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ACTIP Bulletin 32

December 2002

 

In this issue:
ACTIP website remains popular

Governments should fund more basic and long term research
Fewer drug applications: EMEA budget down
FDA restructures new product review centers
The biomanufacturing bottleneck
News on transgenic animals
Genomics and proteomics
Websites related to genomics and proteomics:
Combinatorial chemistry successful
News from the Commission
Innovation in FP6
Business News
Research News
On the web
Publications/reports
Agenda

 

ACTIP website remains popular


The ACTIP website remains popular. For example, in the month of September, the website was visited 12,190 times, with 2,049 requests for pages and 122 requests to download files. The most popular pages visited were respectively publications, interesting links, news, library, organization and members.

 

Governments should fund more basic and long term research


Benchmarking industry-science relationships (ISBN 9264197419, 35 EURO) is a recently published report. It is the result of a 2 year OECD study of relationships between industry and public research (universities and specialized institutes). The report welcomes the widespread transfer of publicly funded research from specialized research institutes to the multidisciplinary setting of universities. However, it also concludes that universities must be given greater autonomy to decide research priorities, and governments should fund more basic and long-term research, because industry never will.

 

Fewer drug applications: EMEA budget down


For its budget, the EMEA relies on fees charged to drug companies, along with funding from Brussels. Early October, the EMEAs management board said it was downscaling the regulatory body's 2002 budget from 70 mEURO to 65.4 mEURO to reflect the decrease in the number of applications for the authorization of new medicines. By late September, only 25 applications had been received, down from 58 for the previous year, confirming a trend observed by other international regulators. The same pattern is evident at the FDA: last year, the FDA received a mere 16 applications. Observers from different companies and bodies all give different reasons for this decline. Taken together, the following picture emerges:

recent mega-mergers might quash new developments on the research front;
manufacturers move away from developing new products to a more complex generation of preventative remedies;
science is getting more exciting, more complex and more expensive (average R&D cost for a single new medicine in 2001 was 895 mEURO, compared to 200 mEURO in 1991);
many companies pull out in the latest stage of R&D (bad luck, fear of litigation);
on a positive note: there is a steady flow of applications for so-called orphan drugs.
Source: European Voice October 23, 2002.

 

FDA restructures new product review centers


On September 6, the US FDA announced that it will consolidate its two product review centers in an effort to speed up and harmonize the drug review and development process. Under the restructuring, the Center for Drug Evaluation and Research (CDER) will have sole authority over all new pharmaceutical products (including peptide therapeutics). The Center for Biologics Evaluation and Research (CBER) will continue to oversee approvals of tissue and blood products, cell and gene therapies, and vaccines.

Source: Nature Biotechnology, Vol 20, October 2002.

 

The biomanufacturing bottleneck


At the Amsterdam meeting (June 2002), the ACTIP members decided to focus for the next meeting on issues related to biomanufacturing capacity and optimization issues. Since then, a number of articles have appeared addressing this topic. Here is a selection to prepare for the ACTIP meeting in Gent, where this topic will be addressed.
Biologics (including protein hormones, protein-based vaccines, monoclononal antibodies and recombinant proteins, are fast becoming victims of their own success. A looming deficit in biomanufacturing capacity threatens to restrict the expansion of the commercialization of this group of products. Here a summary of some facts:

The sales of recombinant proteins for human applications were better than US $14 billion in 1998 and increased to an estimated 19 billion USD in 2000;
It is estimated that between 500-900 therapeutic proteins are in development;
There are 10 antibodies on the market today, and they are essentially consuming all the manufacturing capacity that is available. The problem is that protein drugs, because of being broken down in the bloodstream, tend to be given in larger doses than small molecule drugs.
There may be as many as 500 antibodies in development. It is foreseen that 20-100 new monoclonal antibodies will reach the drug market in the next ten years, necessitating at least a 200% increase in manufacturing capacity for that class of proteins alone.
Based on the above it has been estimated that by 2010 an additional 16 new production plants with at least the same capacity as the largest one in existence now (90,000 liter Genentech, Vacaville) would be required;
The current standard technology (using CHO cells in bioreactors) presents problems with respect to scaling up and the size of the bioreactors. Building more bioreactors multiplies costs linearly. A CHO-cell based manufacturing plant can cost upwards of 250 million USD. Errors in predicting demand for capacity have the habit of being incredibly costly.
Regulators in both Europe and the USA demand that drugs being produced for the market are produced in the same system as the ones used to produce the drug for the final round of clinical trials. Thus, companies have to build facilities for drugs that might not be approved.
Possible solutions:

