Proposed reduction of the in vivo pyrogen test by the in vitro LAL assay for the quality control of anticrotallic, antiscorpion, antirabies and antitetanus sera

2019 ◽  
Vol 59 ◽  
pp. 292-299
Author(s):  
Fernando F. Fingola ◽  
Sheila R.G. Albertino ◽  
Shirley de M.P. Abrantes ◽  
Helena P.S. Zamith
Keyword(s):  
Quality Control ◽  
Lal Assay

Animal Models and Alternatives in the Quality Control of Vaccines: Are In Vitro Methods or In Vivo Methods the Scientific Equivalent of the Emperor's New Clothes?

1995 ◽  
Vol 23 (1) ◽  
pp. 61-73
Author(s):  
Coenraad Hendriksen ◽  
Johan van der Gun
Keyword(s):  
Quality Control ◽  
Test Methods ◽  
In Vitro Test ◽  
Large Numbers ◽  
Alternative Approach ◽  
Inactivated Vaccines ◽  
Vitro Test

In the quality control of vaccine batches, the potency testing of inactivated vaccines is one of the areas requiring very large numbers of animals, which usually suffer significant distress as a result of the experimental procedures employed. This article deals with the potency testing of diphtheria and tetanus toxoids, two vaccines which are used extensively throughout the world. The relevance of the potency test prescribed by the European Pharmacopoeia monographs is questioned. The validity of the potency test as a model for the human response, the ability of the test to be standardised, and the relevance of the test in relation to the quality of the product are discussed. It is concluded that the potency test has only limited predictive value for the antitoxin responses to be expected in recipients of these toxoids. An alternative approach for estimating the potency of toxoid batches is discussed, in which a distinction is made between estimation of the immunogenic potency of the first few batches obtained from a seed lot and monitoring the consistency of the quality of subsequent batches. The use of animals is limited to the first few batches. Monitoring the consistency of the quality of subsequent batches is based on in vitro test methods. Factors which hamper the introduction and acceptance of the alternative approach are considered. Finally, proposals are made for replacement, reduction and/or refinement (the Three Rs) in the use of animals in the routine potency testing of toxoids.

scholarly journals Humanin is an endogenous activator of chaperone-mediated autophagy

2017 ◽  
Vol 217 (2) ◽  
pp. 635-647 ◽  
Author(s):  
Zhenwei Gong ◽  
Inmaculada Tasset ◽  
Antonio Diaz ◽  
Jaime Anguiano ◽  
Emir Tas ◽  
...  
Keyword(s):  
Quality Control ◽  
Cell Death ◽  
Heat Shock Protein 90 ◽  
Protective Effects ◽  
Neuroprotective Effects ◽  
Selective Degradation ◽  
Cytosolic Side ◽  
Chaperone Mediated Autophagy

Chaperone-mediated autophagy (CMA) serves as quality control during stress conditions through selective degradation of cytosolic proteins in lysosomes. Humanin (HN) is a mitochondria-associated peptide that offers cytoprotective, cardioprotective, and neuroprotective effects in vivo and in vitro. In this study, we demonstrate that HN directly activates CMA by increasing substrate binding and translocation into lysosomes. The potent HN analogue HNG protects from stressor-induced cell death in fibroblasts, cardiomyoblasts, neuronal cells, and primary cardiomyocytes. The protective effects are lost in CMA-deficient cells, suggesting that they are mediated through the activation of CMA. We identified that a fraction of endogenous HN is present at the cytosolic side of the lysosomal membrane, where it interacts with heat shock protein 90 (HSP90) and stabilizes binding of this chaperone to CMA substrates as they bind to the membrane. Inhibition of HSP90 blocks the effect of HNG on substrate translocation and abolishes the cytoprotective effects. Our study provides a novel mechanism by which HN exerts its cardioprotective and neuroprotective effects.

Preparation of fibrils for intracerebral injection v1

2021 ◽  
Author(s):  
The Michael J Fox Foundation Pff Standardization Consortium
Keyword(s):  
Quality Control ◽  
Best Practices ◽  
External Validation ◽  
Alpha Synuclein ◽  
Intracerebral Injection ◽  
Primary Neuron ◽  
Consensus Protocol ◽  
Reference Section

This is a consensus protocol developed through discussions with Laura Volpicelli-Daley, Caryl Sortwell, Kelvin Luk, Lindsey Gottler, and Virginia Lee. This protocol is intended for research purposes only, using specially-formulated monomeric alpha-synuclein protein available for purchase at Proteos, Inc as the result of efforts by The Michael J. Fox Foundation (MJFF). Each batch of the “Alpha-Synuclein Monomer Protein for Making Pre- Formed Fibrils” has undergone internal purification and quality control at Proteos in addition to external validation to confirm successful generation of pathogenic aSyn PFFs. See Reference section for methods and results from application of alpha-synuclein pre-formed fibrils (aSyn PFFs) in primary neuron cultures in vitro or in mice in vivo. This protocol is referenced in the Polinski et al 2018 paper entitled "Best Practices for Generating and Using Alpha-Synuclein Pre-Formed Fibrils to Model Parkinson's Disease in Rodents" (doi: 10.3233/JPD-171248).

