Insights into exosomes role in disease transmission gained using NanoSight from Malvern

Data measured using the NanoSight NS300 from Malvern Panalytical is providing new insights into the role of exosomes in diseases such as cancer, arthritis, Alzheimer’s disease and cystic fibrosis, in pioneering research at the University of Alabama - Birmingham (UAB).

“Early research on exosomes, or extracellular vesicles, focused on their role in excreting unwanted waste from cells,” said Shawn Williams, facility manager and imaging specialist. “However, we’re now starting to understand their wider functionality in intercellular communication and the transmission of disease. The NanoSight system is playing a central role in helping us advance in this new and exciting area of research, by enabling us to size, count and track the movement of these crucial molecules in their native state.”

NanoSight Malvern Panalytical

Malvern Panalytical’s NanoSight NS300 provides new insights into the role of exosomes in disease transmission for researchers at the University of Alabama

The NanoSight NS300 system is sited within an analytical service facility for the university, which also extends support to external customers on a contract basis. Around 85% of the analysis being carried out in the lab is associated with disease-related research. The NanoSight system is used by several hundred different researchers, so rapid training and robust measurement protocols are essential. Mr. Williams and his team have developed a series of simple procedures to support new researchers in their use of the instrument, to enable the efficient generation of accurate particle size and count data, and video footage of the exosomes within a sample.

There are very few instruments that can do what the NanoSight does in terms of providing us with statistically significant size and count data in a cost-efficient way,” said Mr. Williams.

“The sample preparation required is negligible, which is a major advantage relative to Transmission Electron Microscopy (TEM), and we find that the instrument has a really short learning curve. All our users are able to access the data they need, presented in a great and easy to digest format, with minimal training.”

The NanoSight NS300 measures particle size in the range 10 nm to 2000 nm, comfortably spanning the 30 nm – 100 nm size range of primary interest for exosomes. NanoSight software is designed to make it easy for researchers to use the instrument, access the data required and present it in an easily assimilated form.

“I really appreciate the fact that Malvern Panalytical continues to enhance the functionality of the NanoSight system through software upgrades which add relevant and valuable capabilities. For us it is all about gathering information that will give us new understanding.  The more quickly and easily we can measure relevant parameters, the greater our productivity and the faster we can progress our research. This is crucial as demand for our services increases to meet the growing need to develop new and successful drug products.”

Malvern Panalytical, Malvern Panalytical, NanoSight are registered trademarks of Malvern Panalytical

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Comments

  1. Terry Singeltary Terry Singeltary United States says:

    I would kindly like to comment further on ;


    Alzheimer-type brain pathology may be transmitted by grafts of dura mater

    26/01/2016 By Karl Frontzek, et al.:

    blog.smw.ch/.../#respond


    Original article | Published 26 January 2016, doi:10.4414/smw.2016.14287

    Cite this as: Swiss Med Wkly. 2016;146:w14287

    Amyloid-β pathology and cerebral amyloid angiopathy are frequent in iatrogenic Creutzfeldt-Jakob disease after dural grafting

    http://www.smw.ch/content/smw-2016-14287/


    MY comment as follows ;

    Evidence for human transmission of amyloid-β pathology and cerebral amyloid angiopathy

    07 02:27 AM

    Terry S. Singeltary Sr. said:

    re-Evidence for human transmission of amyloid-β pathology and cerebral amyloid angiopathy

    2015-12-07 02:27 AM

    Terry S. Singeltary Sr. said: re-Evidence for human transmission of amyloid-? pathology and cerebral amyloid angiopathy Nature 525, 247?250 (10 September 2015) doi:10.1038/nature15369 Received 26 April 2015 Accepted 14 August 2015 Published online 09 September 2015 Updated online 11 September 2015 Erratum (October, 2015)

    www.nature.com/.../nature15369.html


    I would kindly like to comment on the Nature Paper, the Lancet reply, and the newspaper articles.

    First, I applaud Nature, the Scientist and Authors of the Nature paper, for bringing this important finding to the attention of the public domain, and the media for printing said findings.

    Secondly, it seems once again, politics is getting in the way possibly of more important Transmissible Spongiform Encephalopathy TSE Prion scientific findings. findings that could have great implications for human health, and great implications for the medical surgical arena. but apparently, the government peer review process, of the peer review science, tries to intervene again to water down said disturbing findings.

    where have we all heard this before? it’s been well documented via the BSE Inquiry. have they not learned a lesson from the last time?

    we have seen this time and time again in England (and other Country’s) with the BSE mad cow TSE Prion debacle.


    web.archive.org/.../16005001.pdf


    collections.europarchive.org/.../http://www.bseinquiry.gov.uk/.../04001001.pdf

    www.plosone.org/.../listThread.action?root=82860


    Terry S. Singeltary Sr. Bacliff, Texas USA 77518

    snip...see Singeltary comment ;

    www.nature.com/.../comments


    Subject: 1992 IN CONFIDENCE TRANSMISSION OF ALZHEIMER TYPE PLAQUES TO PRIMATES POSSIBILITY ON A TRANSMISSIBLE PRION REMAINS OPEN

    BSE101/1 0136

    IN CONFIDENCE

    CMO

    From: . Dr J S Metiers DCMO

    4 November 1992

    TRANSMISSION OF ALZHEIMER TYPE PLAQUES TO PRIMATES

    1. Thank you for showing me Diana Dunstan's letter. I am glad that MRC have recognised the public sensitivity of these findings and intend to report them in their proper context. 'This hopefully will avoid misunderstanding and possible distortion by the media to portray the results as having more greater significance than the findings so far justify.