A number of possible solutions are available. All are costly and most are difficult to realize on the short term. Here is a list of possibilities:

  • Build your own pilot-scale manufacturing facility;
  • Expanding capacity, either in house or by acquiring contract manufacturing capacity;
  • Use high yield expression systems;
  • Rethink scale-up practices and improve upstream and downstream purification processes;
  • investigate the possibility of using non-standard expression systems. For example:
transgenic mammals may express therapeutic proteins in their milk (mammary bioreactor) (dairy animals, rabbits). Costs approx 1-2 USD/g raw material)
transgenic chickens may also express therapeutic proteins in the eggs (costs approx 1-2 USD /g raw material)
transgenic plants may express the desired protein in seeds (approx costs 0.05 USD/g raw material)
In all these systems, correct folding and glycosylation may present problems. An alternative solution is developing a generic glycosylation process, that can be used to glycosylate proteins derived from any biomanufacturing process.

Interesting reading ......
http://www.chireports.com/content/reports/MAB-222.ASP Prospects for Commercialization of Human Monoclonal Antibodies
This report, based on interviews with leading experts in the field, examines business and
technological trends fueling the use of monoclonal antibodies in drug discovery and development.Ý
For more information, visit http://www.chiresource.com/genomelink.asp or http://www.CHIreports.com.

 


News on transgenic animals


Humantherapeutic proteins that have been expressed in transgenic animals such as pigs, cows, sheep, and goats include: collagen to treat burns and bone fractures, fibrinogen to treat burns, human fertility hormones and anti-clotting proteins.
Mammals are considered the best-suited animals to produce recombinant proteins, as the complexity of the structure of these proteins often requires mammal cell systems to reach industrial scale at reasonable costs. Mammary glands in females and seminal vesicles in males generate kilograms of very potent products to be sold as pharmaceuticals. Recently, a Canadian group patented a technology whereby genetically engineered pigs produce recombinant proteins in their seminal fluid. The semen is collected and the specific protein is purified. Advantages are: a short generation time of the pigs (four months), animals reach sexual maturity at six months and produce 10-12 piglets per litter; there is a minimal risk of zoonosis; each animal produces 250-500 ml of semen per day, year-round; and the technology allows the production of complex recombinant proteins (carboxylation, glycosylation, methylation) difficult to produce in other systems.

Pharming to continue
Pharming, troubled by financial debts, has been able to raise new capital, drawn up schemes to pay off its debtors, and restructured its employee base (from 230 to 40) and activities. Pharming's current focus is on human C1-esterase inhibitor produced in transgenic rabbits. A phase 2 study has been planned. For phase 3 studies, Pharming is looking for a partner, i.e. a large pharmaceutical company.

Source: Bionieuws October 5, 2002

 

Genomics and proteomics


Proteomic techniques
Genomics is producing a gigantic number of nucleotide sequences that need to undergo ëproof of principleí by translating them to a functional protein and in vivo testing. Here a review of some of the methods used:
Generating proteins
fast and easy: production of target proteins from isolated mRNA or intron-less cDNA, cloning into a plasmid via a cell free system. Suitable for first screening experiments of polypeptides up to 20KDa;
employing bacteria (mainly E.coli) for the translation of novel sequences. Only intronfree cDNA and not genomic DNA can be handled by bacteria, and no post-translational modifications, so limited set of proteins;
eukaryotic expression systems (yeast, insect cells, mammalian cells). High costs, and use of FCS might present problems (and lowers yield). New cell lines adapted to serum and protein free media. Lengthy procedures, but novel cell based technologies continuously discovered;
DNA/RNA, peptide and protein biochips. Next step will be adhered whole cells or tissues equipped with inducible reporters.

