Re: adult mito/meta doctors/RH

[Guest guest]

Okay, now I really am muddled up, yes I know that is easy with me

but have just got my report out...and I quote..

The conclusion of the muscle biopsy which was taken from the left

quadriceps shows an excess of cytochrome oxidase negative fibres. It

was felt that these were relatively mild abnormalities but did raise

the possibility of a mitochondrial cytopathy. A piece of FROZEN

muscle was then analysed for enzyme activity and the data was

consistent with deficiencies in Complex 1 and 1V...

So would the enzyme reading be moderately right on frozen muscle and

as a general query to the group does anyone know if the labs are

told of any prescription meds you have been taking which could

affect the readings..I am thinking here of statins and other basic

drugs...just me thinking...

Gillian

> > > > >

> > > > > Kim,

> > > > > Hi! Where about are you in Illinois? I am in

Cedar

> > > Rapids,

> > > > > Iowa, but have family spread throughout Iowa and Illinois,

> > plus

> > > a few

> > > > > other states. Dr. Peltier up in Milwaukee, does work with

> > adult

> > > mito

> > > > > patients. There are a few on this list that have seen

her,

> but

> > > > > experiences have been different for each one. You can e-

mail

> > me

> > > off list

> > > > > if you have any questions about her.

> > > > >

> > > > > Smiles,

> > > > > a

> > > > >

> > > > > On Mon, 11 Apr 2005 21:40:45 -0000 " klaga5 "

> > writes:

> > > > >

> > > > > I'm in Illinois.

> > > > > Kim

> > > > > --- In , Laurie Fitzgerald

> > > > >

> > > > >

[Guest guest]

Gillian,

Reposting what I posted Dec 29, 2004 on fresh vs. frozen. I'm not an expert,

so I can only quote experts. According to these experts, one can measure the

activity of individual respiratory chain enzymes in frozen tissue. What

cannot be done in frozen tissue is look at overall OXPHOS capacity. This

latter testing requires live cells. According to the Ann Clin Biochem

abstract, approx 25% cases of mito manifest only as reduced overall OXPHOS

capacity. In these cases, the enzyme activity in individual complexes is

normal, even though there is reduced overall OXPHOS capacity. Therefore, the

Ann Clin Biochem paper suggests that using frozen tissue for diagnosis would

yield a diagnosis for approx 75% of mito cases, but miss 25% of

cases---those in which complex activity is normal but overall OXPHOS

capacity is reduced. These numbers presuppose viable tissue and valid lab

protocols. Note my original comments and disclaimers below.

Gillian, re your question on drugs: Unfortunately, not all labs require that

a history accompany specimens. A good lab will require detailed history and

design testing accordingly.

B

Some of you might be interested in this info from DiMauro on the MDA site:

QUESTION: Can you tell if a mitochondrial myopathy is or is not maternally

inherited by a frozen muscle biopsy alone? What information does a fresh

biopsy give as opposed to a frozen biopsy?

REPLY from MDA: Salvatore DiMauro, M.D., Prof. of Neurology, New York

A frozen muscle biopsy can provide clues suggesting a mitochondrial DNA

(mtDNA) mutation and, therefore, maternal inheritance of the related

disorder. The clues are of two types: (i) if the biopsy is adequate for

histochemistry, evidence of scattered cytochrome c oxidase-deficient

(COX-negative) fibers, having a " mosaic " distribution among COX-positive

(normal) fibers, is a good indication of a mtDNA mutation, probably

affecting mtDNA genes involved in protein synthesis (such as tRNA genes);

(ii) biochemical analysis of respiratory chain enzymes may reveal decreased

activities of those complexes that contain mtDNA-encoded subunits, i.e.

complexes I, III, and IV (complex V is not assayed routinely in frozen

tissue). This is additional circumstantial evidence of a mtDNA mutation

affecting mitochondrial protein synthesis.

A fresh muscle biopsy allows careful isolation of a mitochondrial fraction;

the isolated mitochondria can be used to assess respiration and

phosphorylation using different substrates and an oxygen electrode

( " polarographic analyses " ), or to study ATP synthesis from ADP and

radio-labeled inorganic phosphate (Pi). These studies can provide

information on the site of a metabolic block in the electron transport chain

(complexes I to IV) and on the function of ATP synthetase (complex V).

