stratton and lerner protocols

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Hi:

Can anyone post their experiences with either of these protocols.

What does each protocol entail.

Searching for some relief.

Thanks,

[Guest guest]

>

> Hi:

>

> Can anyone post their experiences with either of these protocols. 

> What does each protocol entail.

>

> Searching for some relief.

>

> Thanks,

Diseases Associated with Chlamydial Infection

An association has been discovered between chronic Chlamydia infection

of body fluids and/or tissues with several disease syndromes of

previously unknown etiology in humans which respond to unique

antichlamydial regimens described herein. To date, these diseases

include Multiple Sclerosis (MS), Rheumatoid Arthritis (RA),

Inflammatory Bowel Disease (IBD), Interstitial Cystitis (IC),

Fibromyalgia (FM), Autonomic nervous dysfunction (AND, neural-mediated

hypotension); Pyoderma Gangrenosum (PG), Chronic Fatigue (CF) and

Chronic Fatigue Syndrome (CFS). Other diseases are under

investigation. Correlation between Chlamydia infection and these

diseases has only recently been established as a result of the

diagnostic methodologies and combination therapies described herein.

The therapies described herein can thus be used for the treatment of

acute and chronic immune and autoimmune diseases when patients are

demonstrated to have a Chlamydia load by the diagnostic procedures

described herein which diseases include, but are not limited to,

chronic hepatitis, systemic lupus erythematosus, arthritis,

thyroidosis, scleroderma, diabetes mellitus, Graves' disease,

Beschet's disease and graft versus host disease (graft rejection). The

therapies of this invention can also be used to treat any disorders in

which a chlamydial species is a factor or co-factor.

Among the various inflammatory diseases, there are certain features of

the inflammatory process that are generally agreed to be

characteristic. These include fenestration of the microvasculature,

leakage of the elements of blood into the interstitial spaces, and

migration of leukocytes into the inflamed tissue. On a macroscopic

level, this is usually accompanied by the familiar clinical signs of

erythema, edema, tenderness (hyperalgesia), and pain. Inflammatory

diseases, such as chronic inflammatory pathologies and vascular

inflammatory pathologies, including chronic inflammatory pathologies

such as aneurysms, hemorrhoids, sarcoidosis, chronic inflammatory

bowel disease, ulcerative colitis, and Crohn's disease and vascular

inflammatory pathologies, such as, but not limited to, disseminated

intravascular coagulation, atherosclerosis, and Kawasaki's pathology

are also suitable for treatment by methods described herein. The

invention can also be used to treat inflammatory diseases such as

coronary artery disease, hypertension, stroke, asthma, chronic

hepatitis, multiple sclerosis, peripheral neuropathy, chronic or

recurrent sore throat, laryngitis, tracheobronchitis, chronic vascular

headaches (including migraines, cluster headaches and tension

headaches) and pneumonia when demonstrated to be pathogenically

related to Chlamydia infection.

The invention is based upon the discovery that current susceptibility

testing methods for Chlamydiae do not accurately predict the ability of

antimicrobial agents to successfully and totally eradicate chronic

chlamydial infections. This is because the current susceptibility

testing methods measure only replication of chlamydia and ignores the

well-known " cryptic phase " in which intracellular Chlamydiae are not

actively replicating. Moreover, it has also been discovered that the

so-called " cryptic phase " of Chlamydiae includes multiple and

different sub-phases. The following are some of the phases of the

chlamydial life cycle in which the intracellular Chlamydiae are not

replicating: an initial intranuclear phase in which elementary bodies

(EBs) transition to reticulate bodies (RBs), an intracytoplasmic phase

in which there is a transition of the RB phenotype to the EB

phenotype, an intracytoplasmic phase with a nonreplicating, but

metabolizing RB, and intracellular/extracellular EB phases, including

endocytotic and exocytotic phases, in which there is neither

replication nor metabolism. In order to assess the cumulative and long

term effect of antimicrobial therapy on these multiple life phases,

unique in vitro and in vivo susceptibility test methods have been

developed and are described herein.

The subject invention also pertains to a method for treating porphyria

caused by Chlamydia in an individual in need thereof, comprising

reducing the levels of active stage, latent stage and elementary

bodies of the pathogen from the individual and administering one or

more compounds which reduce adverse effects associated with secondary

porphyria. In one embodiment, the method additionally comprises

administering a compound which reduces the adverse effects of

porphyrins associated with porphyria. In a particular embodiment, the

compound is cimetidine. This method can also be valuably combined with

additional steps, including at least one of administering

antioxidants; orally administering activated charcoal; administering a

high carbohydrate diet regimen; administering hydroxychloroquine;

administering benzodiazepine drugs; performing hemodialysis;

performing plasmaphoresis; and administering chelating agents; and

administering intravenous hematin.

The invention also pertains to a method of detecting elevated porphyrin

levels in an individual by testing that individual for antibodies to

porphyrins. The invention also pertains to diagnosing deficiency by

detecting antibodies to B-12. Monoclonal and polyclonal antibodies to

prophyrins and/or Vitamin B12 can be produced.

This method is unique because it measures the complete eradication of

all life forms of chlamydiae in known murine target organs for

chlamydial infection. This in vivo susceptibility method has revealed,

for example, that antimicrobial therapy with the triple agents, INH,

metronidazole and penicillamine, can completely eradicate C. pneumoniae

from infected mice in four months. Moreover, following complete

eradication of chlamydiae, multiple attempts to reinfect these cured

mice via intranasal inoculation have proven unsuccessful. This

suggests that effective management and complete [eradiaction]

eradication results in the development of protective immunity, and

that effective management is therefore a way to create effective

immunity.

Examples of suitable nitroimidazoles include, but are not limited to,

metronidazole, tinidazole, bamnidazole, benznidazole, flunidazole,

ipronidazole, misonidazole, moxnidazole, ronidazole, sulnidazole, and

their metabolites, analogs and derivatives thereof. Metronidazole is

most preferred. Examples of nitrofurans that can be used include, but

are not limited to, nitrofurantoin, nitrofurazone, nifurtimox,

nifuratel, nifuradene, nifurdazil, nifurpirinol, nifuratrone,

furazolidone, and their metabolites, analogs and derivatives thereof.

Nitrofurantoin is preferred within the class of nitrofurans.

INH and its congeners can be used to clear infection from monocytes

and/or macrophages. When monocytes and macrophages are infected by

Chlamydia, they become debilitated and cannot properly or effectively

fight infection. It is believed that, if the chlamydial infection, per

se, is cleared from these cells, then the monocytes and macrophages

can resume their critical roles fighting chlamydial or other

infection(s). Thus, patient responsiveness to combination therapy can

be optimized by the inclusion of isonicotinic acid congeners.

Accordingly, one aspect of the invention provides a specific method

for reempowering monocytes or macrophages that have been compromised

by a Chlamydia infection and, in turn, comprise treating the infection

in other sites. Such compromised macrophages or monocytes can be

activated by treating the chlamydial infection by contacting the

infected macrophages and/or monocytes with an antichlamydial agent.

Cells to be treated can already be cryptically infected or they can be

subjected to stringent metabolic or environmental conditions which

cause or induce the replicating phase to enter the cryptic phase. Such

stringent conditions can include changing environmental/culturing

conditions in the instance where the infected cells are exposed to

..gamma.-interferon; or by exposing cells to conventional antimicrobial

agents (such as macrolides and tetracyclines) which induce this

cryptic phase of chlamydial infection in human host cells.

Therapy Directed Toward Elementary Bodies of Chlamydia

As discussed above, it has been discovered that adverse conditions,

such as limited nutrients, antimicrobial agents, and the host immune

response, produce a stringent response in Chlamydia. Such adverse

conditions are known to induce stringent responses in other

microorganisms (C. W. Stratton, In: Antibiotics in Laboratory

Medicine, Fourth Edition. Lorian V (ed) & Wilkins, Baltimore,

pp 579-603 (1996)) and not surprisingly induce a stringent response in

Chlamydia. This stringent response in Chlamydia alters the

morphological state of the intracellular microorganism and creates

dormant forms, including the intracellular EB, which then can

cryptically persist until its developmental cycle is reactivated.

Conversely, the host cell may lyse and allow the EBs to reach the

extracellular milieu. Thus, it is necessary to utilize a combination

of agents directed toward the various life stages of Chlamydia and, in

particular, against the elementary body for successful management of

infection.

As described in previous sections, it is also believed that

[persistance] persistence of chlamydial infections, in part, may be

due to the presence of cryptic forms of Chlamydia within the cells.

This cryptic intracellular chlamydial form apparently can be activated

by certain host factors such as cortisone (Yang et al., Infection and

Immunity, 39:655-658 (1983); and Malinverni et al., The Journal of

Infectious Diseases, 172:593-594 (1995)). Antichlamydial therapy for

chronic Chlamydia infections must be continued until any intracellular

EBs or other intracellular cryptic forms have been activated and

extracellular EBs have infected host cells. This

reactivation/reinfection by chlamydial EBs clearly is undesirable as it

prolongs the therapy of chlamydial infections, as well as increases

the opportunity for antimicrobial resistance to occur.

Physiochemical agents have been identified that can inactivate

chlamydial EBs in their respective hosts by reducing disulfide bonds

which maintain the integrity of the outer membrane proteins of the

EBs. For Chlamydia, disruption of the outer membrane proteins of EBs

thereby initiates the transition of the EB form to the RB form. When

this occurs in the acellular milieu where there is no available energy

source, the nascent RB perishes or falls victim to the immune system.

Thus, disulfide reducing agents that can interfere with this process

are suitable as compounds for eliminating EBs.

One such class of disulfide reducing agents are thiol-disulfide

exchange agents. Examples of these include, but are not limited to,

2,3-dimercaptosuccinic acid (DMSA; also referred to herein as

" succimer " ); D,L,-.beta.,.beta.-dimethylcysteine (also known as

penicillamine); .beta.-lactam agents (e.g., penicillins, penicillin G,

ampicillin and amoxicillin, which produce penicillamine as a

degradation product), cycloserine, dithiotreitol, mercaptoethylamine

(e.g., mesna, cysteiamine, dimercaptol), N-acetylcysteine, tiopronin,

and glutathione. A particularly effective extracellular antichlamydial

agent within this class is DMSA which is a chelating agent having four

ionizable hydrogens and two highly charged carboxyl groups which

prevent its relative passage through human cell membranes. DMSA thus

remains in the extracellular fluid where it can readily encounter

extracellular EBs. The two thiol (sulflhydryl) groups on the succimer

molecule (DMSA) are able to reduce disulfide bonds in the MOMP of EBs

located in the extracellular milieu.

As chlamydial RBs transform into EBs, they begin to utilize active

transcription of chlamydial DNA and translation of the resulting mRNA.

As such, these forms of Chlamydia are susceptible to currently used

antimicrobial agents. The antichlamydial effectiveness of these agents

can be significantly improved by using them in combination with other

agents directed at different stages of Chlamydia life cycle, as

discussed herein.

Classes of suitable antimicrobial agents include, but are not limited

to, rifamycins (also known as ansamacrolides), quinolones,

fluoroquinolones, chloramphenicol, sulfonamides/sulfides, azalides,

cycloserine, macrolides and tetracyclines. Examples of these agents

which are members of these classes, as well as those which are

preferred, are illustrated below in Table 5.

Agents Effective Against the Replicating

Phase of Chlamydia

Drug Class Examples Preferred

Quinolones/ Ofloxacin Levofloxacin

Fluoroquinolones Levofloxacin

Trovafloxacin

Sparfloxacin

Norfloxacin

Lomefloxacin

Cinoxacin

Enoxacin

Nalidixic Acid

Fleroxacin

Ciprofloxacin

Sulfonamides Sulfamethoxazole Sulfamethoxazole/

Trimethoprim

Azalides Azithromycin Azithromycin

Macrolides Erythromycin Clarithromycin

Clarithromycin

Lincosamides Lincomycin

Clindamycin

Tetracyclines Tetracycline Minocycline

Doxycycline

Minocycline

Methacycline

Oxytetracyline

Rifamycins Rifampin Rifampin

(Ansamacrolides) Rifabutin

All members of the Chlamydia species, including C. pneumoniae, are

considered to be inhibited, and some killed, by the use of a single

agent selected from currently used antimicrobial agents such as those

described above. However, using the new susceptability test, the

inventors have found complete eradication of Chlamydia cannot be

achieved by the use of any one of these agents alone because none are

efficacious against all phases of the Chlamydia life cycle and appear

to induce a stringent response in Chlamydia causing the replicating

phase to transform into cryptic forms. This results in a persistent

infection in vivo or in vitro that can be demonstrated by PCR

techniques which assess the presence or absence of chlamydial DNA.

Nevertheless, one or more of these currently used agents, or a new

agent directed against the replicating phase of Chlamydia, should be

included as one of the chlamydial agents in a combination therapy in

order to slow or halt the transition of the EB to the RB as well as to

inhibit chlamydial replication.

Methodology for Selecting Potential Agent Combinations

In attempting to manage or eradicate a systemic infection, it is

critical to target multiple phases in the life cycle of Chlamydia,

otherwise viable Chlamydia in the untargeted phases will remain after

therapy and result in continued, chronic infection. This fundamental

insight is at the core of this invention.

A preferred method for selecting an appropriate combination of agents

that satisfies the requirements of this strategy comprises a plurality

of steps as follows:

1. Identify the phases of the chlamydial life cycle. For example, the

following phases are currently known:

a. Elementary Body ( " EB " )--Extracellular or Intracellular.

Intracellular EBs may represent a type of " cryptic phase " .

b. EB to Reticulate Body ( " RB " ) transition phase.

c. Stationary RB phase. This is what is traditionally thought of as the

" cryptic phase " .

d. Replicating RB phase.

e. RB to EB transition phase (also called " condensation " ).

