissolved in PBS, the peak of NPH4 gradually decreased and almost disappeared 90 min after the dissolution. In contrast, another peak of an unidentified substance (one of dihydroneopterins), appeared from the first time, gradually increased and became most dominant among various peaks 60 min after. The peak of 7,8-dihydroneopterin (7,8-NPH2) appeared 30 min after and gradually increased, but its height was less than that of the above-mentioned peak from first to last. These results indicate that NPH4 is readily oxidized by dissolved oxygen in PBS to produce 7,8-NPH2 and other dihydroneopterins before the reaction with oxygen free radicals and that the substances which actually serve as antioxidants in vivo are these dihydroneopterins.

scholarly journals Screening and Identification of the Main Metabolites of Schisantherin a In Vivo and In Vitro by Using UHPLC-Q-TOF-MS/MS

Molecules ◽  
2020 ◽  
Vol 25 (2) ◽  
pp. 258 ◽  
Author(s):  
Wei Feng ◽  
Ling-Yu Zhou ◽  
Rui-Feng Mu ◽  
Le Gao ◽  
Bing-Yuan Xu ◽  
...  
Keyword(s):  
Metabolic Pathways ◽  
High Performance ◽  
Liver Microsomes ◽  
Rat Liver Microsomes ◽  
Sprague Dawley ◽  
First Time ◽  
Schisantherin A ◽  
Tof Ms

Schisantherin A is an active ingredient originating from Schisandra chinensis (Turcz.) which has hepatoprotective and anti-oxidation activities. In this study, in vitro metabolisms investigated on rat liver microsomes (RLMs) and in vivo metabolisms explored on male Sprague Dawley rats of Schisantherin A were tested, respectively. The metabolites of Schisantherin A were identified using ultra-high-performance liquid chromatography coupled with hybrid triple quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS/MS). Based on the method, 60 metabolites were successfully identified and structurally characterized including 48 phase-I and 12 phase-II metabolites. Among the metabolites, 45 metabolites were reported for the first time. Moreover, 56 and eight metabolites were detected in urine and bile and 19 metabolites were identified in rats’ plasma. It demonstrated that hepatic and extra-hepatic metabolic pathways were both involved in Schisantherin A biotransformation in rats. Five in vitro metabolites were structurally characterized for the first time. The results indicated that the metabolic pathways mainly include oxidation, reduction, methylation, and conjugation with glucuronide, taurine, glucose, and glutathione groups. This study provides a practical strategy for rapidly screening and identifying metabolites, and the results provide basic data for future pharmacological and toxicology studies of Schisantherin A and other lignin ingredients.

Pyclen Based Ln(III) Complexes as Highly Luminescent Bioprobes for in Vitro and in Vivo 1P and 2P Bioimaging Applications

2020 ◽  
Author(s):  
Nadège Hamon ◽  
Amandine Roux ◽  
Maryline Beyler ◽  
Jean-Christophe Mulatier ◽  
Chantal Andraud ◽  
...  
Keyword(s):  
High Performance ◽  
Breast Cancer Cells ◽  
Positive Impact ◽  
Quantum Yields ◽  
Human Breast ◽  
Zebrafish Model ◽  
High Brightness ◽  
First Time

Two pyclen based lanthanide chelators, <b>L<sup>4b</sup> </b>and <b>L<sup>4c</sup>, </b>bearing two specific picolinate 2P antennas (tailor-made for each targeted metal) and one acetate arm arranged in a dissymmetrical manner, have been synthesized to form, with the already described ligand <b>L<sup>4a</sup>,</b> a complete family of lanthanide luminescent bioprobes: [Eu<b>L<sup>4a</sup></b>], [Sm<b>L<sup>4a</sup></b>], [Yb<b>L<sup>4b</sup></b>], [Tb<b>L<sup>4c</sup></b>] and [Dy<b>L<sup>4c</sup></b>]. Additionally, symmetrically arranged regioisomer <b>L<sup>4a’</sup> </b>was also synthesized as well as its [Eu<b>L<sup>4a’</sup></b>] complex to highlight the astonishing positive impact of the dissymmetrical <i>N</i>-distribution of the functional chelating arm. The investigation clearly shows the high performance of each bioprobe, which, depending on the complexed lanthanide, could be used in various applications. Each presents high brightness, quantum yields and lifetimes. Staining of the complexes into living human breast cancer cells was observed. In addition, <i>in vivo</i> 2P-microscopy was performed for the first time on a living Zebrafish model with [Eu<b>L<sup>4a</sup></b>]. No apparent toxicity was detected on the growth of the zebrafish and images of high quality were obtained.

scholarly journals In Vitro and In Vivo Evaluation of Two Hydroxychloroquine Tablet Formulations: HPLC Assay Development

