Red and near-infrared light evokes Ca2+ influx, endoplasmic reticulum release and membrane depolarization in neurons and cancer cells.

Low level light therapy uses light of specific wavelengths in red and near-infrared spectral range to treat various pathological conditions. This light is able to modulate biochemical cascade reactions in cells that can have important health implications. In this study, the effect of low intensity light at 650, 808 and 1064 nm on neurons and two types of cancer cells (neuroblastoma and HeLa) is reported, with focus on the photoinduced change of intracellular level of Ca2+ ions and corresponding signaling pathways. The obtained results show that 650 and 808 nm light promotes intracellular Ca2+ elevation regardless of cell type, but with different dynamics due to the specificities of Ca2+ regulation in neurons and cancer cells. Two origins responsible for Ca2+ elevation are determined to be: influx of exogenous Ca2+ ions into cells and Ca2+ release from endoplasmic reticulum. Our investigation of the related cellular processes shows that light-induced membrane depolarization is distinctly involved in the mechanism of Ca2+ influx. Ca2+ release from endoplasmic reticulum activated by reactive oxygen species generation is considered as a possible light-dependent signaling pathway. In contrast to the irradiation with 650 and 808 nm light, no effects are observed under 1064 nm irradiation. We believe that the obtained insights are of high significance and can be useful for the development of drug-free phototherapy.

Low-power laser irradiation promotes the proliferation and osteogenic differentiation of human periodontal ligament cells via cyclic adenosine monophosphate.

Retaining or improving periodontal ligament (PDL) function is crucial for restoring periodontal defects. The aim of this study was to evaluate the physiological effects of low-power laser irradiation (LPLI) on the proliferation and osteogenic differentiation of human PDL (hPDL) cells. Cultured hPDL cells were irradiated (660 nm) daily with doses of 0, 1, 2 or 4 J⋅cm(-2). Cell proliferation was evaluated by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay, and the effect of LPLI on osteogenic differentiation was assessed by Alizarin Red S staining and alkaline phosphatase (ALP) activity. Additionally, osteogenic marker gene expression was confirmed by real-time reverse transcription-polymerase chain reaction (RT-PCR). Our data showed that LPLI at a dose of 2 J⋅cm(-2) significantly promoted hPDL cell proliferation at days 3 and 5. In addition, LPLI at energy doses of 2 and 4 J⋅cm(-2) showed potential osteogenic capacity, as it stimulated ALP activity, calcium deposition, and osteogenic gene expression. We also showed that cyclic adenosine monophosphate (cAMP) is a critical regulator of the LPLI-mediated effects on hPDL cells. This study shows that LPLI can promote the proliferation and osteogenic differentiation of hPDL cells. These results suggest the potential use of LPLI in clinical applications for periodontal tissue regeneration.

Serum 25-hydroxyvitamin D in erythropoietic protoporphyria.

BACKGROUND: Vitamin D, produced by the action of sunlight on skin, is an important hormone for calcium homeostasis and has been implicated as tumour-protective agent. Some previous studies of photosensitive patients who actively avoid sunlight have failed to show convincing evidence of vitamin D insufficiency. OBJECTIVES: The aim of this study was to characterize the vitamin D status of a large cohort of patients with erythropoietic protoporphyria (EPP). METHODS: U.K. patients with EPP were recruited prospectively and seen locally by a single study investigator. A blood sample was taken for vitamin D assay. All blood analyses were performed in the same laboratory. RESULTS: A cohort of 201 patients with known EPP was seen over a 7-month period between January and July. Thirty-four patients (17%) were deficient in vitamin D and 126 (63%) had insufficient vitamin D. Both insufficiency and deficiency were significantly associated with the total erythrocyte protoporphyrin concentration and inversely with the time in minutes to the onset of symptoms following sunlight exposure. CONCLUSIONS: This is the first report of significant levels of vitamin D deficiency and insufficiency in a large cohort of patients with a photodermatosis. Such individuals are at risk of associated adverse events. In future, clinicians should consider monitoring 25-hydroxyvitamin D levels and instigating oral supplementation or dietary advice if appropriate.

Two-photon fluorescent probes for long-term imaging of calcium waves in live tissue.

2-Acetyl-6-(dimethylamino)naphthalene-derived two-photon fluorescent Ca2+ probes (ACa1-ACa3) are reported. They can be excited by a 780 nm laser beam, show 23-50-fold enhancement in one- and two-photon excited fluorescence in response to Ca2+, emit fourfold stronger two-photon excited fluorescence than Oregon Green 488 BAPTA-1 upon complexation with Ca2+, and can selectively detect intracellular free Ca2+ ions in live cells and living tissues with minimum interference from other metal ions and membrane-bound probes. Moreover, these probes are capable of monitoring calcium waves at a depth of 120-170 microm in live tissues for 1100-4000 s using two-photon microscopy with no artifacts of photobleaching.

