Biphasic dose response in low level light therapy - an update

Low-level laser (light) therapy (LLLT) has been known since 1967 but still remains controversial due to incomplete understanding of the basic mechanisms and the selection of inappropriate dosimetric parameters that led to negative studies. The biphasic dose-response or Arndt-Schulz curve in LLLT has been shown both in vitro studies and in animal experiments. This review will provide an update to our previous (Huang et al. 2009) coverage of this topic. In vitro mediators of LLLT such as adenosine triphosphate (ATP) and mitochondrial membrane potential show biphasic patterns, while others such as mitochondrial reactive oxygen species show a triphasic dose-response with two distinct peaks. The Janus nature of reactive oxygen species (ROS) that may act as a beneficial signaling molecule at low concentrations and a harmful cytotoxic agent at high concentrations, may partly explain the observed responses in vivo. Transcranial LLLT for traumatic brain injury (TBI) in mice shows a distinct biphasic pattern with peaks in beneficial neurological effects observed when the number of treatments is varied, and when the energy density of an individual treatment is varied. Further understanding of the extent to which biphasic dose responses apply in LLLT will be necessary to optimize clinical treatments.     

Peptides and Hormesis

The article provides a broad assessment of the occurrence of hormetic-like biphasic dose-response relationships by over 30 peptides representing many major peptide classes. These peptide-induced biphasic dose responses were observed to occur in a extensive range of tissues, affecting an diverse range of biological endpoints. Despite diversity of peptides, models and endpoints, the quantitative features of the biphasic dose responses are remarkably similar with respect to the amplitude and width of the stimulatory response. These findings strongly suggest that hormetic-like biphasic dose responses represent a broadly generalizable biological phenomenon.     

Peptides and hormesis

The article provides a broad assessment of the occurrence of hormetic-like biphasic dose-response relationships by over 30 peptides representing many major peptide classes. These peptide-induced biphasic dose responses were observed to occur in a extensive range of tissues, affecting an diverse range of biological endpoints. Despite diversity of peptides, models and endpoints, the quantitative features of the biphasic dose responses are remarkably similar with respect to the amplitude and width of the stimulatory response. These findings strongly suggest that hormetic-like biphasic dose responses represent a broadly generalizable biological phenomenon.     

Cancer biology and hormesis: commentary

The observation of biphasic dose-response relationships following exposure to pharmacological and toxicological agents has been well documented. In this review Dr. Calabrese, using published data on human tumor cell lines treated with a variety of agents has provided additional support for the demonstration of hormesis in the cancer process. While this review has restricted the examination to human tumor cell lines, this limitation dose not take away from the value of the treatise and helps to point out the need for further analysis of the biphasic does response in other cancer models including in vivo carcinogenesis studies and human cancer epidemiology. This issue is further enhanced when the potential mechanisms for hormetic responses in the cancer cells are discussed, since the same mechanisms participate in the carcinogenesis process. Overall, this review provides an excellent opening examination into the definition of biphasic dose-response effects of toxic and pharmacological agents in cancer cells.     

Cancer Biology and Hormesis: Commentary

The observation of biphasic dose-response relationships following exposure to pharmacological and toxicological agents has been well documented. In this review Dr. Calabrese, using published data on human tumor cell lines treated with a variety of agents has provided additional support for the demonstration of hormesis in the cancer process. While this review has restricted the examination to human tumor cell lines, this limitation dose not take away from the value of the treatise and helps to point out the need for further analysis of the biphasic does response in other cancer models including in vivo carcinogenesis studies and human cancer epidemiology. This issue is further enhanced when the potential mechanisms for hormetic responses in the cancer cells are discussed, since the same mechanisms participate in the carcinogenesis process. Overall, this review provides an excellent opening examination into the definition of biphasic dose-response effects of toxic and pharmacological agents in cancer cells.     

