An integrated WRF/HYSPLIT modeling approach for the assessment of PM(2.5) source regions over the Mississippi Gulf Coast region.

Fine particulate matter (PM(2.5)) is majorly formed by precursor gases, such as sulfur dioxide (SO(2)) and nitrogen oxides (NO(x)), which are emitted largely from intense industrial operations and transportation activities. PM(2.5) has been shown to affect respiratory health in humans. Evaluation of source regions and assessment of emission source contributions in the Gulf Coast region of the USA will be useful for the development of PM(2.5) regulatory and mitigation strategies. In the present study, the Hybrid Single-Particle Lagrangian Integrated Trajectory (HYSPLIT) model driven by the Weather Research & Forecasting (WRF) model is used to identify the emission source locations and transportation trends. Meteorological observations as well as PM(2.5) sulfate and nitric acid concentrations were collected at two sites during the Mississippi Coastal Atmospheric Dispersion Study, a summer 2009 field experiment along the Mississippi Gulf Coast. Meteorological fields during the campaign were simulated using WRF with three nested domains of 36, 12, and 4 km horizontal resolutions and 43 vertical levels and validated with North American Mesoscale Analysis. The HYSPLIT model was integrated with meteorological fields derived from the WRF model to identify the source locations using backward trajectory analysis. The backward trajectories for a 24-h period were plotted at 1-h intervals starting from two observation locations to identify probable sources. The back trajectories distinctly indicated the sources to be in the direction between south and west, thus to have origin from local Mississippi, neighboring Louisiana state, and Gulf of Mexico. Out of the eight power plants located within the radius of 300 km of the two monitoring sites examined as sources, only Watson, Cajun, and Morrow power plants fall in the path of the derived back trajectories. Forward dispersions patterns computed using HYSPLIT were plotted from each of these source locations using the hourly mean emission concentrations as computed from past annual emission strength data to assess extent of their contribution. An assessment of the relative contributions from the eight sources reveal that only Cajun and Morrow power plants contribute to the observations at the Wiggins Airport to a certain extent while none of the eight power plants contribute to the observations at Harrison Central High School. As these observations represent a moderate event with daily average values of 5-8 µg m(-3) for sulfate and 1-3 µg m(-3) for HNO(3) with differences between the two spatially varied sites, the local sources may also be significant contributors for the observed values of PM(2.5).

Age-related retention of fiber cell nuclei and nuclear fragments in the lens cortices of multiple species.

PURPOSE: To determine the differences between species in the retention of lens fiber cell nuclei and nuclear fragments in the aging lens cortex and the relationship of nuclear retention to lens opacity. For this purpose old human, monkey, dog, and rat lenses were compared to those of three strains of mouse. We also investigated possible mechanisms leading to nuclear retention. METHODS: Fixed specimens of the species referred to above were obtained from immediate on site sacrifice of mice and rats, or from recently fixed lenses of other species, dogs, monkeys, and humans, obtained from collaborators. The retention of undegraded nuclei and nuclear fragments was graded 1-4 from histologic observation. All species lenses were examined microscopically in fixed sections stained with hematoxylin and eosin (H&E) or 4',6-diamidino-2-phenylindole (DAPI). Slit lamp observations were made only on the mice and rats before sacrifice and lens fixation. Values of 0 to 4 (clear lens to cataract) were given to degree of opacity. MRNA content in young versus old C57BL/6 mouse lenses was determined by quantitative PCR (qPCR) for DNase II-like acid DNase (DLAD) and other proteins. DLAD protein was determined by immunofluorescence of fixed eye sections. RESULTS: In old C57BL/6 and DBA mice and, to a lesser degree, in old CBA mice and old Brown Norway (BN) rats lenses were seen to contain a greatly expanded pool of unresolved whole nuclei or fragments of nuclei in differentiating lens fiber cells. This generally correlated with increased slit lamp opacities in these mice. Most old dog lenses also had an increase in retained cortical nuclei, as did a few old humans. However, a second rat strain, BNF1, in which opacity was quite high had no increase in retained nuclei with age nor did any of the old monkeys, indicating that retained nuclei could not be a cause of opacity in these animals. The nuclei and nuclear fragments were located at all levels in the outer cortex extending inward from the lens equator and were observable by the DAPI. These nuclei and nuclear fragments were seen from 12 months onward in all C57BL/6 and DBA/2 mice and to a lesser degree in the CBA, increasing in number and in space occupancy with increasing age. Preliminary results suggest that retention of nuclei in the C57BL/6 mouse is correlated with an age-related loss of DLAD from old lenses. CONCLUSIONS: A very marked apparently light refractive condition caused by retained cortical nuclei and nuclear fragments is present in the lens cortices, increasing with age in the three strains of mice examined and in one of two strains of rats (BN). This condition was also seen in some old dogs and a few old humans. It may be caused by an age-related loss of DLAD, which is essential for nuclear DNA degradation in the lens. However, this condition does not develop in old BNF1 rats, or old monkeys and is only seen sporadically in humans. Thus, it can not be a universal cause for age related lens opacity or cataract presence, although it develops concurrently with opacity in mice. This phenomenon should be considered when using the old mouse as a model for human age-related cataract.

