Help needed!!

[Guest guest]

What are the ages of her kids and sizes? Sex? My nanny has some

clothes for a girl that is 4 and my kids are girl size 10-12 and boy size

12-14, boy size 18mo-2t.

From:

[mailto: ] On Behalf Of Nina

Sent: Tuesday, July 10, 2007 2:10 PM

Subject: HELP NEEDED!!

I am very sad to tell you that one of our

patients is in great need.

She went home after surgery to find out her house had burned down. She

has 5 children ages 4 thru 14. She lives in Georgia.

She needs clothes for herself, she is wearing size 22/24 right now but

is losing weight very rapidly since her bypass surgery 2 weeks ago. She

lost everything needed to recovery as well: vitmains, protein powders,

everything.

Any help is greatly appreciate it. We are all very sad and want to

reach out in any way we can. I have asked her for a mailing address so

we can send her things for her and her kids. If anyone can help in any

way let me know and I can give you her address.

Nina

[Guest guest]

nina

Please send me the address, I have lots of clothes and will send other things.

Amy in NY

HELP NEEDED!!

I am very sad to tell you that one of our patients is in great need. She went home after surgery to find out her house had burned down. She has 5 children ages 4 thru 14. She lives in Georgia.She needs clothes for herself, she is wearing size 22/24 right now but is losing weight very rapidly since her bypass surgery 2 weeks ago. She lost everything needed to recovery as well: vitmains, protein powders, everything. Any help is greatly appreciate it. We are all very sad and want to reach out in any way we can. I have asked her for a mailing address so we can send her things for her and her kids. If anyone can help in any way let me know and I can give you her address. Nina

[Guest guest]

nina i live in north carolina not far from georgia what can i do do help i hae alot of 20-24 size clothes and anything else for the home i can look for

molly marshall

HELP NEEDED!!

I am very sad to tell you that one of our patients is in great need. She went home after surgery to find out her house had burned down. She has 5 children ages 4 thru 14. She lives in Georgia.She needs clothes for herself, she is wearing size 22/24 right now but is losing weight very rapidly since her bypass surgery 2 weeks ago. She lost everything needed to recovery as well: vitmains, protein powders, everything. Any help is greatly appreciate it. We are all very sad and want to reach out in any way we can. I have asked her for a mailing address so we can send her things for her and her kids. If anyone can help in any way let me know and I can give you her address. Nina

[Guest guest]

Nina,

Please let us know where we can send things to the woman you are referring to, and her name. Also, outside of clothes and after surgery recovery products is there anything special she and her family may want or need.

Angel

HELP NEEDED!!

I am very sad to tell you that one of our patients is in great need. She went home after surgery to find out her house had burned down. She has 5 children ages 4 thru 14. She lives in Georgia.She needs clothes for herself, she is wearing size 22/24 right now but is losing weight very rapidly since her bypass surgery 2 weeks ago. She lost everything needed to recovery as well: vitmains, protein powders, everything. Any help is greatly appreciate it. We are all very sad and want to reach out in any way we can. I have asked her for a mailing address so we can send her things for her and her kids. If anyone can help in any way let me know and I can give you her address. Nina

[Guest guest]

I asked the same thing, here is the list of ages and sex, her name is Dawn, I just sent her the link to the group.

Nina

- age 4 1/2 - boy - size 6-7 with size 1 shoes

Isaac - age 10 - boy - size 14/16 with size 4 shoes

PJ - age 12 - boy - size 32waist 30 length XL shirt

- age 13 - girl - size 0 womens/juniors size XS shirt with size 7 shoes

Kati - age 14 - girl - she is a size 14 womens XL shirt size 10 shoes

Dawn (myself) size 20,22 or 24 with size 10 shoes

Maxx (my hubby) size 40 waist 34 inseam XXLT shirt

We are in need or sheets, bedding, blankets, towels, any household items.....

And honestly I simply need friends!!

>> What are the ages of her kids and sizes? Sex? My nanny has some clothes for> a girl that is 4 and my kids are girl size 10-12 and boy size 12-14, boy> size 18mo-2t.> > > > From: > [mailto: ] On Behalf Of Nina> Sent: Tuesday, July 10, 2007 2:10 PM> > Subject: HELP NEEDED!!> > > > I am very sad to tell you that one of our patients is in great need. > She went home after surgery to find out her house had burned down. She > has 5 children ages 4 thru 14. She lives in Georgia.> > She needs clothes for herself, she is wearing size 22/24 right now but > is losing weight very rapidly since her bypass surgery 2 weeks ago. She > lost everything needed to recovery as well: vitmains, protein powders, > everything. > > Any help is greatly appreciate it. We are all very sad and want to > reach out in any way we can. I have asked her for a mailing address so > we can send her things for her and her kids. If anyone can help in any > way let me know and I can give you her address. > > Nina>

[Guest guest]

Great!