Enhancement of macrosphelide-induced apoptosis by mild hyperthermia.

Hyperthermia is a useful adjunct in cancer therapy as it can increase the effectiveness and decrease the toxicity of currently available cancer treatments such as chemotherapy and radiation. In the present study, we investigated whether 41 degrees C hyperthermia (mild HT) for 20 min can enhance macrosphelide (MS5)-induced apoptosis in human lymphoma U937 cells. Our results revealed that, compared with MS5 (5 microM) and mild HT alone, the combined treatment exhibited significant enhancement in apoptosis at 6 h, which was evaluated by observing morphological changes and DNA fragmentation. Marked increase in the reactive oxygen species (ROS) generation was observed immediately after the combined treatment. Significant increase in Fas externalization, caspase-8 and caspase-3 activation, and loss of mitochondrial membrane potential (MMP) was found after the combined treatment compared with MS5 and mild HT alone. Moreover, this combination can also alter the expression of apoptosis-related proteins as evident by the cleavage of Bid and down-regulation of Bcl-2 while no change in the expression of Bax was observed. Furthermore, an immediate rise in the intracellular calcium ion ([Ca(2+)]i) concentration was observed after the combined treatment, which continuously increased in a time-dependent manner. In addition, mild HT treatment alone also increases [Ca(2+)]i concentration without inducing apoptosis. Our data indicate that early increase in ROS generation is mainly responsible for the enhancement of apoptosis after the combined treatment.

Subcellular translocation of the eGFP-tagged TRPL channel in Drosophila photoreceptors requires activation of the phototransduction cascade.

Signal-mediated translocation of transient receptor potential (TRP) channels is a novel mechanism to fine tune a variety of signaling pathways including neuronal path finding and Drosophila photoreception. In Drosophila phototransduction the cation channels TRP and TRP-like (TRPL) are the targets of a prototypical G protein-coupled signaling pathway. We have recently found that the TRPL channel translocates between the rhabdomere and the cell body in a light-dependent manner. This translocation modifies the ion channel composition of the signaling membrane and induces long-term adaptation. However, the molecular mechanism underlying TRPL translocation remains unclear. Here we report that eGFP-tagged TRPL expressed in the photoreceptor cells formed functional ion channels with properties of the native channels, whereas TRPL-eGFP translocation could be directly visualized in intact eyes. TRPL-eGFP failed to translocate to the cell body in flies carrying severe mutations in essential phototransduction proteins, including rhodopsin, Galphaq, phospholipase Cbeta and the TRP ion channel, or in proteins required for TRP function. Our data, furthermore, show that the activation of a small fraction of rhodopsin and of residual amounts of the Gq protein is sufficient to trigger TRPL-eGFP internalization. In addition, we found that endocytosis of TRPL-eGFP occurs independently of dynamin, whereas a mutation of the unconventional myosin III, NINAC, hinders complete translocation of TRPL-eGFP to the cell body. Altogether, this study revealed that activation of the phototransduction cascade is mandatory for TRPL internalization, suggesting a critical role for the light induced conductance increase and the ensuing Ca2+ -influx in the translocation process. The critical role of Ca2+ influx was directly demonstrated when the light-induced TRPL-eGFP translocation was blocked by removing extracellular Ca2+.

50-Hertz magnetic field and calcium transients in Jurkat cells: results of a research and public information dissemination (RAPID) program study.

