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MELISA® A revolutionary blood test that detects type-IV allergy to metals and other common substances from a single blood sample FACILITATING THE DIAGNOSIS Recently, more and more research has been done on the effects of metals on human health. A connection between metals and the onset or worsening of symptoms has been established by many scientists. Previously, metal allergy has been difficult to diagnose. But this has changed with the MELISA® test - the only scientifically validated metal allergy blood test developed for screening of many metals and other antigens at the same time. METAL EXPOSURE AND HEALTH Chronic exposure to metals in jewellery, dental implants and restorations, cosmetics or joint prostheses can sensitize genetically predisposed individuals and induce a so-called Type IV allergic reaction. Metals are potential allergens. The binding of metal ions to cellular proteins can alter the activity of these enzymes or the presentation of these antigens and thereby induce local or systemic symptoms characteristic of allergic reactions or autoimmune diseases. In contrast to a type I allergy, which is mediated by IgE antibodies, a type IV allergic reaction is mediated by T lymphocytes that have had prior contact to a given allergen (memory lymphocytes). Following renewed contact with the sensitizing allergen, these memory lymphocytes respond by enlarging (lymphoblast transformation) and dividing (proliferation). The newly formed cells (effector cells) together with their secreted cytokines mediate the resulting allergic reaction. METAL ALLERGY VERSUS TOXICITY It is important to differentiate between metal allergy and metal toxicity. The MELISA® test does not measure the levels of metals in a patient’s

body; it measures whether the patient is allergic to metals. For example, hair samples may show levels of mercury or other substances which are below the official „safe limit” – but the patient can still be allergic. For allergic individuals, there is no such thing as a „safe” limit. Even trace amounts of a substance pose danger if the substance triggers an allergic reaction. ADVANTAGES FOR IMPLANTOLOGY Metals are found in body implants such as hip joints, knee prosthesis, screws, pacemakers, silicone breast implants, cochlear implants and dental implants. In the majority of patients, these implants are biocompatible. However, in sensitive patients, the implants may induce side-effects such as pain, dermatitis, cutaneous swelling, impaired wound healing, bone infections and implant loosening. Although bacterial infection is often the cause of implant rejection, sometimes no bacteria are detected despite of inflammation. These adverse effects are mediated by so-called wearing products of implant materials. Nickel, cobalt and chromium are known metal allergens. Recently, the cases of titanium allergy have also been described in the literature. Titanium is a transition metal and thus may function as a hapten and trigger cellular hypersensitivity. Since titanium is used as white pigment in tooth-pastes, cosmetics and medicaments, the latent sensitization of susceptible individuals is possible.

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MELISA® CAN PREVENT PROBLEMS Case reports convey disappearance of symptoms after the removal of the implant and positive delayed-type reactions are confirmed by patch-testing or by decreased lymphocyte proliferation to specific metal allergens in vitro. MELISA® screening prior implantation can detect possible existence of sensitization and allow the use of biocompatible implants on the individual basis.

pen if asymptomatic beryllium-sensitized workers are relocated to a beryllium-free environment. In addition to lifelong suffering, considerable savings in sick leave costs are obtained. Certain occupations pose a special risk for inducing metal allergy. These include construction workers, miners, electricians, rubber/wood/paper/ textile industry workers, dentists, hairdressers and painters.

ADVANTAGES FOR DENTISTRY Mucosal changes around dental metal fillings such as oral lichen may be due to an allergic reaction to the metal content. Replacement of fillings in metalsensitive patients leads to disappearance of the symptoms. Allergic reactions to metals used in dental prosthesis and braces can be measured in the MELISA® test. Symptoms may not always be local but can be systemic and include inflammation in different body parts. Patients with known clinical metal sensitivity such as adverse reactions to gold and nickel-containing earrings should consider nonmetallic restorative materials.

APPLICATION IN GENERAL MEDICINE Metal allergy is common in patients with various diseases such as skin diseases (psoriasis, eczema), autoimmune diseases (Multiple Sclerosis, thyroiditis, Sjögren’s disease) and gastrointestinal diseases. Patients with symptoms of profound fatigue of unclear etiology (Chronic Fatigue Syndrome, Myalgic Encephalitis) often suffer from metal allergy induced by dental metals. Candida allergy, drug allergy (such as penicillin allergy) and allergy to chemicals (such as formaldehyde) can also be tested. Furthermore, metal allergy has been implicated in autistic disorders. Allergy against thimerosal (a mercury preservative in vaccines) can also be detected by the MELISA® test. Avoidance of allergens often results in significant health improvement of the patient. Follow-up MELISA® testing usually shows reduced lymphocyte reactivity as well. A new development of the MELISA® test involves diagnosis of Lyme disease / active Borreliosis. Standard laboratory testing includes the serological test ELISA, which has very low specificity, and the Western blot, which is very specific. These tests can give false negative results in patients with early disease and therefore a misdiagnosis can occur.

OCCUPATIONAL MEDICINE AND ENVIRONMENTAL HEALTH The MELISA® test can be used for screen workers who are exposed to metals, chemicals or other allergens at work. If sensitivity is detected, the worker should be placed in an allergen-free environment to avoid the development of sometimes serious diseases. In the case of berylliosis, a sarcoidosis-like disease, a beryllium-specific blood test is used as a golden standard by US industry to screen workers for allergy against beryllium. It has been found that asymptomatic workers with memory cells to beryllium will - if beryllium exposure continues - develop a serious lung disease. This will not hap-

MELISA® detects cellular immunoreactivity of active infections of Borrelia burgdorferi (see picture). The test improves laboratory diagnosis by confirming an active infection in patients with or without any clinical symptoms. Finally, MELISA® can also test for allergy to gluten and gliadin, and testing to other food substances is being developed.