I just sent her the link to the group, her name is Dawn. I cannot

give you more details but am waiting for the mailing address. Her

email is jellybean2141973@... in case you want to email her

personally, she needs support, emotional too.

Nina

>

> nina i live in north carolina not far from georgia what can i do

do help i hae alot of 20-24 size clothes and anything else for the

home i can look for

> molly marshall

> HELP NEEDED!!

>

>

> I am very sad to tell you that one of our patients is in great

need.

> She went home after surgery to find out her house had burned

down. She

> has 5 children ages 4 thru 14. She lives in Georgia.

>

> She needs clothes for herself, she is wearing size 22/24 right

now but

> is losing weight very rapidly since her bypass surgery 2 weeks

ago. She

> lost everything needed to recovery as well: vitmains, protein

powders,

> everything.

>

> Any help is greatly appreciate it. We are all very sad and want

to

> reach out in any way we can. I have asked her for a mailing

address so

> we can send her things for her and her kids. If anyone can help

in any

> way let me know and I can give you her address.

>

> Nina

>

[Guest guest]

I have denim scrubs of various sizes. They are great because they have a drawstring and last for a few sizes. ;o) I have lots of protein shakes and assorted stuff as well. I also have some dress clothes that are 1X & 2X, would that work? I have a bunch of new nail polishes and such that I haven't even opened, she can have them.

252/172/150

Banded 12/06

On 7/10/07, Nina <nina_eguia@...> wrote:

I am very sad to tell you that one of our patients is in great need. She went home after surgery to find out her house had burned down. She has 5 children ages 4 thru 14. She lives in Georgia.She needs clothes for herself, she is wearing size 22/24 right now but is losing weight very rapidly since her bypass surgery 2 weeks ago. She lost everything needed to recovery as well: vitmains, protein powders, everything. Any help is greatly appreciate it. We are all very sad and want to reach out in any way we can. I have asked her for a mailing address so we can send her things for her and her kids. If anyone can help in any way let me know and I can give you her address. Nina

[Guest guest]

I just spoke to Dawn and I have a better idea of what she needs.

She needs protein products but is allergic to NutraSweet. I'm sending her some Matrix since it no longer contains NutraSweet. I am sending denim scrubs because they have a drawstring and will work for a few sizes.

She is currently a 3X but is losing very fast. By the time you send her a 3X it will be too big. She needs ALL sizes. All their clothing was lost in the fire and as you know, bypass patients lose quickly. She needs casual clothes and dressy outfits. She wears a size 10 shoe, if you have any extras.

I'm sending her a bottle of chewable vitamins. But if you have any others that would be great, not everyone likes my vitamins. ;o)

She could use lots of emails as well. She was serious, she needs some friends right now. She just had life changing surgery and now a house fire. Things are rough right now.

252/172/150

Banded 12/06

On 7/10/07, Nina <nina_eguia@...> wrote:

I am very sad to tell you that one of our patients is in great need. She went home after surgery to find out her house had burned down. She has 5 children ages 4 thru 14. She lives in Georgia.She needs clothes for herself, she is wearing size 22/24 right now but is losing weight very rapidly since her bypass surgery 2 weeks ago. She lost everything needed to recovery as well: vitmains, protein powders, everything. Any help is greatly appreciate it. We are all very sad and want to reach out in any way we can. I have asked her for a mailing address so we can send her things for her and her kids. If anyone can help in any way let me know and I can give you her address. Nina

[Guest guest]

Where in Georgia is Dawn and her family? I will be traveling to Georgia next week so if it is close to where I am going, I would gladly take her some things. What a sad thing for her and her family. Jenni CurrieNina <nina_eguia@...> wrote: I am very sad to tell you that one of our patients is in great need. She went home after surgery to find out her house had burned down. She has 5 children ages 4 thru 14. She lives in Georgia.She needs clothes for

herself, she is wearing size 22/24 right now but is losing weight very rapidly since her bypass surgery 2 weeks ago. She lost everything needed to recovery as well: vitmains, protein powders, everything. Any help is greatly appreciate it. We are all very sad and want to reach out in any way we can. I have asked her for a mailing address so we can send her things for her and her kids. If anyone can help in any way let me know and I can give you her address. Nina

Be a better Globetrotter. Get better travel answers from someone who knows. Answers - Check it out.

[Guest guest]

I can just kick myself I got rid of alot of clothes to a church sale

for a friend. However, if you can send me the address I'd like to

send a gift card from Walmart so they can pick up whatever is not

given to them.