Protocol for Generation of Pre-Formed Fibrils from Alpha-Synuclein Monomer v1

2021 ◽  
Author(s):  
The Michael J Fox Foundation Pff Standardization Consortium
Keyword(s):  
Parkinson’S Disease ◽  
Quality Control ◽  
Best Practices ◽  
External Validation ◽  
Alpha Synuclein ◽  
Primary Neuron ◽  
Consensus Protocol ◽  
Reference Section

This is a consensus protocol developed through discussions with Laura Volpicelli-Daley, Caryl Sortwell, Kelvin Luk, Lindsey Gottler, and Virginia Lee. This protocol is intended for research purposes only, using specially-formulated monomeric alpha-synuclein protein available for purchase at Proteos, Inc as the result of efforts by The Michael J. Fox Foundation (MJFF). Each batch of the “Alpha-Synuclein Monomer Protein for Making Pre- Formed Fibrils” has undergone internal purification and quality control at Proteos in addition to external validation to confirm successful generation of pathogenic aSyn PFFs. See Reference section for methods and results from application of alpha-synuclein pre-formed fibrils (aSyn PFFs) in primary neuron cultures in vitro or in mice in vivo. This protocol is referenced in the Polinski et al 2018 paper entitled "Best Practices for Generating and Using Alpha-Synuclein Pre-Formed Fibrils to Model Parkinson's Disease in Rodents" (doi: 10.3233/JPD-171248).

Marsdenia tenacissima: A Review of Traditional Uses, Phytochemistry and Pharmacology

2018 ◽  
Vol 46 (07) ◽  
pp. 1449-1480 ◽  
Author(s):  
Peile Wang ◽  
Jing Yang ◽  
Zhenfeng Zhu ◽  
Xiaojian Zhang
Keyword(s):  
Quality Control ◽  
Multidrug Resistance ◽  
Control Method ◽  
Food Poisoning ◽  
Bioactive Constituents ◽  
Resistance Reversal ◽  
Wide Clinical Application ◽  
Marsdenia Tenacissima

The stems and roots of Marsdenia tenacissima (Roxb.) Wight et Arn., a traditional Chinese medicine and Dai herbal medicine, have been widely used for the treatment of asthma, trachitis, tonsillitis, pharyngitis, cystitis, pneumonia and drug or food poisoning. Nowadays, the extract of Marsdenia tenacissima, under the trademark of “Xiao-ai-ping”, is widely used in clinic for the treatment of different cancers in China. To date, approximately 196 chemical ingredients covering steroids, triterpenes and organic acids have been identified from different parts of this plant. Steroids are the major characteristic and bioactive constituents of this plant. Modern pharmacology has demonstrated that the crude extracts and steroids have various in vitro and in vivo pharmacological activities, such as multidrug resistance reversal, antitumor, anti-angiogenic, immunomodulation and anti-HIV activities. The multidrug resistance reversal of steroids provided evidence for the use of this herb in clinic. However, despite wide clinical application, clinical trials, quality control method, pharmacokinetic and toxicity research on Marsdenia tenacissima were seldom reported and deserved further efforts. The present review aimed to achieve a comprehensive and up-to-date investigation in ethnopharmacology, phytochemistry, pharmacology, clinical study, pharmacokinetics, toxicology and quality control of Marsdenia tenacissima. In addition, the possible perspectives and trends for future studies of Marsdenia tenacissima have also been put forward. It is believed that this review would provide a theoretical basis and valuable data for future in-depth studies and applications.

Chemical assays--new opportunities.

1987 ◽  
Vol 33 (9) ◽  
pp. 1574-1578 ◽  
Author(s):  
A E Burkhardt
Keyword(s):  
Quality Control ◽  
Solid Phase ◽  
Clinical Chemistry ◽  
Test Format ◽  
Home Testing

Abstract The acceptance of the solid-phase format in various areas of clinical chemistry is the consequence of the advantages of this test format, which include stability of the reagents, unitized packaging, convenient and small instruments, and minimal preparations by users before testing. Overall, these advantages provide very convenient tests. Future successful uses of solid-phase reagents depend upon how well these features meet the needs of the users. Needs for systems to be used in the decentralized laboratory include even less cost, even more convenience, and improved quality control. Needs for home testing include convenient tests with clinically useful accuracy, improved quality control, and improved recording systems to overcome user bias in recording results. New solid-phase technologies being developed include noncolorimetric systems suitable for use with miniature probes, for in vitro or in vivo use, and spectrophotometric systems for determinations of analytes directly in capillaries of the skin without invasive sampling.

