Renewing potency assays: Moving forward from traditional methods

2020 ◽  
Author(s):  
Francesco Nevelli
Keyword(s):  
Growth Hormone ◽  
Physiological Mechanism ◽  
Global Market ◽  
Hormone Deficiency ◽  
In Vitro Assays ◽  
In Vitro Methods ◽  
In Vivo Assays ◽  
Market Leader

Merck is global market leader in the fertility and growth hormone deficiency treatment. The quality control analytical panels for each new produced batch envisage the potency quantification that is estimated using a dedicated in vivo assay. Indeed, no in vitro methods for gonadotropin potency quantification are available in any pharmacopoeia. Merck Ivrea started a project to replace the in vivo assays with in vitro assays able to mimic the physiological mechanism of action of each gonadotropin and growth hormone.

scholarly journals Preliminary Study and Improve the Production of Metabolites with Antifungal Activity by a Bacillus Sp Strain IBA 33

2009 ◽  
Vol 2 ◽  
pp. MBI.S995 ◽  
Author(s):  
María Antonieta Gordillo ◽  
Antonio Roberto Navarro ◽  
Lidia María Benitez ◽  
Marta Inés Torres De Plaza ◽  
Maria Cristina Maldonado
Keyword(s):  
Pathogenic Fungi ◽  
Geotrichum Candidum ◽  
Culture Conditions ◽  
Penicillium Expansum ◽  
In Vitro Assays ◽  
Bacillus Sp ◽  
In Vivo Assays ◽  
Metabolites Production

Bacillus sp strain IBA 33 metabolites, isolated from decaying lemon fruits, were evaluated for the control of pathogenic and non-pathogenic fungi ( Penicillium digitatum, Geotrichum candidum, Penicillium expansum, Aspergillus clavatus, Aspergillus flavus, Aspergillus niger, and Fusarium moniliforme). These metabolites were recovered from Landy medium (LM) without aminoacids. In order to optimize metabolites production the LM was modified by adding different concentrations and sources of amino acids and carbohydrates at different culture conditions. Bacillus sp strain IBA 33 metabolites efficacy to control fungi were evaluated with in vitro and in vivo assays. A. flavus growth inhibition was 52% with the metabolites of Bacillus sp strain IBA 33 recovered from LM (MBLM) in vitro assays. MBLM supplemented with 0.5% glutamic acid, inhibited the growth of P. digitatum, G. candidum, A. clavatus, A. niger and F. moniliforme by 65%, 88.44%, 84%, 34% and 92% respectively. The highest inhibition of P. expansum was 45% with MBLM supplemented with 0.5% aspartic acid. Similar results were obtained in vivo assays. These results showed that Bacillus sp strain IBA 33 metabolites specificity against fungi depended on the composition of the LM.

Polystachyasaponin with Adjuvant Activity from Entada polystachya

2010 ◽  
Vol 65 (5) ◽  
pp. 628-634 ◽  
Author(s):  
Bernadete P. da Silva ◽  
José P. Parente
Keyword(s):  
2D Nmr ◽  
In Vitro Assays ◽  
Triterpenoid Saponin ◽  
Spectroscopic Techniques ◽  
Adjuvant Activity ◽  
In Vivo Assays ◽  
Cellular Immune ◽  
C Nmr

A new complex triterpenoid saponin, polystachyasaponin, was isolated from leaves of Entada polystachya (L.) DC. (Leguminosae) by using chromatographic methods. Its structure was established as 15,16-dihydroxy-3-[[O-β -D-xylopyranosyl-(1→2)-O-α-L-arabinopyranosyl-(1→6)-2- (acetylamino)-2-deoxy-β -D-glucopyranosyl]oxy]-(3β ,15α,16α)-olean-12-en-28-oic acid O-D-apio- β -D-furanosyl-(1→3)-O-β -D-xylopyranosyl-(1→2)-O-[β -D-glucopyranosyl-(1→4)]-6-O-[(2E,6R)- 6-hydroxy-2,6-dimethyl-1-oxo-2,7-octadienyl]-β -D-glucopyranosyl ester. Structural elucidation was performed using detailed analyses of 1H and 13C NMR spectra including 2D NMR spectroscopic techniques and chemical conversions. The hemolytic activity of the saponin was evaluated using in vitro assays, and its adjuvant potential on the cellular immune response against ovalbumin antigen was investigated using in vivo assays.

scholarly journals Recent Reduction in Thrombogenic Enzyme Content of Prothrombin Complex Concentrates