THanks

Kelli

> >

> > What are the ages of her kids and sizes? Sex? My nanny has some

> clothes for

> > a girl that is 4 and my kids are girl size 10-12 and boy size 12-

14,

> boy

> > size 18mo-2t.

> >

> >

> >

> > From:

> > [mailto: ] On Behalf Of Nina

> > Sent: Tuesday, July 10, 2007 2:10 PM

> >

> > Subject: HELP NEEDED!!

> >

> >

> >

> > I am very sad to tell you that one of our patients is in great

need.

> > She went home after surgery to find out her house had burned

down. She

> > has 5 children ages 4 thru 14. She lives in Georgia.

> >

> > She needs clothes for herself, she is wearing size 22/24 right

now but

> > is losing weight very rapidly since her bypass surgery 2 weeks

ago.

> She

> > lost everything needed to recovery as well: vitmains, protein

powders,

> > everything.

> >

> > Any help is greatly appreciate it. We are all very sad and want to

> > reach out in any way we can. I have asked her for a mailing

address so

> > we can send her things for her and her kids. If anyone can help

in any

> > way let me know and I can give you her address.

> >

> > Nina

> >

>

[Guest guest]

I am getting ready to have a killer yard sale but I would rather get the things to Dawn if at all possible. I have tons of kitchen stuff including dishes, glasses, toaster oven, etc. I don't know about clothing. I have several household items they can have....I wish I could figure out how to get some furniture to them. Anyway, if someone can just get me some contact information. Jenni CurrieBipley <Bipley@...> wrote: I have denim scrubs

of various sizes. They are great because they have a drawstring and last for a few sizes. ;o) I have lots of protein shakes and assorted stuff as well. I also have some dress clothes that are 1X & 2X, would that work? I have a bunch of new nail polishes and such that I haven't even opened, she can have them. 252/172/150 Banded 12/06 On 7/10/07, Nina <nina_eguia > wrote: I am very sad to tell you that one of our patients is in great need. She went home after surgery to find out her house had burned down. She has 5 children ages 4 thru

14. She lives in Georgia.She needs clothes for herself, she is wearing size 22/24 right now but is losing weight very rapidly since her bypass surgery 2 weeks ago. She lost everything needed to recovery as well: vitmains, protein powders, everything. Any help is greatly appreciate it. We are all very sad and want to reach out in any way we can. I have asked her for a mailing address so we can send her things for her and her kids. If anyone can help in any way let me know and I can give you her address. Nina

Get the toolbar and be alerted to new email wherever you're surfing.

[Guest guest]

Great idea, Kelli! I have given away all my stuff too so the gift

card is the way to go! I'd like the address too.

Thanks Nina,

Penny in Colorado

> > >

> > > What are the ages of her kids and sizes? Sex? My nanny has some

> > clothes for

> > > a girl that is 4 and my kids are girl size 10-12 and boy size

12-

> 14,

> > boy

> > > size 18mo-2t.

> > >

> > >

> > >

> > > From:

> > > [mailto: ] On Behalf Of Nina

> > > Sent: Tuesday, July 10, 2007 2:10 PM

> > >

> > > Subject: HELP NEEDED!!

> > >

> > >

> > >

> > > I am very sad to tell you that one of our patients is in great

> need.

> > > She went home after surgery to find out her house had burned

> down. She

> > > has 5 children ages 4 thru 14. She lives in Georgia.

> > >

> > > She needs clothes for herself, she is wearing size 22/24 right

> now but

> > > is losing weight very rapidly since her bypass surgery 2 weeks

> ago.

> > She

> > > lost everything needed to recovery as well: vitmains, protein

> powders,

> > > everything.

> > >

> > > Any help is greatly appreciate it. We are all very sad and want

to

> > > reach out in any way we can. I have asked her for a mailing

> address so

> > > we can send her things for her and her kids. If anyone can help

> in any

> > > way let me know and I can give you her address.

> > >

> > > Nina

> > >

> >

>

[Guest guest]

Nina,I live in NC and may be able to help. Does she have an email address where we could contact her and see exactly what she might need. Also, what part of Georgia? I may be able to drive some stuff down. Please let us know what you know and if there are any specifics that she might need.

ColleenOn 7/11/07, Penny Manville <pmanvill@...> wrote:

Great idea, Kelli! I have given away all my stuff too so the gift

card is the way to go! I'd like the address too.

Thanks Nina,

Penny in Colorado

> > >

> > > What are the ages of her kids and sizes? Sex? My nanny has some

> > clothes for

> > > a girl that is 4 and my kids are girl size 10-12 and boy size

12-

> 14,

> > boy

> > > size 18mo-2t.

> > >

> > >

> > >

> > > From:

> > > [mailto: ] On Behalf Of Nina

> > > Sent: Tuesday, July 10, 2007 2:10 PM

> > >

> > > Subject: HELP NEEDED!!