------------

There is also an interesting 2003 abstract on the subject:

Ann Clin Biochem. 2003 Jan;40(Pt 1):3-8.

http://www.ncbi.nlm.nih.gov/entrez/utils/lofref.fcgi?PrId=3152&uid=12542905

&db=PubMed&url=http://www.ingentaselect.com/rpsv/cgi-bin/cgi?body=linker&ini

=nlm&reqidx=issn=0004-5632vl=40is=Pt%201yr=2003mn=Janpg=3> Click here to

read

Some practical aspects of providing a diagnostic service for respiratory

chain defects.

Janssen AJ, Smeitink JA, van den Heuvel LP.

Nijmegen Centre for Mitochondrial Disorders, Department of Pediatrics,

University Medical Centre Nijmegen, Geert Grooteplein 10, PO Box 9101, 6500

HB Nijmegen, The Netherlands.

The oxidative phosphorylation system (OXPHOS) is organized into five

multi-protein complexes, comprising four complexes (I-IV) of the respiratory

chain and ATP synthase (complex V). OXPHOS has a vital role in cellular

energy metabolism and ATP production. Enzyme analysis of individual OXPHOS

complexes in a skeletal muscle biopsy remains the mainstay of the diagnostic

process for patients suspected of mitochondrial cytopathy. Practical

guidelines are presented to provide optimal conditions for performance of

laboratory investigations and a reliable diagnosis. A fresh muscle biopsy is

preferable to a frozen muscle sample because the ****overall capacity of the

OXPHOS system**** can be measured in a fresh biopsy. In about 25% of

patients referred for muscle biopsy to our centre, reduced substrate

oxidation rates and ATP+creatine phosphate production rates were found

****without any defect in complexes I-V**** and the pyruvate dehydrogenase

complex. Investigation of frozen muscle biopsy alone may lead to

false-negative diagnoses in many patients. In some patients, it is necessary

to investigate fibroblasts for prospective diagnostic purposes. An exact

diagnosis of respiratory chain defects is a prerequisite for rational

therapy and genetic counselling. Provided guidelines for specimen collection

are followed, there are now reliable methods for identifying respiratory

chain defects.

As I understand it, this says something similar to DiMauro but may be a

little easier to understand. They both say that complexes I-IV can be

assayed in frozen tissue, but to look at ****overall OXPHOS capacity*** one

needs

fresh tissue. According to this abstract of a study in the Netherlands,

about 25% of their mito cases do not manifest as specific deficiencies in

one of the complexes, but only show up as reduced overall OXPHOS

capacity--which can only be measured in fresh tissue. If correct, this would

mean that 25% of mito patients could only get a diagnosis from fresh muscle

tissue. I don't know if all experts agree with these authors (or their

percentages) that a person can have normal activity in complexes I-V and

still have reduced overall OXPHOS capacity that causes disease. But this

distinction may explain why Dr. Korson and others feel some of their

patients with typical mito symptoms do have mito in spite of normal complex

I-V activity in muscle.

B

_____

From: gillianstumps

Sent: Tuesday, April 12, 2005 11:08 AM

To:

Subject: Re: adult mito/meta doctors/RH

Okay, now I really am muddled up, yes I know that is easy with me

but have just got my report out...and I quote..

The conclusion of the muscle biopsy which was taken from the left

quadriceps shows an excess of cytochrome oxidase negative fibres. It

was felt that these were relatively mild abnormalities but did raise

the possibility of a mitochondrial cytopathy. A piece of FROZEN

muscle was then analysed for enzyme activity and the data was

consistent with deficiencies in Complex 1 and 1V...

So would the enzyme reading be moderately right on frozen muscle and

as a general query to the group does anyone know if the labs are

told of any prescription meds you have been taking which could

affect the readings..I am thinking here of statins and other basic

drugs...just me thinking...

Gillian

> > > > >

> > > > > Kim,

> > > > > Hi! Where about are you in Illinois? I am in

Cedar

> > > Rapids,

> > > > > Iowa, but have family spread throughout Iowa and Illinois,

> > plus

> > > a few

> > > > > other states. Dr. Peltier up in Milwaukee, does work with

> > adult

> > > mito

> > > > > patients. There are a few on this list that have seen

her,

> but

> > > > > experiences have been different for each one. You can e-

mail

> > me

> > > off list

> > > > > if you have any questions about her.