scholarly journals Quality Control of a Transcriptional Regulator by SUMO-Targeted Degradation

2009 ◽  
Vol 29 (7) ◽  
pp. 1694-1706 ◽  
Author(s):  
Zheng Wang ◽  
Gregory Prelich
Keyword(s):  
Quality Control ◽  
Protein Quality ◽  
Temperature Sensitive ◽  
Wild Type ◽  
Physiological Importance ◽  
Sensitive Allele ◽  
Quality Control Mechanism ◽  
Mutant Phenotypes

ABSTRACT Slx5 and Slx8 are heterodimeric RING domain-containing proteins that possess SUMO-targeted ubiquitin ligase (STUbL) activity in vitro. Slx5-Slx8 and its orthologs are proposed to target SUMO conjugates for ubiquitin-mediated proteolysis, but the only in vivo substrate identified to date is mammalian PML, and the physiological importance of SUMO-targeted ubiquitylation remains largely unknown. We previously identified mutations in SLX5 and SLX8 by selecting for suppressors of a temperature-sensitive allele of MOT1, which encodes a regulator of TATA-binding protein. Here, we demonstrate that Mot1 is SUMOylated in vivo and that disrupting the Slx5-Slx8 pathway by mutation of the target lysines in Mot1, by deletion of SLX5 or the ubiquitin E2 UBC4, or by inhibition of the proteosome suppresses mot1-301 mutant phenotypes and increases the stability of the Mot1-301 protein. The Mot1-301 mutant protein is targeted for proteolysis by SUMOylation to a much greater extent than wild-type Mot1, suggesting a quality control mechanism. In support of this idea, growth of Saccharomyces cerevisiae in the presence of the arginine analog canavanine results in increased SUMOylation and Slx5-Slx8-mediated degradation of wild-type Mot1. These results therefore demonstrate that Mot1 is an in vivo STUbL target in yeast and suggest a role for SUMO-targeted degradation in protein quality control.

In vitro and in vivo assays to assess the functions of calnexin and calreticulin in ER protein folding and quality control

Methods ◽  
2005 ◽  
Vol 35 (4) ◽  
pp. 338-347 ◽  
Author(s):  
Marie-Eve Paquet ◽  
Michael R. Leach ◽  
David B. Williams
Keyword(s):  
Quality Control ◽  
Protein Folding ◽  
In Vivo Assays

scholarly journals Identifying of Anti-Thrombin Active Components From Curcumae Rhizoma by Affinity-Ultrafiltration Coupled With UPLC-Q-Exactive Orbitrap/MS

2021 ◽  
Vol 12 ◽  
Author(s):  
Zhenwei Lan ◽  
Ying Zhang ◽  
Yue Sun ◽  
Lvhong Wang ◽  
Yuting Huang ◽  
...  
Keyword(s):  
Quality Control ◽  
Molecular Docking ◽  
Targeted Therapy ◽  
Screening Method ◽  
Active Ingredients ◽  
Active Compounds ◽  
Q Exactive ◽  
Orbitrap Ms

Recent studies concerning products that originate from natural plants have sought to clarify active ingredients, which both explains the mechanisms of the function and aids in quality control during production. As a traditional functional plant, Curcumae Rhizoma (CR) has been proven to be effective in promoting blood circulation and removing blood stasis. However, the components that play a role in its huge compound library are still unclear. The present study aimed to develop a high-throughput screening method to identify thrombin inhibitors in CR and validate them by in vitro and in vivo experiments. The effect of CR on thrombin in HUVECs cells was determined by ELISA, then an affinity-ultrafiltration-UPLC-Q-Exactive Orbitrap/MS approach was applied. Agatroban and adenosine were used as positive and negative drugs respectively to verify the reliability of the established method. The in vitro activity of the compounds was determined by specific substrate S-2238. The in vivo effect of the active ingredients was determined using zebrafish. Molecular docking was used to understand the internal interactions between compounds and enzymes. ELISA results showed that CR had an inhibitory effect on thrombin. The screening method established in this paper is reliable, by which a total of 15 active compounds were successfully identified. This study is the first to report that C7, 8, and 11 have in vitro thrombin-inhibitory activity and significantly inhibit thrombosis in zebrafish models at a safe dose. Molecular docking studies were employed to analyze the possible active binding sites, with the results suggesting that compound 16 is likely a better thrombin inhibitor compared with the other compounds. Based on the affinity-ultrafiltration-UPLC-Q-Exactive Orbitrap/MS approach, a precisely targeted therapy method using bio-active compounds from CR might be successfully established, which also provides a valuable reference for targeted therapy, mechanism exploration, and the quality control of traditional herbal medicine.

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