X-ray induced cataract is preceded by LEC loss, and coincident with accumulation of cortical DNA, and ROS; similarities with age-related cataracts.

PURPOSE: To compare age-related cataractous (ARC) changes in unirradiated mice lenses to those induced by head-only X-irradiation of 3 month-old mice. METHODS: lens epithelial cells (LECs) as well as partially degraded cortical DNA were visualized in fixed sections using 4',6-diamidino-2-phenylindole (DAPI) staining, and in fresh lenses using the vital stain Hoechst 33342. reactive oxygen species (ROS) activity was also visualized directly in fresh lenses using the vital dye Dihydrorhodamine (DHR). In fixed lenses an antibody specific for 8-OH Guanosine (8-OH-G) lesions was used to visualize DNA oxidative adducts from ROS damage. Alpha smooth muscle actin was visualized using specific antibodies to determine if myofibroblasts were present. Fluorescence was quantified using Laser Scanning Confocal Microscopy (LSCM). The degree of lens opacity and cataract formation was determined by slit lamp, or from digitalized images of light reflections taken with a low magnification light microscope. RESULTS: Using DNA- and ROS-specific vital fluorescent dyes, and laser scanning confocal microscopy we have previously described 4 changes in the aging rodent lenses: 1) a significantly decreased density of surface LECs in lenses from old compared to younger mice and rats; 2) a very large increase in retained cortical nuclei and DNA fragments in the secondary lens fibers of old rodent lenses; 3) increased cortical ROS in old rodent lenses; 4) increased cataract concomitantly with the cortical DNA and ROS increases. In the current study we report that these same 4 changes also occur in an accelerated fashion in mice given head-only X-irradiation at 3 months of age. In addition to vital staining of fresh lenses, we also examined sections from fixed eyes stained with DAPI or hematoxylin and eosin (H&E) and found the same loss of surface LECs and accumulation of undigested nuclei and debris in secondary lens fibers occur with age or following X-irradiation. In addition sections from fixed-eyes were examined for ROS damage to DNA with antibodies specific for 8-OH-G lesions. The frequency of 8-OH-G lesions increased dramatically in lenses from old unirradiated mice over 24 months of age, and similarly in X-irradiated lenses by 9-11 months post irradiation. The accumulation of cortical nuclei was not the result of conversion or invasion by myofibroblasts as tested by antibodies to a marker for such cells, alpha smooth muscle actin. CONCLUSIONS: X-irradiation damage induces a large decrease in surface LECs over a period of 3-11 months post X-irradiation of young mice. These changes are similar in extent to those seen in 24-29 months-old control mouse lenses with age-related cataracts. In 24+ month-old unirradiated mice the secondary lens fibers are not able to degrade nuclei or nuclear DNA efficiently and accumulate large numbers of cortical nuclei and nuclear fragments as well as ROS and 8-OHG lesions. X-irradiated lenses develop the same abnormalities in a more accelerated fashion. The extensive loss of LECS and accumulation of undegraded nuclei, ROS, and ROS damage may play a causal role in cataract generation in both unirradiated old mice and in previously irradiated young adult mice.

Dietary restriction suppresses proteotoxicity and enhances longevity by an hsf-1-dependent mechanism in Caenorhabditis elegans.