2. Evaluate the relative importance of targeting each particular phase

in eradicating reservoirs of Chlamydia from the host organism. For

example, the life-cycle stages listed in step 1 can be prioritized

based on the following assumptions:

a. In the host, [Extracellular] extracellular and intracellular EBs

represent a very important reservoir of infectious agents that result

in chronic and persistent infection.

b. Most intracellular RBs in chronic infections are non-replicating.

The 3-4 day reproduction cycle seen in cycloheximide-treated

eukaryotic cells is an artifact of an atypical, cell culture

environment designed primarily to propagate Chlamydia.

c. The transition phases represent only a small portion of Chlamydia in

chronic infections.

3. Identify " targets " for each phase of the selected life cycle phases.

A target is an attribute of Chlamydia which is vulnerable during a

particular life cycle phase. For example, the disulfide bonds in MOMP

are a target during the EB phase.

4. Identify agents with known or theoretical mechanism(s) of action

against those targets.

5. Estimate whether those agents would be merely inhibitory or,

preferably, cidal, through an understanding of their mechanism of

action.

6. Confirm the estimate by using the following approaches:

a. In the case of anti-EB agents, treat EBs with the agent, then

attempt to infect cells with the treated EBs. If the cells do not

become infected, the agent is EB-cidal.

b. In the case of other agents, use the susceptibility tests disclosed

elsewhere herein, to determine whether the agent, either alone or in

combination with other agents, is chlamydicidal.

7. Select a combination of agents that, through their individual

effects, provide activity against targets for the most important

phases within the chlamydial life cycle. Preferably, a combination

should target as many phases of the life cycle as possible, seeking to

maximize the total of the relative important scores of the phases

targeted while minimizing the number of drugs involved.

8. Test the combination using the susceptibility testing procedures

described elsewhere. This step is necessary because the selected

combination may or may not be chlamydicidal for various reasons such as

intracellular penetration and/or efflux.

9. Set initial dosages based on clinical standards which consider the

pharmacokenetics and pharmacodynamics for the drugs prescribed

individually; modifications, if needed, are based on results of

susceptibility testing and in vivo efficacy.

Table 6 provides an example of how the foregoing methodology can be

used. The preferred embodiment includes agents which:

a) Target disulfide bonds in the EB and condensation phases;

Target non-oxidative metabolism in the stationary/cryptic phase;

c) Target constitutive production of peroxidases and catalyses in the

stationary and replicative phases;

d) In the latter two cases, work through physio-chemical disruption of

the organism through free radicals, which are very difficult for

organism to develop resistance to; and

e) Optionally adds an agent to target DNA-dependent RNA polymerase in

the EB->RB phase.

The foregoing methodology for selecting combination therapies can be

automated (e.g., by a computer system) at one or a combination of the

steps described above. This methodology is applicable even after

greater understanding of the chlamydial life cycle leads to a

re-prioritization or even sub-division of the life-cycle phases, new

theoretical targets within Chlamydia are identified, or new drugs are

developed which attack currently known or new targets within

Chlamydia. For example, the phases of the life cycle could be further

sub-classified based on the type of host cell the phase is in. Thus,

stationary phase RBs in macrophages could be considered a separate

phase than stationary phase RBs in hepatocytes. This allows the

methodology to be used to design a single or multi-tissue specific

combination of agents.

Vitamin C (2 gms bid) has also been introduced based on the report that

Vitamin C (ascorbic acid) at moderate intracellular concentrations

stimulates replication of C. trachomatis (Wang et al., J. Clin. Micro.

30:2551-2554 (1992)) as well as its potential effect on biofilm charge

and infectivity of the bacterium and specifically the EB (Hancock, R.

E. W., Annual Review in Microbiology, 38:237-264 (1984)).

Diagnosis and Treatment of Secondary Porphyria

Chlamydia is a parasite of normal energy production in infected

eukaryotic cells. As a result, host cells have insufficient energy

available for their normal functioning. The energy shortage also

causes the host cell mitochondria to attempt to synthesize certain

critical enzymes involved in energy production in order to increase

energy production. Because Chlamydia also prevents this synthesis from

completing, these enzyme's precursors, called porphyrins, build up in

cell and often escape into the intracellular [mileau] milieu.

Porphyrins readily form free-radicals, that, in turn, damage cells.

Thus, there is an obligate secondary porphyria that accompanies many

chlamydial infections. Therapy for this secondary porphyria, which is

adjunct to anti-chlamydial therapy, involves at least three

strategies: a) supplement the cellular energy supply to mitigate cell

malfunction and the formation of porphyrins; reduce the levels of

systemic porphyrins; and c) mitigate the harmful effects of the

porphyrins.

The pathogenesis of chronic/systemic chlamydial infection is unique in

that the intracellular infection by this parasite results in a number

of heretofore unrecognized concomitant and obligatory

metabolic/autoimmune disorders including secondary porphyria with

associated autoantibodies against the porphyrins. Cross reaction with

Vitamin B12 can result in a subclinical autoimmune-mediated Vitamin

B12 deficiency. These associated disorders often require diagnosis and

preventive and/or specific adjunctive therapy.

The inventors have discovered the existence of antibodies to the

various metabolites of heme biosynthesis, as well as Vitamin B12

(cobalamin), which is molecularly similar to these metabolites, in

patients with active systemic infection with C. pneumoniae. The

antibodies are primarily IgM; this is similar to the antibody

responses to the MOMP of C. pneumoniae in severely symptomatic

patients. Example 8 illustrates titers in symptomatic patients with

systemic C. pneumoniae infections. The presence of antibodies to

Vitamin B12 may have functional significance by decreasing the amount

of bioavailable Vitamin B12. Thus, a Chlamydia infection may cause a

previously unrecognized secondary Vitamin B12 deficiency.

Administration (e.g., intramuscular) of large quantities of Vitamin B12

(1000 to 5000 .mu.g) (e.g., parenteral cobalamin therapy) creates

large amounts of Vitamin B12 available for binding to the native

receptors of antibodies with an affinity for Vitamin B12, thereby

saturating these anti-Vitamin B12 antibodies and increasing the amount

of bioavailable circulating Vitamin B12.

Glucose is an important source of cellular energy. Glucose levels can

be enhanced by diet and through vitamin supplements as described

below.

A high carbohydrate diet should be maintained to promote production of

glucose (Pierach et al., Journal of the American Medical Association,

257:60-61 (1987)). Approximately 70% of the caloric intake should be in

the form of complex carbohydrates such as bread, potato, rice and

pasta. The remaining 30% of the daily diet should comprise protein and

fat, which should ideally be in the form of fish or chicken. Red

meats, including beef, dark turkey, tuna and salmon, contain

[tryprophan] tryptophan. Increased levels of tryptophan in the liver

inhibit the activity of phosphoenol pyruvate carboxykinase with

consequent disruption of gluconeogenesis. This accounts for the

abnormal glucose tolerance seen in porphyria. Increased plasmic

concentrations of tryptophan also enhances tryptophan transport into

the brain. The concentration of tryptophan in the brain is the

rate-limiting factor for the synthesis of the neurotransmitter

5-hydroxytryptamine (5-HT, serotonin). Serotonin is synthesized by the

endothelium of brain capillaries for circulating tryptophan. Thus,

increased concentrations of tryptophan in the brain would be expected

to enhance production of serotonin and its metabolic,

5-hydroxyindole-acetic acid (5HIAA). Acute increases in serotonin

turnover in the brain are followed by vascular and metabolic changes

which include decreases in glucose consumption, disturbances in EEG

tracings, and decreases in the postischemic neurological score. In

addition, while serotonin increases brain perfusion on a single

injection, repetitive administration initially opens the blood-brain

barrier and subsequently induces vasoconstriction. It is likely that

any transient opening of the blood-brain barrier by serotonin could

allow circulating substrates such as ALA and PBG, if present, to enter

the central nervous system. As would be expected from the location of

serotonin receptors and from the barrier function of the endothelium

of cerebral arteries, the constricting effect of serotonin is

amplified in cerebral arteries where endothelium is damage or removed.

Damaged endothelial cells, as would be expected with chlamydial

infection, would no longer have operational catabolic processes for

serotonin. This would be particularly true in the event of depleted

ATP as caused by chlamydial infection. This means that increased

concentrations of serotonin will reach the smooth muscle layer of the

cerebral vessels and cause more constriction. Finally, serotonin is

also stored in blood platelets. Because blood platelets do not adhere

and aggregate under normal conditions, they do not release serotonin

when the vessel lumen is intact. However, if the vessel lumen is

altered by chlamydial infection, platelet deposition and release of

serotonin can occur.

Another adverse effect of increased serotonin levels due to porphyria

is seen with nervous tissues. Sympathetic nerve endings store

serotonin taken up from the circulation. These serotonergic neurons

form plexuses around brain vessels where they are likely to liberate

their serotonin contents when subjected to cellular lysis from any

cause including ischemia, free-radical ionizing damage to cell

membranes, and/or chlamydial infection.

In rats, elevated circulating tryptophan has been shown to produce

structural alteration of brain astrocytes, oligodendroglia, and

neurons, as well as degeneration of Purkinje cells and wasting of

axons. Similar neurohistological alterations have been reported in

patients with acute porphyria. Elevated tryptophan levels in plasma

and brain have been associated with human encepholopathy. Finally,

serotonin is also recognized as an active neurotransmitter in the

gastrointestinal tract. The pharmacologic effects of serotonin in the

central nervous system and gastrointestinal tract resemble the

neurological manifestations of acute porphyric attacks. In fact,

administration of either tryptophan or serotonin to humans have been

reported to cause severe abdominal pain, psychomotor disturbances,

nausea, and dysuria; all of which are symptoms of acute porphyria.

Sucrose and fructose should be avoided (Bottomly et al., American

Journal of Clinical Pathology, 76:133-139 (1981)) because the

ingestion of large amounts of fructose trigger hepatic gluconeogenesis

which then decreases the available glucose which is derived from

glycogen breakdown within the liver. It is recommended that sport

drinks which contain glucose be consumed.

It is recommended that a patient suffering from porphyria avoid milk

products. Milk products contain lactose and lactoferrin, and have been

empirically shown to make symptoms of porphyria worse.

Multivitamins containing the B complex vitamins should be administered

daily (e.g., one or multiple times), preferably in excess of RDA, to

enhance glucose availability. Hepatic breakdown of glycogen with

generation of glucose is assisted by taking these multivitamins that

contain the B complex vitamins. Pyridoxine minimizes the porphyrin

related porphyrial neuropathy. B complex vitamins include folic acid

(e.g., 400 .mu.g per dosage; 1200 .mu.g daily maximum); vitamin B-1

(thiamin; e.g., 10 mg per dosage; 30 mg daily maximum); B-2

(riboflavin; e.g., 10 mg per dosage; 30 mg daily maximum); B-5

(panothenate; e.g., 100 mg per dosage; 300 mg daily maximum); B-6

(pyridoxine; e.g., 100 mg per dosage; 300 mg daily maximum) or

pyridoxal-5-phosphate (e.g., 25 mg per dosage; 100 mg daily maximum)

and B-12 (e.g., 500 .mu.g per dosage; 10,000 .mu.g daily maximum). The

preferred method of administration is oral for the majority of these

vitamins (twice daily), except for B-12 for which sublingual

administration (three-times daily) is preferred. It has been discovered

that one important effect of this secondary porphyria in some patients

is the production of IgM and IgG antibodies against

coproporphyrinogen-III. These antibodies cross-react with Vitamin B12

(cobalamin) and can thus cause a deficiency. Vitamin B12

supplementation (e.g., parenteral cobalamin therapy) can remedy the

deficiency.

D. Reducing Porphyrin Levels

Dietary and pharmaceutical methods can be used to reduce systemic

porphyrin levels (both water-soluble and fat-soluble).

Plenty of oral fluids in the form of bicarbonated water or " sports

drinks " (i.e., water with glucose and salts) should be incorporated

into the regimen. This flushes water-soluble porphyrins from the

patient's system. Drinking seltzer water is the easiest way to achieve

this goal. The color of the urine should always be almost clear

instead of yellow. It is noted that dehydration concentrates

prophyrins and makes patients more symptomatic.

Activated charcoal can be daily administered in an amount sufficient to

absorb fat-soluble porphyrins from the enterohepatic circulation.

Treatment with activated oral is charcoal, which is nonabsorbable and

binds porphyrins in the gastrointestinal tract and hence interrupts

their enterohepatic circulation, has been associated with a decrease

of plasma and skin porphyrin levels. Charcoal should be taken between

meals and without any other oral drugs or the charcoal will absorb the

food or drugs rather than the porphyrins. For those who have

difficulty taking the charcoal due to other medications being taken

during the day, the charcoal can be taken all at one time before bed.

Taking between 2 and 20 grams, preferably at least 6 grams

(24.times.250 mg capsules) of activated charcoal per day (Perlroth et

al., Metabolism, 17:571-581 (1968)) is recommended. Much more charcoal

can be safely taken; up to 20 grams six times a day for nine months has

been taken without any side effects.

Antioxidants at high dosages (preferably taken twice per day) help to

mitigate the effects of free radicals produced by porphyrins. Examples

of suitable antioxidants include but are not limited to Vitamin C

(e.g., 1 gram per dosage; 10 g daily maximum); Vitamin E (e.g., 400

units per dosage; 3000 daily maximum); L-Carnitine (e.g., 500 mg per

dosage; 3 g daily maximum); coenzyme Q-10 (uniquinone (e.g., 30 mg per

dosage; 200 mg daily maximum); biotin (e.g., 5 mg per dosage; 20 mg

daily maximum); lipoic acid (e.g., 400 mg per dosage; 1 g daily

maximum); selenium (e.g., 100 .mu.g per dosage; 300 .mu.g daily

maximum); gultamine (e.g., from 2 to about 4 g per dosage);

glucosamine (e.g., from about 750 to about 1000 mg per dosage); and

chondroitin sulfate (e.g., from about 250 to about 500 mg per dosage).