> > >

> > >

> > >

> > > I am very sad to tell you that one of our patients is in great

> need.

> > > She went home after surgery to find out her house had burned

> down. She

> > > has 5 children ages 4 thru 14. She lives in Georgia.

> > >

> > > She needs clothes for herself, she is wearing size 22/24 right

> now but

> > > is losing weight very rapidly since her bypass surgery 2 weeks

> ago.

> > She

> > > lost everything needed to recovery as well: vitmains, protein

> powders,

> > > everything.

> > >

> > > Any help is greatly appreciate it. We are all very sad and want

to

> > > reach out in any way we can. I have asked her for a mailing

> address so

> > > we can send her things for her and her kids. If anyone can help

> in any

> > > way let me know and I can give you her address.

> > >

> > > Nina

> > >

> >

>

[Guest guest]

hi gaby can u please send us a list of fill Dr.sNina <nina_eguia@...> wrote: I am very sad to tell you that one of our patients is in great need. She went home after surgery to find out her house had burned down. She has 5 children ages 4 thru 14. She lives in Georgia.She needs clothes for herself, she is wearing size 22/24 right now but is losing weight very rapidly since her bypass surgery 2 weeks ago. She lost everything needed to recovery as well: vitmains, protein powders, everything. Any help is greatly appreciate it. We are all very sad and want to reach out in any way we can. I have asked her for a mailing address so we can send her things for her and her kids. If anyone can help in any way let me know and I can give you her address.

Nina

[Guest guest]

Gaby doesn't read the boards. You might just want to go to the data base. Go to the bottom of this email and see the links. Click on Database and you can find it from there.

252/170/150

Banded 12/06

On 7/12/07, Chantel Godbey <chantelgodbey@...> wrote:

hi gaby

can u please send us a list of fill Dr.sNina <nina_eguia@...> wrote:

I am very sad to tell you that one of our patients is in great need. She went home after surgery to find out her house had burned down. She has 5 children ages 4 thru 14. She lives in Georgia.

She needs clothes for herself, she is wearing size 22/24 right now but is losing weight very rapidly since her bypass surgery 2 weeks ago. She lost everything needed to recovery as well: vitmains, protein powders, everything. Any help is greatly appreciate it. We are all very sad and want to reach out in any way we can. I have asked her for a mailing address so we can send her things for her and her kids. If anyone can help in any way let me know and I can give you her address. Nina

[Guest guest]

Hi Chantel

There is a list in the database to the left on the message board.

Like I told you there is a good one in Aurora Co. His name is Dr.

Kiechenbaum and his phone number is in the data base. I don't think

there is anyone on the Western Slope but I know there is a Dr. Green

in Salt Lake City. He has gotten very expensive but is good and does

flouro. I will email you Dr. Kirchenbaum's phone number if I can

find it.

Talk to you soon,

Penny up in Walden

>

> hi gaby

> can u please send us a list of fill Dr.s

>

>>

>

>

>

>

>

> ---------------------------------

> Be a PS3 game guru.

> Get your game face on with the latest PS3 news and previews at

Games.

>

[Guest guest]

Hey Chantel

I did look at the database and found a Dr. Brown in Canon

City, too. His number is 866-333-band. Dr. Kirshenbaum's number is

303-695-1034. There is a Dr. in Greeley too but that would probably

be farther away than the other 2.

Keep me posted on what you find out!

Penny in Walden

> I am very sad to tell you that one of our patients is in great

need.

> She went home after surgery to find out her house had burned down.

She

> has 5 children ages 4 thru 14. She lives in Georgia.

>

> She needs clothes for herself, she is wearing size 22/24 right now

but

> is losing weight very rapidly since her bypass surgery 2 weeks ago.

She

> lost everything needed to recovery as well: vitmains, protein

powders,

> everything.

>

> Any help is greatly appreciate it. We are all very sad and want to

> reach out in any way we can. I have asked her for a mailing address

so

> we can send her things for her and her kids. If anyone can help in

any

> way let me know and I can give you her address.