1977 ◽  
Author(s):  
H.S. Kingdon
Keyword(s):  
Calcium Ions ◽  
Calcium Ion ◽  
Deae Cellulose ◽  
In Vitro Assays ◽  
Prothrombin Complex Concentrates ◽  
In Vivo Assays ◽  
Prolonged Contact ◽  
Plasma Fractions

In previous studies of prothrombin complex concentrates, it was demonstrated that there was a high content of potentially thrombogenic enzymes in the products from certain manufacturers, and that the enzyme conteat correlated closely with in vivo assays for thrombogenicity and with the observed incidence of thrombotic episodes following infusion (Thromb. Diath. Haemorrh. 33, 617, 1 975\Subsequently, it was shown that the thrombogenic enzymes could be generated by prolonged contact with DEAE-cellulose or calcium ions during preparation or later handling (Blood, 49, 159, 1977). In view of. these observations, efforts have been made to reduce the contact time of plasma fractions with either DEAE-cellulose or calcium ion during purification of these fractions for therapeutic use. In vitro assays for thrombogenic enzymes using the nonactivated partial thromboplastin time (NAPTT) were recently repeated on some of the currently available therapeutic materials. Assays were standardized and compared with previous results by using a provisional standard provided by Dr. David Aronson, Bureau of Biologies, USFDA. In the 1975 study, one manufacturer was identified as making a concentrate virtually free of thrombogenic enzyme. The concentrate currently being made by this manufacturer still does not significantly shorten the NAPTT. In the 1975 study, 2 manufacturers were shown to be making concentrates with high titers of thrombogenic enzyme. The current products of these two manufacturers contain detectable but significantly lower levels of thrombogenic enzymes. Thus it appears that for these two manufacturers, minor changes in production procedures have led to a product containing less potentially thrombogenic material.

scholarly journals In vitro assays misrepresent in vivo lineage potentials of murine lymphoid progenitors

Blood ◽  
2011 ◽  
Vol 117 (9) ◽  
pp. 2618-2624 ◽  
Author(s):  
Lauren I. Richie Ehrlich ◽  
Thomas Serwold ◽  
Irving L. Weissman
Keyword(s):  
T Cell ◽  
The Other ◽  
In Vitro Assays ◽  
In Vivo Assays ◽  
Other Hand ◽  
Multipotent Progenitors ◽  
Lymphoid Progenitors

Abstract The identity of T-cell progenitors that seed the thymus has remained controversial, largely because many studies differ over whether these progenitors retain myeloid potential. Contradictory reports diverge in their use of various in vitro and in vivo assays. To consolidate these discordant findings, we compared the myeloid potential of 2 putative thymus seeding populations, common lymphoid progenitors (CLPs) and multipotent progenitors (MPPs), and the earliest intrathymic progenitor (DN1), using 2 in vitro assays and in vivo readouts. These assays gave contradictory results: CLP and DN1 displayed surprisingly robust myeloid potential on OP9-DL1 in vitro stromal cocultures but displayed little myeloid potential in vivo, as well as in methylcellulose cultures. MPP, on the other hand, displayed robust myeloid potential in all settings. We conclude that stromal cocultures reveal cryptic, but nonphysiologic, myeloid potentials of lymphoid progenitors, providing an explanation for contradictory findings in the field and underscoring the importance of using in vivo assays for the determination of physiologic lineage potentials.

Evaluation of antioxidant activity and toxicity of sulfur- or selenium-containing 4-(arylchalcogenyl)-1H-pyrazoles

2020 ◽  
Vol 98 (7) ◽  
pp. 441-448
Author(s):  
Daniela Hartwig de Oliveira ◽  
Fernanda Severo Sabedra Sousa ◽  
Paloma Taborda Birmann ◽  
Ana Paula Pesarico ◽  
Diego Alves ◽  
...  
Keyword(s):  
Lipid Peroxidation ◽  
Mouse Brain ◽  
Aminolevulinic Acid ◽  
In Vitro Assays ◽  
Scavenging Activity ◽  
In Vivo Assays ◽  
Reducing Ability ◽  
Liver And Kidney