> > > > >

> > > > > Smiles,

> > > > > a

> > > > >

> > > > > On Mon, 11 Apr 2005 21:40:45 -0000 " klaga5 "

> > writes:

> > > > >

> > > > > I'm in Illinois.

> > > > > Kim

> > > > > --- In , Laurie Fitzgerald

> > > > >

> > > > >

[Guest guest]

Gillian

Some people can be diagnosed with frozen muscle tissue, but others,

like me, require the fresh.

As far as I know labs are not told of the drugs being taken. It is up

to the doctor to interpret what the lab has found in light of the

meds. This is why many doctors want you all but critical meds for

several weeks before the biopsy.

laurie

On Tue, 12 Apr 2005 11:57:30 -0500, Barbara Seaman

wheatchild@...> wrote:

>

> Gillian,

>

>

>

> Reposting what I posted Dec 29, 2004 on fresh vs. frozen. I'm not an

> expert,

> so I can only quote experts. According to these experts, one can measure

> the

> activity of individual respiratory chain enzymes in frozen tissue. What

> cannot be done in frozen tissue is look at overall OXPHOS capacity. This

> latter testing requires live cells. According to the Ann Clin Biochem

> abstract, approx 25% cases of mito manifest only as reduced overall OXPHOS

> capacity. In these cases, the enzyme activity in individual complexes is

> normal, even though there is reduced overall OXPHOS capacity. Therefore,

> the

> Ann Clin Biochem paper suggests that using frozen tissue for diagnosis

> would

> yield a diagnosis for approx 75% of mito cases, but miss 25% of

> cases---those in which complex activity is normal but overall OXPHOS

> capacity is reduced. These numbers presuppose viable tissue and valid lab

> protocols. Note my original comments and disclaimers below.

>

>

>

> Gillian, re your question on drugs: Unfortunately, not all labs require

> that

> a history accompany specimens. A good lab will require detailed history and

> design testing accordingly.

>

>

>

> B

>

>

>

>

>

> Some of you might be interested in this info from DiMauro on the MDA site:

>

> QUESTION: Can you tell if a mitochondrial myopathy is or is not maternally

> inherited by a frozen muscle biopsy alone? What information does a fresh

> biopsy give as opposed to a frozen biopsy?

>

> REPLY from MDA: Salvatore DiMauro, M.D., Prof. of Neurology, New York

>

> A frozen muscle biopsy can provide clues suggesting a mitochondrial DNA

> (mtDNA) mutation and, therefore, maternal inheritance of the related

> disorder. The clues are of two types: (i) if the biopsy is adequate for

> histochemistry, evidence of scattered cytochrome c oxidase-deficient

> (COX-negative) fibers, having a " mosaic " distribution among COX-positive

> (normal) fibers, is a good indication of a mtDNA mutation, probably

> affecting mtDNA genes involved in protein synthesis (such as tRNA genes);

> (ii) biochemical analysis of respiratory chain enzymes may reveal decreased

> activities of those complexes that contain mtDNA-encoded subunits, i.e.

> complexes I, III, and IV (complex V is not assayed routinely in frozen

> tissue). This is additional circumstantial evidence of a mtDNA mutation

> affecting mitochondrial protein synthesis.

>

> A fresh muscle biopsy allows careful isolation of a mitochondrial fraction;

> the isolated mitochondria can be used to assess respiration and

> phosphorylation using different substrates and an oxygen electrode

> ( " polarographic analyses " ), or to study ATP synthesis from ADP and

> radio-labeled inorganic phosphate (Pi). These studies can provide

> information on the site of a metabolic block in the electron transport

> chain

> (complexes I to IV) and on the function of ATP synthetase (complex V).

>

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

> There is also an interesting 2003 abstract on the subject:

>

>

>

>

> Ann Clin Biochem. 2003 Jan;40(Pt 1):3-8.

