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From: TSS ()
Subject: Animal Compound–Free Medium and Poloxamer for Human Corneal Organ Culture and Deswelling
Date: February 23, 2005 at 1:55 pm PST

-------- Original Message --------
Subject: Animal Compound–Free Medium and Poloxamer for Human Corneal Organ Culture and Deswelling
Date: Wed, 23 Feb 2005 10:25:01 -0600
From: "Terry S. Singeltary Sr."
Reply-To: Bovine Spongiform Encephalopathy

##################### Bovine Spongiform Encephalopathy #####################

Animal Compound–Free Medium and Poloxamer for Human Corneal Organ
Culture and Deswelling

Gilles Thuret,1,2 Chloe Manissolle,2 Lydia Campos-Guyotat,2 Denis
Guyotat,2 and Philippe Gain1,2

1From the Ophthalmology Department, University Hospital of
Saint-Etienne, France; and the 2Cell Survival and Adherence in Cancers
and Grafts, Laboratory EA3063, Faculty of Medicine, University Jean
Monnet, Saint-Etienne, France.

PURPOSE. Eliminating fetal calf serum (FCS) from corneal organ culture
(OC) media has long been a challenge. This study was an assessment of a
new animal compound–free (ACF) medium for corneal storage and of its
combination with poloxamer for end-of-storage corneal deswelling.

METHODS. A randomized controlled study with masked assessment compared
the ACF medium to standard commercialized media containing 2% FCS and
their combination with dextran for deswelling. Paired human corneas were
randomly allocated at procurement, one to the ACF medium and the other
to the FCS media, and then assessed at day (D)2 and D30 of OC storage
and after 48 hours of deswelling. Comparison criteria were endothelial
cell density (ECD) and morphometry by a corneal analyser, quality of
endothelial visualization (using saline), EC mortality (trypan blue),
corneal thickness, corneal transparency, and folding. Fifty-six corneas
(28 pairs) with ECD of 2000 cells/mm2 or more were enrolled. Data were
compared using paired tests with P < 0.01 deemed significant.

RESULTS. Parameters were similar at baseline (D2) between groups. Daily
EC loss during the 30 days of storage was reduced with the ACF compared
with standard (–0.31% ± 0.30% vs. –0.88% ± 0.38%, P < 0.001). With
poloxamer 188 (Lutrol F68; BASF, Ludwigshafen, Germany), EC loss was
substantially reduced (–1.43% ± 3.60 vs. –15.41% ± 10.13%, P < 0.001)
and morphometry better preserved, despite thickness reduction,
transparency improvement and folding reduction comparable to dextran.
After 30 days of storage in ACF medium and deswelling in poloxamer 188,
ECD was 30% higher (2466 ± 447 cells/mm2 vs. 1729 ± 281 cells/mm2, P <
0.001). ACF medium alone and combined with poloxamer 188 considerably
facilitated EC visualization at D30 and after deswelling.

CONCLUSIONS. The ACF medium combined with poloxamer 188 for deswelling
showed superiority over standard FCS medium in its ability to preserve
EC viability and facilitate endothelial visualization. This innovative
use of poloxamer for deswelling appears far less toxic than does dextran.


Greetings list members,

nothing like injecting some TSE directly through the pathway to the soul,
via the eyes, also a direct route to the brain ;


The public summary of SEAC's meeting on 17 December 2001 has highlighted
an investigation into variant Creutzfeldt Jakob disease (vCJD) cases in
Southampton as part of the routine exploration of geographical patterns
of vCJD. This showed that two cases of vCJD had received the same batch
of oral polio vaccine in 1994. The batch associated with the two cases
was numbered APTF181/01 and was in use in the fourth quarter of 1994.

If any parents express concern they can be strongly reassured that:

o This particular oral polio vaccine was withdrawn from use in
October 2000.
o The Committee on Safety of Medicines concluded it was very
unlikely that there would be any BSE risks from the use of
the UK-sourced fetal calf serum that was involved in this
vaccine manufacture.
o SEAC stressed that any historical theoretical risk must be
balanced against the overwhelming benefits that would have
resulted from the vaccination programme.

Detailed question and answer material is attached below.

If you require further information you may contact Dr David Salisbury,

tel: 0207 972 1522. Department of Health, 607A Skipton House, 80 London
Road, London SE1 6LH

full text ;

29. The Committee concluded that the observation that two vCJD cases had
received OPV from the same batch did not provide sufficient reason for the
Committee on the Safety of Medicines (CSM) to re-consider its recent
assessment. The CSM considered that it was very unlikely there would be
any BSE risks from the use of UK-sourced fetal calf serum (which had been
used in the manufacture of OPV). The Committee suggested it might be of
value to determine if the MRC-5 cell line used to generate the vaccine could
replicate TSE agents. The Committee stressed the benefits that accrued
from vaccination and the important role they had in controlling the common
infectious diseases of childhood.

If vaccines are safe why did the UK recall their polio vaccine?