An effect on intracellular calcium continues to be proposed as a biochemical pathway for the mediation of biologic effects of electrical-power-frequency magnetic fields (MF). However, reproducible results among laboratories are difficult to attain and the characteristics of magnetic field effects on intracellular free calcium ([Ca(2+)](i)) are not well understood. We attempted to repeat the studies of Lindström et al. [Intracellular Calcium Oscillations in a T-Cell Line by a Weak 50 Hz Magnetic Field. J Cell Physiol 156:395-398 (1993)] by investigating the effect of a 1.5-G 50-Hz MF on [Ca(2+)](i) in the Jurkat lymphocyte T-cell line. Changes in [Ca(2+)](i) were determined using microscopic imaging of fura-2 loaded Jurkat cells on poly-l-lysine-coated glass coverslips. The MF was generated by a single coil constructed with bifilar wire and located in the same plane as the cells. Cells were randomly exposed for 8 min to MF, sham field (SF), or no field (NF) conditions. The exposure condition remained coded until data analysis was complete. Each exposure period was preceded by an 8-min data collection to establish a baseline for [Ca(2+)](i). After each exposure condition, cells were exposed to anti-CD3 antibody that induced a rapid increase in [Ca(2+)](i) in responsive cells; this provided a positive control. [Ca(2+)](i) was analyzed for individual cells as spatially-averaged background-corrected 340/380 nm ratios, and a [Ca(2+)](i) transient was considered significant for positive deviations from baseline of 3 [multiple] an estimate of noise in the baseline. Typically, 25-50 cells/field were viewed and approximately 50% had no [Ca(2+)](i) transients in the baseline period and also responded to positive control. Only cells responding to positive control and lacking changes in [Ca(2+)](i) during the baseline period were considered qualified for assessment during the exposure period. The incidences of [Ca(2+)](i) transients during the exposure period for two experiments (40 [multiple] objective) were 16.5, 14.6, and 14.2% for MF, SF, and NF, respectively, and were not statistically significantly different. Previous studies by Lindström et al. [Intracellular Calcium Oscillations in a T-Cell Line after Exposure to Extremely-Low-Frequency Magnetic Fields with Variable Frequencies and Flux Densities. Bioelectromagnetics 16:41-47 (1995)] showed a high response rate (92%) for exposure to 1. 5-G 50-Hz MF when individual cells were preselected for investigation. We found no such effect when examining many cells simultaneously in a random and blind fashion. These results do not preclude an effect of MF on [Ca(2+)](i), but suggest that responsive cells, if they exist, were not identified using the approaches that we used in this study.

Particle-induced X-ray emission and scanning electron microscopic analyses of the effects of CO2 laser irradiation on dentinal structure.

OBJECTIVES: The purpose of this study was to analyze the physico-chemical changes present on the dentinal surface after using CO2 laser irradiation, and to determine whether or not it is possible to seal the dentinal tubules. METHODS: Thirty human-extracted first premolars were obtained for this study. A Class V cavity was prepared on the buccal surface of all the specimens with a carbide pear-shaped bur, using a conventional high speed handpiece. Fifteen premolars (experimental group) were irradiated with a CO2 laser (with a wavelength of 10.6 microm, 2 W, 10 J, 0.2 s, 25 pulses). The remaining 15 premolars were used as the control group. RESULTS: Scanning electron microscopy showed that the effect of laser energy on dentin varied from charring, cratering, poring, fissuring, fracturing and cracking up to melting; also, the dentinal tubules were not sealed, in contrast with the control group in which the dentinal surfaces were more homogeneous. Particle-induced X-ray emission results showed that the irradiated dentinal surface presented a decrease in calcium content and an increase in phosphorous content, possibly due to a vaporization process which occurred during the irradiation. CONCLUSION: The physicochemical changes observed on the irradiated dentinal surface suggest that changes in the hydroxyapatite crystal structure take place, and that these structural changes may be responsible for the observed effects.

The effect of normal pulsed Nd-YAG laser irradiation on pits and fissures in human teeth.

The effects of normal pulsed Nd-YAG laser irradiation on the acid resistance of human dental enamel of pits and fissures, the cleaning of the pit and fissure contents and fluoride uptake into deep pits and fissures were examined. The acid resistance of the pit and fissure enamel was evaluated by the amount of dissolved calcium per square millimeter of the surface area. The pit and fissure enamel treated with laser irradiation obtained an acid resistance 30% higher than that of the unlased controls. The cleaning effect of laser irradiation on the pit and fissure contents was compared with chemicomechanical and mechanical methods. The laser irradiation was found to clean the pits and fissures to a greater depth without alterating the shape of pits and fissures, compared with the other two methods. The distribution of calcium, phosphorus and fluoride in the enamel of the pits and fissures was then measured by electron probe microanalyzer. At the entrance and in the deep part of the pits and fissures, the fluoride content of the enamel treated with acidulated phosphate fluoride after laser irradiation was higher than that of the enamel treated with acidulated phosphate fluoride alone. These results thus suggest that Nd-YAG laser irradiation might be effective in increasing the acid resistance of the pit and fissure enamel, while removing the pit and fissure debris contents and increasing the fluoride uptake into the pit and fissure enamel.

[The effect of high-dose ultraviolet irradiation on sodium, calcium and aldosterone in the blood of calves]. / Pôsobenie nadmerného ultrafialového ziarenia na sodík, vápnik a aldosteron v plazme teliat.

