A prospective, non-randomised, transversal and comparative study, carried out in INOVA Vision Institute and Autonomous University of Aguascalientes. Pterygium is an important illness that affects 22% people from tropic and equatorial zones. Is an inflammatory process caused by UV rays, and it has a behavior similar to a neoplasm. For this study was taken into consideration 191 samples from the INOVA Vision Institute, Aguascalientes, Mexico. Include 73 pterygia samples, which were obtained during resection under sterile conditions. 44 normal conjunctiva samples were obtained from the same patients when harvesting the conjunctival autograft, or from other patients undergoing extracapsular cataract extraction from the superior bulbar region. Tears from patients with pterygium (n = 50) and normal volunteers (n = 24) were obtained using a calibrated glass micro capillary tube. The surgical conjunctiva and pterygia samples were subjected to reverse-transcription polymerase chain reaction (RT-PCR), western blot, and immunohistochemistry. Tears were analyzed by enzyme-linked immunosorbent assays.This was a prospective, non-randomised study involving 191 biological samples taken from patients with pterygium and normal volunteers, whom were operated under local anaesthesia by either complete resection of the lesion with primary closure, or resection with conjunctival autograft. Tissue samples were fixed in 10% formaldehyde. Sections were routinely stained with hematoxylin and eosin. HCC expression was evaluated by reverse-transcription polymerase chain reaction (RT-PCR), immunohistochemistry, and by western blotting. All tears samples were analyzed by enzyme-linked immunosorbent assays (ELISA).Expression levels and distribution patterns of HCC in normal conjunctiva and pterygium. Higher levels of HCC mRNAs and proteins were detected in pterygium compared with a normal conjunctiva. Immunohistochemistry revealed that HCC was localized in the apical cells of the epithelium in the normal conjunctiva. In contrast, HCC was detected in all extension of epithelial tissue, from apical to basal cells in pterygia. The concentration of HCC protein in tears was higher in patients with pterygium versus controls.HCC may play an important role in protecting normal conjunctiva, and regulating inflammatory conditions of the anterior ocular surface. A pterygium is a fibrovascular lesion of the ocular surface that can display aggressive clinical behavior and occasionally threaten vision . The lesThis was a prospective, non-randomised study involving 191 biological samples taken from patients with pterygium and normal volunteers. HCC expression was evaluated using several techniques. Written informed consent was obtained for each patient. Biological samples were taken during surgery (pterygia and conjunctivas) or consultation (tears) from the anterior segment department of INOVA Vision Institute, Aguascalientes, Mexico. All experiments were carried out in accordance with the Helsinki Declaration and approved by the medical ethics committee of the INOVA Vision Institute. 52 surgical of primary pterygia samples from persons its age ranging between 55 to 65 years old (median age 55.21 ± 10 years), the 21 recurrent pterygia samples were obtained from persons which ages ranging between 58 to 63 years old (median age 65.12 ± 14 years old), the samples were obtained during resection under sterile conditions. 44 normal conjunctiva samples from persons which ages ranging between 52 to 68 years old (median age 62.57 ±6 years) the conjunctivas were obtained from the same patients when conjunctival autografts were harvested, or from other patients undergoing extracapsular cataract extraction from the superior bulbar region. Tears from 50 patients with pterygium and 24 normal volunteers who ages ranging between 51 to 67 years old (median age 61.31 ±9 years) were obtained using a calibrated glass micro capillary tube. One-third of tissue samples (pterygia and conjunctivas) were analyzed by reverse-transcription polymerase chain reaction (RT-PCR), one-third by immunohistochemistry, and the remainder by western blotting. All tears samples were analyzed by enzyme-linked immunosorbent assays (ELISA).Patients with pterygium were operated under local anaesthesia by either complete resection of the lesion with primary closure, or resection with conjunctival autograft . The ptePatients were evaluated with a slit lamp to exclude any disease of the anterior segment. Tear samples were collected using a 5-μl calibrated glass micro capillary tube  without touching the eye or eyelids. Samples were taken at different times of the day to avoid diurnal variations . One samPterygia and normal conjunctivas were fixed in 10% formaldehyde. Sections were routinely stained with hematoxylin and eosin . Each seWe used an Axioscop 40 (Carl Zeiss) light microscope equipped with a digital image acquisition system. For each sample examined, 10 randomly chosen fields from epithelial tissue represented by continuous lines squares in Figure ′-GCG GCG TGC ACT GGA CTT TG-3′ (HCC primer forward) and 5′-GCC GCC