The UK recalled the Evans/Medeva Oral Polio Vaccine in October, 2000.
This vaccine has never been licensed for use in the US. The Medicines
Control Agency (MCA) had requested and received assurances from drug
companies that they were implementing guidance not to use UK-sourced
bovine materials in the manufacture of injectable medicinal products.
The recall was prompted by evidence that the Evans/Medeva vaccine was
manufactured using fetal calf serum from the UK at a time when there was
a risk of BSE in that country. This is in contravention of European
Union guidelines. According to a statement from the Chief Medical
Officer at the UK Dept. of Health (10.20.00) the company had assured the
MCA of the UK that UK-sourced bovine materials were not used in the
manufacture of the vaccine. However, these assurances were inaccurate,
thus the vaccine was withdrawn. (

d) Assumed that the Canadian feed ban has had and will have the same
effect of
reducing the incidence of BSE in Canada as the European feed ban has had
in Europe,
despite the fact that European countries also prohibit the use of
ruminant blood products
and rendered animal fat in cattle feed, which Canada does not. This
distinction is a
critical one, given that blood has now been scientifically identified as
a vector for
transmitting prions such as those that can cause BSE, and given that
acknowledged that: ?Based on scientific evidence currently available, it
is not possible to
dismiss the possibility that ingestion of tallow infected with BSE
creates a risk of the
transmission of BSE.? 70 Fed. Reg. at 501.,%20R-CALF%20USA%20Complaint%20against%20Final%20Rule.pdf

however, bovine ingredients can be used as formulation ingredients in
any of the 12 cosmetic product categories, and a small subset of animal
ingredients may be derived from high risk tissues.

3002.10.0040: FETAL BOVINE SERUM (FBS)
U.S. Imports for Consumption: December 1998 and 1998 Year-to-Date
(Customs Value, in Thousands of Dollars) (Units of Quantity: Kilograms)

<--- Dec 1998 --- <--- 1998 YTD ---
Country Quantity Value Quantity Value
WORLD TOTAL . . . . . . . 2,727 233 131,486 8,502
Belgium . . . . . . . . . --- --- 17 32
United Kingdom . . . . . 329 82 743 756


Subject: MBM/U.K. imports of MBM to the U.S./BSE Inquiry
Date:Mon, 10 Apr 2000 15:14:21 -0700
From: "Terry S. Singeltary Sr."

69. On 14 February 1990, Mr Meldrum wrote a letter to the
Chief Veterinary Officers of a number of countries. [76] On 15
February 1990, Mrs Attridge and other officials were sent a
copy of the letter of 14 February 1990 and a list of the
countries to which it had been sent. They were stated to be
the countries which had imported ruminant based meat
and bone meal from the United Kingdom. The countries listed
were Norway; Sweden, Switzerland, Czechoslovakia,
Hungary, Nigeria, Thailand, South Africa, Malaysia, Taiwan,
Hong Kong, South Korea, Japan, Canada, USA,
Turkey, Kenya, Malta, Libera, Lebanon, Saudi Arabia, Sri
Lanka, Puerto Rico, Curacao, Finland.[77] The letter from
Mr Meldrum included the following:
Although we have kept the Office Internationale des Epizooties (OIE)
fully informed about this new disease, and they will
shortly be disseminating information and recommendations to member
countries, I am writing to you on a personal basis to
ensure that you are aware of all the developments in relation to BSE,
including its likely cause. The majority of our findings
have now been published in the Veterinary Record.í[78]
70. On 20 February 1990, Dr Pickles wrote to Ms Verity
(APS/CMO). Dr Picklesí minute included the following:
1. Mr Meldrum is arguing that MAFF have already taken all the
necessary and responsible steps to warn importing countries
of the BSE dangers in UK meat and bone meal. Yet the action taken
so far overseas suggest the message has not got
through, or where it has this has been late. The first nation
that woke up to the danger did so a year after our own feed
ban. It seems even now several EC countries neither ban our
imports or the general feeding of ruminant protein. It also
seems the OIE and CVO have yet to inform the rest of the world.
2. I do not see how this can be claimed to be responsibleí. We
do not need an expert group of the Scientific Veterinary
Committee to tell us British meat and bone meal is unsafe for
ruminants. I fail to understand why this cannot be tackled
from the British end which seems to be the only sure way of doing
it, preferably by banning exports. As CMO says in his
letter of 3 January surely it is short sighted for us to risk
being seen in future as having been responsible for the
introduction of BSE to the food chain in other countries.íí[79]

>As CMO says in his
> letter of 3 January surely it is short sighted for us to risk
>being seen in future as having been responsible for the
> introduction of BSE to the food chain in other countries.íí[79]

around the globe, will make for an interesting recipe.

WITH the recent findings of transmission of TSE via blood,
THE findings of new strains of TSE in cattle and the lack of knowledge
of tissue infectivity distribution (BASE which is very similar to
sporadic CJD
as opposed to the nvCJD), the findings in Japan of Tissue infectivity of BSE
of the peripheral nerve tissue, suprarenal gland first time from
non-Specified Risk Material or SRM, plus the new test from Prusiner
et al that are some 1,000 to 100,000 times more sensitive, with all this,
i would never say never with any assumption about human animal TSE and
any potential risk. Too many times to date they have all been proven


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