Deceleration of carcinogenic potential by adaptation with low dose gamma irradiation

Animals that have been exposed to a very low dose of radiation are known to have many physiological benefits. Very low dose of ionizing radiation also induces mechanisms whereby cell or tissue become better fit to cope with subsequent exposures of high doses. This phenomenon of low dose radiation is termed 'adaptive response'. This response has been reported to be true in many biological systems and confirmed by experiments on chromosomal and chromatid aberrations, micronucleus formation, sister chromatid exchange tests, DNA mutation and cell survival study and using many other biological end points, although there are quite a few exceptions. The adaptation induced by low doses of radiation has been attributed to the induction of an efficient chromosome break repair mechanism at molecular and biochemical level. It is also substantiated in whole animal systems. When mice are initially conditioned with very small adapting doses, incidence of a challenging dose induced thymic lymphoma is recorded, with delayed latency and reduced frequency. Similarly, appearance of a transplanted barcl-95 thymic tumor has been delayed when mice are preconditioned with a small dose of radiation. Appearance and development of a tumour following transplantation of in vitro irradiated barcl-95 tumour cells with a small dose of 1 cGy are also delayed and volume of the tumour is reduced. Latency period of radiation-induced leukemia is modified by prior treatment with an adapting dose of radiation. Neoplastic transformation of several human cultured cells is also significantly decreased by prior low dose exposure of radiation compared to non-exposed cells. These results indicate that an earlier exposure to a small dose of radiation also reduces the radiation-induced carcinogenesis. Various aspects of molecular mechanism underlying the radio-adaptation have been explained. However, the mechanism underlying the inhibition of carcinogenesis by low dose radiation is yet to be fully resolved.     

Systems Cancer Biology and the Controlling Mechanisms for the J-Shaped Cancer Dose Response: Towards Relaxing the LNT Hypothesis

The hormesis phenomena or J-shaped dose response have been accepted as a common phenomenon regardless of the involved biological model, endpoint measured and chemical class/physical stressor. This paper first introduced a mathematical dose response model based on systems biology approach. It links molecular-level cell cycle checkpoint control information to clonal growth cancer model to predict the possible shapes of the dose response curves of Ionizing Radiation (IR) induced tumor transformation frequency. J-shaped dose response curves have been captured with consideration of cell cycle checkpoint control mechanisms. The simulation results indicate the shape of the dose response curve relates to the behavior of the saddle-node points of the model in the bifurcation diagram. A simplified version of the model in previous work of the authors was used mathematically to analyze behaviors relating to the saddle-node points for the J-shaped dose response curve. It indicates that low-linear energy transfer (LET) is more likely to have a J-shaped dose response curve. This result emphasizes the significance of systems biology approach, which encourages collaboration of multidiscipline of biologists, toxicologists and mathematicians, to illustrate complex cancer-related events, and confirm the biphasic dose-response at low doses.     

Apoptosis: Biphasic Dose Responses

The features of apoptotic dose responses typically have been evaluated over a broad dose-response continuum to differentiate apoptotic and necrotic responses. The present article specifically addresses the dose-response relationship for apoptosis in detail. It was determined that a number of well-documented cases exist in which the dose-response relationship for apoptosis is biphasic. The underlying mechanisms and biomedical implications of this biphasic dose-response phenomenon are discussed.     

Apoptosis: biphasic dose responses

The features of apoptotic dose responses typically have been evaluated over a broad dose-response continuum to differentiate apoptotic and necrotic responses. The present article specifically addresses the dose-response relationship for apoptosis in detail. It was determined that a number of well-documented cases exist in which the dose-response relationship for apoptosis is biphasic. The underlying mechanisms and biomedical implications of this biphasic dose-response phenomenon are discussed.     

Nanoparticle-emitted light attenuates amyloid-β-induced superoxide and inflammation in astrocytes

Alzheimer’s disease (AD) is the sixth leading cause of age-related death with no effective intervention yet available. Our previous studies have demonstrated the potential efficacy of Low Level Laser Therapy (LLLT) in AD cell models by mitigating amyloid-β peptide (Aβ)-induced oxidative stress and inflammation. However, the penetration depth of light is still the major challenge for implementing LLLT in animal models and in the clinical settings. In this study, we present the potential of applying Bioluminescence Resonance Energy Transfer to Quantum Dots (BRET-Qdots) as an alternative near infrared (NIR) light source for LLLT. Our results show that BRET-Qdot-emitted NIR suppresses Aβ-induced oxidative stress and inflammatory responses in primary rat astrocytes. These data provide a proof of concept for a nanomedicine platform for LLLT.From the Clinical EditorLow Level Laser Therapy has already been demonstrated to mitigate amyloid-β peptide induced oxidative stress and inflammation, a key driver of Alzheimer's disease. The major issue in moving this forward from cell cultures to live animals and potentially to human subjects is light penetration depth. In this novel study, BRET-Qdots were used as an alternative near infrared light source with good efficacy, paving the way to the development of a nanomedicine platform.     