>

> Nina

>

>

>

>

>

>

>

[Guest guest]

Colleen

WHere in NC do you live IM in flat Rock near Asheville maybe we can work together to help her

MOlly Marshall

HELP NEEDED!!> > >> > >> > >> > > I am very sad to tell you that one of our patients is in great > need.> > > She went home after surgery to find out her house had burned > down. She> > > has 5 children ages 4 thru 14. She lives in Georgia.> > >> > > She needs clothes for herself, she is wearing size 22/24 right > now but> > > is losing weight very rapidly since her bypass surgery 2 weeks > ago.> > She> > > lost everything needed to recovery as well: vitmains, protein > powders,> > > everything.> > >> > > Any help is greatly appreciate it. We are all very sad and want to> > > reach out in any way we can. I have asked her for a mailing > address so> > > we can send her things for her and her kids. If anyone can help > in any> > > way let me know and I can give you her address.> > >> > > Nina> > >> >>

[Guest guest]

Please send me an address to send help. Molly <msmolly6255@...> wrote: Colleen WHere in NC do you live IM in flat Rock near Asheville maybe we can work together to help her MOlly Marshall HELP NEEDED!!> > >> > >> > >> > > I am very sad to tell you that one of our patients is in great > need.> > > She went home after surgery to find out her house had burned > down. She> > > has 5 children ages 4 thru 14. She lives in Georgia.> > >> > > She needs clothes for herself, she is wearing size 22/24 right > now but> > > is losing weight very rapidly since her bypass surgery 2 weeks > ago.> > She> > > lost everything needed to recovery as well: vitmains, protein > powders,> > > everything.> > >> > > Any

help is greatly appreciate it. We are all very sad and want to> > > reach out in any way we can. I have asked her for a mailing > address so> > > we can send her things for her and her kids. If anyone can help > in any> > > way let me know and I can give you her address.> > >> > > Nina> > >> >>

Get the free toolbar and rest assured with the added security of spyware protection.

[Guest guest]

That is such a great idea Kelli! I went this morning and picked up

a Walmart gift card and will send out pronto. Has anyone heard how

Dawn and her family are doing?

Nadine

DOB 3/24/07

362(346)/308/150

1 fill .5cc

> > >

> > > What are the ages of her kids and sizes? Sex? My nanny has some

> > clothes for

> > > a girl that is 4 and my kids are girl size 10-12 and boy size

12-

> 14,

> > boy

> > > size 18mo-2t.

> > >

> > >

> > >

> > > From:

> > > [mailto: ] On Behalf Of Nina

> > > Sent: Tuesday, July 10, 2007 2:10 PM

> > >

> > > Subject: HELP NEEDED!!

> > >

> > >

> > >

> > > I am very sad to tell you that one of our patients is in great

> need.

> > > She went home after surgery to find out her house had burned

> down. She

> > > has 5 children ages 4 thru 14. She lives in Georgia.

> > >

> > > She needs clothes for herself, she is wearing size 22/24 right

> now but

> > > is losing weight very rapidly since her bypass surgery 2 weeks

> ago.

> > She

> > > lost everything needed to recovery as well: vitmains, protein

> powders,

> > > everything.

> > >

> > > Any help is greatly appreciate it. We are all very sad and

want to

> > > reach out in any way we can. I have asked her for a mailing

> address so

> > > we can send her things for her and her kids. If anyone can

help

> in any

> > > way let me know and I can give you her address.

> > >

> > > Nina

> > >

> >

>

[Guest guest]

Dawn ~

Hello Dawn, so glad you found the group ! I am so sorry

you are going thru so much.

Your symptoms are ditto many gals with implants and

implant related issues. There are doctors that have mentioned

that as soon as the implants are out, they can actually see

the heart calm down.......

It is not unusual for the adrenals to be compromised when

your body is fighting foreign bodies, and it just so happens

that the ones your body is fighting are also made up of many

neuro-toxic chemicals, so you are dealing with multiple chemical

sensitivities as well.

All implants are made out of silicone. They are just filled with

different things.....

Here is a partial list of the chemicals used in the manufacturing

of the silicone used in ALL implants, remember all are known

neuro-toxins ! :

1)Methyl Ethyl Ketone

2)Cyclobexanone

3)Isopropyl Alcohol

4)Denatured Alcohol

5)Acetone

6)Urethane

7)Poly vinyl Chloride

8)Lacquer Thinner

9)Ethyl Acetate

10)Epoxy Resin

11)Epoxy Hardener

12)Amine

13)Printing Ink

14)Toluene

15)Freon

16)Silica

17)Flux

18)Solder

]19)Chlorplatinic Acid

20)Metal Cleaning Acid

21)Formaldehyde

22)Talcum Powder

23)Color Pigmentation (Printers Ink)

24)Oakite

25)Cyanoacyrylates

26)Ethylene Oxide

27)Carob Black

28)Xylene

29)Hexone

30)Benzene

31)Hexanone 2

32)Thixon-OSN-2

33)Rubber

34)Acid Stearic

35)Zinc Oxide

36)Naptha

37)Phenol

38)Methylene Chloride

39)Platinum Salt

In the FDA website under immuno-toxicology, it discusses how silicone disrupts the entire endocrin system. It is not however under the implant section. Well, I was gonna post the link, but

it is no longer there, but here is some of it that I had copied into

an email a couple years ago. Part of it is in symbol for some reason...