The above-mentioned therapeutic diets can be combined with traditional

or currently recognized drug therapies for porphyria. In one

embodiment, benzodiazapine drugs, such as but not limited to valium,

klonafin, flurazepam hydrochloride (e.g., Dalmanc.TM., Roche) and

alprazolam (e.g., Xanax), can be administered. Preferably, sedatives,

such as alprazolam (e.g., Xanax; 0.5 mg per dosage for 3 to 4 times

daily), can be prescribed for panic attacks and flurazepam

hydrochloride (e.g., Dalmane.TM., Roche or Restoril.TM. (e.g., 30 mg

per dosage)) can be prescribed for sleeping. The rationale is based

upon the presence of peripheral benzodiazepine receptors in high

quantities in phagocytic cells known to produce high levels of radical

oxygen species. A protective role against hydrogen peroxide has been

demonstrated for peripheral benzodiazipine receptors. This suggests

that these receptors may prevent mitochondria from radical damages and

thereby regulate apoptosis in the hematopoietic system.

Benzodiazepines have also been shown to interfere with the

intracellular circulation of heme and porphyrinogens (Scholnick et

al., Journal of Investigative Dermatology, 1973, 61:226-232). This is

likely to decrease porphyrins and their adverse effects. The specific

benzodiazipine will depend on the porphyrin-related symptoms.

Table 12 provides the results of an expanded study of antimicrobial

susceptibilities at two different concentrations of antimicrobial

agents, used alone and in combination, when exposed to the

antimicrobial agents for two weeks. In addition to the agents already

mentioned, minocycline, doxycycline, rifampin and

sulfamethoxizole/trimethoprim, at all concentrations tested, failed to

clear the PCR signal for chlamydial MOMP. Only the triple combination

of isoniazid, metronidazole and penicillamine cleared the PCR signal

in two weeks. The triple combination was effective at both low and

high concentrations. Table 12 also demonstrates the effect of a 4 week

exposure with the same expanded series of antimicrobial agents alone

and in combination. A number of triple combinations of antimicrobial

agents resulted in cell cultures in which the PCR signal for the

chlamydial MOMP gene could not be detected at four weeks. The most

effective combinations to have the greatest impact on the important

life cycle phases are those predicted according to the methodology

described in the section above entitled " Methodology for Selecting

Potential Agent Combinations " .

TABLE 11

Susceptibility to Antibiotics for

Cryptic C. pneumoniae Cultured in HeLa Cells.sup.a

Antibiotic Conc (.mu.g/ml) PCR.sup.b

Ofloxacin 1 positive

Clarithromycin 1 positive

INH 1 positive

Metronidazole 1 positive

Penicillamine 1/1 positive

INH + Metronidazole + 1/1/4 negative

Penicillamine

Control 0 positive

.sup.a Cultured in the presence of the indicated antibiotic(s), but

with no

cycloheximide. Media changes at 48-72 hours.

.sup.b Analysis following 2 weeks exposure to antimicrobioal agents.

TABLE 12

Susceptibility to

Antibiotics by PCR

for

Cryptic Chlamydia pneumoniae Cultured

in HeLa

Cells.sup.1

Phase of the Chlamydial Life Cycle

EB (Extracellular EB->RB Stationary Phase RB RB->EB

Transition Concentration PCR PCR

or Intracellular) Transition ( " Cryptic phase " ) Replicating RB

( " Condensation " ) (.mu.g/ml) 2 week 4 week Comment

0 + + Control

Minocycline

0.25 + + One drug

Minocycline

1 + + One drug

Doxycycline

0.25 + + One drug

Doxycycline

1 + + One drug

TMP/SMZ.sup.2

25 + + One drug

TMP/SMZ.sup.2

100 + + One drug

Clarithromycin

0.25.sup.1 + + One drug

Ofloxacin

0.25 + + One drug

Metronidazole

0.25 + + One drug

Rifampin

0.25 + + One drug

Isoniazide Isoniazide

1 + + Misses EB phase

Metronidazole TMP/SMZ.sup.1

25/0.25 + + Misses EB phase

Metronidazole Ofloxacin

0.25/0.25 + + Misses EB phase

Rifampin Metronidazole

0.25/0.25 + + Misses EB phase

penicillamine Rifampin Metronidazole

penicillamine

4/.25/.25 + + Misses replicating

phase

Rifampin Metronidazole Ofloxacin

.25/.25/.25 + + Misses EB phase

penicillamine Metronidazole Doxycyclin

penicillamine

1/.25/.25 + + Concentration too low

penicillamine Metronidazole Doxycyclin

penicillamine

4/1/1 + - Covers key phases

penicillamine Metronidazole Minocycline

penicillamine

1/.25/.25 + - Covers key phases

penicillamine Metronidazole Minocycline

penicillamine

4/1/1 + - Original report of 4

week positive was a

typo

penicillamine Metronidazole TMP/SMZ

penicillamine

4/1/100 + - Covers key phases

penicillamine Metronidazole Clarithromycin

penicillamine

1/.25/.25 + - Covers key phases

penicillamine Isoniazid Isoniazid

penicillamine

1/.25/.25 - - Covers key phases

Metronidazole

penicillamine Isoniazid Isoniazid

penicillamine

4/1/1 - - Covers key phases

Metronidazole

.sup.1 Cultured in the presence of the indicated antibiotics, but

with no

cycloheximide. Media changes at 48-72 hours. + = positive; - =

negative

.sup.2 TMP/SMX = trimethoprim/sulfamethoxazole

>

>

[Guest guest]

I found this really fascinating. I have been taking Doxy on and off

for ten months for persistent Chlamydia infections. Chlamydia is

nasty. It is known to cause chronic bronchitis and heart disease.

I never have a cough when I take Doxy. Herxes are rough.

Taking Flagyl now for ten days (2nd course). Mild herx happening.

I believe that Chlamydia is at the root of my CFS/ME and fibro, even

possibly the osteo arthritis.

Bugs, Bugs, everywhere and in every form, it seems. The trick is to

diagnose who is what, where, when and then kill 'em.

Cheers,

*Sue*

> >

> > Hi:

> >

> > Can anyone post their experiences with either of these

protocols. 

> > What does each protocol entail.

> >

> > Searching for some relief.

> >

> > Thanks,

>

>

> Diseases Associated with Chlamydial Infection

>

> An association has been discovered between chronic Chlamydia

infection

> of body fluids and/or tissues with several disease syndromes of

> previously unknown etiology in humans which respond to unique

> antichlamydial regimens described herein. To date, these diseases

> include Multiple Sclerosis (MS), Rheumatoid Arthritis (RA),

> Inflammatory Bowel Disease (IBD), Interstitial Cystitis (IC),

> Fibromyalgia (FM), Autonomic nervous dysfunction (AND, neural-

mediated

> hypotension); Pyoderma Gangrenosum (PG), Chronic Fatigue (CF) and

> Chronic Fatigue Syndrome (CFS). Other diseases are under

> investigation. Correlation between Chlamydia infection and these

> diseases has only recently been established as a result of the

> diagnostic methodologies and combination therapies described

herein.

>

> The therapies described herein can thus be used for the treatment

of

> acute and chronic immune and autoimmune diseases when patients

are

> demonstrated to have a Chlamydia load by the diagnostic

procedures

> described herein which diseases include, but are not limited to,

> chronic hepatitis, systemic lupus erythematosus, arthritis,

> thyroidosis, scleroderma, diabetes mellitus, Graves' disease,

> Beschet's disease and graft versus host disease (graft

rejection). The

> therapies of this invention can also be used to treat any

disorders in

> which a chlamydial species is a factor or co-factor.

>

> Among the various inflammatory diseases, there are certain

features of

> the inflammatory process that are generally agreed to be

> characteristic. These include fenestration of the

microvasculature,

> leakage of the elements of blood into the interstitial spaces,

and

> migration of leukocytes into the inflamed tissue. On a

macroscopic

> level, this is usually accompanied by the familiar clinical signs

of

> erythema, edema, tenderness (hyperalgesia), and pain.

Inflammatory

> diseases, such as chronic inflammatory pathologies and vascular

> inflammatory pathologies, including chronic inflammatory

pathologies

> such as aneurysms, hemorrhoids, sarcoidosis, chronic inflammatory

> bowel disease, ulcerative colitis, and Crohn's disease and

vascular

> inflammatory pathologies, such as, but not limited to,

disseminated

> intravascular coagulation, atherosclerosis, and Kawasaki's

pathology

> are also suitable for treatment by methods described herein. The

> invention can also be used to treat inflammatory diseases such as

> coronary artery disease, hypertension, stroke, asthma, chronic

> hepatitis, multiple sclerosis, peripheral neuropathy, chronic or

> recurrent sore throat, laryngitis, tracheobronchitis, chronic

vascular

> headaches (including migraines, cluster headaches and tension

> headaches) and pneumonia when demonstrated to be pathogenically

> related to Chlamydia infection.

>

>

> The invention is based upon the discovery that current

susceptibility

> testing methods for Chlamydiae do not accurately predict the

ability of

> antimicrobial agents to successfully and totally eradicate

chronic

> chlamydial infections. This is because the current susceptibility

> testing methods measure only replication of chlamydia and ignores

the

> well-known " cryptic phase " in which intracellular Chlamydiae are

not

> actively replicating. Moreover, it has also been discovered that

the

> so-called " cryptic phase " of Chlamydiae includes multiple and

> different sub-phases. The following are some of the phases of the

> chlamydial life cycle in which the intracellular Chlamydiae are

not

> replicating: an initial intranuclear phase in which elementary

bodies

> (EBs) transition to reticulate bodies (RBs), an intracytoplasmic

phase

> in which there is a transition of the RB phenotype to the EB

> phenotype, an intracytoplasmic phase with a nonreplicating, but

> metabolizing RB, and intracellular/extracellular EB phases,

including

> endocytotic and exocytotic phases, in which there is neither

> replication nor metabolism. In order to assess the cumulative and

long

> term effect of antimicrobial therapy on these multiple life

phases,

> unique in vitro and in vivo susceptibility test methods have been

> developed and are described herein.

>

>

> The subject invention also pertains to a method for treating

porphyria

> caused by Chlamydia in an individual in need thereof, comprising

> reducing the levels of active stage, latent stage and elementary

> bodies of the pathogen from the individual and administering one

or

> more compounds which reduce adverse effects associated with

secondary

> porphyria. In one embodiment, the method additionally comprises

> administering a compound which reduces the adverse effects of

> porphyrins associated with porphyria. In a particular embodiment,

the

> compound is cimetidine. This method can also be valuably combined

with

> additional steps, including at least one of administering

> antioxidants; orally administering activated charcoal;

administering a

> high carbohydrate diet regimen; administering hydroxychloroquine;

> administering benzodiazepine drugs; performing hemodialysis;

> performing plasmaphoresis; and administering chelating agents; and

> administering intravenous hematin.

>

> The invention also pertains to a method of detecting elevated

porphyrin

> levels in an individual by testing that individual for

antibodies to

> porphyrins. The invention also pertains to diagnosing deficiency

by

> detecting antibodies to B-12. Monoclonal and polyclonal antibodies

to

> prophyrins and/or Vitamin B12 can be produced.

>

> This method is unique because it measures the complete

eradication of

> all life forms of chlamydiae in known murine target organs for

> chlamydial infection. This in vivo susceptibility method has

revealed,

> for example, that antimicrobial therapy with the triple agents,

INH,

> metronidazole and penicillamine, can completely eradicate C.

pneumoniae

> from infected mice in four months. Moreover, following complete

> eradication of chlamydiae, multiple attempts to reinfect these

cured

> mice via intranasal inoculation have proven unsuccessful. This

> suggests that effective management and complete [eradiaction]

> eradication results in the development of protective immunity,

and

> that effective management is therefore a way to create effective

> immunity.

>

> Examples of suitable nitroimidazoles include, but are not limited

to,

> metronidazole, tinidazole, bamnidazole, benznidazole,

flunidazole,

> ipronidazole, misonidazole, moxnidazole, ronidazole, sulnidazole,

and

> their metabolites, analogs and derivatives thereof. Metronidazole

is

> most preferred. Examples of nitrofurans that can be used include,

but

> are not limited to, nitrofurantoin, nitrofurazone, nifurtimox,

> nifuratel, nifuradene, nifurdazil, nifurpirinol, nifuratrone,

> furazolidone, and their metabolites, analogs and derivatives

thereof.

> Nitrofurantoin is preferred within the class of nitrofurans.

>

> INH and its congeners can be used to clear infection from

monocytes

> and/or macrophages. When monocytes and macrophages are infected

by

> Chlamydia, they become debilitated and cannot properly or

effectively

> fight infection. It is believed that, if the chlamydial

infection, per

> se, is cleared from these cells, then the monocytes and

macrophages

> can resume their critical roles fighting chlamydial or other

> infection(s). Thus, patient responsiveness to combination therapy

can

> be optimized by the inclusion of isonicotinic acid congeners.

> Accordingly, one aspect of the invention provides a specific

method

> for reempowering monocytes or macrophages that have been

compromised

> by a Chlamydia infection and, in turn, comprise treating the

infection

> in other sites. Such compromised macrophages or monocytes can be

> activated by treating the chlamydial infection by contacting the

> infected macrophages and/or monocytes with an antichlamydial

agent.

>

>

> Cells to be treated can already be cryptically infected or they

can be

> subjected to stringent metabolic or environmental conditions which

> cause or induce the replicating phase to enter the cryptic phase.

Such

> stringent conditions can include changing environmental/culturing

> conditions in the instance where the infected cells are exposed

to

> .gamma.-interferon; or by exposing cells to conventional

antimicrobial

> agents (such as macrolides and tetracyclines) which induce this

> cryptic phase of chlamydial infection in human host cells.