Dietary restriction increases lifespan and slows the onset of age-associated disease in organisms from yeast to mammals. In humans, several age-related diseases are associated with aberrant protein folding or aggregation, including neurodegenerative disorders such as Alzheimer's, Parkinson's, and Huntington's diseases. We report here that dietary restriction dramatically suppresses age-associated paralysis in three nematode models of proteotoxicity. Similar to its longevity-enhancing properties, dietary restriction protects against proteotoxicity by a mechanism distinct from reduced insulin/IGF-1-like signaling. Instead, the heat shock transcription factor, hsf-1, is required for enhanced thermotolerance, suppression of proteotoxicity, and lifespan extension by dietary restriction. These findings demonstrate that dietary restriction confers a general protective effect against proteotoxicity and promotes longevity by a mechanism involving hsf-1.

Radiation cataracts: mechanisms involved in their long delayed occurrence but then rapid progression.

PURPOSE: This study was directed to assess the DNA damage and DNA repair response to X-ray inflicted lens oxidative damage and to investigate the subsequent changes in lens epithelial cell (LEC) behavior in vivo that led to long delayed but then rapidly developing cataracts. METHODS: Two-month-old C57Bl/6 female mice received 11 Grays (Gy) of soft x-irradiation to the head only. The animals' eyes were examined for cataract status in 30 day intervals by slit lamp over an 11 month period post-irradiation. LEC migration, DNA fragment, free DNA retention, and reactive oxygen species (ROS) presence were established in the living lenses with fluorescent dyes using laser scanning confocal microscopy (LSCM). The extent and removal of initial LEC DNA damage were determined by comet assay. Immunohistochemistry was used to determine the presence of oxidized DNA and the response of a DNA repair protein in the lenses. RESULTS: This treatment resulted in advanced cortical cataracts that developed 5-11 months post-irradiation but then appeared suddenly within a 30 day period. The initially incurred DNA strand breaks were repaired within 30 min, but DNA damage remained as shown 72 h post-irradiation by the presence of the DNA adduct, 8-hydroxyguanosine (8-OHG), and a DNA repair protein, XRCC1. This was followed months later by abnormal behavior by LEC descendant cells with abnormal differentiation and migration patterns as seen with LSCM and fluorescent dyes. CONCLUSIONS: The sudden development of cortical cataracts several months post-irradiation coupled with the above findings suggests an accumulation of damaged descendants from the initially x-irradiated LECs. As these cells migrate abnormally and leave acellular lens surface sites, eventually a crisis point may arrive for lens entry of environmental O(2) with resultant ROS formation that overwhelms protection by resident antioxidant enzymes and results in the coagulation of lens proteins. The events seen in this study indicate the retention and transmission of progenitor cell DNA damage in descendant LEC. The cellular and molecular events parallel those previously reported for LSCM observations in age-related cataracts.

Cellular debris and ROS in age-related cortical cataract are caused by inappropriate involution of the surface epithelial cells into the lens cortex.

PURPOSE: To quantify changes in the lens epithelial cells and underlying lens cortex responsible for age-related cortical cataract (ARCC) in the rat. METHODS: Freshly isolated lenses were stained vitally for DNA with Hoechst 33342. Reactive oxygen species (ROS) and mitochondria were visualized and quantified by dihydrorhodamine 123 (DHR). The fluorescence was quantified using Laser Scanning Confocal Microscopy (LSCM) of vitally stained lenses. Cortical DNA was verified as such by DNAse I digestion. Cataract reflections were determined from digitalized images of light reflections taken with a low magnification light microscope, or with the LSCM. RESULTS: The anterior surface epithelia of old rat lenses were full of gaps and ragged in appearance with a decrease of over 50% in lens epithelial cell (LEC) density. The surface LECs were frequently seen to have involuted into the cortex at inappropriate sites, forming deposits full of DNA, nuclear and mitochondrial debris, and abundant ROS. These involutions frequently originated near open gaps in the surface epithelia, where they appear to have detached from the capsular membrane. Cortical cataracts in the rat lenses were seen to co-localize with these LEC involutions, as had been seen previously in mice with ARCC. CONCLUSIONS: ARCC in rats co-localized with inappropriate accumulations of nuclei, mitochondria, DNA, and expression of ROS in debris filled foci. These were the result of both involution of surface LECs into areas of cortical ARCC, and by an extension of the normal bow region deep into the anterior and posterior of cataractous lenses. These results were in complete agreement with our previous studies on ARCC in mice.