Below is parts of the FDA article that I had copied into an email a long time ago.:

as you can see at the end, is in symbol or something.....

Endocrine Disruption by Medical Device Materials

Key words: estrogen disruption, uterotrophic assay, heat shock proteins,

bisphenol A

CDRH is concerned with the potential for certain medical device materials to mimic or interfere with endogenous hormone actions. Because the hormone estrogen is a potent molecule having profound effects at remarkably low doses, assays are needed to assess the potential for harm from materials that may induce unwanted effects due to interference with normal estrogen homeostasis. OST scientists are collaborating with researchers at the Department of Biology, Washington University, on projects focused on improving the use of a key biomarker of exposure to estrogenic compounds and determining the characteristics of the estrogenic activity of bisphenol A, a plasticizer found in some medical devices.OST scientists are modifying and enhancing the traditional assay for estrogenic effects. The traditional assay utilizes the ability of estrogen to cause a hypertrophy of the uterus in ovariectomized or immature female mice. The purpose of these modifications is to provide mechanistic information that will reduce the number of uncertainties in assessing risk from exposure to estrogenic materials. OST scientists are developing an assay that promises to do just that: obtain mechanistic information while examining estrogenic responses in the whole animal. This is accomplished by a side-by-side comparison of the traditional uterotrophic assay and a specific cellular event, the induction of stress proteins, that occurs when estrogen or estrogen-mimicking materials bind to its specific cellular receptor.Stress, or heat shock, proteins are synthesized rapidly by most cells in response to various chemical and physical stressors, especially heat. This response is thought to serve a cellular protective function. Other functions have been found, and heat shock proteins are called "chaperones" because they appear to associate with other proteins and hold them in proper conformations. This appears to be their function in relation to steroid hormone receptors: they appear to hold the receptor molecule in an open position, so that the receptor can bind to ligand easily. When the ligand binds, the heat shock protein dissociates; the receptor is free to assume a new shape, one that retains the bound ligand but which also enables the receptor to bind the correct sequence on the genomic DNA, called the hormone responsive element. Thus, OST has tried to identify changes in heat shock proteins as a marker of estrogen receptor binding in tissues from whole animals treated with estrogens.OST scientists have shown that such changes can be detected in a number of heat shock proteins in the uterus in response to estrogen and that the response is specific to uterine and estrogenic compounds. The effect of exposure time on changes in heat shock proteins was assessed in ovariectomized mice treated with estradiol, the primary estrogen form in the body. Increased relative uterus weight increased linearly between the 4-hour , 8-hour, and 12-hour post-exposure time points, and remained for 1 and 2 days. The histopathology of the uterine lining showed a similar pattern. Changes in the pattern of heat shock protein expression were observed between 6 and 12 hours post-treatment. Thus all three endpoints examined appear at approximately the same time after treatment. This is consistent with the idea that the heat shock protein changes are related specifically to the estradiol effects.Other steroid and steroid-like compounds were used in order to examine the specificity of the response. The compounds included an androgen, an androgen-receptor antagonist, a progestin, and a partial estrogen. These compounds induced changes in heat shock protein expression, but did not induce uterine swelling. This result demonstrates that the heat shock protein changes are not the result of the stresses to the cells caused by uterine swelling, and the heat shock response may be more sensitive than the uterine response. Compound ICI 182,780, a potent anti-estrogen, had no effect on uterine weights and did not induce changes in heat shock proteins. However, the highest dose of the ICI compound blocked the uterine and heat shock protein effects of co-administered estradiol. Thus heat shock protein induction: 1) is an estrogenic effect independent of uterine swelling; and 2) is a more sensitive indicator of estrogenic effect than is uterine swelling. The histological changes occur in parallel with the uterine swelling effects.The three endpoints (uterine hypertrophy, histology, and stress protein expression, were used to examine the estrogenic response of the plasticizer bisphenol A. Bisphenol A was determined to be less estrogenic that estradiol (by more than 1000-fold), but caused similar effects. Histological changes in the uterine epithelia were observed at doses 40-fold lower than doses that caused uterine swelling. Although responses were variable, heat shock protein induction was also more sensitive than the uterotrophic response. Immunological Responses to Silicone Breast ImplantsKey words: autoantibodies, silicone breast implants, immunopathologyThe breast implant experiments were accomplished in two parts: first to demonstrate immune responses to silicone gel implants in an animal model; and second, to apply the experimental protocols to samples from women with/without breast implants. The objective of these studies was to determine which autoimmune-like symptoms and other symptoms are associated with implanted materials.OST has developed a rat model to study and understand immune responses to silicone gel and oil used in breast