>

> Therapy Directed Toward Elementary Bodies of Chlamydia

>

> As discussed above, it has been discovered that adverse

conditions,

> such as limited nutrients, antimicrobial agents, and the host

immune

> response, produce a stringent response in Chlamydia. Such adverse

> conditions are known to induce stringent responses in other

> microorganisms (C. W. Stratton, In: Antibiotics in Laboratory

> Medicine, Fourth Edition. Lorian V (ed) & Wilkins,

Baltimore,

> pp 579-603 (1996)) and not surprisingly induce a stringent

response in

> Chlamydia. This stringent response in Chlamydia alters the

> morphological state of the intracellular microorganism and

creates

> dormant forms, including the intracellular EB, which then can

> cryptically persist until its developmental cycle is reactivated.

> Conversely, the host cell may lyse and allow the EBs to reach the

> extracellular milieu. Thus, it is necessary to utilize a

combination

> of agents directed toward the various life stages of Chlamydia

and, in

> particular, against the elementary body for successful management

of

> infection.

>

> As described in previous sections, it is also believed that

> [persistance] persistence of chlamydial infections, in part, may

be

> due to the presence of cryptic forms of Chlamydia within the

cells.

> This cryptic intracellular chlamydial form apparently can be

activated

> by certain host factors such as cortisone (Yang et al., Infection

and

> Immunity, 39:655-658 (1983); and Malinverni et al., The Journal

of

> Infectious Diseases, 172:593-594 (1995)). Antichlamydial therapy

for

> chronic Chlamydia infections must be continued until any

intracellular

> EBs or other intracellular cryptic forms have been activated and

> extracellular EBs have infected host cells. This

> reactivation/reinfection by chlamydial EBs clearly is undesirable

as it

> prolongs the therapy of chlamydial infections, as well as

increases

> the opportunity for antimicrobial resistance to occur.

>

> Physiochemical agents have been identified that can inactivate

> chlamydial EBs in their respective hosts by reducing disulfide

bonds

> which maintain the integrity of the outer membrane proteins of

the

> EBs. For Chlamydia, disruption of the outer membrane proteins of

EBs

> thereby initiates the transition of the EB form to the RB form.

When

> this occurs in the acellular milieu where there is no available

energy

> source, the nascent RB perishes or falls victim to the immune

system.

> Thus, disulfide reducing agents that can interfere with this

process

> are suitable as compounds for eliminating EBs.

>

> One such class of disulfide reducing agents are thiol-disulfide

> exchange agents. Examples of these include, but are not limited

to,

> 2,3-dimercaptosuccinic acid (DMSA; also referred to herein as

> " succimer " ); D,L,-.beta.,.beta.-dimethylcysteine (also known as

> penicillamine); .beta.-lactam agents (e.g., penicillins,

penicillin G,

> ampicillin and amoxicillin, which produce penicillamine as a

> degradation product), cycloserine, dithiotreitol,

mercaptoethylamine

> (e.g., mesna, cysteiamine, dimercaptol), N-acetylcysteine,

tiopronin,

> and glutathione. A particularly effective extracellular

antichlamydial

> agent within this class is DMSA which is a chelating agent having

four

> ionizable hydrogens and two highly charged carboxyl groups which

> prevent its relative passage through human cell membranes. DMSA

thus

> remains in the extracellular fluid where it can readily encounter

> extracellular EBs. The two thiol (sulflhydryl) groups on the

succimer

> molecule (DMSA) are able to reduce disulfide bonds in the MOMP of

EBs

> located in the extracellular milieu.

>

>

> As chlamydial RBs transform into EBs, they begin to utilize

active

> transcription of chlamydial DNA and translation of the resulting

mRNA.

> As such, these forms of Chlamydia are susceptible to currently

used

> antimicrobial agents. The antichlamydial effectiveness of these

agents

> can be significantly improved by using them in combination with

other

> agents directed at different stages of Chlamydia life cycle, as

> discussed herein.

>

> Classes of suitable antimicrobial agents include, but are not

limited

> to, rifamycins (also known as ansamacrolides), quinolones,

> fluoroquinolones, chloramphenicol, sulfonamides/sulfides,

azalides,

> cycloserine, macrolides and tetracyclines. Examples of these

agents

> which are members of these classes, as well as those which are

> preferred, are illustrated below in Table 5.

>

> Agents Effective Against the Replicating

> Phase of Chlamydia

> Drug Class Examples Preferred

> Quinolones/ Ofloxacin Levofloxacin

> Fluoroquinolones Levofloxacin

> Trovafloxacin

> Sparfloxacin

> Norfloxacin

> Lomefloxacin

> Cinoxacin

> Enoxacin

> Nalidixic Acid

> Fleroxacin

> Ciprofloxacin

> Sulfonamides Sulfamethoxazole Sulfamethoxazole/

> Trimethoprim

> Azalides Azithromycin Azithromycin

> Macrolides Erythromycin Clarithromycin

> Clarithromycin

> Lincosamides Lincomycin

> Clindamycin

> Tetracyclines Tetracycline Minocycline

> Doxycycline

> Minocycline

> Methacycline

> Oxytetracyline

> Rifamycins Rifampin Rifampin

> (Ansamacrolides) Rifabutin

>

>

>

> All members of the Chlamydia species, including C. pneumoniae,

are

> considered to be inhibited, and some killed, by the use of a

single

> agent selected from currently used antimicrobial agents such as

those

> described above. However, using the new susceptability test, the

> inventors have found complete eradication of Chlamydia cannot be

> achieved by the use of any one of these agents alone because none

are

> efficacious against all phases of the Chlamydia life cycle and

appear

> to induce a stringent response in Chlamydia causing the

replicating

> phase to transform into cryptic forms. This results in a

persistent

> infection in vivo or in vitro that can be demonstrated by PCR

> techniques which assess the presence or absence of chlamydial

DNA.

> Nevertheless, one or more of these currently used agents, or a

new

> agent directed against the replicating phase of Chlamydia, should

be

> included as one of the chlamydial agents in a combination therapy

in

> order to slow or halt the transition of the EB to the RB as well

as to

> inhibit chlamydial replication.

>

> Methodology for Selecting Potential Agent Combinations

>

> In attempting to manage or eradicate a systemic infection, it is

> critical to target multiple phases in the life cycle of

Chlamydia,

> otherwise viable Chlamydia in the untargeted phases will remain

after

> therapy and result in continued, chronic infection. This

fundamental

> insight is at the core of this invention.

>

> A preferred method for selecting an appropriate combination of

agents

> that satisfies the requirements of this strategy comprises a

plurality

> of steps as follows:

>

> 1. Identify the phases of the chlamydial life cycle. For example,

the

> following phases are currently known:

>

> a. Elementary Body ( " EB " )--Extracellular or Intracellular.

> Intracellular EBs may represent a type of " cryptic phase " .

>

> b. EB to Reticulate Body ( " RB " ) transition phase.

>

> c. Stationary RB phase. This is what is traditionally thought of

as the

> " cryptic phase " .

>

> d. Replicating RB phase.

>

> e. RB to EB transition phase (also called " condensation " ).

>

>

> 2. Evaluate the relative importance of targeting each particular

phase

> in eradicating reservoirs of Chlamydia from the host organism.

For

> example, the life-cycle stages listed in step 1 can be

prioritized

> based on the following assumptions:

>

> a. In the host, [Extracellular] extracellular and intracellular

EBs

> represent a very important reservoir of infectious agents that

result

> in chronic and persistent infection.

>

> b. Most intracellular RBs in chronic infections are non-

replicating.

> The 3-4 day reproduction cycle seen in cycloheximide-treated

> eukaryotic cells is an artifact of an atypical, cell culture

> environment designed primarily to propagate Chlamydia.

>

> c. The transition phases represent only a small portion of

Chlamydia in

> chronic infections.

>

> 3. Identify " targets " for each phase of the selected life cycle

phases.

> A target is an attribute of Chlamydia which is vulnerable during

a

> particular life cycle phase. For example, the disulfide bonds in

MOMP

> are a target during the EB phase.

>

> 4. Identify agents with known or theoretical mechanism(s) of

action

> against those targets.

>

> 5. Estimate whether those agents would be merely inhibitory or,

> preferably, cidal, through an understanding of their mechanism of

> action.

>

> 6. Confirm the estimate by using the following approaches:

>

> a. In the case of anti-EB agents, treat EBs with the agent, then

> attempt to infect cells with the treated EBs. If the cells do not

> become infected, the agent is EB-cidal.

>

> b. In the case of other agents, use the susceptibility tests

disclosed

> elsewhere herein, to determine whether the agent, either alone or

in

> combination with other agents, is chlamydicidal.

>

> 7. Select a combination of agents that, through their individual

> effects, provide activity against targets for the most important

> phases within the chlamydial life cycle. Preferably, a

combination

> should target as many phases of the life cycle as possible,

seeking to

> maximize the total of the relative important scores of the phases

> targeted while minimizing the number of drugs involved.

>

> 8. Test the combination using the susceptibility testing

procedures

> described elsewhere. This step is necessary because the selected

> combination may or may not be chlamydicidal for various reasons

such as

> intracellular penetration and/or efflux.

>

> 9. Set initial dosages based on clinical standards which consider

the

> pharmacokenetics and pharmacodynamics for the drugs prescribed

> individually; modifications, if needed, are based on results of

> susceptibility testing and in vivo efficacy.

>

> Table 6 provides an example of how the foregoing methodology can

be

> used. The preferred embodiment includes agents which:

>

> a) Target disulfide bonds in the EB and condensation phases;

>

> Target non-oxidative metabolism in the stationary/cryptic phase;

>

> c) Target constitutive production of peroxidases and catalyses in

the

> stationary and replicative phases;

>

> d) In the latter two cases, work through physio-chemical

disruption of

> the organism through free radicals, which are very difficult for

> organism to develop resistance to; and

>

> e) Optionally adds an agent to target DNA-dependent RNA polymerase

in

> the EB->RB phase.

>

> The foregoing methodology for selecting combination therapies can

be

> automated (e.g., by a computer system) at one or a combination of

the

> steps described above. This methodology is applicable even after

> greater understanding of the chlamydial life cycle leads to a

> re-prioritization or even sub-division of the life-cycle phases,

new

> theoretical targets within Chlamydia are identified, or new drugs

are

> developed which attack currently known or new targets within

> Chlamydia. For example, the phases of the life cycle could be

further

> sub-classified based on the type of host cell the phase is in.

Thus,

> stationary phase RBs in macrophages could be considered a

separate

> phase than stationary phase RBs in hepatocytes. This allows the

> methodology to be used to design a single or multi-tissue

specific

> combination of agents.

>

>

> Vitamin C (2 gms bid) has also been introduced based on the report

that

> Vitamin C (ascorbic acid) at moderate intracellular

concentrations

> stimulates replication of C. trachomatis (Wang et al., J. Clin.

Micro.

> 30:2551-2554 (1992)) as well as its potential effect on biofilm

charge

> and infectivity of the bacterium and specifically the EB

(Hancock, R.

> E. W., Annual Review in Microbiology, 38:237-264 (1984)).

>

>

> Diagnosis and Treatment of Secondary Porphyria

>

> Chlamydia is a parasite of normal energy production in infected

> eukaryotic cells. As a result, host cells have insufficient

energy

> available for their normal functioning. The energy shortage also

> causes the host cell mitochondria to attempt to synthesize

certain

> critical enzymes involved in energy production in order to

increase

> energy production. Because Chlamydia also prevents this synthesis

from

> completing, these enzyme's precursors, called porphyrins, build

up in

> cell and often escape into the intracellular [mileau] milieu.

> Porphyrins readily form free-radicals, that, in turn, damage

cells.

> Thus, there is an obligate secondary porphyria that accompanies

many

> chlamydial infections. Therapy for this secondary porphyria,

which is

> adjunct to anti-chlamydial therapy, involves at least three

> strategies: a) supplement the cellular energy supply to mitigate

cell

> malfunction and the formation of porphyrins; reduce the levels

of

> systemic porphyrins; and c) mitigate the harmful effects of the

> porphyrins.

>

> The pathogenesis of chronic/systemic chlamydial infection is

unique in

> that the intracellular infection by this parasite results in a

number

> of heretofore unrecognized concomitant and obligatory

> metabolic/autoimmune disorders including secondary porphyria with

> associated autoantibodies against the porphyrins. Cross reaction

with

> Vitamin B12 can result in a subclinical autoimmune-mediated

Vitamin

> B12 deficiency. These associated disorders often require

diagnosis and

> preventive and/or specific adjunctive therapy.

>

>

> The inventors have discovered the existence of antibodies to the

> various metabolites of heme biosynthesis, as well as Vitamin B12

> (cobalamin), which is molecularly similar to these metabolites,

in

> patients with active systemic infection with C. pneumoniae. The

> antibodies are primarily IgM; this is similar to the antibody

> responses to the MOMP of C. pneumoniae in severely symptomatic

> patients. Example 8 illustrates titers in symptomatic patients

with

> systemic C. pneumoniae infections. The presence of antibodies to

> Vitamin B12 may have functional significance by decreasing the

amount

> of bioavailable Vitamin B12. Thus, a Chlamydia infection may

cause a

> previously unrecognized secondary Vitamin B12 deficiency.

> Administration (e.g., intramuscular) of large quantities of

Vitamin B12

> (1000 to 5000 .mu.g) (e.g., parenteral cobalamin therapy)

creates

> large amounts of Vitamin B12 available for binding to the native

> receptors of antibodies with an affinity for Vitamin B12, thereby

> saturating these anti-Vitamin B12 antibodies and increasing the

amount

> of bioavailable circulating Vitamin B12.