TGC TGC CTT CTC TG-3′ (HCC primer reverse). The PCR consisted of 1 μl of cDNA, 1.5 mM MgCl2, 0.2 mM dNTP, 0.5 μM each primer and 2.5 U of Taq DNA polymerase. Thermal cycling involved a denaturing step at 94°C for 5 min, followed by 25 cycles of denaturation  and annealing , then a final extension step at 72°C for 2 min. All reactions were performed in duplicate. Amplicons were visualized by electrophoresis on 1.5% (w/v) agarose gels stained with ethidium bromide.RNA was extracted from individual pterygium samples and pooled normal conjunctivas. RNA was extracted from 5 mg of tissue using a total RNA Isolation System  according to the manufacturer’s instructions. Purity and quality of RNA were assessed with the Nanodrop 1000 system . We obtained 90 ± 10 ng of RNA for each pterygium, and 100 ± 10 ng of RNA for pooled conjunctiva samples. Synthesis of cDNA was conducted using SuperScript™ first strand synthesis for RT-PCR  at 55°C for 45 min. Primers for the PCR were designed using DNAstar 4.02 software. Primer sequences were 5Of the 74 tear samples we analyzed, 28 were from patients with primary pterygium; 22 were from recurrent pterygium, and 24 were from a normal conjunctiva. In the wells of a 96-well plate, 50 μl of a tear sample was added to 50 μl of carbonate/bicarbonate buffer (pH 10) to coat each well. Plates were incubated at 4°C overnight. We added 0.1% (w/v) BSA in PBS (pH 7.4) to each well for 45 min at 37°C, then performed three 5-min washes with PBS containing 0.05% (v/v) Tween-20 (PBS-Tw). Rabbit anti-HCC (diluted 1:1000 in PBS-Tw) was added to wells and incubated for 1.5 h at 37°C. Wells were washed with PBS-Tw, and then incubated with goat anti-rabbit IgG conjugated to horseradish peroxidase (diluted 1:2500 in PBS-Tw) for 1 h at room temperature. Wells were again washed with PBS-Tw, and ortho phenylenediamine added for color development. Absorbance values for each well were determined at 490 nm in a iMark (Bio Rad) spectrophotometer .Frozen tissue sections from pterygium and conjunctiva were placed in a glass tissue macerator (Kontes 15 ml) with lysis buffer -1-chloro-3-tosylamido-7-amino-2-heptanone hydrochloride TLCK and 1 mM phenylmethylsulfonyl fluoride PMSF). The proportion of tissue to lysis buffer was 1 g/ml. Tissues were lysed with 100 strokes, and samples kept on ice; however, it was necessary to centrifuge samples at 15,000 rpm for 10 min at 4°C as they became dense. We obtained two phases: the bottom phase corresponded to membrane contents, and the supernatant contained the contents of the cytosol. These phases were stored at −20°C until required. We subjected human serum to an LC10 column  according to the manufacturer’s instructions. Briefly, 100 μL of serum was diluted 5-fold with buffer A and injected onto the column in buffer A at a flow rate of 0.5 ml/min for 25 min, then 2.0 ml/min for 5 min on a Shimadzu LC10A VP system . After collection of the flow-through fraction containing unbound proteins, the column was washed and bound proteins were eluted with buffer B (stripping buffer) at a flow rate of 2.0 ml/min for 18 min. Fractions were collected into 1.5 ml micro centrifuge tubes.A standard curve was constructed using BSA (range 0–1 mg/ml). Samples (1 μl of membrane or cytosol contents) were added to 1 ml of Bradford's solution . The absA positive control was taken from the serum patients with chronic renal failure. Pterygia and conjunctiva tissues containing 100 μg of protein were mixed with an equal volume of buffer and then boiled for 5 min. Samples from human serum were dissolved in 15 μl of sample preparation buffer, mixed well, and subjected to sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis according to Laemmli protocol . SamplesDifferential expression of HCC was observed, with an over expression in recurrent and primary pterygia compared with a normal conjunctiva Figure B.In the healthy conjunctiva, we observed non-keratinized stratified squamous epithelium with four to eight layers of cells Figure A. These In the normal conjunctiva, we detected HCC in most apical epithelial cells . A comparison of recurrent pterygia and normal conjunctivas, or of primary and recurrent pterygia, indicated no significance.From the images that we obtained we saw that samples in the pterygia groups contained larger positive areas in terms of distribution regarding the conjunctiva Figure . We compHCC expression levels were increased in active primary and active recurrent pterygia Figure  as deterOur analysis showed that HCC molecules were present in the tear film of all patients and controls. In healthy people, the concentration of HCC was lower (0.05 UA ± 0.0003) than in patients with pterygium  obtained from surgery, and tears from patients, with and without pterygium. All patients who underwent surgery had active pterygium. Conversely, outpatients with pterygium were classified as active if they presented with vascularization, edema and active hyperaemia, or as inactive if they showed a fibrotic component. Using RT-PCR we found that HCC was expressed in both normal conjunctiva and pterygium, supporting