The effect of extracellular Ca2+ and related ions on the cardiac action of milrinone

The biphasic single dose, dose-response curve of milrinone was sensitive to [Ca2+]0. At concentrations of 1.8 nM Ca2+ or less this biphasic response is observed but at [Ca2+]0 of 4.5 mM or more the dose response curve becomes monotonic. The inotropic response to milrinone in contrast to norepinephrine is highly sensitive to the extracellular [Ca2+]0. At low [Ca2+]0 of 0.15 mM milrinone could produce a negative inotropic effect. The positive inotropic effect of milrinone was proportional to [Ca2+]0 up to 2.7 mM. With [Ca2+]0 above 3.6 mM and low [Na+]0, the inotropic response to milrinone was reduced. These effects were due to increased [Ca2+]i and not due to the increase in contractile force produced by Ca2+. The positive inotropic effect of milrinone in contrast to norepinephrine is increased with an increase in [K+]0 possibly due to the depolarization produced by K+. The positive inotropic response to 10 micrograms of milrinone when [Ka+]0 = 4 mM was not significantly changed by Ca2+ channel blocking agents. In depolarized tissue (20 mM K+) the electropharmacological and contractile effects of milrinone are blocked by verapamil and ruthenium red. This suggests that under these conditions different mechanisms of Ca2+ channel activation are operative. Substitution of Sr2+ for Ca2+ increased contractile force and prolonged time to peak tension and relaxation time. Milrinone decreased time to peak tension but had no detectable effect on relaxation time. The results are discussed and it is suggested that milrinone acts on Ca2+ channels in the sarcolemma and intracellularly by increasing cyclic AMP which activates Ca2+ release and uptake from the sarcoplasmic reticulum.     

Differential effects of dopamine D2 agonist quinpirole upon the dorsal immobility response in rats.

The effects of various dose levels of systemically injected quinpirole upon the dorsal immobility response (DIR) over a time course was investigated in male rats. A low dose of quinpirole (0.01 mg/kg) significantly attenuated the duration of the DIR following the 10-min interval, whereas the highest dose (1.0 mg/kg) had a biphasic effect so that at the 10-min interval the duration of the DIR was significantly potentiated and at the 60-min interval the duration of the DIR was significantly attenuated. The intermediate dose (0.1 mg/kg) had intermediate behavioral effects. The data support the growing evidence that quinpirole has differential effects upon behavior over time as a function of the dose levels. The present data were discussed in reference to presynaptic and postsynaptic dopamine D2 receptor theory.     

Evocation of a biphasic febrile response in the rhesus monkey by intracerebral injection of bacterial endotoxins

Intravenous injection of 2.0 ml of a suspension of killed E. coli cell bodies produced a robust biphasic febrile response in the rhesus monkey. Intracerebral injection of the same pyrogen at 1/4000 of the intravenous dose produced a quite similar biphasic febrile effect. When administered i.v., this low dose had no effect on body temperature. Thirteen of the 16 sites (81 %) whose stimulation with endotoxin evoked a biphasic fever were located within 4 mm of the midpoint between the optic chiasm and the decussation of the anterior commissure. The second peak of the biphasic response to i.v. administered endotoxin is generally considered to be due to an endogenous pyrogen, released from leukocytes in the periphery, which penetrates the blood-brain barrier to act directly upon central thermoregulatory mechanisms. The present findings indicate that the entire biphasic response can be reproduced by an action of endotoxin within the CNS. Thus, it is possible that a portion of the second phase of a fever evoked by endotoxin injected i.v. may be due to a release of endogenous pyrogen within the brain, rather than within the bloodstream.     