Source: Michael Zang-Gandor, Eugenex biotechnologies, in European Biopharmaceutical Review, Summer 2002, pp 94-96

Analyzing proteins
The key technology platforms, which is mostly used in proteomics research, uses 2D-PAGE for separation and quantification, followed by identification of the proteins in the spots, by mass fingerprint or partial amino acid sequence information using MS. This approach has various limitations, and as a result various approaches are being taken to develop ëgel-freeí systems:
- High Performance liquid chromatography (HPLC) is generally used for the analysis of peptides. Not applicable to proteins or peptides with a higher molecular weight, and has a limited resolution;
- Forms and variations of one and two-dimensional capillary electrophoresis (CE), with or without online coupling to mass spectrometry. Wide range of proteins can be analyzed, but limitation is its resolution, allowing a separation of only several hundred proteins and peptides;
- Isotope coded affinity tag, aims to directly compare protein quantities between samples. Only cysteine-containing peptides can be seen. Mostly applicable for the analysis of samples with a limited number of proteins;
- Protein sequence tagging (PST) selects and analyses proteolytically-generated N-terminal peptides. Overcomes limitations other methods. Allows a sensitive study of physiological changes on the basis of primary protein structure information.

Source: Van Rooijen/Schwarz, Xzillion Gmbh, in European Biopharmaceutical Review, Summer 2002, pp 74-77

 

Websites related to genomics and proteomics:


Websites for proteomic research. All are publicly accessible.
Peptide fragment databases:
http://prospector.ucsf.edu

http://www.wolf.bms.umist.ac.uk/mapper
http://www.molgen.mpg.de/~mass-spec
http://www.hgmp.mrc.ac.uk
2-DE databases
http://toothprint.otago.ac.nz

http://www.expasy.ch/ch2d
Protocols for sample preparation
http://www.expasy.ch/ch2

http://www.pierroton.inra.fr/genetics/2D
Genomic databases:
http://www.ncbi.nlm.nih.gov/Entrez

http://www.tigr.org/tdb
http://www.expasy.ch

(Source: IVD Technology, September 2002, pp 38)
'Pat Brown approach': http://cmgm.stanford.edu/pbrown

Crosslinker to bind DNA fragments to non-modified glass: http://imgen.bcm.tmc.edu/molgen/facultyaz/cai.html
NIH microarray project:
http://www.nhgri.nih.gov/DIR/Microarray/main.html
Microfluidics, example to reduce hybridization variation:
http://www.biomicro.com/technology/hfd_mix.html
Totally new approach: glass fibre arrays:
http://ase.tufts.edu/chemistry/walt/
Nanotechnological approaches:
http://www.technologyreview.com/articles/stikeman0502.asp

http://www.gene-chips.com/Leming.html
Canadian programme (580 mCAN$) on genomics:
http://www.genomecanada.ca
Cellular signaling and protein-protein interactions:
http://www.cellularsignaling.org

 

Combinatorial chemistry successful

To be able to use combinatorial chemistry, a company must be able to combine informatics, engineering, micro-electronics, material sciences and wet chemistry. Application of combinatorial chemistry is very successful in the search for new drug candidates, but so far the costs are still extremely high. Also, it should be applied for process-based applications and to explore complex systems.
See also:
A combi web portal: http://www.combi-web.com
Combinatorial index website: http://www.combinatorial.com
Combichemnet, portal: http://www.combichem.net/home/index.html
High throughput experimentation, portal: http://www.highthroughputexperimentation.com
Microfluidics: http://www.engineering.ucsb.edu/~microflu ; http://microfluidics.stanford.edu ;
http://www.aip.org/pt/vol-54/iss-6/p42.html
Labs on a chip:
Portal: http://www.lab-on-a-chip.com/home/index.html
Lab on a chip journal: http://www.rsc.org/is/journals/current/loc/locpub.htm

 