the findings of Chang et al., who found HCC in another mucosa . AbrahamUsing immunohistochemistry, we found that normal conjunctiva contained HCC in the most superficial epithelial layers. In contrast, HCC was expressed in all epithelial tissues of pterygia, from basal epithelial cells to the most superficial layer of the epithelium. It is interesting to note that this compartment has a high proliferatives rate in comparison with a normal conjunctiva, such as morphological changes we observed in a picture from two tissues stained with H&E technique Figures A and B. Our ELISA and western blot results indicate low levels of HCC in the tears and tissues of patients with a normal conjunctiva. The amount and concentration of HCC were increased in the tears and tissues from patients with pterygium, with the highest levels observed in patients with active recurrent pterygium. Sharman et al., assess the concentration of HCC in gingival crevicular fluid (GCF) and serum from patients with various periodontal diseases, the mean HCC concentration in GCF and serum was observed to be highest in patients with periodontitis, and lowest in healthy individuals, they suggested that HCC levels increased with disease progression to prevent further periodontal degeneration; HCC levels then decrease after treatment to maintain bone metabolic homeostasis . LertnawWe found that HCC was present to a great extent in epithelial cells of pterygium samples. Underlying cells in CT did not exhibit strong staining for HCC, suggesting that the production of HCC is confined to epithelial cells. This further indicates that pterygium does not involve a clear inflammatory process. Based on these findings, we can deduce that HCC acts as an epithelial proliferatives stimulant, and at the same time as a modulator of inflammation in underlying CT. This is similar to what was outlined by Koroloenko et al., (2008)  and TaveOur study has established that HCC is expressed in normal conjunctiva and that this protein is up regulated in proliferatives diseases of the ocular surface, such as pterygium. This suggests that HCC may play a key role(s) in the protection of a conjunctiva against aggressors, and may regulate the inflammatory ocular response. The molecular mechanism and potential of HCC as a prognostic biomarker for activity or recurrence of pterygium remains an area that requires further investigation.HCC: Human cystatin C; RT-PCR: Reverse-transcription polymerase chain reaction; ELISA: Enzyme-linked immunosorbent assay; ECM: Extracellular matrix; MMP: Matrix metalloproteinase; kDa: Kilodaltons; PBS: Phosphate-buffered saline; BSA: Bovine serum albumin; RNA: Ribonucleic acid; DNA: Deoxyribonucleic acid; PCR: Polymerase chain reaction; GCF: Gingival crevicular fluid; SLE: Systemic lupus erythematosus; ESR: Erythrocyte sedimentation rate; TNF-α: Tumor necrosis factor-alpha; IL-6: Interleukin 6; Pri: Primary; Rec: Recurrent; Pt: Pterygia.The authors declare that they have no financial interest in any of the products used in this research.BGLF carried out the molecular genetic and immunohistochemistry studies, participated in the design of the study, and drafted the manuscript. VJJ carried out the immunohistochemistry studies, participated in data analysis and helped draft the manuscript. KD critically revised the manuscript. GCR carried out molecular genetic studies and participated in the design of the study. TBE carried out the immunoassays and their statistical analysis. TBLF conceived of the study, helped design and coordinate experiments, and helped to draft the manuscript. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2415/13/6/prepub


Background: Physical inactivity and exposure to air pollution are important risk factors for death and disease globally. The built environment may influence exposures to these risk factors in different ways and thus differentially affect the health of urban populations.Objective: We investigated the built environment’s association with air pollution and physical inactivity, and estimated attributable health risks.2.5), nitrogen oxides (NOx), and ozone (O3)] for 30,007 individuals in southern California. We then estimated the resulting risk for ischemic heart disease (IHD) using literature-derived dose–response values. Using a cross-sectional approach, we compared estimated IHD mortality risks among neighborhoods based on “walkability” scores.Methods: We used a regional travel survey to estimate within-urban variability in physical inactivity and home-based air pollution exposure [particulate matter with aerodynamic diameter ≤ 2.5 μm , suggesting that population health benefits from increased physical activity in high-walkability neighborhoods may be offset by adverse effects of air pollution exposure.Results: The proportion of physically active individuals was higher in high- versus low-walkability neighborhoods (24.9% vs. 12.5%); however, only a small proportion of the population was physically active, and between-neighborhood variability in estimated IHD mortality attributable to physical inactivity was modest . Between-neighborhood