>

>

>

>

> http://www.ncbi.nlm.nih.gov/entrez/utils/lofref.fcgi?PrId=3152&uid=12542905

> &db=PubMed&url=http://www.ingentaselect.com/rpsv/cgi-bin/cgi?body=linker&ini

> =nlm&reqidx=issn=0004-5632vl=40is=Pt%201yr=2003mn=Janpg=3> Click here to

> read

> Some practical aspects of providing a diagnostic service for respiratory

> chain defects.

>

> Janssen AJ, Smeitink JA, van den Heuvel LP.

>

> Nijmegen Centre for Mitochondrial Disorders, Department of Pediatrics,

> University Medical Centre Nijmegen, Geert Grooteplein 10, PO Box 9101, 6500

> HB Nijmegen, The Netherlands.

>

> The oxidative phosphorylation system (OXPHOS) is organized into five

> multi-protein complexes, comprising four complexes (I-IV) of the

> respiratory

> chain and ATP synthase (complex V). OXPHOS has a vital role in cellular

> energy metabolism and ATP production. Enzyme analysis of individual OXPHOS

> complexes in a skeletal muscle biopsy remains the mainstay of the

> diagnostic

> process for patients suspected of mitochondrial cytopathy. Practical

> guidelines are presented to provide optimal conditions for performance of

> laboratory investigations and a reliable diagnosis. A fresh muscle biopsy

> is

> preferable to a frozen muscle sample because the ****overall capacity of

> the

> OXPHOS system**** can be measured in a fresh biopsy. In about 25% of

> patients referred for muscle biopsy to our centre, reduced substrate

> oxidation rates and ATP+creatine phosphate production rates were found

> ****without any defect in complexes I-V**** and the pyruvate dehydrogenase

> complex. Investigation of frozen muscle biopsy alone may lead to

> false-negative diagnoses in many patients. In some patients, it is

> necessary

> to investigate fibroblasts for prospective diagnostic purposes. An exact

> diagnosis of respiratory chain defects is a prerequisite for rational

> therapy and genetic counselling. Provided guidelines for specimen

> collection

> are followed, there are now reliable methods for identifying respiratory

> chain defects.

>

>

>

>

>

>

>

>

>

> As I understand it, this says something similar to DiMauro but may be a

> little easier to understand. They both say that complexes I-IV can be

> assayed in frozen tissue, but to look at ****overall OXPHOS capacity*** one

> needs

> fresh tissue. According to this abstract of a study in the Netherlands,

> about 25% of their mito cases do not manifest as specific deficiencies in

> one of the complexes, but only show up as reduced overall OXPHOS

> capacity--which can only be measured in fresh tissue. If correct, this

> would

> mean that 25% of mito patients could only get a diagnosis from fresh muscle

> tissue. I don't know if all experts agree with these authors (or their

> percentages) that a person can have normal activity in complexes I-V and

> still have reduced overall OXPHOS capacity that causes disease. But this

> distinction may explain why Dr. Korson and others feel some of their

> patients with typical mito symptoms do have mito in spite of normal complex

> I-V activity in muscle.

>

> B

>

>

>

>

>

>

>

>

>

>

>

>

>

> _____

>

> From: gillianstumps

> Sent: Tuesday, April 12, 2005 11:08 AM

> To:

> Subject: Re: adult mito/meta doctors/RH

>

>

>

>

> Okay, now I really am muddled up, yes I know that is easy with me

> but have just got my report out...and I quote..

> The conclusion of the muscle biopsy which was taken from the left

> quadriceps shows an excess of cytochrome oxidase negative fibres. It

> was felt that these were relatively mild abnormalities but did raise

> the possibility of a mitochondrial cytopathy. A piece of FROZEN

> muscle was then analysed for enzyme activity and the data was

> consistent with deficiencies in Complex 1 and 1V...

>

> So would the enzyme reading be moderately right on frozen muscle and

> as a general query to the group does anyone know if the labs are

> told of any prescription meds you have been taking which could

> affect the readings..I am thinking here of statins and other basic

> drugs...just me thinking...

