RE: ANT--- PS

June 21, 2005

Oh, one further question. Do you know if the ANT testing was done on fresh

or frozen tissue? I know Cleveland does some testing on fresh and some

testing on frozen from each biopsy. I just wondered which was used for ANT.

Barbara

_____

From: [mailto: ] On Behalf

Of Joanne Kocourek

Sent: Tuesday, June 21, 2005 9:52 AM

To:

Subject: RE: ANT

Dr. Hoppel (Cleveland CIDDM) did the ANT diagnosis. He was so surprised

that he actually has rechecked the study at least three times. It was been

a positive study each time. Dr. Wallace is the ANT expert in the US. So

given the girls also have identified defects in Complex I, III and IV, I

decided to discuss this atypical mix with him at the conference. (He was

very approachable). He said that, from his perspective, this mix is unusual

and is likely the result of severely abnormal dysfunction both within the

mitochondria and across the inner mitochondrial membrane. He is positive

that in my girls their base defect lies within mtDNA. Dr. Shoffner is doing

an evaluation of their entire mitochondrial genone and, to date, has not

identified any " known " mutations or deletions, etc. One curious

comment/questions he posed was, " Your daughter's are significantly

affected, aren't they? " How did he know??? Funny, they look " fine " but

medically are quite compromised with major organ

system involvement.

Barbara Seaman wheatchild@...> wrote:Joanne, Do you know if

there is any genetic connection between the two

defects your daughter has? In other words, is it possible that the ANT

defect is secondarily causing the deficiencies found in the respiratory

chain or are there two separate mutations, one for each defect? Or has this

even been defined? I ask because I also have two mitochondrial defects,

though different from your daughter's. You are the only other person I can

remember talking about two defects in one patient, so I am interested and

curious. Do you mind sharing who did the ANT defect diagnosis?

Thanks.

Barbara

_____

From: [mailto: ] On Behalf

Of Joanne Kocourek

Sent: Tuesday, June 21, 2005 9:11 AM

To:

Subject: Re: ANT

Adenine nucleotide translocators

General characteristics

Most abundant mitochondrial proteins

Integral components of inner mitochondrial membrane

Facilitate exchange of ADP and ATP between cytosol & mitochondria

ohgminion rakshasis@...> wrote:

Sorry, what is ANT?

Thanks,

RH

Joanne Kocourek (mom to , lies, and )

visit us at: http://www.caringbridge.org/il/annakris

June 21, 2005

I believe that the ANT studies are performed after the tissue has been frozen

(given Dr. Hoppel repeated the studies over a year later, some of the study was

definitely done using frozen tissue). The RTC studies are completed when the

sample is fresh.

As for ANT genetics....

increased expression of ANT-1 and ATP synthase: subunit mRNAs have been

described in patients with respiratory defects resulting from mtDNA point

mutations and increased mRNAs encoding cytochrome oxidase (COX IV and COX VI)

were also observed. While eukaryotic cells can respond to mitochondrial

dysfunction by a modulation of nuclear gene expression, evidence for signaling

pathways and transcriptional mechanisms are lacking in most instances. Several

studies have also questioned mt tRNA abnormalities. I honestly believe that

there can be multiple genetic reasons for a particluar abnormality or mix of

abnormalities. The science is just too young for a complete body of knowledge.

We are positive mtDNA is responsible in my girls we just don't know where/how.

Dr. Wallace recently identified a novel homogygous mt related defect for another

family (with an ANT defect and FOD), so his work is continuing.

Barbara Seaman wheatchild@...> wrote:

Oh, one further question. Do you know if the ANT testing was done on fresh

or frozen tissue?

Joanne Kocourek (mom to , lies, and )

visit us at: http://www.caringbridge.org/il/annakris

June 21, 2005

VLCAD too? Wow. Now that IS interesting. I have several papers on GA II. Are

your daughters by any chance riboflavin-responsive? Some patients with GA II

are, and my son and I are very riboflavin-responsive. Riboflavin saved my

functional life 17 years ago. I have carnitine palmitoyl transferase II

deficiency (CPT deficiency), a long-chain defect in fat metabolism, 10% of

normal, in homozygote range. Also every other FOD enzyme we have measured

(which is many) is reduced to 20-50% of normal, both in muscle and

fibroblasts. Complex I, II, III, IV all fall in similar range, 20% to 50% of

normal and citrate synthase in the Krebs cycle is at 30%. (Citrate synthase

is a marker for mitochondrial function.) We have identified the mutation in

the CPT2 gene, but not the underlying cause of all the other deficiencies,

which have been labeled (for lack of a better term) " global mitochondrial

defect. " Most likely cause is thought to be a defect in mitochondrial

transport and I have been screened for several heat shock protein mutations

(HSPs are mitochondrial chaperones) but no found mutations found so far. I

always keep my eyes and ears open for any new findings that might lead to

further definition and understanding of the second defect. When I have time,

I will do some PubMed research and see what I can learn about ANT defects.

My main question at this point would be whether they are associated with

broad secondary deficiencies in many mitochondrial enzymes, or whether their

biochemical effects are seen primarily in reduced ANT activity. I will see

what I can learn and if it seems relevant to my case, will pursue with my

doctors.

Thanks again.

PS Is this Dr. Kendall Wallace or Dr. Wallace? Both are on UMDF

Advisory Board.

Barbara

_____

From: [mailto: ] On Behalf

Of Joanne Kocourek

Sent: Tuesday, June 21, 2005 10:54 AM

To:

Subject: RE: ANT--- PS

If you don't mind my asking, which FOD? My girls seem to have a VLCFAD

(testing still in process). Dr. Cohen is also highly suspicious of GAII.

Barbara Seaman wheatchild@...> wrote:Oh, this is most

interesting, as I also have an FOD. I will pass on

something about Dr. Wallace's work to my doctors and see how they respond.

Thanks again. Much appreciated.

Barbara

_____

From: [mailto: ] On Behalf

Of Joanne Kocourek

Sent: Tuesday, June 21, 2005 10:31 AM

To:

Subject: RE: ANT--- PS

I believe that the ANT studies are performed after the tissue has been

frozen (given Dr. Hoppel repeated the studies over a year later, some of the

study was definitely done using frozen tissue). The RTC studies are

completed when the sample is fresh.

As for ANT genetics....

increased expression of ANT-1 and ATP synthase: subunit mRNAs have been

described in patients with respiratory defects resulting from mtDNA point

mutations and increased mRNAs encoding cytochrome oxidase (COX IV and COX

VI) were also observed. While eukaryotic cells can respond to mitochondrial

dysfunction by a modulation of nuclear gene expression, evidence for

signaling pathways and transcriptional mechanisms are lacking in most

instances. Several studies have also questioned mt tRNA abnormalities. I

honestly believe that there can be multiple genetic reasons for a particluar

abnormality or mix of abnormalities. The science is just too young for a

complete body of knowledge. We are positive mtDNA is responsible in my

girls we just don't know where/how. Dr. Wallace recently identified a novel

homogygous mt related defect for another family (with an ANT defect and

FOD), so his work is continuing.

Barbara Seaman wheatchild@...> wrote:

Oh, one further question. Do you know if the ANT testing was done on fresh

or frozen tissue?

Joanne Kocourek (mom to , lies, and )

visit us at: http://www.caringbridge.org/il/annakris

June 21, 2005