Pyrazoles represent a significant class of heterocyclic compounds that exhibit pharmacological properties. The present study aimed to investigate the antioxidant potential of pyrazol derivative compounds in brain of mice in vitro and the effect of pyrazol derivative compounds in the oxidative damage and toxicity parameters in mouse brain and plasma of mice. The compounds tested were 3,5-dimethyl-1-phenyl-4-(phenylselanyl)-1H-pyrazol (1a), 3,5-dimethyl-4-(phenylselanyl)-1H-pyrazole (2a), 4-((4-methoxyphenyl)selanyl)-3,5-dimethyl-1-phenyl-1H-pyrazole (3a), 4-((4-chlorophenyl)selanyl)-3,5-dimethyl-1-phenyl-1H-pyrazole (4a), 3,5-dimethyl-1-phenyl-4-(phenylthio)-1H-pyrazole (1b), 3,5-dimethyl-4-(phenylthio)-1H-pyrazole (2b), 4-((4-methoxyphenyl)thio)-3,5-dimethyl-1-phenyl-1H-pyrazole (3b), 4-((4-chlorophenyl)thio)-3,5-dimethyl-1-phenyl-1H-pyrazole (4b), and 3,5-dimethyl-1-phenyl-1H-pyrazole (1c). In vitro, 4-(arylcalcogenyl)-1H-pyrazoles, at low molecular range, reduced lipid peroxidation and reactive species in mouse brain homogenates. The compounds also presented ferric-reducing ability as well nitric oxide-scavenging activity. Especially compounds 1a, 1b, and 1c presented efficiency to 1,1-diphenyl-2-picryl-hydrazyl-scavenging activity. Compounds 1b and 1c presented 2,20 -azino-bis(3-ethylbenzthiazoline-6-sulfonic acid)-scavenging activity. In vivo assays demonstrated that compounds 1a, 1b, and 1c (300 mg/kg, intragastric, a single administration) did not cause alteration in the of δ-aminolevulinic acid dehydratase activity, an enzyme that exhibits high sensibility to prooxidants situations, in the brain, liver, and kidney of mice. Compound 1c reduced per se the lipid peroxidation in liver and brain of mice. Toxicological assays demonstrate that compounds 1a, 1b, and 1c did not present toxicity in the aspartate aminotransferase, alanine aminotransferase, urea, and creatinine levels in the plasma. In conclusion, the results demonstrated the antioxidant action of pyrazol derivative compounds in in vitro assays. Furthermore, the results showed low toxicity of compounds in in vivo assays.

scholarly journals Cytochrome P450-mediated warfarin metabolic ability is not a critical determinant of warfarin sensitivity in avian species: In vitro assays in several birds and in vivo assays in chicken

2015 ◽  
Vol 34 (10) ◽  
pp. 2328-2334 ◽  
Author(s):  
Kensuke P. Watanabe ◽  
Minami Kawata ◽  
Yoshinori Ikenaka ◽  
Shouta M. M. Nakayama ◽  
Chihiro Ishii ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Avian Species ◽  
In Vitro Assays ◽  
Critical Determinant ◽  
In Vivo Assays

scholarly journals Phosphorylation of RhoGDI by Src Regulates Rho GTPase Binding and Cytosol-Membrane Cycling

2006 ◽  
Vol 17 (11) ◽  
pp. 4760-4768 ◽  
Author(s):  
Céline DerMardirossian ◽  
Gabriel Rocklin ◽  
Ji-Yeon Seo ◽  
Gary M. Bokoch
Keyword(s):  
Rho Gtpases ◽  
Cell Function ◽  
Rho Gtpase ◽  
Physiological Mechanism ◽  
In Vitro Assays ◽  
Cortical Actin ◽  
Membrane Ruffling ◽  
Reactive Oxidant

Rho GTPases (Rac, Rho, and Cdc42) play important roles in regulating cell function through their ability to coordinate the actin cytoskeleton, modulate the formation of signaling reactive oxidant species, and control gene transcription. Activation of Rho GTPase signaling pathways requires the regulated release of Rho GTPases from RhoGDI complexes, followed by their reuptake after membrane cycling. We show here that Src kinase binds and phosphorylates RhoGDI both in vitro and in vivo at Tyr156. Analysis of Rho GTPase–RhoGDI complexes using in vitro assays of complexation and in vivo by coimmunoprecipitation analysis indicates that Src-mediated phosphorylation of Tyr156 causes a dramatic decrease in the ability of RhoGDI to form a complex with RhoA, Rac1, or Cdc42. Phosphomimetic mutation of Tyr156→Glu results in the constitutive association of RhoGDIY156E with the plasma membrane and/or associated cortical actin. Substantial cortical localization of tyrosine-phosphorylated RhoGDI is also observed in fibroblasts expressing active Src, where it is most evident in podosomes and regions of membrane ruffling. Expression of membrane-localized RhoGDIY156E mutant is associated with enhanced cell spreading and membrane ruffling. These results suggest that Src-mediated RhoGDI phosphorylation is a novel physiological mechanism for regulating Rho GTPase cytosol membrane–cycling and activity.

