RE: High Prevalence Mycoplasmal Infections -CFS & GWS

[Guest guest]

Hi. Where do you get Acclydine.

High Prevalence Mycoplasmal

Infections -CFS & GWS

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Send an Email for free membership

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>>>> Help ME Circle <<<<

2 January 2003

Editorship : j.van.roijen@...

Outgoing mail scanned by Norton AV

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~JvR: I was tested for Mycoplasmal Infections via PCR in 2000;

the result was positive. I started using an antibiotic treatment

in

cycle: 6 weeks on, 2 weeks off. I didn't feel much change in the

beginning, perhaps some more worse. But after about 6 months

I got a big change for the best.

For example after 18 years of diarrhea my defecation became

normal again. But the biggest change was in my head. The

terrible and frustrating " brainfog " disappeared for 80%. I could

read a text in one or two, instead of ten, twenty times. (only

short

ones; I can't read books anymore)

There also came a big change in my personality: a little of the

'old'

Jan came back again: the joker, the story-teller, the actor.

I come out of my house two times a week; in the neighbourhood

for the necessary shopping. I started again 'small talking' to

neighbours, to people in the shops, or the streets, like I did

in

my former life, twenty years ago; to a mother with a beautiful

laughing child, or something like that.

Although the disease itself was not disappeared, (for example: I

got a relapse for 3 - 4 days after my physician's advise to

walk on tiptoe several times a day in connection with oedema in

my legs, which disappeared later on after the use of potassium),

but I was really happy with the result. I became hopeful again

after many years.

Because my level of Growth Hormone (GH) was very low, my

physician advised me to try Acclydine, which is a new

supplement from vegetable origin. It stimulates the hypothalamus

to produce growth hormone releasing hormone, and this

stimulates the pituitary to produce growth hormone; this

stimulates the liver to produce IGF-1 (insulinelike growth

factor-1). (it is rather expensive)

(see abstract: " Abstracts Sydney Conf. (27-35) = P De Becker,

et al. " - Help ME Circle, 30 Dec. 2001; at Co-Cure:

http://listserv.nodak.edu/scripts/wa.exe?A2=ind0112e & L=co-cure & F= & S= & P=1

260

In this research is demonstrated that 54% of the ME/CFS

patients with low GH, will get a big change for the best after

using this preparation.

But in the insert of this medicine, graded exercise was advised

from the first day, starting with 5 minutes walking, building up

till

at least 30 minutes.

I became ill after the third day of 'exercise'; but I wanted to

be

hardy and went on with it. After two weeks, I was so terrible

sick,

that I had to stop.

My physician supposed that the HHV-6 virus was reactivated

again. This virus was found to be positive by PCR testing about

a year for that. This was in June/July 2001.

And indeed after several weeks, an anti-herpes medicine was

very sweet for me again; I became much better. But after this

" graded exercise " adventure, I never came on the same health

level as before. I'm thankful to my physician, who prescribed me

the anti-biotic, because I'm still much better then before this

cure.

But sometimes I feel sad, because I can't find back the 'old'