News from the Commission

On November 11-13, the European Commission launched its Framework Programme 6 in the Heysel conference center in Brussels. The launch was attended by more than 8,000 people, and there were numerous sessions explaining the rules and regulations (or: how to participate) of FP6. There were also information stands where Commission officials could be approached for information.
The Specific Programmes were adopted by the Council on September 30 and are available from the cordis website. Unfortunately, the Work Programmes were not available yet. This is expected to happen around December 17, together with the first call for proposals. It will be worthwhile to check the CORDIS website around that date. The deadline for the first call for proposals will be the End of March 2003. The official text of FP6 and of the Specific Programme (in English) can also be found on the secure page of the ACTIP website ( http://www.actip.org/secure)
At the ACTIP meeting in Gent, an overview of the launch of FP6 will be presented. As soon as the Work Programme will be published, we will prepare a special ACTIP Newsbulletin on FP6 and the Work Programme 2003. For the time being, we prefer to refer to the following websites:
Up to date information on calls for proposals, work programmes, guides for participants etc, see:
http://www.cordis.lu
For information on genomics, see pp 11-12 of the Specific Programme for RTD for FP6:
http://europa.eu.int/comm/research/fp6
Rules of participation in FP6:
http://europa.eu.int/comm/research/fp6/pdf/rules_en.pdf
Instruments for implementing the thematic priorities of FP6:
http://europa.eu.int/comm/research/fp6/networks-ip.html

 

Innovation in FP6

Unlike previous Framework Programmes, FP6 does not include a specific 'innovation programme'. Instead, innovation-related activities are distributed throughout Work Programmes 1 and 2 &endash; integrating and strengthening the European Research Area (WP1) and Structuring the European Research Area (WP2). They remain under the management of the Directorate-General for Enterprise, and will build on the equivalent actions of FP5. A significant proportion of FP6s innovation-related work concerns the trans-regional, trans-sectoral and cross-disciplinary networking of public authorities, economic actors and social partners ñ the stakeholders who together form Europeís innovation community. DG Enterprise has recently published a directory on all aspects of innovation policy in ëInnovation and Technology Transfer(October 2002). The directory covers:
the principal services of CORDIS;
the other main innovation-related resources provided by the Commission;
national authorities responsible for innovation support;
key networks of regional innovation service providers.
The directory is intended as a resource for innovation actors for the lifetime of FP6. Warmly recommended. See also http://www.cordis.lu/rtd2002/home.html . Below we already give some of the resources to be found in this directory:
Trend chart on innovation in Europe: http://trendchart.cordis.lu
National R&D and Innovation Information services (websites for each member state): http://www.cordis.lu/national_service/
CORDIS Wire (where the business and research community can publish press releases): mailto:wire@corids.lu" wire@corids.lu
News services for FP6 participants: http://www.cordis/lu/fp6
Cordis News with daily updates on programme calls, events, policy developments etc: http://dbs.cordis.lu/news/en . CORDIS Express - http://www.cordis.lu/express/ contains summaries of the News services most recent items on research and innovation policy. You can submit news items to: mailto:news@cordis.lu
CORDIS Technology Marketplace with clearly written digests of the latest research results: http://www.cordis.lu/marketplace/
Euroabstracts with brief summaries of the key facts and ideas on innovation and scientific research from the Commission and international bodies: http://www.cordis.lu/euroabstracts
A Europe wide community for investors and service providers to share knowledge and good practice and with specific networks for early stage venture capital investors, industrial liaison officers etc: http://www.gate2gorwth.com/

 

Business News


Battle in UN over cloning ban
In the United Nations a battle is raging about an international ban on cloning of human beings and human cells. A total ban on reproductive as well as therapeutic cloning is proposed by the United States, Spain, and the Philippines, supported by the Vatican and a number of strict catholic countries such as Italy, Argentina and Costa Rica. Opposite are France and Germany, which have launched a proposal supported by most members of the UN: these countries all support a ban on reproductive cloning, and further negotiations regarding therapeutic cloning. The latter countries hold the opinion that a all-or-nothing approach as advocated by the United States will impede the decision making process, meaning that international regulation will not materialize at all, with all associated risks of the technology proceeding and reproductive cloning being carried out anyway.

Source:
U.N. Tries to Break Deadlock on Human Cloning Treaty, Inter Press Service, October 17, 2002;
Legal Committee calls for working groups on human cloning, http://www.health.fgov.be/WHI3/krant/krantarch2001/kranttekstnov1/011120m09un.htm
International Convention against the reproductive cloning of human beings, United Nations General Assembly, Sixth Committee, Fifty-seventh session,
http://daccess-ods.un.org/doc/UNDOC/LTD/N02/615/01/PDF/N0261501.pdf?OpenElement
Lion Biosciences drops drug discovery

On September 6, Lion Bioscience announced that it would discontinue its iD3-drug discovery business to focus on its core bioinformatics business. Many genomics companies that delved into drug discovery are finding the risks too high in the current economic climate &endash; drug discovery is a luxury that a lot of them can't afford right now (Steve Burrell).