>

>

> Glucose is an important source of cellular energy. Glucose levels

can

> be enhanced by diet and through vitamin supplements as described

> below.

>

> A high carbohydrate diet should be maintained to promote

production of

> glucose (Pierach et al., Journal of the American Medical

Association,

> 257:60-61 (1987)). Approximately 70% of the caloric intake should

be in

> the form of complex carbohydrates such as bread, potato, rice

and

> pasta. The remaining 30% of the daily diet should comprise

protein and

> fat, which should ideally be in the form of fish or chicken. Red

> meats, including beef, dark turkey, tuna and salmon, contain

> [tryprophan] tryptophan. Increased levels of tryptophan in the

liver

> inhibit the activity of phosphoenol pyruvate carboxykinase with

> consequent disruption of gluconeogenesis. This accounts for the

> abnormal glucose tolerance seen in porphyria. Increased plasmic

> concentrations of tryptophan also enhances tryptophan transport

into

> the brain. The concentration of tryptophan in the brain is the

> rate-limiting factor for the synthesis of the neurotransmitter

> 5-hydroxytryptamine (5-HT, serotonin). Serotonin is synthesized

by the

> endothelium of brain capillaries for circulating tryptophan.

Thus,

> increased concentrations of tryptophan in the brain would be

expected

> to enhance production of serotonin and its metabolic,

> 5-hydroxyindole-acetic acid (5HIAA). Acute increases in serotonin

> turnover in the brain are followed by vascular and metabolic

changes

> which include decreases in glucose consumption, disturbances in

EEG

> tracings, and decreases in the postischemic neurological score.

In

> addition, while serotonin increases brain perfusion on a single

> injection, repetitive administration initially opens the blood-

brain

> barrier and subsequently induces vasoconstriction. It is likely

that

> any transient opening of the blood-brain barrier by serotonin

could

> allow circulating substrates such as ALA and PBG, if present, to

enter

> the central nervous system. As would be expected from the

location of

> serotonin receptors and from the barrier function of the

endothelium

> of cerebral arteries, the constricting effect of serotonin is

> amplified in cerebral arteries where endothelium is damage or

removed.

> Damaged endothelial cells, as would be expected with chlamydial

> infection, would no longer have operational catabolic processes

for

> serotonin. This would be particularly true in the event of

depleted

> ATP as caused by chlamydial infection. This means that increased

> concentrations of serotonin will reach the smooth muscle layer of

the

> cerebral vessels and cause more constriction. Finally, serotonin

is

> also stored in blood platelets. Because blood platelets do not

adhere

> and aggregate under normal conditions, they do not release

serotonin

> when the vessel lumen is intact. However, if the vessel lumen is

> altered by chlamydial infection, platelet deposition and release

of

> serotonin can occur.

>

> Another adverse effect of increased serotonin levels due to

porphyria

> is seen with nervous tissues. Sympathetic nerve endings store

> serotonin taken up from the circulation. These serotonergic

neurons

> form plexuses around brain vessels where they are likely to

liberate

> their serotonin contents when subjected to cellular lysis from

any

> cause including ischemia, free-radical ionizing damage to cell

> membranes, and/or chlamydial infection.

>

> In rats, elevated circulating tryptophan has been shown to

produce

> structural alteration of brain astrocytes, oligodendroglia, and

> neurons, as well as degeneration of Purkinje cells and wasting of

> axons. Similar neurohistological alterations have been reported

in

> patients with acute porphyria. Elevated tryptophan levels in

plasma

> and brain have been associated with human encepholopathy.

Finally,

> serotonin is also recognized as an active neurotransmitter in the

> gastrointestinal tract. The pharmacologic effects of serotonin in

the

> central nervous system and gastrointestinal tract resemble the

> neurological manifestations of acute porphyric attacks. In fact,

> administration of either tryptophan or serotonin to humans have

been

> reported to cause severe abdominal pain, psychomotor

disturbances,

> nausea, and dysuria; all of which are symptoms of acute porphyria.

>

> Sucrose and fructose should be avoided (Bottomly et al., American

> Journal of Clinical Pathology, 76:133-139 (1981)) because the

> ingestion of large amounts of fructose trigger hepatic

gluconeogenesis

> which then decreases the available glucose which is derived from

> glycogen breakdown within the liver. It is recommended that sport

> drinks which contain glucose be consumed.

>

> It is recommended that a patient suffering from porphyria avoid

milk

> products. Milk products contain lactose and lactoferrin, and have

been

> empirically shown to make symptoms of porphyria worse.

>

> Multivitamins containing the B complex vitamins should be

administered

> daily (e.g., one or multiple times), preferably in excess of RDA,

to

> enhance glucose availability. Hepatic breakdown of glycogen with

> generation of glucose is assisted by taking these multivitamins

that

> contain the B complex vitamins. Pyridoxine minimizes the porphyrin

> related porphyrial neuropathy. B complex vitamins include folic

acid

> (e.g., 400 .mu.g per dosage; 1200 .mu.g daily maximum); vitamin B-

1

> (thiamin; e.g., 10 mg per dosage; 30 mg daily maximum); B-2

> (riboflavin; e.g., 10 mg per dosage; 30 mg daily maximum); B-5

> (panothenate; e.g., 100 mg per dosage; 300 mg daily maximum); B-6

> (pyridoxine; e.g., 100 mg per dosage; 300 mg daily maximum) or

> pyridoxal-5-phosphate (e.g., 25 mg per dosage; 100 mg daily

maximum)

> and B-12 (e.g., 500 .mu.g per dosage; 10,000 .mu.g daily

maximum). The

> preferred method of administration is oral for the majority of

these

> vitamins (twice daily), except for B-12 for which sublingual

> administration (three-times daily) is preferred. It has been

discovered

> that one important effect of this secondary porphyria in some

patients

> is the production of IgM and IgG antibodies against

> coproporphyrinogen-III. These antibodies cross-react with Vitamin

B12

> (cobalamin) and can thus cause a deficiency. Vitamin B12

> supplementation (e.g., parenteral cobalamin therapy) can remedy

the

> deficiency.

>

> D. Reducing Porphyrin Levels

>

> Dietary and pharmaceutical methods can be used to reduce systemic

> porphyrin levels (both water-soluble and fat-soluble).

>

> Plenty of oral fluids in the form of bicarbonated water or " sports

> drinks " (i.e., water with glucose and salts) should be

incorporated

> into the regimen. This flushes water-soluble porphyrins from the

> patient's system. Drinking seltzer water is the easiest way to

achieve

> this goal. The color of the urine should always be almost clear

> instead of yellow. It is noted that dehydration concentrates

> prophyrins and makes patients more symptomatic.

>

>

> Activated charcoal can be daily administered in an amount

sufficient to

> absorb fat-soluble porphyrins from the enterohepatic

circulation.

> Treatment with activated oral is charcoal, which is nonabsorbable

and

> binds porphyrins in the gastrointestinal tract and hence

interrupts

> their enterohepatic circulation, has been associated with a

decrease

> of plasma and skin porphyrin levels. Charcoal should be taken

between

> meals and without any other oral drugs or the charcoal will

absorb the

> food or drugs rather than the porphyrins. For those who have

> difficulty taking the charcoal due to other medications being

taken

> during the day, the charcoal can be taken all at one time before

bed.

> Taking between 2 and 20 grams, preferably at least 6 grams

> (24.times.250 mg capsules) of activated charcoal per day

(Perlroth et

> al., Metabolism, 17:571-581 (1968)) is recommended. Much more

charcoal

> can be safely taken; up to 20 grams six times a day for nine

months has

> been taken without any side effects.

>

>

> Antioxidants at high dosages (preferably taken twice per day) help

to

> mitigate the effects of free radicals produced by porphyrins.

Examples

> of suitable antioxidants include but are not limited to Vitamin C

> (e.g., 1 gram per dosage; 10 g daily maximum); Vitamin E (e.g.,

400

> units per dosage; 3000 daily maximum); L-Carnitine (e.g., 500 mg

per

> dosage; 3 g daily maximum); coenzyme Q-10 (uniquinone (e.g., 30

mg per

> dosage; 200 mg daily maximum); biotin (e.g., 5 mg per dosage; 20

mg

> daily maximum); lipoic acid (e.g., 400 mg per dosage; 1 g daily

> maximum); selenium (e.g., 100 .mu.g per dosage; 300 .mu.g daily

> maximum); gultamine (e.g., from 2 to about 4 g per dosage);

> glucosamine (e.g., from about 750 to about 1000 mg per dosage);

and

> chondroitin sulfate (e.g., from about 250 to about 500 mg per

dosage).

>

>

> The above-mentioned therapeutic diets can be combined with

traditional

> or currently recognized drug therapies for porphyria. In one

> embodiment, benzodiazapine drugs, such as but not limited to

valium,

> klonafin, flurazepam hydrochloride (e.g., Dalmanc.TM., Roche) and

> alprazolam (e.g., Xanax), can be administered. Preferably,

sedatives,

> such as alprazolam (e.g., Xanax; 0.5 mg per dosage for 3 to 4

times

> daily), can be prescribed for panic attacks and flurazepam

> hydrochloride (e.g., Dalmane.TM., Roche or Restoril.TM. (e.g., 30

mg

> per dosage)) can be prescribed for sleeping. The rationale is

based

> upon the presence of peripheral benzodiazepine receptors in high

> quantities in phagocytic cells known to produce high levels of

radical

> oxygen species. A protective role against hydrogen peroxide has

been

> demonstrated for peripheral benzodiazipine receptors. This

suggests

> that these receptors may prevent mitochondria from radical

damages and

> thereby regulate apoptosis in the hematopoietic system.

> Benzodiazepines have also been shown to interfere with the

> intracellular circulation of heme and porphyrinogens (Scholnick

et

> al., Journal of Investigative Dermatology, 1973, 61:226-232).

This is

> likely to decrease porphyrins and their adverse effects. The

specific

> benzodiazipine will depend on the porphyrin-related symptoms.

>

>

>

> Table 12 provides the results of an expanded study of

antimicrobial

> susceptibilities at two different concentrations of antimicrobial

> agents, used alone and in combination, when exposed to the

> antimicrobial agents for two weeks. In addition to the agents

already

> mentioned, minocycline, doxycycline, rifampin and

> sulfamethoxizole/trimethoprim, at all concentrations tested,

failed to

> clear the PCR signal for chlamydial MOMP. Only the triple

combination

> of isoniazid, metronidazole and penicillamine cleared the PCR

signal

> in two weeks. The triple combination was effective at both low

and

> high concentrations. Table 12 also demonstrates the effect of a 4

week

> exposure with the same expanded series of antimicrobial agents

alone

> and in combination. A number of triple combinations of

antimicrobial

> agents resulted in cell cultures in which the PCR signal for the

> chlamydial MOMP gene could not be detected at four weeks. The

most

> effective combinations to have the greatest impact on the

important

> life cycle phases are those predicted according to the

methodology

> described in the section above entitled " Methodology for Selecting

> Potential Agent Combinations " .

>

>

> TABLE 11

> Susceptibility to Antibiotics for

> Cryptic C. pneumoniae Cultured in HeLa Cells.sup.a

> Antibiotic Conc (.mu.g/ml) PCR.sup.b

> Ofloxacin 1 positive

> Clarithromycin 1 positive

> INH 1 positive

> Metronidazole 1 positive

> Penicillamine 1/1 positive

> INH + Metronidazole + 1/1/4 negative

> Penicillamine

> Control 0 positive

> .sup.a Cultured in the presence of the indicated antibiotic

(s), but

> with no

> cycloheximide. Media changes at 48-72 hours.

> .sup.b Analysis following 2 weeks exposure to antimicrobioal

agents.

>

>

> TABLE 12

> Susceptibility to

> Antibiotics by PCR

> for

> Cryptic Chlamydia pneumoniae

Cultured

> in HeLa

> Cells.sup.1

> Phase of the Chlamydial Life Cycle

> EB (Extracellular EB->RB Stationary Phase RB RB-

>EB

> Transition Concentration PCR PCR

> or Intracellular) Transition ( " Cryptic phase " ) Replicating RB

> ( " Condensation " ) (.mu.g/ml) 2 week 4 week Comment

> 0 + + Control

> Minocycline

> 0.25 + + One drug

> Minocycline

> 1 + + One drug

> Doxycycline

> 0.25 + + One drug

> Doxycycline

> 1 + + One drug

> TMP/SMZ.sup.2

> 25 + + One drug

> TMP/SMZ.sup.2

> 100 + + One drug

> Clarithromycin

> 0.25.sup.1 + + One drug

> Ofloxacin

> 0.25 + + One drug

> Metronidazole

> 0.25 + + One drug

> Rifampin

> 0.25 + + One drug

> Isoniazide Isoniazide

> 1 + + Misses EB phase

> Metronidazole TMP/SMZ.sup.1

> 25/0.25 + + Misses EB phase

> Metronidazole Ofloxacin

> 0.25/0.25 + + Misses EB phase

> Rifampin Metronidazole

> 0.25/0.25 + + Misses EB phase

> penicillamine Rifampin Metronidazole

> penicillamine

> 4/.25/.25 + + Misses replicating

> phase

> Rifampin Metronidazole Ofloxacin

> .25/.25/.25 + + Misses EB phase

> penicillamine Metronidazole Doxycyclin

> penicillamine

> 1/.25/.25 + + Concentration too low

> penicillamine Metronidazole Doxycyclin

> penicillamine

> 4/1/1 + - Covers key phases

> penicillamine Metronidazole Minocycline

> penicillamine

> 1/.25/.25 + - Covers key phases

> penicillamine Metronidazole Minocycline

> penicillamine

> 4/1/1 + - Original report of 4

> week positive was a

> typo

> penicillamine Metronidazole TMP/SMZ

> penicillamine

> 4/1/100 + - Covers key phases

> penicillamine Metronidazole Clarithromycin

> penicillamine

> 1/.25/.25 + - Covers key phases

> penicillamine Isoniazid Isoniazid

> penicillamine

> 1/.25/.25 - - Covers key phases

> Metronidazole

> penicillamine Isoniazid Isoniazid

> penicillamine

> 4/1/1 - - Covers key phases

> Metronidazole

> .sup.1 Cultured in the presence of the indicated antibiotics,

but

> with no

> cycloheximide. Media changes at 48-72 hours. + = positive; -

=

> negative

> .sup.2 TMP/SMX = trimethoprim/sulfamethoxazole

>

>

>

>

>

>

> >

> >

[Guest guest]

yes I think cnp is one of my major players in crime too. I have a few

pockets left that I need to clear, and the flagyl has always been the

likely player but I find it really difficult to tolerate,

>

> I found this really fascinating. I have been taking Doxy on and off

> for ten months for persistent Chlamydia infections. Chlamydia is

> nasty. It is known to cause chronic bronchitis and heart disease.