2020 ◽  
Vol 59 (1) ◽  
pp. 71-78
Author(s):  
Jaber Emami ◽  
Moloud Kazemi ◽  
Anahita Salehi
Keyword(s):  
High Performance ◽  
Internal Standard ◽  
Reversed Phase ◽  
Hplc Method ◽  
In Vivo Studies ◽  
Crossover Fashion ◽  
Content Uniformity ◽  
In Vivo Evaluation

Abstract The relative in vitro and in vivo evaluation of two hydroxychloroquine (HCQ) products was conducted. In vitro studies involved assay, content uniformity and dissolution test, and a two-way crossover fashion were used for in vivo studies. Blood samples were collected at appropriate intervals and HCQ levels were measured using a validated reversed-phase high-performance liquid chromatography (HPLC) method. The drug and the internal standard, chloroquine (CQ), were extracted from blood with diethyl ether, separated and dried under nitrogen gas. Residues were reconstituted in the mobile phase and analyzed at 340 nm on a μ-bondapack C18 (250 × 4.6 mm) HPLC column with acetonitrile:methanol:KH2PO4 (10:10:80) mixture containing 0.01% triethylamine. The standard curve was linear within 50–1,500 ng/mL HCQ (R2 = 0.9996), relative errors were 1.6 to 5%, and the CV% ranged from 7 to 15.4. The resolution factor and RSD were 1.62 and 0.35% and in vitro data of both products met the USP requirements. The 90% confidence intervals for the ratios of the AUC0–96, Cmax and Tmax and their corresponding logarithmically transformed values of generic product over those of Plaquenil® were within the acceptable limit of 0.80–1.20 and 0.80–1.25, respectively. Therefore, the generic HCQ was bioequivalent to the innovator formulation.

Polyphenols from Euphorbia pekinensis Inhibit AGEs Formation In Vitro and Vessel Dilation in Larval Zebrafish In Vivo

Planta Medica ◽  
2017 ◽  
Vol 84 (03) ◽  
pp. 176-181 ◽  
Author(s):  
Ik-Soo Lee ◽  
Seung-Hyun Jung ◽  
Jin Kim
Keyword(s):  
Ellagic Acid ◽  
High Performance ◽  
Spectroscopic Studies ◽  
Reversed Phase ◽  
Control Group ◽  
Retinal Vessels ◽  
Etoh Extract ◽  
Larval Zebrafish

AbstractTo identify active compounds in the roots of Euphorbia pekinensis for treatment of diabetic complications, an active column fraction from a 70% EtOH extract of E. pekinensis root was purified by preparative reversed-phase high-performance liquid chromatography, leading to the isolation of a new ellagic acid derivative, 3,3′-di-O-methylellagic acid 4-O-(6ʺ-O-galloyl)-β-D-galactopyranoside (1), along with three known compounds, geraniin (2), 3,3′-di-O-methylellagic acid 4-O-β-D-xylopyranoside (3), and ellagic acid 3,3′-dimethyl ether (4). The structure of the new compound was established by extensive spectroscopic studies and chemical evidence. The inhibitory effects of isolated compounds 1–4 on advanced glycation end-products (AGEs) formation were examined. All compounds exhibited considerable inhibition of AGEs formation and IC50 values of 0.41 – 12.33 µM, compared with those of the positive controls aminoguanidine (IC50 = 1122.34 µM) and quercetin (IC50 = 27.80 µM). In addition, the effects of 2 and 4 on the dilation of hyaloid-retinal vessels induced by high glucose (HG) in larval zebrafish were investigated; both compounds significantly reduced the HG-induced dilation of hyaloid-retinal vessels relative to the HG-treated control group.

Coronatine production in vitro and in vivo and its relation to symptom development in bacterial blight of soybean

1982 ◽  
Vol 60 (5) ◽  
pp. 645-650 ◽  
Author(s):  
S. S. Gnanamanickam ◽  
A. N. Starratt ◽  
E. W. B. Ward
Keyword(s):  
Bacterial Blight ◽  
High Performance ◽  
High Performance Liquid Chromatographic ◽  
Reversed Phase ◽  
Culture Filtrates ◽  
Soybean Plants ◽  
Systemic Symptoms ◽  
Soybean Leaves

Coronatine was detected in culture filtrates of 12 out of 19 pathogenic strains of Pseudomonas glycinea examined, but not in culture filtrates of strains of P. phaseolicola, P. syringae, or P. tabaci. A reversed-phase high-performance liquid chromatographic procedure was developed for coronatine quantitation. Generally, coronatine production by strains in culture correlated with their ability to induce systemic symptoms (chlorosis and stunting) in inoculated soybean plants. Application of purified preparations of coronatine to unifoliate leaves of soybean plants resulted in localized chlorosis, development of chlorosis in subsequently developing trifoliate leaves, and stunting of plant growth, similar to symptoms induced by infection. Coronatine was demonstrated in soybean leaves infected with P. glycinea but was not detected in healthy leaves. The results indicate that coronatine can play an important role in the development of symptoms of bacterial blight of soybean, but the demonstration that some pathogenic strains do not produce coronatine indicates that it may not be essential for pathogenicity.

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