Source: Nature Biotechnology, Vol 20, October 2002, pp 965

Approval for human activatedprotein C
Human activated protein C has been approved by the European Commission on August 26 as anti-sepsis drug. Activated protein C is produced by Eli Lilly in bioreactors with human kidney cells. The kidney cells glycosylate the protein not 100% correctly, but this does not induce any immune reactions. To save one life, 16 treatments of 7,000 USD each are needed. It is expected that one in 5 sepsis patients might benefit from the treatment. Source: Bionieuws, September 7, 2002.

New chairman EuropaBio
Drs. Feike Sijbesma has been elected new chairman of EuropaBio. Mr. Sijbesma is member of the Board of Directors of DSM Life Science Products. The preceding two years he was vice-chairman of EurpaBio.

 

Research News


Stem cells
To keep human embryonic stem (HES) cells in an undifferentiated state, they need to be grown on a layer of mouse embryonic fibroblasts (MEF), or in MEF-conditioned medium. This introduces risks of contamination by infectious agents, precluding their use in patents. Recently, a group of researchers in Singapore have cultured HES cells in an animal-free system that uses a feeder layer of human fetal or adult cells and medium untainted by animal protein. The cells grown on human feeders are undistinguishable from those grown on MEF feeders.

Source: Nature Biotechnology, Vol 20, Sept 2002; pp 933-936.

Prophylactic for prions
CpGs stimulate innate immunity in the host and could therefore slow the spread of an infection. A German group of researchers have recently shown that CpG oligodeoxynucleotides might prolong prion incubation time and extend life. The researchers injected mice with a mouse variant of scrapie and within 7 hours treated the mice with saline or CpG daily for 4 or 20 days. All control mouse died of scrapie within 183 days; those given CpG daily for 4 days survived 38% longer. All mice treated for 20 days were still alive after 330 days.

Source: Lancet 360, 229-230, 2002.

 

Light-switchable gene promoter system
Regulatable transgene expression systems providing easily controlled, conditional induction or repression of expression are indispensable tools in biomedical research and biotechnology. Several such systems have been developed for eukaryotes, most of these relying on administration of either exogenous chemicals or heat shock. American researchers recently reported that they have developed a promoter system that can be induced, rapidly and reversibly, by short pulses of light. This system is based on the known red-light induced binding of the plant photoreceptor phytochrome to the protein PIF3 and the reversal of this binding by far-red light. The extent of induction can be controlled precisely by titration of the number of photons delivered to the cells by the light pulse. Thus, this system has the potential to provide rapid, non-invasive, switchable control of expression of a desired gene to a preselected level in any suitable cell by simple exposure to a light signal.

Source: Nature Biotechnology, Vol 20, October 2002, pp 1041-1044

Cloned zebrafish
Cloning technology is becoming a potent force in the laboratory. Model laboratory animals that have been cloned by nuclear transfer now include the mouse and two small laboratory fish, medaka and zebrafish. For the latter, a US group chose an approach using long-term cultured donor cells and nuclear transfer. A different strategy was used by a Dutch group, yielding a mutant zebrafish using targeting-induced local lesions in genome (TILLING) technology. This technology has been used in other organisms, such as the plant Arabidopsis thaliana. The major use of the transgenic model animals will be to test gene function by gene disruption.

Source: various, see also Nature Biotechnology Volume 20, August 2002.

 

On the web


Chemical abstracts
Chemical Abstracts Service (CAS) is the premier information source for (bio)chemists. With the Chemical Abstracts (CAplus) database CAS discloses 8.000 scientific, technical journals and patent publications from 32 patent issuing organizations in the area of chemistry and life science (http://www.cas.org/chemplus/chemplus1.html). With the CAS Registry file CAS offers access to more than 39 million compounds with their CAS Registry Number ( http://www.cas.org/casdb.html#regdb ).