> I never have a cough when I take Doxy. Herxes are rough.

> Taking Flagyl now for ten days (2nd course). Mild herx happening.

> I believe that Chlamydia is at the root of my CFS/ME and fibro, even

> possibly the osteo arthritis.

> Bugs, Bugs, everywhere and in every form, it seems. The trick is to

> diagnose who is what, where, when and then kill 'em.

> Cheers,

> *Sue*

>

>

>

> > >

> > >  Hi:

> > >

> > >  Can anyone post their experiences with either of these

> protocols. 

> > >  What does each protocol entail.

> > >

> > >  Searching for some relief.

> > >

> > >  Thanks,

> >

> >

> > Diseases Associated with Chlamydial Infection

> >

> > An association has been discovered between chronic Chlamydia

> infection

> > of  body fluids and/or tissues with several disease syndromes of

> > previously unknown etiology in humans which respond to unique

> > antichlamydial regimens  described herein. To date, these diseases

> > include Multiple Sclerosis (MS), Rheumatoid Arthritis (RA),

> > Inflammatory Bowel Disease (IBD), Interstitial  Cystitis (IC),

> > Fibromyalgia (FM), Autonomic nervous dysfunction (AND,  neural-

> mediated

> > hypotension); Pyoderma Gangrenosum (PG), Chronic Fatigue  (CF) and

> > Chronic Fatigue Syndrome (CFS). Other diseases are under 

> > investigation. Correlation between Chlamydia infection and these

> > diseases  has only recently been established as a result of the

> > diagnostic  methodologies and combination therapies described

> herein.

> >

> > The therapies described herein can thus be used for the treatment

> of

> > acute  and chronic immune and autoimmune diseases when patients

> are

> > demonstrated  to have a Chlamydia load by the diagnostic

> procedures

> > described herein  which diseases include, but are not limited to,

> > chronic hepatitis,  systemic lupus erythematosus, arthritis,

> > thyroidosis, scleroderma,  diabetes mellitus, Graves' disease,

> > Beschet's disease and graft versus  host disease (graft

> rejection). The

> > therapies of this invention can also  be used to treat any

> disorders in

> > which a chlamydial species is a factor  or co-factor.

> >

> > Among the various inflammatory diseases, there are certain

> features of

> > the  inflammatory process that are generally agreed to be

> > characteristic. These  include fenestration of the

> microvasculature,

> > leakage of the elements of  blood into the interstitial spaces,

> and

> > migration of leukocytes into the  inflamed tissue. On a

> macroscopic

> > level, this is usually accompanied by  the familiar clinical signs

> of

> > erythema, edema, tenderness (hyperalgesia),  and pain.

> Inflammatory

> > diseases, such as chronic inflammatory pathologies  and vascular

> > inflammatory pathologies, including chronic inflammatory 

> pathologies

> > such as aneurysms, hemorrhoids, sarcoidosis, chronic  inflammatory

> > bowel disease, ulcerative colitis, and Crohn's disease and 

> vascular

> > inflammatory pathologies, such as, but not limited to, 

> disseminated

> > intravascular coagulation, atherosclerosis, and Kawasaki's 

> pathology

> > are also suitable for treatment by methods described herein. The

> > invention can also be used to treat inflammatory diseases such as

> > coronary  artery disease, hypertension, stroke, asthma, chronic

> > hepatitis, multiple  sclerosis, peripheral neuropathy, chronic or

> > recurrent sore throat,  laryngitis, tracheobronchitis, chronic

> vascular

> > headaches (including  migraines, cluster headaches and tension

> > headaches) and pneumonia when  demonstrated to be pathogenically

> > related to Chlamydia infection.

> >

> >

> > The invention is based upon the discovery that current

> susceptibility 

> > testing methods for Chlamydiae do not accurately predict the

> ability of

> >   antimicrobial agents to successfully and totally eradicate

> chronic 

> > chlamydial infections. This is because the current susceptibility

> > testing  methods measure only replication of chlamydia and ignores

> the

> > well-known  " cryptic phase " in which intracellular Chlamydiae are

> not

> > actively  replicating. Moreover, it has also been discovered that

> the

> > so-called  " cryptic phase " of Chlamydiae includes multiple and

> > different sub-phases.  The following are some of the phases of the

> > chlamydial life cycle in which  the intracellular Chlamydiae are

> not

> > replicating: an initial intranuclear  phase in which elementary

> bodies

> > (EBs) transition to reticulate bodies  (RBs), an intracytoplasmic

> phase

> > in which there is a transition of the RB  phenotype to the EB

> > phenotype, an intracytoplasmic phase with a  nonreplicating, but

> > metabolizing RB, and intracellular/extracellular EB  phases,

> including

> > endocytotic and exocytotic phases, in which there is  neither

> > replication nor metabolism. In order to assess the cumulative and 

> long

> > term effect of antimicrobial therapy on these multiple life

> phases, 

> > unique in vitro and in vivo susceptibility test methods have been 

> > developed and are described herein.

> >

> >

> > The subject invention also pertains to a method for treating

> porphyria 

> > caused by Chlamydia in an individual in need thereof, comprising

> > reducing  the levels of active stage, latent stage and elementary

> > bodies of the  pathogen from the individual and administering one

> or

> > more compounds which  reduce adverse effects associated with

> secondary

> > porphyria. In one  embodiment, the method additionally comprises

> > administering a compound  which reduces the adverse effects of

> > porphyrins associated with porphyria.  In a particular embodiment,

> the

> > compound is cimetidine. This method can  also be valuably combined

> with

> > additional steps, including at least one of  administering

> > antioxidants; orally administering activated charcoal; 

> administering a

> > high carbohydrate diet regimen; administering  hydroxychloroquine;

> > administering benzodiazepine drugs; performing  hemodialysis;

> > performing plasmaphoresis; and administering chelating agents; and

> > administering intravenous hematin.

> >

> > The invention also pertains to a method of detecting elevated

> porphyrin

> >   levels in an individual by testing that individual for

> antibodies to 

> > porphyrins. The invention also pertains to diagnosing deficiency

> by 

> > detecting antibodies to B-12. Monoclonal and polyclonal antibodies

> to 

> > prophyrins and/or Vitamin B12 can be produced.

> >

> >   This method is unique because it measures the complete 

> eradication of

> > all life forms of chlamydiae in known murine target organs  for

> > chlamydial infection. This in vivo susceptibility method has

> revealed, 

> > for example, that antimicrobial therapy with the triple agents,

> INH, 

> > metronidazole and penicillamine, can completely eradicate C.

> pneumoniae

> >   from infected mice in four months. Moreover, following complete 

> > eradication of chlamydiae, multiple attempts to reinfect these

> cured

> > mice  via intranasal inoculation have proven unsuccessful. This

> > suggests that  effective management and complete [eradiaction]

> > eradication results in the  development of protective immunity,

> and

> > that effective management is  therefore a way to create effective

> > immunity.

> >

> > Examples of suitable nitroimidazoles include, but are not limited

> to, 

> > metronidazole, tinidazole, bamnidazole, benznidazole,

> flunidazole, 

> > ipronidazole, misonidazole, moxnidazole, ronidazole, sulnidazole,

> and 

> > their metabolites, analogs and derivatives thereof. Metronidazole

> is

> > most  preferred. Examples of nitrofurans that can be used include,

> but

> > are not  limited to, nitrofurantoin, nitrofurazone, nifurtimox,

> > nifuratel,  nifuradene, nifurdazil, nifurpirinol, nifuratrone,

> > furazolidone, and their  metabolites, analogs and derivatives

> thereof.

> > Nitrofurantoin is preferred  within the class of nitrofurans.

> >

> > INH and its congeners can be used to clear infection from

> monocytes

> > and/or  macrophages. When monocytes and macrophages are infected

> by

> > Chlamydia, they become debilitated and cannot properly or

> effectively

> > fight  infection. It is believed that, if the chlamydial

> infection, per

> > se, is  cleared from these cells, then the monocytes and

> macrophages

> > can resume  their critical roles fighting chlamydial or other

> > infection(s). Thus,  patient responsiveness to combination therapy

> can

> > be optimized by the  inclusion of isonicotinic acid congeners.

> > Accordingly, one aspect of the  invention provides a specific

> method

> > for reempowering monocytes or  macrophages that have been

> compromised

> > by a Chlamydia infection and, in  turn, comprise treating the

> infection

> > in other sites. Such compromised  macrophages or monocytes can be

> > activated by treating the chlamydial  infection by contacting the

> > infected macrophages and/or monocytes with an  antichlamydial

> agent.

> >

> >

> > Cells to be treated can already be cryptically infected or they

> can be 

> > subjected to stringent metabolic or environmental conditions which

> > cause  or induce the replicating phase to enter the cryptic phase.

> Such

> > stringent  conditions can include changing environmental/culturing

> > conditions in the  instance where the infected cells are exposed

> to

> > .gamma.-interferon; or by  exposing cells to conventional

> antimicrobial

> > agents (such as macrolides  and tetracyclines) which induce this

> > cryptic phase of chlamydial infection  in human host cells.

> >

> > Therapy Directed Toward Elementary Bodies of Chlamydia

> >

> > As discussed above, it has been discovered that adverse

> conditions,

> > such as  limited nutrients, antimicrobial agents, and the host

> immune

> > response,  produce a stringent response in Chlamydia. Such adverse

> > conditions are  known to induce stringent responses in other

> > microorganisms (C. W.  Stratton, In: Antibiotics in Laboratory

> > Medicine, Fourth Edition. Lorian V  (ed) & Wilkins,

> Baltimore,

> > pp 579-603 (1996)) and not  surprisingly induce a stringent

> response in

> > Chlamydia. This stringent  response in Chlamydia alters the

> > morphological state of the intracellular  microorganism and

> creates

> > dormant forms, including the intracellular EB,  which then can

> > cryptically persist until its developmental cycle is  reactivated.

> > Conversely, the host cell may lyse and allow the EBs to reach the

> > extracellular milieu. Thus, it is necessary to utilize a

> combination 

> > of agents directed toward the various life stages of Chlamydia

> and, in 

> > particular, against the elementary body for successful management

> of 

> > infection.

> >

> > As described in previous sections, it is also believed that

> > [persistance]  persistence of chlamydial infections, in part, may

> be

> > due to the presence  of cryptic forms of Chlamydia within the

> cells.

> > This cryptic intracellular  chlamydial form apparently can be

> activated

> > by certain host factors such  as cortisone (Yang et al., Infection

> and

> > Immunity, 39:655-658 (1983); and  Malinverni et al., The Journal

> of

> > Infectious Diseases, 172:593-594  (1995)). Antichlamydial therapy

> for

> > chronic Chlamydia infections must be  continued until any

> intracellular

> > EBs or other intracellular cryptic forms  have been activated and

> > extracellular EBs have infected host cells. This 

> > reactivation/reinfection by chlamydial EBs clearly is undesirable

> as it

> >   prolongs the therapy of chlamydial infections, as well as

> increases

> > the  opportunity for antimicrobial resistance to occur.

> >

> > Physiochemical agents have been identified that can inactivate

> > chlamydial  EBs in their respective hosts by reducing disulfide

> bonds

> > which maintain  the integrity of the outer membrane proteins of

> the

> > EBs. For Chlamydia,  disruption of the outer membrane proteins of

> EBs

> > thereby initiates the  transition of the EB form to the RB form.

> When

> > this occurs in the  acellular milieu where there is no available

> energy

> > source, the nascent RB  perishes or falls victim to the immune

> system.

> > Thus, disulfide reducing  agents that can interfere with this

> process

> > are suitable as compounds for  eliminating EBs.

> >

> > One such class of disulfide reducing agents are thiol-disulfide

> > exchange  agents. Examples of these include, but are not limited

> to, 

> > 2,3-dimercaptosuccinic acid (DMSA; also referred to herein as

> > " succimer " );  D,L,-.beta.,.beta.-dimethylcysteine (also known as

> > penicillamine);  .beta.-lactam agents (e.g., penicillins,

> penicillin G,

> > ampicillin and  amoxicillin, which produce penicillamine as a

> > degradation product),  cycloserine, dithiotreitol,

> mercaptoethylamine

> > (e.g., mesna, cysteiamine,  dimercaptol), N-acetylcysteine,

> tiopronin,

> > and glutathione. A particularly  effective extracellular

> antichlamydial

> > agent within this class is DMSA which is a chelating agent having

> four

> > ionizable hydrogens and two highly  charged carboxyl groups which

> > prevent its relative passage through human  cell membranes. DMSA

> thus

> > remains in the extracellular fluid where it can  readily encounter

> > extracellular EBs. The two thiol (sulflhydryl) groups on  the

> succimer

> > molecule (DMSA) are able to reduce disulfide bonds in the  MOMP of

> EBs

> > located in the extracellular milieu.