    2. Using a highly unusual route of transmission (intra-cerebral injection) the researchers have demonstrated the transmission of a pathological process from two cases one of severe Alzheimer's disease the other of Gerstmann-Straussler disease to marmosets. However they have not demonstrated the transmission of either clinical condition as the "animals were behaving normally when killed". As the report emphasises the unanswered question is whether the disease condition would have revealed itself if the marmosets had lived longer. They are planning further research to see if the conditions, as opposed to the partial pathological process, is transmissible.

    what are the implications for public health?

    3. The route 'of transmission is very specific and in the natural state of things highly unusual. However it could be argued that the results reveal a potential risk, in that brain tissue from these two patients has been shown to transmit a pathological process. Should therefore brain tissue from such cases be regarded as potentially infective? Pathologists, morticians, neuro surgeons and those assisting at neuro surgical procedures and others coming into contact with "raw" human brain tissue could in theory be at risk. However, on a priori grounds given the highly specific route of transmission in these experiments that risk must be negligible if the usual precautions for handling brain tissue are observed.

    1

    92/11.4/1.1

    BSE101/1 0137

    4. The other dimension to consider is the public reaction. To some extent the GSS case demonstrates little more than the transmission of BSE to a pig by intra-cerebral injection. If other prion diseases can be transmitted in this way it is little surprise that some pathological findings observed in GSS were also transmissible to a marmoset. But the transmission of features of Alzheimer's pathology is a different matter, given the much greater frequency of this disease and raises the unanswered question whether some cases are the result of a transmissible prion. The only tenable public line will be that "more research is required’’ before that hypothesis could be evaluated. The possibility on a transmissible prion remains open. In the meantime MRC needs carefully to consider the range and sequence of studies needed to follow through from the preliminary observations in these two cases. Not a particularly comfortable message, but until we know more about the causation of Alzheimer's disease the total reassurance is not practical.

    J S METTERS Room 509 Richmond House Pager No: 081-884 3344 Callsign: DOH 832 llllYc!eS 2 92/11.4/1.2

    collections.europarchive.org/.../http://www.bseinquiry.gov.uk/.../04001001.pdf

    >>> The only tenable public line will be that "more research is required’’ <<<

    >>> possibility on a transmissible prion remains open<<<

    O.K., so it’s about 23 years later, so somebody please tell me, when is "more research is required’’ enough time for evaluation ?

    Self-Propagative Replication of Ab Oligomers Suggests Potential Transmissibility in Alzheimer Disease

    *** Singeltary comment PLoS ***

    Alzheimer’s disease and Transmissible Spongiform Encephalopathy prion disease, Iatrogenic, what if ?

    Posted by flounder on 05 Nov 2014 at 21:27 GMT

    www.plosone.org/.../listThread.action?root=82860

    Sunday, November 22, 2015

    *** Effect of heating on the stability of amyloid A (AA) fibrils and the intra- and cross-species transmission of AA amyloidosis Abstract

    Amyloid A (AA) amyloidosis is a protein misfolding disease characterized by extracellular deposition of AA fibrils. AA fibrils are found in several tissues from food animals with AA amyloidosis. For hygienic purposes, heating is widely used to inactivate microbes in food, but it is uncertain whether heating is sufficient to inactivate AA fibrils and prevent intra- or cross-species transmission. We examined the effect of heating (at 60 °C or 100 °C) and autoclaving (at 121 °C or 135 °C) on murine and bovine AA fibrils using Western blot analysis, transmission electron microscopy (TEM), and mouse model transmission experiments. TEM revealed that a mixture of AA fibrils and amorphous aggregates appeared after heating at 100 °C, whereas autoclaving at 135 °C produced large amorphous aggregates. AA fibrils retained antigen specificity in Western blot analysis when heated at 100 °C or autoclaved at 121 °C, but not when autoclaved at 135 °C. Transmissible pathogenicity of murine and bovine AA fibrils subjected to heating (at 60 °C or 100 °C) was significantly stimulated and resulted in amyloid deposition in mice. Autoclaving of murine AA fibrils at 121 °C or 135 °C significantly decreased amyloid deposition. Moreover, amyloid deposition in mice injected with murine AA fibrils was more severe than that in mice injected with bovine AA fibrils. Bovine AA fibrils autoclaved at 121 °C or 135 °C did not induce amyloid deposition in mice. These results suggest that AA fibrils are relatively heat stable and that similar to prions, autoclaving at 135 °C is required to destroy the pathogenicity of AA fibrils. These findings may contribute to the prevention of AA fibril transmission through food materials to different animals and especially to humans.

    Purchase options Price * Issue Purchase USD 511.00 Article Purchase USD 54.00

    www.tandfonline.com/.../13506129.2015.1095735

    betaamyloidcjd.blogspot.com/.../...ability-of.html


    Transmission of Creutzfeldt-Jakob disease to a chimpanzee by electrodes contaminated during neurosurgery.

    Gibbs CJ Jr, Asher DM, Kobrine A, Amyx HL, Sulima MP, Gajdusek DC. Laboratory of Central Nervous System Studies, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD 20892.

    Stereotactic multicontact electrodes used to probe the cerebral cortex of a middle aged woman with progressive dementia were previously implicated in the accidental transmission of Creutzfeldt-Jakob disease (CJD) to two younger patients. The diagnoses of CJD have been confirmed for all three cases. More than two years after their last use in humans, after three cleanings and repeated sterilisation in ethanol and formaldehyde vapour, the electrodes were implanted in the cortex of a chimpanzee. Eighteen months later the animal became ill with CJD. This finding serves to re-emphasise the potential danger posed by reuse of instruments contaminated with the agents of spongiform encephalopathies, even after scrupulous attempts to clean them.

    www.ncbi.nlm.nih.gov/.../query.fcgi

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