Jan

again; he has hide from me somewhere deep in my brain. I'm

longing for him so badly.

````````````

" ...If a patient improves with exercise, that person does not

have ME... "

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http//www.immunesupport.com/library/bulletinarticle.cfm?ID=4186 & PROD=PH1

34

ImmuneSupport

1 January 2003

High Prevalence of Mycoplasmal Infections in

Symptomatic (Chronic Fatigue Syndrome) Family

Members of Mycoplasma-Positive Gulf War Illness Patients

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

---------

Garth L. Nicolson,(1) PhD, Marwan Y. Nasralla,2 PhD, L.

Nicolson,(1) PhD, Joerg Haier,(3) MD, PhD

(1) The Institute for Molecular Medicine, Huntington Beach,

California, USA, (2) International Molecular Diagnostics, Inc.,

Huntington Beach, California, USA, (3)Department of Surgery,

University Hospital Muenster, Germany Correspondence: Prof.

Garth L. Nicolson, Office of the President, The Institute for

Molecular Medicine, 15162 Triton Lane, Huntington Beach,

California 92649. Tel: 714-903-2900; Fax: 714-379-2082;

Website: http://www.immed.org; Email: gnicolson@...

SUMMARY

Immediate family members of veterans diagnosed with Gulf War

Illnesses (GWI) often complain of fatiguing illnesses, and upon

analysis they report similar signs and symptoms as their veteran

family members. Since a relatively common finding in GWI

patients is a bacterial infection due to Mycoplasma species, we

examined military families (149 patients: 42 veterans, 40

spouses, 32 other relatives and 35 children with at least one

family complaint of illness) selected from a group of 110

veterans with GWI who tested positive (~41%) for at least one of

four Mycoplasma species: M. fermentans, M. hominis, M.

pneumoniae or M. genitalium. Consistent with previous results,

over 80% of GWI patients who were positive for blood

mycoplasmal infections had only one Mycoplasma species, M.

fermentans.

In healthy control subjects, the incidence of mycoplasma1

infection was ~8.5% and none were found to have multiple

mycoplasmal species (P The results suggest that a subset of

GWI patients have mycoplasmal infections, possibly obtained as

contaminants from multiple vaccines given during deployment,

and these infections can be transmitted to immediate family

members who subsequently display similar signs and symptoms

and are diagnosed with CFS and/or Fibromyalgia Syndrome.

KEYWORDS. Gulf War Syndrome, bacterial infections, Chronic

Fatigue Syndrome, Myalgic Encephalomyelitis, Fibromyalgia

Syndrome, infectious disease

INTRODUCTION

A sizable fraction (estimated at over 16%) of the veterans of

the

1991 Persian Gulf War from the United States, Great Britain,

Canada, Australia and other nations slowly developed, usually

with a lag of 6-18 months or more after the conflict, a variety

of

complex chronic signs and symptoms. These were

characterized by disabling fatigue, intermittent low-grade

fevers,

night sweats, arthralgias, myalgias, short-term memory loss,

confusion, irritability, depression, headaches, skin rashes,

intermittent diarrhea, respiratory complaints, photophobia and

other signs and symptoms (1-4). These illnesses have been

called Gulf War Syndrome or Gulf War Illnesses (GWI) (5).

Since routine laboratory test results on GWI patients are

usually

not consistent with a single, specific diagnosis, veterans often

did not receive a diagnosis for their condition (illness of

unknown

origin), or they received a diagnosis of somatoform disorder

(1).

The signs and symptoms of GWI do not easily fit into ICD-10

diagnostic categories, but they loosely fall into the category

of

fatiguing illnesses (2-4), and the signs and symptoms of GWI

resemble Chronic Fatigue Syndrome or Myalgic

Encephalomyelitis (CSF/ME) (2, 6-9).

We (6-9) found that most of the signs and symptoms in a large

subset of GWI patients could be explained by chronic

pathogenic bacterial infections. Evidence for infectious agents

has been found in GWI patients' urine (11) and blood (6-10). In

studies on hundreds of U.S. and British veterans with GWI,

approximately 40-50% of GWI patients show evidence of

mycoplasmal infections compared to 6-9% in non-deployed,

healthy subjects (6-10). This has also been confirmed in a large

study of 1,600 veterans at over 30 veterans' and defense

medical centers (VA ative Clinical Study Program #475)

in the United States (12).

Mycoplasmas are small intracellular bacteria lacking cell walls

and extensive intracellular organells. Historically, mycoplasmal

infections were thought to produce relatively mild diseases

limited to particular tissues or organs, such as urinary tract

or

respiratory infections. However, the Mycoplasma species

detected in GWI patients with molecular techniques are highly

virulent, colonize a wide variety of organs and tissues, and are

difficult to treat (13-15). The mycoplasma most commonly

detected in GWI, Mycoplasma fermentans, was found in the

overwhelming majority of those GWI patients positive for any

mycoplasma (7-10).

M. fermentans is an intracellular Mycoplasma, and it is unlikely

that this type of infection will result in a strong antibody

response.

This may explain the lack of a high prevalence of serologic

evidence for these types of intracellular infections in GWI

patients compared to nondeployed controls (15).

When examining GWI patients for illness, we noticed that

immediate family members often had similar complaints (9, 16).