> >

> >

> > As chlamydial RBs transform into EBs, they begin to utilize

> active 

> > transcription of chlamydial DNA and translation of the resulting

> mRNA.

> > As  such, these forms of Chlamydia are susceptible to currently

> used 

> > antimicrobial agents. The antichlamydial effectiveness of these

> agents

> > can  be significantly improved by using them in combination with

> other

> > agents  directed at different stages of Chlamydia life cycle, as

> > discussed herein.

> >

> > Classes of suitable antimicrobial agents include, but are not

> limited

> > to,  rifamycins (also known as ansamacrolides), quinolones,

> > fluoroquinolones, chloramphenicol, sulfonamides/sulfides,

> azalides,

> > cycloserine, macrolides  and tetracyclines. Examples of these

> agents

> > which are members of these  classes, as well as those which are

> > preferred, are illustrated below in  Table 5.

> >

> >        Agents Effective Against the Replicating

> >                          Phase of Chlamydia

> >          Drug Class         Examples          Preferred

> >          Quinolones/        Ofloxacin         Levofloxacin

> >          Fluoroquinolones   Levofloxacin

> >                             Trovafloxacin

> >                             Sparfloxacin

> >                             Norfloxacin

> >                             Lomefloxacin

> >                             Cinoxacin

> >                             Enoxacin

> >                             Nalidixic Acid

> >                             Fleroxacin

> >                             Ciprofloxacin

> >          Sulfonamides       Sulfamethoxazole  Sulfamethoxazole/

> >                                               Trimethoprim

> >          Azalides           Azithromycin      Azithromycin

> >          Macrolides         Erythromycin      Clarithromycin

> >                             Clarithromycin

> >          Lincosamides       Lincomycin

> >                             Clindamycin

> >          Tetracyclines      Tetracycline      Minocycline

> >                             Doxycycline

> >                             Minocycline

> >                             Methacycline

> >                             Oxytetracyline

> >          Rifamycins         Rifampin          Rifampin

> >          (Ansamacrolides)   Rifabutin

> >

> >

> >

> > All members of the Chlamydia species, including C. pneumoniae,

> are 

> > considered to be inhibited, and some killed, by the use of a

> single

> > agent  selected from currently used antimicrobial agents such as

> those

> > described  above. However, using the new susceptability test, the

> > inventors have  found complete eradication of Chlamydia cannot be

> > achieved by the use of  any one of these agents alone because none

> are

> > efficacious against all  phases of the Chlamydia life cycle and

> appear

> > to induce a stringent  response in Chlamydia causing the

> replicating

> > phase to transform into  cryptic forms. This results in a

> persistent

> > infection in vivo or in vitro  that can be demonstrated by PCR

> > techniques which assess the presence or  absence of chlamydial

> DNA.

> > Nevertheless, one or more of these currently  used agents, or a

> new

> > agent directed against the replicating phase of  Chlamydia, should

> be

> > included as one of the chlamydial agents in a combination therapy

> in

> > order to slow or halt the transition of the EB to  the RB as well

> as to

> > inhibit chlamydial replication.

> >

> > Methodology for Selecting Potential Agent Combinations

> >

> > In attempting to manage or eradicate a systemic infection, it is

> > critical  to target multiple phases in the life cycle of

> Chlamydia,

> > otherwise viable  Chlamydia in the untargeted phases will remain

> after

> > therapy and result in  continued, chronic infection. This

> fundamental

> > insight is at the core of  this invention.

> >

> > A preferred method for selecting an appropriate combination of

> agents

> > that  satisfies the requirements of this strategy comprises a

> plurality

> > of steps  as follows:

> >

> > 1. Identify the phases of the chlamydial life cycle. For example,

> the 

> > following phases are currently known:

> >

> > a. Elementary Body ( " EB " )--Extracellular or Intracellular.

> > Intracellular  EBs may represent a type of " cryptic phase " .

> >

> > b. EB to Reticulate Body ( " RB " ) transition phase.

> >

> > c. Stationary RB phase. This is what is traditionally thought of

> as the

> >   " cryptic phase " .

> >

> > d. Replicating RB phase.

> >

> > e. RB to EB transition phase (also called " condensation " ).

> >

> >

> > 2. Evaluate the relative importance of targeting each particular

> phase

> > in  eradicating reservoirs of Chlamydia from the host organism.

> For

> > example,  the life-cycle stages listed in step 1 can be

> prioritized

> > based on the  following assumptions:

> >

> > a. In the host, [Extracellular] extracellular and intracellular

> EBs 

> > represent a very important reservoir of infectious agents that

> result

> > in  chronic and persistent infection.

> >

> > b. Most intracellular RBs in chronic infections are non-

> replicating.

> > The  3-4 day reproduction cycle seen in cycloheximide-treated

> > eukaryotic cells  is an artifact of an atypical, cell culture

> > environment designed primarily  to propagate Chlamydia.

> >

> > c. The transition phases represent only a small portion of

> Chlamydia in

> >   chronic infections.

> >

> > 3. Identify " targets " for each phase of the selected life cycle

> phases.

> > A  target is an attribute of Chlamydia which is vulnerable during

> a 

> > particular life cycle phase. For example, the disulfide bonds in

> MOMP

> > are  a target during the EB phase.

> >

> > 4. Identify agents with known or theoretical mechanism(s) of

> action

> > against  those targets.

> >

> > 5. Estimate whether those agents would be merely inhibitory or,

> > preferably,  cidal, through an understanding of their mechanism of

> > action.

> >

> > 6. Confirm the estimate by using the following approaches:

> >

> > a. In the case of anti-EB agents, treat EBs with the agent, then

> > attempt to  infect cells with the treated EBs. If the cells do not

> > become infected,  the agent is EB-cidal.

> >

> > b. In the case of other agents, use the susceptibility tests

> disclosed 

> > elsewhere herein, to determine whether the agent, either alone or

> in 

> > combination with other agents, is chlamydicidal.

> >

> > 7. Select a combination of agents that, through their individual

> > effects,  provide activity against targets for the most important

> > phases within the  chlamydial life cycle. Preferably, a

> combination

> > should target as many  phases of the life cycle as possible,

> seeking to

> > maximize the total of the  relative important scores of the phases

> > targeted while minimizing the  number of drugs involved.

> >

> > 8. Test the combination using the susceptibility testing

> procedures 

> > described elsewhere. This step is necessary because the selected 

> > combination may or may not be chlamydicidal for various reasons

> such as

> >   intracellular penetration and/or efflux.

> >

> > 9. Set initial dosages based on clinical standards which consider

> the 

> > pharmacokenetics and pharmacodynamics for the drugs prescribed 

> > individually; modifications, if needed, are based on results of 

> > susceptibility testing and in vivo efficacy.

> >

> > Table 6 provides an example of how the foregoing methodology can

> be

> > used.  The preferred embodiment includes agents which:

> >

> > a) Target disulfide bonds in the EB and condensation phases;

> >

> > Target non-oxidative metabolism in the stationary/cryptic phase;

> >

> > c) Target constitutive production of peroxidases and catalyses in

> the 

> > stationary and replicative phases;

> >

> > d) In the latter two cases, work through physio-chemical

> disruption of

> > the  organism through free radicals, which are very difficult for

> > organism to  develop resistance to; and

> >

> > e) Optionally adds an agent to target DNA-dependent RNA polymerase

> in

> > the  EB->RB phase.

> >

> > The foregoing methodology for selecting combination therapies can

> be 

> > automated (e.g., by a computer system) at one or a combination of

> the 

> > steps described above. This methodology is applicable even after

> > greater  understanding of the chlamydial life cycle leads to a

> > re-prioritization or  even sub-division of the life-cycle phases,

> new

> > theoretical targets within  Chlamydia are identified, or new drugs

> are

> > developed which attack  currently known or new targets within

> > Chlamydia. For example, the phases  of the life cycle could be

> further

> > sub-classified based on the type of  host cell the phase is in.

> Thus,

> > stationary phase RBs in macrophages could  be considered a

> separate

> > phase than stationary phase RBs in hepatocytes.  This allows the

> > methodology to be used to design a single or multi-tissue 

> specific

> > combination of agents.

> >

> >

> > Vitamin C (2 gms bid) has also been introduced based on the report

> that

> >   Vitamin C (ascorbic acid) at moderate intracellular

> concentrations 

> > stimulates replication of C. trachomatis (Wang et al., J. Clin.

> Micro. 

> > 30:2551-2554 (1992)) as well as its potential effect on biofilm

> charge

> > and  infectivity of the bacterium and specifically the EB

> (Hancock, R.

> > E. W.,  Annual Review in Microbiology, 38:237-264 (1984)).

> >

> >

> > Diagnosis and Treatment of Secondary Porphyria

> >

> > Chlamydia is a parasite of normal energy production in infected

> > eukaryotic  cells. As a result, host cells have insufficient

> energy

> > available for  their normal functioning. The energy shortage also

> > causes the host cell  mitochondria to attempt to synthesize

> certain

> > critical enzymes involved in  energy production in order to

> increase

> > energy production. Because  Chlamydia also prevents this synthesis

> from

> > completing, these enzyme's  precursors, called porphyrins, build

> up in

> > cell and often escape into the  intracellular [mileau] milieu.

> > Porphyrins readily form free-radicals,  that, in turn, damage

> cells.

> > Thus, there is an obligate secondary  porphyria that accompanies

> many

> > chlamydial infections. Therapy for this  secondary porphyria,

> which is

> > adjunct to anti-chlamydial therapy, involves  at least three

> > strategies: a) supplement the cellular energy supply to  mitigate

> cell

> > malfunction and the formation of porphyrins; reduce the  levels

> of

> > systemic porphyrins; and c) mitigate the harmful effects of the 

> > porphyrins.

> >

> > The pathogenesis of chronic/systemic chlamydial infection is

> unique in

> > that  the intracellular infection by this parasite results in a

> number

> > of  heretofore unrecognized concomitant and obligatory

> > metabolic/autoimmune  disorders including secondary porphyria with

> > associated autoantibodies  against the porphyrins. Cross reaction

> with

> > Vitamin B12 can result in a  subclinical autoimmune-mediated

> Vitamin

> > B12 deficiency. These associated  disorders often require

> diagnosis and

> > preventive and/or specific  adjunctive therapy.

> >

> >

> > The inventors have discovered the existence of antibodies to the

> > various  metabolites of heme biosynthesis, as well as Vitamin B12

> > (cobalamin),  which is molecularly similar to these metabolites,

> in

> > patients with active  systemic infection with C. pneumoniae. The

> > antibodies are primarily IgM;  this is similar to the antibody

> > responses to the MOMP of C. pneumoniae in  severely symptomatic

> > patients. Example 8 illustrates titers in symptomatic  patients

> with

> > systemic C. pneumoniae infections. The presence of  antibodies to

> > Vitamin B12 may have functional significance by decreasing  the

> amount

> > of bioavailable Vitamin B12. Thus, a Chlamydia infection may 

> cause a

> > previously unrecognized secondary Vitamin B12 deficiency. 

> > Administration (e.g., intramuscular) of large quantities of

> Vitamin B12

> >   (1000 to 5000 .mu.g) (e.g., parenteral cobalamin therapy)

> creates

> > large  amounts of Vitamin B12 available for binding to the native

> > receptors of  antibodies with an affinity for Vitamin B12, thereby

> > saturating these  anti-Vitamin B12 antibodies and increasing the

> amount

> > of bioavailable  circulating Vitamin B12.

> >

> >

> > Glucose is an important source of cellular energy. Glucose levels

> can

> > be  enhanced by diet and through vitamin supplements as described

> > below.

> >

> > A high carbohydrate diet should be maintained to promote

> production of 

> > glucose (Pierach et al., Journal of the American Medical

> Association, 

> > 257:60-61 (1987)). Approximately 70% of the caloric intake should

> be in

> >   the form of complex carbohydrates such as bread, potato, rice

> and

> > pasta.  The remaining 30% of the daily diet should comprise

> protein and

> > fat, which  should ideally be in the form of fish or chicken. Red

> > meats, including  beef, dark turkey, tuna and salmon, contain

> > [tryprophan] tryptophan.  Increased levels of tryptophan in the

> liver

> > inhibit the activity of  phosphoenol pyruvate carboxykinase with

> > consequent disruption of  gluconeogenesis. This accounts for the

> > abnormal glucose tolerance seen in  porphyria. Increased plasmic

> > concentrations of tryptophan also enhances  tryptophan transport

> into

> > the brain. The concentration of tryptophan in  the brain is the

> > rate-limiting factor for the synthesis of the neurotransmitter

> > 5-hydroxytryptamine (5-HT, serotonin). Serotonin is  synthesized

> by the

> > endothelium of brain capillaries for circulating  tryptophan.

> Thus,

> > increased concentrations of tryptophan in the brain  would be

> expected

> > to enhance production of serotonin and its metabolic, 

> > 5-hydroxyindole-acetic acid (5HIAA). Acute increases in serotonin

> > turnover  in the brain are followed by vascular and metabolic

> changes

> > which include  decreases in glucose consumption, disturbances in

> EEG

> > tracings, and  decreases in the postischemic neurological score.

> In

> > addition, while  serotonin increases brain perfusion on a single

> > injection, repetitive  administration initially opens the blood-

> brain

> > barrier and subsequently  induces vasoconstriction. It is likely

> that

> > any transient opening of the  blood-brain barrier by serotonin

> could

> > allow circulating substrates such  as ALA and PBG, if present, to

> enter

> > the central nervous system. As would  be expected from the

> location of

> > serotonin receptors and from the barrier  function of the

> endothelium

> > of cerebral arteries, the constricting effect  of serotonin is

> > amplified in cerebral arteries where endothelium is damage  or

> removed.

