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Free Radicals and Low-Level Photon Emission in Human Pathogenesis: State of the Art

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Free Radicals and Low-Level Photon Emission in Human Pathogenesis: State of the Art Indian Journal of Experimental Biology Vol. 46, May 2008, pp. 273-309 Review Article Free radicals and low-level photon emission in human pathogenesis: State of the art Roeland Van Wijk1, Eduard P A Van Wijk1, Fred A C Wiegant2 & John Ives3 1International Institute of Biophysics, Neuss, Germany 2Utrecht University, Utrecht, The Netherlands 3Samueli Institute for Information Biology, Alexandria, VA Convincing evidence supports a role for oxidative stress in the pathogenesis of many chronic diseases. The model includes the formation of radical oxygen species (ROS) and the misassembly and aggregation of proteins when three tiers of cellular defence are insufficient: (a) direct antioxidative systems, (b) molecular damage repairing systems, and (c) compensatory chaperone synthesis. The aim of the present overview is to introduce (a) the basics of free radical and antioxidant metabolism, (b) the role of the protein quality control system in protecting cells from free radical damage and its relation to chronic diseases, (c) the basics of the ultraweak luminescence as marker of the oxidant status of biological systems, and (d) the research in human photon emission as a non-invasive marker of oxidant status in relation to chronic diseases. In considering the role of free radicals in disease, both their generation and their control by the antioxidant system are part of the story. Excessive free radical production leads to the production of heat shock proteins and chaperone proteins as a second line of protection against damage. Chaperones at the molecular level facilitate stress regulation vis-à-vis protein quality control mechanisms. The manifestation of misfolded proteins and aggregates is a hallmark of a range of neurodegenerative disorders including Alzheimer’s disease, Parkinson’s disease, amylotrophic lateral sclerosis, polyglutamine (polyQ) diseases, diabetes and many others. Each of these disorders exhibits aging-dependent onset and a progressive, usually fatal clinical course. The second part reviews the current status of human photon emission techniques and protocols for recording the human oxidative status. Sensitive photomultiplier tubes may provide a tool for non-invasive and continuous monitoring of oxidative metabolism. In that respect, recording ultraweak luminescence has been favored compared to other indirect assays. Several biological models have been used to illustrate the technique in cell cultures and organs in vivo. This initiated practical applications addressing specific human pathological issues. Systematic studies on human emission have presented information on: (a) procedures for reliable measurements, and spectral analysis, (b) anatomic intensity of emission and left-right symmetries, (c) biological rhythms in emission, (d) physical and psychological influences on emission, (e) novel physical characteristics of emission, and (f) the identification of ultraweak photon emission with the staging of ROS-related damage and disease. It is concluded that both patterns and physical properties of ultraweak photon emission hold considerable promise as measure for the oxidative status. Keywords: Antioxidants, Chaperones, Chronic disease, Free radicals, Heat shock proteins, Photon count distribution, Ultraweak photon emission Introduction the enzyme, superoxide dismutase (SOD) with the In 1954 Gerschman and Gilbert proposed that most function of catalytically removing a specific free of the damaging effects of elevated oxygen radical2,3. During the 70’s and 80’s, many scientists, concentrations in living organisms might be attributed unfamiliar with free radicals, regarded the field as to the formation of free radicals1. In 1956, Harman highly specialized or irrelevant to mainstream biology, proposed the “free radical theory of aging” which biochemistry and medicine. In fact, however, it is just suggested that free radical damage on cellular the opposite. macromolecules is responsible for the aging process. Much experimental data emphasizes that aerobic life However, this idea did not capture the interest of many is connected with the continuous production of free biologists and clinicians until the discovery in 1969 of radicals, particularly reactive oxygen species (ROS) that may be dangerous for the living organism4-12. The _________ reactive species attack biomolecules producing Telephone: +31345570080 alterations in DNA, proteins and lipids, and were Fax: +31345570110 13 E-mail: [email protected] implicated in the pathogenesis of age-related disease . 