Perchloroethylene-induced rat kidney tumors: an investigation of the mechanisms involved and their relevance to humans

Lifetime exposure to perchloroethylene by inhalation has been shown to cause a low incidence of renal tumors in male rats. The mechanisms responsible for the induction of these tumors have been investigated following exposure of rats to perchloroethylene by oral gavage (1500 mg/kg for up to 42 days) or by inhalation (400 ppm for 28 days). Comparisons have been made between rats and mice in vivo and between rats, mice, and humans in vitro. High doses of perchloroethylene given by gavage have been shown to be toxic to the rat kidney, causing increases in urinary markers of kidney damage. A marked accumulation of protein droplets (alpha-2u-globulin) was seen in the P2 segment of the kidney proximal tubules. This response were not seen after inhalation exposure to 400 ppm perchloroethylene for 28 days and hence may not be associated with the tumors seen at this dose level. Protein droplet formation was seen after exposure to 1000 ppm perchloroethylene, suggesting that 400 ppm is below the threshold dose required to induce this response. Perchloroethylene has been shown to be metabolized by glutathione conjugation in the liver, resulting in the formation of a mutagenic cysteine conjugate which is activated by the kidney enzyme beta-lyase. Levels of the mercapturic acid of perchloroethylene have been compared in rat and mouse urine. The enzyme kinetics of hepatic glutathione conjugation and renal beta-lyase activation have been compared in rat, mouse, and human tissues in vitro. Results of these studies are consistent with the rat being the species susceptible to kidney tumors. Although human kidney was shown to contain beta-lyase, glutathione conjugation of perchloroethylene could not be detected in human liver. Perchloroethylene-induced male rat kidney tumors may be a result of chronic toxicity, protein droplet nephropathy, and genotoxicity from the beta-lyase pathway. These mechanisms appear to have little relevance to humans.     

Radiation-induced neoplastic transformation in vitro, hormesis and risk assessment

Dose-response curves for various low-LET radiation sources have consistently been demonstrated to be J-shaped for the cancer-relevant endpoint of neoplastic transformation in vitro. Most of these studies have been performed where the radiation has been delivered at intermediate to high dose-rates (30-3000 mGy/min), where the threshold dose for induction of neoplastic transformation is around 100-200 mGy. Below these doses, the transformation frequency is less than that seen spontaneously, indicative of a hormetic effect. More recently, data have been obtained for low dose rates (<0.5 mGy/min) of low-LET radiation, and again hormetic effects are apparent but with threshold doses now being >1000 mGy. Similar trends have been reported in animal experiments as well as in human epidemiologic studies. Indeed, the relative risks for induction of neoplastic transformation in vitro in the dose range 1 to 1000 mGy agree well with those for incidence of radiation-induced breast cancer and leukemia in humans. These findings support the notion that the endpoint of neoplastic transformation in vitro is a plausible endpoint to not only study mechanisms involved in response to low doses of radiation, but also to provide information of potential importance to risk assessment.     

Radiation-Induced Bystander Effects: Evidence for an Adaptive Response to Low Dose Exposures?

This paper reviews our current knowledge of the mechanisms underlying the induction of bystander effects by low dose, low-LET ionizing radiation and discusses how they may be related to observed adaptive responses or other protective effects of low dose exposures. Bystander effects appear to be the result of a generalized stress response in tissues or cells. The signals may be produced by all exposed cells, but the response appears to require a quorum in order to be expressed. The major response involving low LET radiation exposure discussed in the existing literature is a death response. This has many characteristics of apoptosis but is p53 independent. While a death response might appear to be adverse, the position is argued in this paper that it is in fact protective and removes damaged cells from the population. Since many cell populations carry damaged cells without being exposed to radiation, so called "background damage", it is possible that low doses exposures cause removal of cells damaged by agents other than the test dose of radiation. This mechanism would lead to the production of "U-shaped" dose response curves. In this scenario, the level of "adaptive" or beneficial response will be related to the background damage carried by the cell population. This model may be important when attempting to predict the consequences of mixed exposures involving radiation and other environmental stressors.