differences in estimated IHD mortality from air pollution were comparable in magnitude (9 more IHD deaths/100,000/year for PMPolicy implications: Currently, planning efforts mainly focus on increasing physical activity through neighborhood design. Our results suggest that differences in population health impacts among neighborhoods are similar in magnitude for air pollution and physical activity. Thus, physical activity and exposure to air pollution are critical aspects of planning for cleaner, health-promoting cities. Physical inactivity is associated with increased risk of several adverse health outcomes including heart disease, type 2 diabetes, colon cancer, breast cancer, and mortality . Active Exposure to outdoor urban air pollution is associated with various adverse health outcomes including heart disease, respiratory disease, lung cancer, asthma, and mortality . ChronicRecently, the World Health Organization (WHO) cited physical inactivity (4th) and exposure to outdoor urban air pollution (14th) among the top 15 risk factors for the Global Burden of Disease (WHO 2009); for high-income countries, these ranks are 4th  and 8th (outdoor air pollution). Urban planning and the built environment may differentially influence exposures to those two risk factors . A small2.5), nitrogen oxides (NOx), and ozone (O3)] to assess relationships between urban form and public health.We used risk assessment to explore urban-scale spatial patterns in exposures associated with the built environment. We investigated differences in urban form that have been associated with physical inactivity and air pollution . To our knowledge, no other metropolitan-scale travel survey has been used to estimate physical activity and exposure to air pollution  geocoded home location , b) home location within the air pollution modeling domain—the South Coast Air Basin , and c) complete demographic information . The survey generally covered 1 weekday per participant. We multiplied each participant’s 1-day physical activity record by 7 to obtain an estimate of weekly minutes of physical activity. This approach assumed that physical activity was constant across all days of the week. Population-average levels of physical activity were similar (< 15% difference) between weekdays and weekends  based on data from a small number of respondents  who participated in an additional weekend survey supplement . The survey recorded total physical activity and separately disaggregated that total into active transport  versus recreational activities .Of the 40,376 survey respondents, 30,007 (74%) met our inclusion criteria: 2.5, NOx, and O3 in 2001. We interpolated concentrations  for PMmonitors ] to eachNeighborhood walkability. We calculated three built environment variables to represent neighborhood type: a) population density, b) intersection density, and c) land use mix. Neighborhoods that were in the upper (lower) tertile of all three built environment variables were defined as high- (low-) walkability neighborhoods. This approach classified 12% of the survey population as living in a high-walkability neighborhood and 18% as living in a low-walkability neighborhood. We used objective measurements of the built environment rather than geographical overlays to match methods commonly used in the urban planning literature. Although no standard measure of walkability exists, most indices include measures of density, connectivity, and land use mix (http://dx.doi.org/10.1289/ehp.1103806)]. Results were similar for both definitions; therefore, we report results using the first definition only. use mix . As a se use mix  we implePopulation density. We used U.S. Census data from the year 2000 to calculate population density at the tract level for each household (U.S. Census Bureau 2000). Population density has been shown to be a predictor of per capita automobile travel  and tripIntersection density. Intersection density was calculated using road TIGER/Line data (U.S. Census Bureau 2000). A 1-km non-freeway network buffer was generated for each household using ArcGIS . Intersections (more than two road segments) were summed within the buffer, yielding a measure of street connectivity. Previous studies show that street connectivity may reduce vehicle travel and increase walking .http://dx.doi.org/10.1289/ehp.1103806)] is a normalized ratio of the mix of four primary land uses  to total land area within the 1-km network buffer. The index ranges from 0 to 1: A value of 1 represents an equal mixture of the four land uses; a value of 0 indicates 100% of land is a single land use. Impacts of land use mix on health include reducing obesity , we estimated relative risks (RRs) attributable to outdoor air pollution and physical inactivity for one important health outcome: IHD. IHD is consistently associated with outdoor air pollution and physical inactivity (WHO 2009), is responsible for a large proportion of deaths in the United States  [Centers for Disease Control and Prevention (CDC) 2009], and has been shown to be an important health outcome for both risk factors when considering