implants. This model included normal rats and a strain of autoimmune rats, that mimicked clinical conditions found in patients with silicone gel breast implants. Mixtures of silicone gel/oil were injected into the mammary area of female rats. Scientists then measured the levels of autoantibodies to collagen and to nuclear proteins developed over 2 years in response to these mixtures. Results from the rat model indicate that specific autoantibodies may be induced by certain biomaterials. The immune system recognizes a biomaterial-connective tissue protein association as altered-self or as foreign. OST scientists have demonstrated that medical grade silicone oil can stimulate serum autoantibodies against collagen and against DNA when this oil is injected into mammary tissues of rats. Autoantibody production against connective tissue proteins is an immune response that is consistent with reports from women with silicone breast implants. The results also demonstrated pathological changes in animals that may result from the autoimmune response and that silicone gel can migrate to distant anatomical sites or localize at the implantation site. This work has been presented at The FDA Science Forum, The American Association of Immunology, and The American Association of Biochemistry and Molecular Biology.The implication from these findings is that leaked oil from a breast implant via leaching or with rupture might provide stimulus for the production of autoantibodies in clinical patients. Therefore, OST scientists evaluated autoantibody levels (in blinded experiments) in serum samples from 150 patients representing four groups: women with silicone implants without connective tissue disease, women with silicone implants with connective tissue disease (CTD), women with connective disease but no implants, and healthy women volunteers. Results from these experiments show in a statistically significant manner that elevated autoantibodies to collagen type I, collagen type II and anti-DNA were detected in serum of patients with CTD, CTD + silicone implants, and silicone implants without CTD. Using two different assays, autoantibodies to connective tissue proteins (e.g., collagen) and to DNA and intranuclear proteins have been detected in women with silicone breast implants. Historically, there is a strong correlation between anti-nuclear antibodies and clinical symptoms of some autoimmune diseases. OST has documented serum immune responses in these patients with the goal of enhancing the ability to predict the likelihood of immunotoxic symptoms occurring in the presence of implants, including breast prostheses. This work has been presented to the Institute of Medicine.OST scientists are continuing to investigate these results by correlating this data to the clinical history of the patients. Researchers plan to determine the clinical significance of the data and to study a larger patient population well defined with regard to implants and autoimmune disease symptoms.Complement ActivationKey words: complement activation, cellulose acetate, dialysis, sepharose, protein-A, perfusion, research, standards.The term complement describes a series of serum proteins involved in mediating immune reactions. Complement activation is a tightly regulated process which, in addition to direct cell cytolysis, can have profound affects on the immune, vascular, and coagulation systems. Though complement activation is an important defense mechanism of the host, particularly against microbial infections, inappropriate activation (such as by implanted or external medical devices which encounter human blood) may result in serious acute or chronic reactions. Examples of devices whose materials might activate complement include perfusion devices, columns for treating blood externally, indwelling artificial vascular grafts, encapsulated drugs or cells, and vascular shunts. At the request of ODE, OST is conducting research to acquire baseline information needed, in particular, for industry standards concerning testing of materials to be used in blood-contacting devices. A microassay was developed for assaying whole-complement depletion by solid materials used in the construction of blood-contacting medical devices. This method has been submitted as a draft Standard Practice to the ASTM Committee F-4 on Medical and Surgical Materials and Devices via Subcommittee F04.16 on Biocompatibility. Cellulose acetate fibers and powders used in the manufacture of dialysis membranes were tested using this standard practice. Although cellulose powder potently activated complement (57% reduction from control levels when exposed to an equal volume of serum for 1 hour at 37oC), there was no difference between molecular weight 50,000 versus 30,000. Cellulose acetate fibers were less potent than the precursor powders (only 19% reduction from control levels), though storage time (from dates of manufacture) did not produce significant differences between the fibers (9/96 versus 2/89.) These results suggest that adverse patient reactions to dialysis by materials could be related to complement activation, which might not be influenced by molecular weight or age of degradation particles.Sepharose is used to immobilize protein-A in perfusion columns for removing antibodies from patient blood. Both raw sepharose and sepharose-conjugated protein-A were tested for whole complement depletion. Both resulted in a 75% or greater reduction in whole complement activity versus controls. No significant difference was seen between raw sepharose and sepharose-conjugated protein-A, indicating that the majority of the complement activation was due to the matrix, rather than to the attached protein-A. The activation of complement by sepharose was by the alternative pathway (as documented by Bb generation), rather than by the classical