STN International offers 3 interfaces to access CAplus and Registry together with 205 other databases in science and technology ( http://www.stn-international.de ). Via ChemPort the bibliographies are linked to the full text (journal list: http://141.66.18.180/html/english/journals_list.html) or you can order them via FIZ AutoDoc ( http://www.fiz-karlsruhe.de/doc_del/autfirst.html)

 

Major medical/pharmaceutical website

Pharma-lexicon.com (http://www.pharma-lexicon.com) is one of the world's major medical/pharmaceutical websites.
The world's largest online database of medical, nursing,Ýpharmaceutical, biotech and agrochemical abbreviations - over 57,000. Free to use. Directory of pharmaceutical companies, pharmacy faculties, government health departments, healthcare job agencies.


Publications/reports

First ever Ernst & Young global report 2002

Recently, Ernst & Young issued its first global biotechnology report 2002, ëBeyond Bordersí. This report combines the successful US and European reports and also reviews Canada, Asia/Pacific countries and some of the worldís developing markets.
The new report is broken into two major sections:
Section 1, the Global Marketplace and Section 2, Regional markets. In the global marketplace, 25 countries, together accounting for 79% of the global gross domestic product, are surveyed. In Section 2, the reports are broken down into main world markets, and resemble more the old US and European reports. Available from Ernst & Young, also visit: www.ey.com/beyondborders

 

The bio-entrepreneur

In July 2002, Nature Biotechnology issued a supplement called 'Bio-entrepreneur &endash; nurturing the business of biotechnology'. The special covers:

bioentrepreneurship (with 'the rise of the European bio-entrepreneur')
business incubation and start-up
strategic planning
legalities and finance
management and communications (including dealing with the media)

 

Swedish Biopharma

The Swedish biopharma industry has more than doubled in the last sixyears, growing from 25 companies in 1996 to the current 52. Next to the medical device industry the biopharma industry is the largest biotech industry segment in Sweden. The predominant disease groups for R&D in
Swedish biopharma are: Inflammation / autoimmune, metabolic, cancer, infectious, and neurological diseases. However, the pipeline of the industry is in its infancy.
Swedish Biopharma Industry - The Next Wave (released Sept, 2002)
http://www.swedishpharma.info

Other interesting reports are:

Ion Channel Assays in the Drug Discovery Process (released June, 2002)
http://www.ion-channels-in-drug-discovery.info

The Competitive Landscape of Cancer Gene Therapy (released May, 2002)
http://www.competitive-landscape-gene-therapy.info

Commercializing on Tumor Antigens: Analytical Perspectives (released Feb, 2002)
http://www.commercializing-tumor-antigens.info

 

AGENDA


A number of interesting conferences and workshops is coming up. Of all the events mentioned here, the detailed programmes and registration/application forms are available from the ACTIP secretariat.

Plenary ACTIP meeting
December 12-13, Gent, Belgium
Organisation: ACTIP Secretariat: mailto:ACTIP@ACTIP.

Standardisation and Quality Control Cell and Genetherapy products
24-25 February 2003, Strasbourg (France)
Organization: EDQM, email: publicrelations@pheur.org

Marine biotechnology: basics and applications
February 25-March 1, 2003, Matalascanas, Spain
Organisation: marine.biot@algemeen.pk.wau.nl

Conference on Plant-Made Pharmaceuticals
16-19 March, 2003, Quebec, Canada
Organization: http://www.cpmp2003.org

BioTechHelsinki 03
March 24-26, 2003, Helsinki, Finland
Organisation: Finish Bioindustries/Sitra/Tekes.; Tel: + 358 917 28 43 14. ; Web: http://www.biotechhelsinki.com

BioVision 2003
April 8-11, 2003, Lyon France
Organisation: www.biovision.org

ESACT meeting: animal cell technology meets genomics
May 11-15, 2003, Granada
Organisation: ESACT, http://www.esact.org or + 34 932 388 777

Genetics. Genomes, the linkage to life
July 6-12, 2003, Melbourne Australia
Organization: http://www.geneticscongress2003.com

11th EuropeanCongress on Biotechnology, 25th anniversary of EFB:
Building bridges between biosciences and bioengineering

24-29 August, 2003, Basel, Switzerland
Organisation: ECB11, Tel + 41 61 686 28 28; fax: +41 61 686 21 85; email: HYPERLINK mailto:info@ecb11.ch ; web: http://www.ecb11.ch

6th Conference on Protein Expression in Animal Cells
7-11 September, 2003 , Quebec, Canada.
Organization: www.bri.nrc.ca/6thPEACe

 

ACTIP bulletin no. 32, December 2002

 

 


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