>

> Gillian

>

>

>

>

> > > > > >

> > > > > > Kim,

> > > > > > Hi! Where about are you in Illinois? I am in

> Cedar

> > > > Rapids,

> > > > > > Iowa, but have family spread throughout Iowa and Illinois,

> > > plus

> > > > a few

> > > > > > other states. Dr. Peltier up in Milwaukee, does work with

> > > adult

> > > > mito

> > > > > > patients. There are a few on this list that have seen

> her,

> > but

> > > > > > experiences have been different for each one. You can e-

> mail

> > > me

> > > > off list

> > > > > > if you have any questions about her.

> > > > > >

> > > > > > Smiles,

> > > > > > a

> > > > > >

> > > > > > On Mon, 11 Apr 2005 21:40:45 -0000 " klaga5 "

> > > writes:

> > > > > >

> > > > > > I'm in Illinois.

> > > > > > Kim

> > > > > > --- In , Laurie Fitzgerald

> > > > > >

> > > > > >

[Guest guest]

The fact that oxphos wasn't done on my first biopsy makes me

question if it was fresh. Fudge, I wish I hadn't done a second

biopsy there.

Kim

> > > > > >

> > > > > > Kim,

> > > > > > Hi! Where about are you in Illinois? I am in

> Cedar

> > > > Rapids,

> > > > > > Iowa, but have family spread throughout Iowa and

Illinois,

> > > plus

> > > > a few

> > > > > > other states. Dr. Peltier up in Milwaukee, does work

with

> > > adult

> > > > mito

> > > > > > patients. There are a few on this list that have seen

> her,

> > but

> > > > > > experiences have been different for each one. You can e-

> mail

> > > me

> > > > off list

> > > > > > if you have any questions about her.

> > > > > >

> > > > > > Smiles,

> > > > > > a

> > > > > >

> > > > > > On Mon, 11 Apr 2005 21:40:45 -0000 " klaga5 "

> > > writes:

> > > > > >

> > > > > > I'm in Illinois.

> > > > > > Kim

> > > > > > --- In , Laurie Fitzgerald

> > > > > >

> > > > > >

[Guest guest]

I was told to stop all prescription drugs and non-prescription

supplements and vitamins before the testing, but I cowed out and kept

taking my multivitamin and methyl sulfonyl methane (supplement for

joint pain).

Take care,

RH

> > > > > >

> > > > > > Kim,

> > > > > > Hi! Where about are you in Illinois? I am in

> Cedar

> > > > Rapids,

> > > > > > Iowa, but have family spread throughout Iowa and

Illinois,

> > > plus

> > > > a few

> > > > > > other states. Dr. Peltier up in Milwaukee, does work

with

> > > adult

> > > > mito

> > > > > > patients. There are a few on this list that have seen

> her,

> > but

> > > > > > experiences have been different for each one. You can e-

> mail

> > > me

> > > > off list

> > > > > > if you have any questions about her.

> > > > > >

> > > > > > Smiles,

> > > > > > a

> > > > > >

> > > > > > On Mon, 11 Apr 2005 21:40:45 -0000 " klaga5 "

> > > writes:

> > > > > >

> > > > > > I'm in Illinois.

> > > > > > Kim

> > > > > > --- In , Laurie Fitzgerald

> > > > > >

> > > > > >

[Guest guest]

I wasn't told this so now I know to ask some questions of my

mito doc...

Gillian

> > > > > > >

> > > > > > > Kim,

> > > > > > > Hi! Where about are you in Illinois? I am in

> > Cedar

> > > > > Rapids,

> > > > > > > Iowa, but have family spread throughout Iowa and

> Illinois,

> > > > plus

> > > > > a few

> > > > > > > other states. Dr. Peltier up in Milwaukee, does work

> with

> > > > adult

> > > > > mito

> > > > > > > patients. There are a few on this list that have seen

> > her,

> > > but

> > > > > > > experiences have been different for each one. You can

e-

> > mail

> > > > me

> > > > > off list

> > > > > > > if you have any questions about her.

> > > > > > >

> > > > > > > Smiles,

> > > > > > > a

> > > > > > >

> > > > > > > On Mon, 11 Apr 2005 21:40:45 -0000 " klaga5 "

> > > > writes:

> > > > > > >

> > > > > > > I'm in Illinois.

> > > > > > > Kim

> > > > > > > --- In , Laurie Fitzgerald

> > > > > > >

> > > > > > >