Preparation of fibrils for intracerebral injection v1

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

10.17504/protocols.io.bws6pehe ◽

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).

Preparation of fibrils and Quality control v1

10.17504/protocols.io.bwswpefe ◽

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).

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?

Alternatives to Laboratory Animals ◽

10.1177/026119299502300110 ◽

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.

Humanin is an endogenous activator of chaperone-mediated autophagy

The Journal of Cell Biology ◽

10.1083/jcb.201606095 ◽

2017 ◽

Vol 217 (2) ◽

pp. 635-647 ◽

Cited By ~ 33

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.

Quality control in the IVF laboratory: in-vitro and in-vivo development of mouse embryos is unaffected by the quality of water used in culture media

Human Reproduction ◽

10.1093/oxfordjournals.humrep.a136994 ◽

1989 ◽

Vol 4 (7) ◽

pp. 826-831 ◽

Cited By ~ 18

Author(s):

Martin A. George ◽

Peter R. Braude ◽

Martin H. Johnson ◽

David G. Sweetnam

Keyword(s):

Quality Control ◽

Culture Media ◽

Mouse Embryos ◽

Quality Of Water

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

The American Journal of Chinese Medicine ◽

10.1142/s0192415x18500751 ◽

2018 ◽

Vol 46 (07) ◽

pp. 1449-1480 ◽

Cited By ~ 3

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.

Clinical Chemistry ◽

10.1093/clinchem/33.9.1574 ◽

1987 ◽

Vol 33 (9) ◽

pp. 1574-1578 ◽

Cited By ~ 2

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.

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

Toxicology in Vitro ◽

10.1016/j.tiv.2019.04.031 ◽

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

C57BL/6JOlaHsd Mice with Tandem Deletion of the Multimerin 1 and Alpha-Synuclein Genes Have Impaired Platelet Function in Vivo and in Vitro That Can Be Corrected by Multimerin 1

Blood ◽

10.1182/blood.v112.11.3926.3926 ◽

2008 ◽

Vol 112 (11) ◽

pp. 3926-3926 ◽

Cited By ~ 1

Author(s):

Subia Tasneem ◽

Adili Reheman ◽

Heyu Ni ◽

Catherine P.M. Hayward

Keyword(s):

Platelet Aggregation ◽

Platelet Function ◽

Knockout Mice ◽

Platelet Adhesion ◽

Thrombus Formation ◽

Alpha Synuclein ◽

Type I ◽

Significant Difference

Abstract Studies of mice with genetic deficiencies have provided important insights on the functions of many proteins in thrombosis and hemostasis. Recently, a strain of mice (C57BL/6JOlaHsd, an inbred strain of C57BL/6J) has been identified to have a spontaneous, tandem deletion of the multimerin 1 and α-synuclein genes, which are also adjacent genes on human chromosome 4q22. Multimerin 1 is an adhesive protein found in platelets and endothelial cells while α-synuclein is a protein found in the brain and in blood that is implicated in neurodegenerative diseases and exocytosis. In vitro, multimerin 1 supports platelet adhesion while α-synuclein inhibits α-granule release. We postulated that the loss of multimerin 1 and α-synuclein would alter platelet function and that recombinant human multimerin 1 might correct some of these abnormalities. We compared platelet adhesion, aggregation and thrombus formation in vitro and in vivo in C57BL/6JOlaHsd and C57BL/6 mice. Thrombus formation was studied by using the ferric-chloride injured mesenteric arteriole thrombosis model under intravital microscopy. We found that platelet adhesion, aggregation and thrombus formation in C57BL/6JOlaHsd were significantly impaired in comparison to control, C57BL/6 mice. The number of single platelets, deposited 3–5 minutes after injury, was significantly decreased in C57BL/6JOlaHsd mice (P <0.05, platelets/min: C57BL/6 = 157 ± 15, n=16; C57BL/6JOlaHsd = 77 ± 13, n=17). Moreover, thrombus formation in these mice was significantly delayed. Thrombi in C57BL/6JOlaHsd were unstable and easily dissolved, which resulted in significant delays (P<0.001) in vessel occlusion (mean occlusion times: C57BL/6 = 15.6 ± 1.2 min, n=16; C57BL/6JOlaHsd = 31.9 ± 2.1 min, n=17). We further tested platelet function in these mice by ADP and thrombin induced platelet aggregation using platelet rich plasma and gel-filtered platelets, respectively. Although no significant differences were seen with ADP aggregation, thrombin-induced platelet aggregation was significantly impaired in C57BL/6JOlaHsd mice. Platelet adhesion to type I collagen (evaluated using microcapillary chambers, perfused at 1500 s−1 with whole blood) was also impaired in C57BL/6JOlaHsd mice. However, platelets from C57BL/6JOlaHsd mice showed a normal pattern of agonist-induced release of α-granule P-selectin. Multimerin 1 corrected the in vitro aggregation and adhesion defects of C57BL/6JOlaHsd platelets. Furthermore, the transfusion of multimerin 1 into C57BL/6JOlaHsd mice corrected the impaired platelet deposition and thrombus formation in vivo. No significant difference was found in tail bleeding time between the two groups of mice. As α-synuclein knockout mice have a shortened time to thrombus formation (Circulation2007;116:II_76), the effects of multimerin 1 on impaired platelet function in C57BL/6JOlaHsd mice provide supportive evidence that multimerin 1 contributes to platelet adhesion and thrombus formation at the site of vessel injury. The findings suggest multimerin 1 knockout mice will be useful to explore platelet function. The first two authors and participating laboratories contributed equally to this study.

P1-036: Seeding of cross linked alpha-synuclein oligomers in vitro and in vivo

Alzheimer s & Dementia ◽

10.1016/j.jalz.2010.05.583 ◽

2010 ◽

Vol 6 ◽

pp. S183-S184

Author(s):

Therese Wahlberg ◽

Thomas Nasstrom ◽

Charlotte Sahlin ◽

Sofie Ingvast ◽

Mikael Karlsson ◽

...

Keyword(s):

Alpha Synuclein