Visual light effects on mitochondria: The potential implications in relation to glaucoma.

Light of different wave-lengths have the potential to interact with four major mitochondrial protein complexes that are involved in the generation of ATP. Neurones of the central nervous system have an absolute dependence on mitochondrial generated ATP. Laboratory studies show that short-wave or blue light (400-480nm) that impinges on the retina affect flavin and cytochrome constituents associated with mitochondria to decrease the rate of ATP formation, stimulate ROS and results in cell death. This suggests that blue light could potentially have a negative influence on retinal ganglion cell (RGC) mitochondria that are abundant and not shielded by macular pigments as occurs for photoreceptor mitochondria. This might be of significance in glaucoma where it is likely that RGC mitochondria are already affected and therefore be more susceptible to blue light. Thus simply filtering out some natural blue light from entering the eye might be beneficial for the treatment of glaucoma. Long-wave or red light (650-800nm) affects mitochondrial complex IV or cytochrome oxidase to increase the rate of formation of ATP and ROS causing the generation of a number of beneficial factors. Significantly, laboratory studies show that increasing the normal amount of natural red light reaching rat RGC mitochondria in situ, subjected to ischemia, proved to be beneficial. A challenge now is to test whether extra red light delivered to the human retina can slow-down RGC loss in glaucoma. Such a methodology has also the advantage of being non-invasive. One very exciting possibility might be in the production of a lens where solar UV light is convertes to add to the amount of natural red light entering the eye.

Flavin-mediated photo-oxidation for the detection of mitochondrial flavins.

We report a new approach for the detection of mitochondrial flavins through photo-oxidation of a probe molecule. Probe 1 showed high brightness (ε × Φf = 6.50 × 103 M-1 cm-1) at long wavelengths (maximum emission wavelength, λmax = 600 nm) upon photo-oxidation, assisted by the strong electron accepting ability of the isoalloxazine moiety in flavins. Probe 1 also exhibited high selectivity for flavins over various biological oxidants, remarkable photo-stability, and mitochondrial localization.

TK1299, a highly thermostable NAD(P)H oxidase from Thermococcus kodakaraensis exhibiting higher enzymatic activity with NADPH.

Seven nicotinamide adenine dinucleotide oxidase homologs have been found in the genome of Thermococcus kodakaraensis. The gene encoding one of them, TK1299, consisted of 1326 nucleotides, corresponding to a polypeptide of 442 amino acids. To examine the molecular properties of TK1299, the structural gene was cloned, expressed in Escherichia coli and the gene product was characterized. Molecular weight of the recombinant protein was 49,375 Da when determined by matrix-assisted laser desorption/ionization time-of-flight and 300 kDa when analyzed by gel filtration chromatography indicating that it existed in a hexameric form. The enzyme was highly thermostable even in boiling water where it exhibited more than 95% of the enzyme activity after incubation of 150 min. TK1299 catalyzed the oxidation of NADH as well as NADPH and predominantly converted O2 to H2O (more than 75%). K(m) value of the enzyme towards NADH and NADPH was almost same (24 ± 2 µM) where as specific activity was higher with NADPH compared to NADH. To our knowledge this is the most thermostable and unique NAD(P)H oxidase displaying higher enzyme activity with NADPH.

Biomarkers spectral subspace for cancer detection.

A novel approach to cancer detection in biomarkers spectral subspace (BSS) is proposed. The basis spectra of the subspace spanned by fluorescence spectra of biomarkers are obtained by the Gram-Schmidt method. A support vector machine classifier (SVM) is trained in the subspace. The spectrum of a sample tissue is projected onto and is classified in the subspace. In addition to sensitivity and specificity, the metrics of positive predictivity, Score1, maximum Score1, and accuracy (AC) are employed for performance evaluation. The proposed BSS using SVM is applied to breast cancer detection using four biomarkers: collagen, NADH, flavin, and elastin, with 340-nm excitation. It is found that the BSS SVM outperforms the approach based on multivariate curve resolution (MCR) using SVM and achieves the best performance of principal component analysis (PCA) using SVM among all combinations of PCs. The descent order of efficacy of the four biomarkers in the breast cancer detection of this experiment is collagen, NADH, elastin, and flavin. The advantage of BSS is twofold. First, all diagnostically useful information of biomarkers for cancer detection is retained while dimensionality of data is significantly reduced to obviate the curse of dimensionality. Second, the efficacy of biomarkers in cancer detection can be determined.

Fate of the herbicides 2,4,5-T, atrazine, and DNOC in a shallow, anaerobic aquifer investigated by in situ passive diffusive emitters and laboratory batch experiments.

The fate of the three herbicides 2,4,5-T (2,4,5-trichlorophenoxyacetic acid), atrazine (6-chloro-N-ethyl-N'-[1-methyl-ethyl]-1,3,5-triazine-2,4-diamine), and DNOC (4,6-dinitro-2-methylphenol) in an anaerobic sandy aquifer was investigated. In the field, each of the herbicides was released simultaneously with tritiated water (HTO) as tracer in the depth interval 3 to 4 mbs (meters below surface) by use of passive diffusive emitters. Atrazine and 2,4,5-T were persistent during the approximately 18 days residence time in the aquifer. In contrast, DNOC was rapidly removed from the water phase following first-order kinetics. The removal mechanism was likely an abiotic reduction. At day 25, the first-order rate constant was 1.47 d(-1), but it decreased with time and seemed to stabilize at 0.35 d(-1) after 150 to 200 days. In the laboratory, batch experiments were conducted with sediments from 3 to 4 mbs and from 8 to 9 mbs. In these incubations, formation of Fe2+ and depletion of sulfate showed iron and sulfate reduction in sediment from 3 to 3.5 mbs and sulfate reduction in 3.5 to 4 mbs sediment. In sediment from 8 to 9 mbs, the dominant redox process was methane formation. In sediment from 3 to 3.5 mbs, only 27% to 52% of the 2,4,5-T remained after 196 days. 2,4,5-trichlorophenol was identified as the major metabolite. A lag period of at least 50 days was observed, and no degradation occurred in HgCl2 amended controls, verifying that the process was microbially mediated. In the other 2,4,5-T incubations and all the atrazine incubations, concentrations decreased linearly, but less than 25% was removed within 200 to 250 days. No degradation products could be detected, and slow sorption was the likely explanation. In all the laboratory incubations DNOC was degraded, following first-order kinetics, and when normalized to the sediment/water-ratio, the field and laboratory derived rate constants compared well. The DNOC degradation in the methanogenic incubations (8 to 9 mbs) was up to 50 times faster than in the sediments from 3 to 4 mbs, likely due to the low redox potential.