Accumulation of DNA, nuclear and mitochondrial debris, and ROS at sites of age-related cortical cataract in mice.

PURPOSE: Lenses from young and old mice were analyzed by laser scanning confocal microscopy (LSCM) with vital dyes, to determine whether age-related subcapsular and cortical cataracts were linked to the failure of lens fiber cells to degrade nuclei, DNA, and mitochondria properly and whether they result in the overproduction of reactive oxygen species (ROS) at the same sites. RESULTS: As opposed to the clear DNA-free subcapsular and cortical areas of young adult mouse lenses, these areas in cataractous old mouse lenses were found to contain accumulations of nuclei, nuclear fragments, aggregated mitochondria, and amorphous DNA as cortical inclusions (P < 0.001 between young and old lenses). These inclusions correlated spatially with age-related cataracts and with the presence of ROS. The source of such undegraded material was a large expansion of transition nuclei in the bow region and also direct involution of surface lens epithelial cells (LECs) into the underlying cortex, frequently leaving bare patches devoid of nuclei on the surface of the anterior epithelium. METHODS: Live lenses were stained vitally for DNA with Hoechst 33342. ROS and mitochondria were stained and quantified with dihydrorhodamine 123 (DHR). In fixed lenses, DNA was stained with propidium iodide (PI) or 4',6-diamidino-2-phenylindole, dihydrochloride (DAPI). The intensity and position of each probe's fluorescence was determined by LSCM. Cataract localization was ascertained by digitalized microscopy of reflected light. CONCLUSIONS: In aged mice, most subcapsular and cortical cataracts colocalize with accumulations of nuclei, mitochondria, and DNA, These effects are accompanied at the same sites by the production of ROS. The condition is due to the failure of lens fiber cells in the bow region to differentiate properly into the clear fiber state and to the improper involution of cells from the anterior epithelium directly into the underlying cortex, resulting in cataractous opacities.

Age-related cataract progression in five mouse models for anti-oxidant protection or hormonal influence.

Five mouse models with known alterations of resistance to oxidative damage were compared by slit lamp examination for the presence and degree of advancement of age-related cataract in young adult and old animals along with wild type controls. A group of young and old normal C57BL/6Jax mice were examined first to constitute a standard, and they were found to exhibit age-related cataract development. Following this, four models on the C57BL/6 background with imposed genetic alterations affecting anti-oxidant enzyme presence or activity, and one outbred model in which a deletion blocked the growth hormone/IGF-1 axis, were similarly examined. There was no evidence of foetal or juvenile cataract development in any of these models, and an age-related severity for lens opacities was shown between young adult and old mice in all groups. Model 1, mice null for the anti-oxidant gene glutathione peroxidase-1 (GPX1) had significantly advanced cataracts in older mice vs. same age controls. In mouse model 2 hemizygous knockout of SOD2 (MnSOD) did not affect age-related cataract development. In model 3 combining the GPX1 and SOD2 deficiencies in the same animal did not advance cataract development beyond that of the GPX1 null alone. In model 4 the addition of anti-oxidant protection in the lens by transfection of human catalase targeted only to the mitochondria resulted in a significant delay in cataract development. The 5th model, growth hormone receptor knockout (GHR-/-) mice, also demonstrated a significant reduction in age-related cataract development, as well as dwarfism. These findings, in general, support the oxidative theory of age-related cataract development. The exception, the partial deletion of SOD2 in the hemizygous KO model, probably did not represent a sufficiently severe deprivation of anti-oxidant protection to produce pathologic changes in the lens.

White light-mediated DNA strand breaks in lens epithelial cells.

It was hypothesized that white light can induce DNA single-strand breaks/alkali labile sites in lens epithelial cells during the dissection process when lenses are being collected for study. Dissection of lenses in white light significantly increased 'Integrated Intensity' (P=0.0216), an index of DNA single strand breaks/alkali labile sites, using the alkaline microgel electrophoresis assay. In addition, we investigated the DNA damaging effect of visible light in a human lymphocyte model in which cells were exposed to white light, red light or minimal indirect light for 5 min. Results of these experiments confirm our conclusion that visible light induces DNA damage in cells. These findings may be relevant in a genotoxicity study on cells ex vivo.