Metal allergy plays an important role in medical fields such as: • • • •

Implantology Dentistry Occupational medicine General medicine

THE MELISA® TEST PROCEDURE Blood in citrate tubes must arrive at the laboratory optimally within 24 hours after the blood has been drawn, maximally within 48 hours. White blood cells (lymphocytes) are isolated and tested against allergens chosen accordingly to the patient’s anamnesis and exposure to metals. The blood is incubated for five days and the lymphocyte reaction is measured by two separate technologies: one based on the uptake of radioisotope by dividing lymphocytes; the other by classical evaluation by microscopy. The level of reactivity is measured as a Stimulation Index (SI). A value over 3 indicates a positive reaction to a given allergen. The results are available in ten days. MELISA® RESULTS A MELISA® test report gives detailed information on the metals tested and possible sources of exposure. In the below case, a patient tested strongly positive to nickel and positive to inorganic mercury – which makes up 50% of dental amalgam fillings. Silver is also a component of amalgam fillings. The patient did not react to methylmercury (organic mercury) found in fish. A doctor may recommend that the patient avoids nickel-containing items (earrings, wristwatches, buttons) and consults an experienced dentist regarding replacement of amalgam „silver” fillings.

MELISA® IN THE WORLD Currently, laboratories performing the test are found Australia, Belgium, Canada, Germany, South Africa, Spain, Switzerland and USA. LICENSING OF THE MELISA® TEST Certified laboratories may perform the MELISA® test under license. The MELISA® test is protected by European patent and trademark in major countries. Standard license agreements consist of a 15% running royalty on the outgoing test price to the patient.

REERENCE LITERATURE • Validity of MELISA® for metal sensitivity testing Valentine-Thon E., et. al, Neuroendocrinology Letters, 2003; 24(1/2):57–64 • A novel lymphocyte transformation test (LTTMELISA®) for Lyme borreliosis Valentine-Thon E, et. Al., Diagn Microbiol Infect Dis. 2006 Jul 27 • The role of metals in autoimmunity Stejskal, J., Stejskal, V., Neuroendocrinology Letters, 1999; 20:351–364 • The beneficial effect of amalgam replacement on health in patients with autoimmunity Prochazkova, J., et. al, Neuroendocrinology Letters, 2004; 25:159–166 • Metal-specific lymphocytes: biomarkers of sensitivity in man Stejskal, V., et al., Neuroendocrinology Letters, 1999; 20:289–298 • MELISA® - an in vitro tool for the study of metal allergy Stejskal, Vera, PhD, et. al., Toxicology In Vitro, 1994,8; 991-1000

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Version: 07.05.2008

Acetylcysteine Acrolein Alcuronium Alprenolol Alprenolol epoxide Ammonium-persulfat Ampicillin Amoxycillin Articain Auranofin Aurothioglucose Azidocillin Bacampicillin Benzalkoniumchlorid Benzimidazol derivatives Benzocain Benzylpenicillin Bupivacain Butanilicain Candida albicans Cefalosporines Chloramine B Chloramine T Cloxacillin Chlorhexidin Cimetidin Cisplatin Clomethiazole Clotrimazol Collagen and derivatives Colophonium Cyclohexidine Epoxidhartz Erythromycin Flucloxacillin Formaldehyde Gentamycine Gluten

Gliadin Griseofulvin Halothan Hemineurin Insect venom: bee, wasp Isoprenaline sulfate Isothiazolones Ispagula (Phyllium) Jod, iopamiro Kathon CG and other thiazolidones Lansoprazol Lidocain Local anaesthetics generally Mercurochrome Mepivacain Metoprolol Metoprolol epoxide Myelin Basic Protein and derivatives Natriumaurothiomalat Natriumsulfit and derivatives Novocain Omeprazol Omnipaque (and other radiocontrast fluids) Pantoprazol Phenoxymethyl-penicillin Phenylendiamime and derivatives Phenytoin Piperacillin Pivampicillin Pivmecillinam Pivoxicam Pollen: birch, grass,

Distribution by: Evomed MedizinService GmbH Germany

pollen Prilocain Quinidine Quinine Ranitidin Ropivacain Salbutamol sulfate Silica and derivatives Silicon Streptokinase (Streptodermase) Streptomycin Sulfa drugs generally Sulfamethoxy-pyridazine Sulfasalazin Sulfathiazol Tetanus toxoid Tetracain Terbutaline and derivatives Theophyllin Trimecain Trimipramin Trimethoprim Zimelidine Metals: Aluminium (Al) Antimony (Sb) Arsenic (As) Barium (Ba) Beryllium (Be) Bismuth (Bi) Cadmium (Cd) Chromium (Cr) Cobalt (Co) Copper (Cu) Gallium (Ga) Gold (Au)

Indium (In) Inorganic mercury (Hg) Iridium (Ir) Iron (Fe) Lead (Pb) Manganese (Mn) Methyl mercury Molybdenum (Mo) Nickel (Ni) Niobium (Nb) Palladium (Pd) Phenyl mercury Platinum (Pt) Rhodium (Rh) Rubidium (Rb) Rutenium (Ru) Silver (Ag) Tantalum (Ta) Thimerosal Tin (Sn) Titanium dioxide (TiO2) Titanium trichloride (TiCl3) Uranium (U) Vanadium (V) Wolfram (W) Zink (Zn) Zirconium (Zr)

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