Previously a U.S. Congressional committee had questioned

approximately 1,200 families where one member was a veteran

with GWI and found that 77% of spouses and 65% of children

born after the Gulf War had similar health complaints as the

veteran (17). This suggested that in these families GWI was

being transmitted from veterans with GWI to immediate family

members. The most likely type of transmitted agent would be the

chronic infections found in Gulf War veterans, such as

mycoplasmal infections (6-10).

Therefore, we examined the family members of Gulf War

veterans who were positive for mycoplasmal infections to see if

their signs and symptoms and chronic infections were similar.

We found that most family members of GWI patients that were

symptomatic with signs and symptoms of CFS/ME also had

mycoplasmal infections.

MATERIALS AND METHODS

Patients

Gulf War veterans with GWI and a positive test for mycoplasmal

infection and their immediate family members (149 patients: 42

veterans, 40 spouses, 32 other relatives and 35 children) were

enrolled (Table 1). Seventy age-matched healthy volunteers

were recruited and used as control subjects. All subjects

underwent a medical history and routine laboratory tests. If

necessary, medical records were also reviewed to determine if

patients suffered from organic or psychiatric illnesses that

could

explain their symptoms (3).

When positive results were found in any of the evaluations of

civilians that met the Fukuda et al. (18) exclusionary criteria

for

CFS, the patients were not included in the study. All subjects

completed an illness survey questionnaire (Illness Survey Form,

www.immed.org/signsympt.htm), which included demographic

information, known environmental exposures, dates of illness

onset, health status before and immediately after the Gulf War

and current health status. Additionally, all subjects were

questioned about medication use during the three months prior

to the study, and they had to be free of antibiotic treatment

for

three months prior to blood collection. Controls had to be free

of

disease for at least three months prior to data collection.

Blood Collection

Blood was collected in EDTA-containing tubes and immediately

brought to ice bath temperature as described previously (19,

20). Samples were shipped with wet ice by air courier to the

Institute for Molecular Medicine and International Molecular

Diagnostics, Inc. for analysis. All blood samples were blinded.

Whole blood (50 mul) was used for preparation of DNA using

Chelex (Biorad, Hercules, USA) as follows. Blood cells were

lysed with nano-pure water (1.3 ml) at room temperature for 30

min. After centrifugation at 13 000 x g for 2 min, the

supernatants

were discarded. Chelex solution (200 ml) was added, and the

samples were incubated at 56C and at 100C for 15 minutes

each. Aliquots from the centrifuged samples were used

immediately for PCR or stored at -70C until use. Multiple

Mycoplasma tests were performed on all patients (19, 20).

Amplification of Gene Sequences

Amplification of the target gene sequences (Table in Nasralla et

al., ref. 19) containing 0.1% Triton X-100, 200 mm each of

dATP, dTTP, dGTP, dCTP, 100 pmol of each primer, and 0.5 - 1

mg of chromosomal DNA. Purified mycoplasmal DNA (0.1 - 1

ng of DNA) was used as a positive control for amplification. The

amplification was carried out for 40 cycles with denaturing at

94C and annealing at 60C (genus-specific primers and M.

penetrans) or 55C (M. pneumoniae, M. hominis, M. fermentans,

M. genitalium). Extension temperature was 72C in all cases.

Finally, product extension was performed at 72C for 10 min.

Negative and positive controls were present in each

experimental run (19, 20).

Southern Blot Confirmation

The amplified samples were run on a 1% agarose gel containing

5 mul/100 ml of ethidium bromide in TAE buffer (0.04 M

Tris-Acetate, 0.001 M EDTA, pH 8.0). After denaturing and

neutralization, Southern blotting was performed as follows. The

PCR product was transferred to a Nytran membrane. After

transfer, UV cross-linking was performed. Membranes were

prehybridized with hybridization buffer consisting of 1x

Denhardt's solution and 1 mg/ml salmon sperm DNA as

blocking reagent. Membranes were then hybridized with

32P-labeled internal probe (107 cpm per bag). After hybrization

and washing to remove unbounded probe, the membranes were

exposed to autoradiography film for 1- 2 days at -70C (19, 20).

Statistics

Subjects' demographic characteristics were assessed using

descriptive statistics and students' t-tests (independent

samples

test, t-test for equality of means, 2-tailed). The 95%

confidence

interval was chosen. Pearson Chi-Square test was performed to

compare prevalence data between patients and control

subjects. Additionally, sex differences were investigated using

Pearson Chi-Square test. Illness survey data were statistically

analyzed using Spearman Rank correlation and Mann-Whitney

tests.

RESULTS

As found in previous studies (9,16), veterans of the Gulf War

with

chronic illnesses (GWI) exhibited multiple signs and symptoms

(Figures 1A, . Upon examination, the signs and symptoms of

GWI were indistinguishable from civilian patients diagnosed with

CFS/ME (Figures 1A, B c.f. ref. 9).

Examination of the signs and symptoms of approximately equal

numbers of male and female GWI patients indicated that there

were no significant differences between their general signs and

symptoms (data not shown). When family members who were