pathway (documented by lack of C4d generation). Washing the sepharose with citrate buffers (used in preparing columns for patient blood) resulted in a temporary blockage of complement activation that was rapidly lost upon sequential exposure to additional serum aliquots. These results indicate the potential for complement activation to be a hazard and/or confounding systemic modifier in therapeutic use of antibody-depleting columns. Particulate Effects on Immunologic FunctionKey words: particles, cytokines, wear and degradation, macrophages, standards, researchWear and corrosion of implanted medical devices, such as dental and orthopedic prostheses, may produce particulate debris which may lead to acute and chronic inflammatory responses in the host. In addition, polymeric particles, such as polytetrafluoroethylene (PTFE), may be injected directly into the patients for clinical indications. When particulates are present, the host monocytes/macrophages are activated and they synthesize or secrete mediators of inflammation, and phagocytize particles. In order to understand the mechanisms underlying the host immune response to particulates and device-associated infections, OST scientists have focused their studies on the impact of these particulates on macrophage function. Macrophages play a pivotal role in the body’s response to foreign bodies and they also interact with other cellular components in the immune system. OST developed an in vitro assay using established murine macrophages. This assay system was incorporated into an ASTM standard on the Biological Responses to Particles (F04.16.01). The inflammatory potential of particles prepared from medical device materials, such as PTFE, titanium oxide, hydroxyapatite (HA), polymethylmethacrylate (PMMA), SiO2 and fumed silica, polystyrene (PS), CdCl2, CdO, Al2O3 , and diamond particle was studied.In the in vitro assay, murine macrophage cells were exposed to particles or chemicals with and without bacterial lipopolysaccharide (LPS), which is a component of bacterial cell walls that mimics bacterial infections. The cells were then evaluated for cytotoxicity, production of nitric oxide (NO), tumor necrosis factor-alpha (TNF-a ) and interleukin-6 (IL-6), both inflammatory cytokines. NO is induced by LPS and is critically important in eradicating microorganisms associated with infections, but it can also be harmful by causing tissue injury and vascular collapse. OST studies showed that minute amounts of LPS, which could be associated with a bacterial infection at the site of an implanted device, induced a significant amount of TNF-a , IL-6 and NO production by macrophages.The in vitro studies showed that TNF-a was induced by PTFE, PMMA, TiO2, HA, SiO2, both forms of cadmium, and Al2O3 particles and by LPS. Polystyrene alone did not stimulate activity. PS in combination with LPS stimulated no activity significantly above the levels in response to LPS alone. Addition of particles to the macrophages did not stimulate NO production. However, addition of LPS to the particles affected the NO production. NO production increased in a dose-response manner with LPS plus PMMA, increased but not in a dose-dependent response with HA, and was inhibited by increasing concentrations of TiO2. Polystyrene particles in combination with LPS produced minimal and variable differences of NO compared to LPS alone.The in vitro system showed that IL-6 is stimulated by LPS. Neither Al2O3 nor PS alone stimulated IL-6 production. IL-6 production was stimulated by Al2O3 in combination with LPS, but PS in combination with LPS did not stimulate above the LPS alone level..Thus far, the OST studies indicate the following: 1) this in vitro system measuring TNF-a , IL-6 and NO responses can detect differences in biological responses to particles; 2) no particle tested by OST thus far has induced NO whereas LPS is a potent stimulator; 3) some particles stimulate TNF-a production and some do not; 4) some particles enhance, some inhibit, and some have no effect on the stimulation of TNF-a and NO by macrophages in response to LPS; and 5) PS particles may serve as a negative control for evaluating the induction of these three biological responses. Decontaminating Particles Exposed to Bacterial LipopolysacchardeKey words: particles, lipopolysaccharide, nitric oxide, macrophage, researchThe presence of lipopolysaccharide (LPS) or endotoxin associated with implanted medical devices can stimulate fever in the host. Manufacturers must submit evidence that their sterilized implanted devices are below a given endotoxin level. The most commonly used assay to test for the presence of endotoxin is the Limulus Amebocyte Lysate (LAL) test. OST scientists have observed, using an in vitro murine macrophage assay, that LPS stimulates nitric oxide (NO) production. Clean, sterilized medical device particles examined thus far do not stimulate NO production, but certain particles that are contaminated with minute amounts of lipopolysaccharide (LPS, endotoxin) do stimulate NO production. Polymethylmethacrylate (PMMA) particles deliberately contaminated with minute amounts of LPS were assayed for the production of NO in the murine macrophage cell system. Treating particles deliberately contaminated with LPS with 70% ethanol at room temperature or higher for more than 24 hours followed by washing three times with endotoxin-free phosphate buffered saline reduced the level of detection of LPS in the in vitro cell system. LPS treated with 70% ethanol also did not stimulate NO production. Both the LAL test and the lack of LPS stimulation of NO production by murine macrophages show that 70% ethanol can inactivate LPS and may be a method to remove LPS from particles that are being tested for inflammatory potential.

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