Five Holstein-Friesian calves, from one sire, with prevalent black hair coat pigmentation were used in the experiment. The mean age was 33 days and the mean live weight 51 kg. The animals were exposed free running without interruption for 12 hours to an artificial ultraviolet light in the range of 280-320 nm. The mean doses of radiation was 179.10(-10) J/h/m. One-spot high-pressure mercury discharge lamps Tesla RVK 400 W were used as a radiation source. The dose rate was estimated from measurements by a spectral photometer with filter UG 2 for absorbtion of visible light located at the height of the back of standing calf. Blood samples were collected immediately before the beginning of treatment and after 5, 12, 24, 48 and 72 hours. The blood plasma aldosterone was measured by radioimmunoassays, the levels of sodium, potassium and calcium in blood plasma by flame spectrophotometry. Double classification variance analysis and evaluation according to the Snedecor F-test, the contrast effect test according to Duncan and regression analysis were used for statistical evaluation. Compared to the first sampling, sodium increased significantly after 5 and 12 hours of exposure (Tab. I) to 138.1 and 138.3 mmol/l, respectively. In the subsequent samplings this trend continued up to 72 hours from the beginning of irradiation (140.5 mmol/l). The potassium level did not change statistically significantly. Owing to an excessive irradiation, the calcium concentration increased significantly. The greatest increase occurred after 12 hours of irradiation (from 2.29 mmol/l to 2.61 mmol/l) and after 36 hours from the end of irradiation (2.70 mmol/l).(ABSTRACT TRUNCATED AT 250 WORDS)

[The effect of the light from a helium-neon laser on normal dental enamel solubility and in caries]. / Vliianie sveta gelii-neonovogo lazera na rastvorimost' émali zubov v norme i pri kariese.

Effects of He-Ne laser radiation at a wavelength 0.63 micron for 90 sec on dental enamel mineral metabolism were studied from Ca, P levels and Ca/P ratio in 67 children aged 3 to 7. Ca and P solubility in surface enamel layers was found reduced after He-Ne laser irradiation in both healthy children and those with caries. Laser was found to normalize the oral fluid mineral composition.

[Experimental research on the effect of ultraviolet irradiation on the phosphorus-calcium metabolic status]. / Eksperimental'noe issledovanie vliianiia ul'trafioletovogo oblucheniia na sostoianie fosforno-kal'tsievogo obmena.

The influence of vitamin D and ultraviolet irradiation (UVI), used for prevention of vitamin D deficiency, on the state of phosphoric-calcium metabolism was studied in experiments on rats. It has been shown that daily injections of 1-5 IU of vitamin D, and UVI in biodoses from 1/8 to 1/4 promote the maintainance of the normal level of phosphoric-calcium metabolism. When UVI was applied according to the widely used "basic scheme" (in biodoses from 1/4 to 2 1/2) the following anomalies were recorded: a three-fold increase of 25-ON-D3 concentration in the blood plasma, hyperphosphatemia, a tendency to hypercalcemia. The use of higher doses of UVI led to a further increase of 25-OH-D3 level, hypophosphatemia, hypercalcemia, kidney and heart calcification. The data obtained have evidenced a possibility of vitamin D intoxication during UVI.

[The biochemical changes of bone collagen after high-dose irradiation (author's transl)].

In our clinic, patients with malignant bone tumors have been treated by high-dose irradiation therapy, 10,000-20,000 rads, for primary lesions. In order to study the biochemical changes of normal bone around tumor tissue, especially bone collagen, after high-dose irradiation, the author performed the following experiments. The right knee joint of rabbits was irradiated with either 6,000, 10,000, or 15,000 rads by 60Co-gamma ray. The cortical bone of the right tibial metaphysis was used for analyses and compared with the left tibia of the same rabbit. These studies were followed for one year after the final irradiation. The calcium, phosphorus and collagen contents of irradiated bone were remarkably changed. These data indicate that collagen biosynthesis of irradiated bone was decreased and the calcification was disturbed. An increase in the amount of total soluble collagen and a decrease in the amount of hydroxylysine bound sugar were observed. The ratio of beta to alpha chains of the collagen molecule was also changed by the irradiation. The amount of reducible cross-links per hydroxyproline residue was strikingly increased three months after the final irradiation. These changes were remarkable especially in the 10,000 and 15,000 rads irradiated group and found to be recovered approximately six months to one year after the final irradiation. These findings indicate that high-dose irradiation reduces the stability of bone collagen both with the destruction of sugar bonds of hydroxylysine residues and the replacement of matured collagen matrix to immatured one which contain mostly labile reducible cross-links.