Brevifoliasaponin with Adjuvant Activity from Calliandra brevifolia

2008 ◽  
Vol 63 (7) ◽  
pp. 894-902 ◽  
Author(s):  
Antony P. Barbosa ◽  
Bernadete P. da Silva ◽  
José P. Parente
Keyword(s):  
2D Nmr ◽  
In Vitro Assays ◽  
Triterpenoid Saponin ◽  
Spectroscopic Techniques ◽  
Adjuvant Activity ◽  
In Vivo Assays ◽  
Cellular Immune ◽  
C Nmr

A new complex triterpenoid saponin, brevifoliasaponin, was isolated from leaves of Calliandra brevifolia Benth. (Leguminosae) by using chromatographic methods. Its structure was established as 3-[(O-α-L-arabinopyranosyl-(1 → 2)-O-α-L-arabinopyranosyl-(1 → 6)-2-(acetylamino)- 2-deoxy-β -D-glucopyranosyl)oxy]-16-hydroxy-(3β ,16α)-olean-12-en-28-oic acid O-β - D-xylopyranosyl-(1 → 3)-O-β -D-xylopyranosyl-(1→4)-O-[β -D-glucopyranosyl-(1 → 3)]-O-6-deoxy- α-L-mannopyranosyl-(1 → 2)-6-O-[(2E,6S)-6-[[2-O-[(2E,6S)-6-[[2-O-[(2E,6S)-2,6-dimethy- l1-oxo-6-(β -D-xylopyranosyloxy)-2,7-octadienyl]-β -D-xylopyranosyl]oxy]-2,6-dimethyl-1-oxo- 2,7-octadienyl]-β -D-xylopyranosyl]oxy]-2,6-dimethyl-1-oxo-2,7-octadienyl]-β -D-glucopyranosyl ester. Its structural elucidation was performed using detailed analyses of 1H and 13C NMR spectra including 2D-NMR spectroscopic techniques and chemical conversions. The hemolytic activity of the saponin was evaluated using in vitro assays, and its adjuvant potential on the cellular immune response against ovalbumin antigen was investigated using in vivo assays.

scholarly journals Antioxidant Activities of Commiphora leptophloeos (Mart.) J. B. Gillett) (Burseraceae) Leaf Extracts Using In Vitro and In Vivo Assays

2021 ◽  
Vol 2021 ◽  
pp. 1-11
Author(s):  
Maria Lúcia da Silva Cordeiro ◽  
Ana Raquel Carneiro Ribeiro ◽  
Luciana Fentanes Moura de Melo ◽  
Lucas Felipe da Silva ◽  
Gabriel Pereira Fidelis ◽  
...  
Keyword(s):  
Liquid Chromatography ◽  
Antioxidant Capacity ◽  
Antioxidant Activities ◽  
Biological Activities ◽  
Radical Scavenging ◽  
In Vitro Assays ◽  
Leaf Extracts ◽  
In Vivo Assays

Commiphora leptophloeos is widely used in folk medicine without any scientific basis. Considering this, the aim of this study was to evaluate the chemical profile and the antioxidant activity of C. leptophloeos leaf extracts using in vitro and in vivo assays. Six extracts were obtained from fresh leaves using a serial extraction (nonpolar to polar solvents). These extracts were first evaluated with the presence of phytochemical compounds using the methods thin layer chromatography (TLC), ultrahigh performance liquid chromatography (UHPLC-DAD), and high performance liquid chromatography, both with diode array detection (HPLC-DAD). Based on the compounds identified, it was used some bioinformatics tools in order to identify possible pathway and gene targets. After that, the antioxidant capacity from these extracts was analysed by in vitro assays and in vivo assays using Caenorhabditis elegans model. Phytochemical analyses showed the presence of polyphenols, such as rutin, vitexin, and quercetin diglycosides in all extracts, especially in ethanol extract (EE) and methanol extract (EM). Bioinformatics analysis showed these polyphenols linked to antioxidant pathways. Furthermore, EE and EM displayed a high antioxidant capacity in DPPH and superoxide radical scavenging assays. They also had no effect on cell viability for 3T3 nontumour cell. However, for B16-F10 tumour cell lines, these extracts had toxicity effect. In vivo assays using C. elegans N2 showed that EE was not toxic, and it did not affect its viability nor its development. Besides, EE increased worm survival under oxidative stress and reduced intracellular reactive oxygen species (ROS) levels by 50%. Thus, C. leptophloeos EE displayed an important in vitro and in vivo antioxidant capacity. The EE extract has polyphenols, suggesting that these compounds may be responsible for a myriad of biological activities having this potential to be used in various biotechnological applications.

Sign in / Sign up

Export Citation Format

Share Document