> > Damaged endothelial cells, as would be expected with  chlamydial

> > infection, would no longer have operational catabolic processes 

> for

> > serotonin. This would be particularly true in the event of

> depleted 

> > ATP as caused by chlamydial infection. This means that increased 

> > concentrations of serotonin will reach the smooth muscle layer of

> the 

> > cerebral vessels and cause more constriction. Finally, serotonin

> is

> > also  stored in blood platelets. Because blood platelets do not

> adhere

> > and  aggregate under normal conditions, they do not release

> serotonin

> > when the  vessel lumen is intact. However, if the vessel lumen is

> > altered by  chlamydial infection, platelet deposition and release

> of

> > serotonin can  occur.

> >

> > Another adverse effect of increased serotonin levels due to

> porphyria

> > is  seen with nervous tissues. Sympathetic nerve endings store

> > serotonin taken  up from the circulation. These serotonergic

> neurons

> > form plexuses around  brain vessels where they are likely to

> liberate

> > their serotonin contents  when subjected to cellular lysis from

> any

> > cause including ischemia,  free-radical ionizing damage to cell

> > membranes, and/or chlamydial  infection.

> >

> > In rats, elevated circulating tryptophan has been shown to

> produce 

> > structural alteration of brain astrocytes, oligodendroglia, and

> > neurons,  as well as degeneration of Purkinje cells and wasting of

> > axons. Similar  neurohistological alterations have been reported

> in

> > patients with acute  porphyria. Elevated tryptophan levels in

> plasma

> > and brain have been  associated with human encepholopathy.

> Finally,

> > serotonin is also  recognized as an active neurotransmitter in the

> > gastrointestinal tract.  The pharmacologic effects of serotonin in

> the

> > central nervous system and  gastrointestinal tract resemble the

> > neurological manifestations of acute  porphyric attacks. In fact,

> > administration of either tryptophan or  serotonin to humans have

> been

> > reported to cause severe abdominal pain,  psychomotor

> disturbances,

> > nausea, and dysuria; all of which are symptoms  of acute porphyria.

> >

> > Sucrose and fructose should be avoided (Bottomly et al., American

> > Journal  of Clinical Pathology, 76:133-139 (1981)) because the

> > ingestion of large  amounts of fructose trigger hepatic

> gluconeogenesis

> > which then decreases  the available glucose which is derived from

> > glycogen breakdown within the  liver. It is recommended that sport

> > drinks which contain glucose be  consumed.

> >

> > It is recommended that a patient suffering from porphyria avoid

> milk 

> > products. Milk products contain lactose and lactoferrin, and have

> been 

> > empirically shown to make symptoms of porphyria worse.

> >

> > Multivitamins containing the B complex vitamins should be

> administered 

> > daily (e.g., one or multiple times), preferably in excess of RDA,

> to 

> > enhance glucose availability. Hepatic breakdown of glycogen with 

> > generation of glucose is assisted by taking these multivitamins

> that 

> > contain the B complex vitamins. Pyridoxine minimizes the porphyrin

> > related  porphyrial neuropathy. B complex vitamins include folic

> acid

> > (e.g., 400  .mu.g per dosage; 1200 .mu.g daily maximum); vitamin B-

> 1

> > (thiamin; e.g.,  10 mg per dosage; 30 mg daily maximum); B-2

> > (riboflavin; e.g., 10 mg per  dosage; 30 mg daily maximum); B-5

> > (panothenate; e.g., 100 mg per dosage;  300 mg daily maximum); B-6

> > (pyridoxine; e.g., 100 mg per dosage; 300 mg  daily maximum) or

> > pyridoxal-5-phosphate (e.g., 25 mg per dosage; 100 mg  daily

> maximum)

> > and B-12 (e.g., 500 .mu.g per dosage; 10,000 .mu.g daily 

> maximum). The

> > preferred method of administration is oral for the majority  of

> these

> > vitamins (twice daily), except for B-12 for which sublingual 

> > administration (three-times daily) is preferred. It has been

> discovered

> >   that one important effect of this secondary porphyria in some

> patients

> > is  the production of IgM and IgG antibodies against

> > coproporphyrinogen-III.  These antibodies cross-react with Vitamin

> B12

> > (cobalamin) and can thus  cause a deficiency. Vitamin B12

> > supplementation (e.g., parenteral  cobalamin therapy) can remedy

> the

> > deficiency.

> >

> > D. Reducing Porphyrin Levels

> >

> > Dietary and pharmaceutical methods can be used to reduce systemic

> > porphyrin  levels (both water-soluble and fat-soluble).

> >

> > Plenty of oral fluids in the form of bicarbonated water or " sports

> > drinks "   (i.e., water with glucose and salts) should be

> incorporated

> > into the  regimen. This flushes water-soluble porphyrins from the

> > patient's system.  Drinking seltzer water is the easiest way to

> achieve

> > this goal. The color  of the urine should always be almost clear

> > instead of yellow. It is noted  that dehydration concentrates

> > prophyrins and makes patients more  symptomatic.

> >

> >

> > Activated charcoal can be daily administered in an amount

> sufficient to

> >   absorb fat-soluble porphyrins from the enterohepatic

> circulation. 

> > Treatment with activated oral is charcoal, which is nonabsorbable

> and 

> > binds porphyrins in the gastrointestinal tract and hence

> interrupts

> > their  enterohepatic circulation, has been associated with a

> decrease

> > of plasma  and skin porphyrin levels. Charcoal should be taken

> between

> > meals and  without any other oral drugs or the charcoal will

> absorb the

> > food or drugs  rather than the porphyrins. For those who have

> > difficulty taking the  charcoal due to other medications being

> taken

> > during the day, the charcoal  can be taken all at one time before

> bed.

> > Taking between 2 and 20 grams,  preferably at least 6 grams

> > (24.times.250 mg capsules) of activated  charcoal per day

> (Perlroth et

> > al., Metabolism, 17:571-581 (1968)) is  recommended. Much more

> charcoal

> > can be safely taken; up to 20 grams six times a day for nine

> months has

> > been taken without any side effects.

> >

> >

> > Antioxidants at high dosages (preferably taken twice per day) help

> to 

> > mitigate the effects of free radicals produced by porphyrins.

> Examples

> > of  suitable antioxidants include but are not limited to Vitamin C

> > (e.g., 1  gram per dosage; 10 g daily maximum); Vitamin E (e.g.,

> 400

> > units per  dosage; 3000 daily maximum); L-Carnitine (e.g., 500 mg

> per

> > dosage; 3 g  daily maximum); coenzyme Q-10 (uniquinone (e.g., 30

> mg per

> > dosage; 200 mg  daily maximum); biotin (e.g., 5 mg per dosage; 20

> mg

> > daily maximum);  lipoic acid (e.g., 400 mg per dosage; 1 g daily

> > maximum); selenium (e.g.,  100 .mu.g per dosage; 300 .mu.g daily

> > maximum); gultamine (e.g., from 2 to  about 4 g per dosage);

> > glucosamine (e.g., from about 750 to about 1000 mg  per dosage);

> and

> > chondroitin sulfate (e.g., from about 250 to about 500 mg  per

> dosage).

> >

> >

> > The above-mentioned therapeutic diets can be combined with

> traditional

> > or  currently recognized drug therapies for porphyria. In one

> > embodiment, benzodiazapine drugs, such as but not limited to

> valium,

> > klonafin,  flurazepam hydrochloride (e.g., Dalmanc.TM., Roche) and

> > alprazolam (e.g.,  Xanax), can be administered. Preferably,

> sedatives,

> > such as alprazolam  (e.g., Xanax; 0.5 mg per dosage for 3 to 4

> times

> > daily), can be prescribed  for panic attacks and flurazepam

> > hydrochloride (e.g., Dalmane.TM., Roche  or Restoril.TM. (e.g., 30

> mg

> > per dosage)) can be prescribed for sleeping.  The rationale is

> based

> > upon the presence of peripheral benzodiazepine  receptors in high

> > quantities in phagocytic cells known to produce high  levels of

> radical

> > oxygen species. A protective role against hydrogen  peroxide has

> been

> > demonstrated for peripheral benzodiazipine receptors.  This

> suggests

> > that these receptors may prevent mitochondria from radical 

> damages and

> > thereby regulate apoptosis in the hematopoietic system. 

> > Benzodiazepines have also been shown to interfere with the

> > intracellular  circulation of heme and porphyrinogens (Scholnick

> et

> > al., Journal of  Investigative Dermatology, 1973, 61:226-232).

> This is

> > likely to decrease porphyrins and their adverse effects. The

> specific

> > benzodiazipine will  depend on the porphyrin-related symptoms.

> >

> >

> >

> > Table 12 provides the results of an expanded study of

> antimicrobial 

> > susceptibilities at two different concentrations of antimicrobial

> > agents,  used alone and in combination, when exposed to the

> > antimicrobial agents  for two weeks. In addition to the agents

> already

> > mentioned, minocycline,  doxycycline, rifampin and

> > sulfamethoxizole/trimethoprim, at all  concentrations tested,

> failed to

> > clear the PCR signal for chlamydial MOMP.  Only the triple

> combination

> > of isoniazid, metronidazole and penicillamine  cleared the PCR

> signal

> > in two weeks. The triple combination was effective  at both low

> and

> > high concentrations. Table 12 also demonstrates the effect  of a 4

> week

> > exposure with the same expanded series of antimicrobial agents 

> alone

> > and in combination. A number of triple combinations of

> antimicrobial 

> > agents resulted in cell cultures in which the PCR signal for the 

> > chlamydial MOMP gene could not be detected at four weeks. The

> most 

> > effective combinations to have the greatest impact on the

> important

> > life  cycle phases are those predicted according to the

> methodology

> > described in the section above entitled " Methodology for Selecting

> > Potential Agent  Combinations " .

> >

> >

> >                               TABLE 11

> >                  Susceptibility to Antibiotics for

> >         Cryptic C. pneumoniae Cultured in HeLa Cells.sup.a

> >          Antibiotic               Conc (.mu.g/ml)   PCR.sup.b

> >          Ofloxacin                1                 positive

> >          Clarithromycin           1                 positive

> >          INH                      1                 positive

> >          Metronidazole            1                 positive

> >          Penicillamine            1/1               positive

> >          INH + Metronidazole +    1/1/4             negative

> >          Penicillamine

> >          Control                  0                 positive

> >      .sup.a Cultured in the presence of the indicated antibiotic

> (s), but

> > with no

> >       cycloheximide. Media changes at 48-72 hours.

> >      .sup.b Analysis following 2 weeks exposure to antimicrobioal

> agents.

> >

> >

> >                                                         TABLE 12

> >                                             Susceptibility to

> > Antibiotics by PCR

> >       for

> >                                    Cryptic Chlamydia pneumoniae

> Cultured

> > in HeLa

> >       Cells.sup.1

> >                        Phase of the Chlamydial Life Cycle

> >      EB (Extracellular EB->RB Stationary Phase RB               RB-

> >EB

> >       Transition Concentration   PCR     PCR

> >      or Intracellular) Transition ( " Cryptic phase " ) Replicating RB

> >       ( " Condensation " )   (.mu.g/ml)   2 week  4 week  Comment

> >             0          +       +    Control

> >                                                  Minocycline

> >             0.25       +       +    One drug

> >                                                  Minocycline

> >             1          +       +    One drug

> >                                                  Doxycycline

> >             0.25       +       +    One drug

> >                                                  Doxycycline

> >             1          +       +    One drug

> >                                                  TMP/SMZ.sup.2

> >             25          +       +    One drug

> >                                                  TMP/SMZ.sup.2

> >             100         +       +    One drug

> >                                                  Clarithromycin

> >         0.25.sup.1      +       +    One drug

> >                                                  Ofloxacin

> >             0.25       +       +    One drug

> >                                Metronidazole

> >             0.25       +       +    One drug

> >                      Rifampin

> >             0.25       +       +    One drug

> >                                Isoniazide        Isoniazide

> >             1          +       +    Misses EB phase

> >                                Metronidazole     TMP/SMZ.sup.1

> >           25/0.25       +       +    Misses EB phase

> >                                Metronidazole     Ofloxacin

> >         0.25/0.25      +       +    Misses EB phase

> >                      Rifampin  Metronidazole

> >         0.25/0.25      +       +    Misses EB phase

> >      penicillamine   Rifampin  Metronidazole                  

> > penicillamine

> >         4/.25/.25      +       +    Misses replicating

> >                                     phase

> >                      Rifampin  Metronidazole     Ofloxacin

> >        .25/.25/.25     +       +    Misses EB phase

> >      penicillamine             Metronidazole     Doxycyclin   

> > penicillamine

> >         1/.25/.25      +       +    Concentration too low

> >      penicillamine             Metronidazole     Doxycyclin   

> > penicillamine

> >           4/1/1        +       -    Covers key phases

> >      penicillamine             Metronidazole     Minocycline  

> > penicillamine

> >         1/.25/.25      +       -    Covers key phases

> >      penicillamine             Metronidazole     Minocycline  

> > penicillamine

> >           4/1/1        +       -    Original report of 4

> >                                     week positive was a

> >                                     typo

> >      penicillamine             Metronidazole     TMP/SMZ      

> > penicillamine

> >          4/1/100       +       -    Covers key phases

> >      penicillamine             Metronidazole     Clarithromycin

> > penicillamine

> >          1/.25/.25      +       -    Covers key phases

> >      penicillamine             Isoniazid         Isoniazid    

> > penicillamine

> >         1/.25/.25      -       -    Covers key phases

> >                                Metronidazole

> >      penicillamine             Isoniazid         Isoniazid    

> > penicillamine

> >           4/1/1        -       -    Covers key phases

> >                                Metronidazole

> >      .sup.1 Cultured in the presence of the indicated antibiotics,

> but

> > with no

> >       cycloheximide. Media changes at 48-72 hours. + = positive; -

> =

> > negative

> >      .sup.2 TMP/SMX = trimethoprim/sulfamethoxazole

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