symptomatic with chronic signs and symptoms and diagnosed

with CFS/ME were examined (Table 1, age range 4-49 years),

they also appeared to have signs and symptoms

indistinguishable from veterans with GWI (Figures 1A, . When

CFS-symptomatic GWI family members were grouped into

male/female or adults/children, there were no significant

differences between the signs and symptoms found in these

groups. The only signs/symptoms differences found were in

symptomatic family members who did not test positive for

mycoplasmal infections (Figures 1A, .

Similar to previous studies (6-10) 45 of 110 GWI patients or

~41% had mycoplasmal infections (Table 2), and almost all of

these (37 out of 45 or ~82%) were single infections (one

species of mycoplasma) (Table 3). M. fermentans was found in

~85% of these single infection cases. When the few multiple

infection cases were examined, most were found to have

combinations of M. fermentans plus either M. pneumoniae or M.

genitalium (Table 3). In contrast, in healthy control subjects

only 6

of 70 subjects (8.5%) were positive for mycoplasmal infections,

and all of these were single species infections of various types

(Table 4).

Comparing GWI patients and non-symptomatic control subjects,

there was a significant difference in the incidence of

mycoplasmal infections (P In family members of Gulf War

veterans with GWI there was evidence of transmission of the

illness. These families were not randomly chosen; they were

families in which one or more veteran members were found to

be positive for a mycoplasmal infection and one or more family

members reported illnesses. We found that 57 out of 107

(53.2%) of these members from families with one or more Gulf

War veteran diagnosed with GWI and with a positive test for a

mycoplasmal infection showed symptoms of CFS/ME.

Among CFS-symptomatic family members, most (40 out of 57

or 70.2%) had mycoplasmal infections compared to the few

non-symptomatic family members who had similar mycoplasmal

infections (6 out of 50 or 12%) (Table 5). When the incidence of

mycoplasmal infection was compared within families, the

CFS-symptomatic family members were more likely to have

mycoplasmal infections compared to non-symptomatic family

members (P The mycoplasma infection types were also similar

between GWI patients and their CFS-symptomatic family

members. In 45 mycoplasma-positive CFS-symptomatic family

members, most (31 out of 40 or 77.5%) had single species

infections, similar to the mycoplasma-positive Gulf War veterans

(37 out of 45 or 82%). Most of the species found in

mycoplasma-positive GWI patients as well as in

mycoplasma-positive family members with CFS were M.

fermentans (Table 5). We did not find differences in the

incidence of infection or type of infections between males and

females, children versus adults or spouses versus other family

members (data not shown).

However, similar to previous reports, the time of onset of CFS

illness after the Gulf War tended to be shorter in spouses than

other family members, but these differences did not achieve

significance (data not shown).

DISCUSSION

There is growing awareness that GWI is a not a unique, new

syndrome caused by exposures that occurred during the Gulf

War (5,9,21-24). GWI is characterized by a diverse collection of

overlapping, persistent signs and symptoms from which several

distinct subclasses of illness have been identified (3,4,21-24).

Chronic infections are suspected as an important source of

morbidity in a rather large subset of these patients

(6-10,22-26).

The most common infections found were intracellular

mycoplasmal infections (6-10), but a few other chronic

infections

have been documented in GWI patients (25-27).

For example, parasitic infections, such as Malaria,

Leishmaniasis and Schistosomiasis, have been found in some

Gulf War veterans. These infections cause characteristic signs

and symptoms and diagnostic tests are available (27). However,

the prevalence of Leishmaniasis has been estimated at less

than 100 cases in Gulf War veterans (28). Chronic infections

may also cause other problems in GWI patients. Activation of the

coagulation system seen in GWI patients could be related to

chronic infections that cause vasculitis and coagulation

disturbances (29).

In addition, approx. one-half of GWI patients were found to have

fragile chromosomes that are more easily degraded by cellular

nucleases, resulting in release of characteristic nucleotide

fragments (30). This might be due to the action of intracellular

mycoplasmas that are known to release chromosome-damaging

chemicals (31).

Similar to previous results (6-10), mycoplasmal infections were

found in ~41% of GWI patients. This has now been confirmed in

a U. S. Department of Veterans' Affairs study of 1,600 veterans

(28). Although mycoplasmal infections were previously thought to

produce relatively mild diseases limited to particular tissues

or

organs, such as urinary tract or respiratory system (13,14,28),

the intracellular mycoplasmas detected in GWI patients with

molecular techniques, primarily Mycoplasma fermentans, are

highly virulent, colonize a wide variety of organs and tissues,

and

are difficult to treat (13,14). Such intracellular mycoplasmal

infections have been successfully treated with long-term

antibiotics, such as doxycycline, along with immune and

nutritional support (6-9,16).

Although the results presented here document that the chronic

infections found in Gulf War veterans with GWI can be found in