274 INDIAN J EXP BIOL, MAY 2008 In recent years a wealth of experimental data was chronic diseases, (c) the basics of the ultraweak collected to clarify mechanisms that are critically luminescence as marker of the oxidant status of involved in free radical damage resulting in biological systems, and (d) the research in human pathologies. The data emphasize the role of heat stress photon emission as a non-invasive marker of oxidant proteins (HSP’s) in protection against damage by free status in relation to chronic diseases. Perspectives in radicals. The HSP’s are also named as to their future research is presented that allow the evaluation function, such as “chaperone proteins”, since they of ultraweak luminescence as a method for recording form complexes with proteinaceous structures in in vivo and noninvasively the state of oxidative stress order to prevent deleterious interactions between in human subjects vis-à-vis the development of proteins. Understanding the molecular mechanisms of chronic disease. cellular protection and recovery from damage in injured cells had steadily increased. In particular, how Free radicals and antioxidants chaperones at the molecular level facilitate stress A “free radical” is defined as any atom, group of regulation vis-à-vis protein quality control atoms or molecules containing one unpaired electron mechanisms, and have become critical in within an outer orbit. Molecular oxygen (O2) is a development of a range of chronic diseases. triplet in its ground state because it contains two unpaired electrons within its outer orbits having To record ROS, many techniques have been made parallel or unpaired spins. Singlet oxygen, by available. Most of these techniques are invasive; they definition, is not a free radical; both electrons occupy require the destruction of living cellular or tissue the same orbit and the electron spins are paired. In O2, structures to estimate either specific ROS species or parallel electron spin prevents the direct addition of products derived from reactions between ROS and electron pairs (this would include electron spins in macromolecules, mostly lipids. Although these both parallel and anti-parallel directions) unless an techniques are available to measure the progress of electron spin inversion occurs. A number of oxidation, none is applicable to all circumstances. In enzymatic systems have evolved that are capable of the present study, attention is drawn to the method of circumventing electron spin restriction by a one- low-level chemiluminescence to detect electronically- electron reduction of O2. This intermediate univalent excited states in biological systems. Low-level pathway is an essential biological process that chemiluminescence was related to the direct 14,15 provides the pairing electron. The cytochrome oxidase utilization of molecular oxygen and the production complex localized at the inner mitochondrial of electronically-excited states in biological membrane tetravalently reduces the majority of O systems16; in particular, the oxygen dependent chain 2 17-19 used by aerobic cells. It appears to be a major reactions involving biological lipids . This earlier - intercellular source of both O2 and H2O2. research on low-level chemiluminescence was largely Apart from the mitochondrial respiratory chain, all unnoticed in America and Europe, notwithstanding the monooxygenases, several dehydrogenases, cyto- the reports by Stauff and Ostrowski on the 20 chrome-P450, prostaglandin synthetase, leucotri-ene chemiluminescence of mitochondria as well as synthetase, vitamin K-dependent enzymes and many Howes and Steele on the chemiluminescence of 21,22 other enzymes normally generate radicals. The body microsomes , both from rat liver. This hesitation not only produces radicals during normal metabolism probably evolved because of earlier reports of the so- 23,24 but it also purposefully produces radicals, designed to called “mutagenic radiation” from living tissue be toxic, during immune and inflammatory responses. which could not be observed with the then These radicals are deliberately generated during the contemporary photon counting equipment25. respiratory burst of a macrophage in order to kill In the meantime, data have demonstrated that invading organisms. spontaneous (natural) ultraweak photon emission In considering the role of free radicals in disease, originating from living organisms may be considered their generation is only part of the story; the other part to reflect the state of oxidative stress in vivo. The aim is their control, containment and safe disposal. of the present overview is to introduce (a) the basics Because radicals and their products are continuously of free radical and antioxidant metabolism, (b) the generated and are so reactive chemically, they must role of the protein quality control system in protecting physiologically be closely controlled and they must be cells from free radical damage and its relation to released in an orderly fashion to avoid damage of vital
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