symptomatic family members, we cannot extrapolate our results

to the entire GWI patient population or their family members.

First, our patient sample was not randomly selected. The

presence of a positive mycoplasma test result on a veteran with

GWI who reported illness in his/her immediate family formed the

criteria for inclusion in the study.

Although CFS illnesses in immediate family members were

commonly seen in our study, which examined families of

mycoplasma-positive GWI patients, these illnesses are

expected to be more difficult to find in the general GWI

population where chemical, radiological and environmental

exposures probably account for the majority of cases.

Non-biological exposures should not be transmitted to family

members, so studies on the entire GWI population may not yield

significant results on transmission of illnesses to family

members. Second, GWI patients and their family members were

recruited from veterans groups, word of mouth, physician

referrals and the Institute for Molecular Medicine website

(www.immed.org); they were not recruited from specific military

units.

Therefore, information on presumed exposures of veterans with

respect to unit locations cannot be determined. Testing was

performed using coded samples. After testing, all patients and

control subjects completed patient Illness Survey Forms.

Although some of these patients were examined by physicians

at our associated clinics, most were seen by their own private

physicians. Fourth, the validity of PCR techniques for

Mycoplasma species detection has been questioned. In our

studies, however, the sensitivity and specificity of the PCR

method for Mycoplasma species detection were determined by

examining serial dilutions of purified DNA from M. fermentans,

M. pneumoniae, M. hominis and M. genitalium.

The primers produced the expected amplification product size in

all test species, which was confirmed by hybridization using the

appropriate 32P-labeled internal probe. Amounts as low as a

few fg of purified DNA were detectable for all species with the

specific internal probes. There was no cross-reactivity between

the internal probes of one species and the PCR product from

another species. In other experiments Mycoplasma species

were added to whole blood at various concentrations. Specific

PCR products down to 10 ccu/ml of blood could be detected

(20).

Symptomatic family members of GWI patients were diagnosed

with CSF/ME or a related fatiguing illness, Fibromyalgia

Syndrome (FMS). In other studies, CFS and/or FMS patients

were examined for systemic mycoplasmal infections, and about

50-60% of these patients were positive for any species of

mycoplasma (9,19,20,32-34), indicating another link between

mycoplasma-positive GWI patients and CFS-symptomatic

family members. In contrast to mycoplasma-positive GWI

patients and their mycoplasma-positive family members with

CFS, several species of mycoplasmas in addition to M.

fermentans were found in CSF/ME and FMS from non-military

families (19,34). This further supports the hypothesis that

mycoplasmal infections were transmitted from GWI patients to

immediate family members who then presented with CFS/ME

and for the most part had the same species of mycoplasmal

infection.

The source of the mycoplasmal infections in GWI patients

remains undetermined (28,35). A possible source for GWI

patients is the multiple vaccines that were administered during

the time of deployment to the Persian Gulf. A strong association

has been found between GWI and the multiple vaccines that

were administered (36-38). For example, Steele (4) found a

three-fold increased incidence of GWI in non-deployed veterans

who had been vaccinated in preparation for deployment,

compared to non-deployed, non-vaccinated veterans, and

Mahan et al. (38) found a two-times higher incidence of GWI

signs and symptoms in veterans who recalled receiving anthrax

vaccinations versus those who thought they had not.

Although the mycoplasmal infections found in GWI patients could

have come from several sources, including offensive Biological

Warfare attacks (35), we consider the most likely sources of the

mycoplasmal infections in GWI patients were the multiple

vaccines administered during deployment. Indeed, the signs and

symptoms that have developed in Armed Forces personnel who

recently received the anthrax vaccine are similar to those found

in GWI patients. On some military bases this has resulted in

chronic illnesses in as many as 7-10% of personnel receiving

the vaccine (12,39).

Although the chronic signs and symptoms associated with

anthrax vaccination are similar to those found in GWI patients,

it

is unlikely that all of the chronic illnesses reported by Gulf

War

veterans were caused by vaccines (39).

Undetectable microorganism contaminants in vaccines could

have resulted in illness, and this may have been more likely in

individuals with compromised immune systems caused by

chemical and other exposures (28). Contamination with

mycoplasmas has been found in commercial vaccines (40).

Thus the vaccines used in the Gulf War should be considered as

a possible source of the chronic infections found in

mycoplasma-positive GWI patients and by airborne

transmission in their mycoplasma-positive, CFS-symptomatic

family members.

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© 2002 Journal of Chronic Fatigue Syndrome

© 2003 Pro Health, Inc.

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