large-scale shifts to active travel (http://dx.doi.org/10.1289/ehp.1103806)] for the range of observed air pollutant concentrations and the referent exposure levels described below. In contrast, WHO (2004) suggests a three-tier dose–response for physical activity: a) active , b) insufficiently active , and c) inactive , allowing for only three possible physical activity RRs for each survey participant. We estimated attributable fractions for outdoor air pollution and physical inactivity using the mean individual RR in high- or low-walkability neighborhoods.e travel . BecauseJournal of the American Medical Association, New England Journal of Medicine, British Medical Journal, Lancet) for the years 2000–2010 for air pollution risk estimates. We also performed a search of key words in Google Scholar and ISI Knowledge, including (in various combinations) “air pollution,” “O3/NOx/PM2.5,” “ischemic heart disease,” “cardiovascular disease,” “cardiopulmonary disease,” “respiratory disease,” “mortality,” “health effects,” “chronic/acute,” and “dose–response.” We used the “cited by” function in Google Scholar to explore subsequent studies related to each article. Through this process, we identified 62 articles. We then selected studies that focused on within-city variation and included IHD as a health outcome : 1.06, 1.11]. 3 increase in PM2.5 . 2.5 in Los Angeles, but the RR estimate is roughly consistent with two between-city studies that did report statistically significant RRs: 3 increase in PM2.5; 95% CI: 1.14, 1.23) and 3 increase in O3  was based on between-city variation (ACS cohort) in 96 U.S. metropolitan statistical areas generated from a one-pollutant model. However, it is important to note that 3 based on a two-pollutant model adjusted for PM2.5 , and overall there is less evidence in the literature for O3 associations with IHD compared with those for PM2.5. A within-city study of O3 and IHD was not available.Each RR for air pollution was estimated from cohort studies of long-term exposures; however, these estimates differed in important ways. For example, 3 for PM2.5, 39.8 μg/m3 for NOx, 80.3 μg/80.3 μg/m3 for O3) for air pollution, consistent with exposures in a relatively clean neighborhood in the study area. Each survey participant’s air pollution RR was estimated based on the difference between their home-location air pollution exposure and the referent exposure level. For example, for PM2.5, an individual whose home-location exposure estimate was 23.6 μg/m3 (10 μg/m3 above the referent level) would be assigned an RR of 1.25.The referent exposure levels used to estimate individuals’ RRs were “active” for physical inactivity (> 150 min of moderate-vigorous activity per week), and the 10th percentile of exposure  and estimated attributable IHD mortality rates for each risk factor in high- and low-walkability neighborhoods. PAF for a neighborhood was calculated based on the proportion of individuals exposed to each risk factor and average RR among all individuals in a neighborhood . We used the 2000–2001 age-adjusted IHD mortality rate in California . Attributable mortality due to physical inactivity, PM2.5, NOx, and O3 cannot be summed because of confounding among the risk factors and overlap of at-risk populations. Therefore, we report attributable mortality due to the different factors separately.Here, RR is the mean individual RR in each group  and risk factor, and http://dx.doi.org/10.1289/ehp.1103806)] Results based on this alternative method were similar to those reported below.We separately calculated PAF using a method with multiple exposure levels instead of the dichotomous exposure levels implicit in Sensitivity analyses. To explore the robustness of our estimates, we used three sensitivity analyses to assess a) different methods of scaling minutes of physical activity, b) alternate modeling approaches for air pollution, and c) stepwise versus linear dose–response for physical activity.a) every other day or b) every third day. The Monte Carlo simulation distributes total minutes of physical activity accordingly, stratifying by age, sex, and ethnicity. The resulting distributions of physical activity better approximate national estimates on the prevalence of physical inactivity (WHO 2004).Sensitivity analysis 1: scaling method for minutes of physical activity. Our approach requires extrapolating weekly exercise rates based on the 1-day travel diary because most physical activity epidemiological literature employs the metric “minutes of physical activity per week.” To test the limitations of this extrapolation for our analysis, we developed a Monte Carlo simulation that relaxes our base-case assumption , by employing two alternative assumptions: that people who are nonsedentary are physically active http://www.camx.com; nitrous oxide (NO), nitrogen dioxide (NO2), O3] and land-use regression ; Sensitivity analysis 3: physical activity dose–response. We tested the sensitivity of our results to the dose–response curve for physical inactivity. Our base case used the stepwise dose–response from WHO (2004) . For thi3 for NO2 . PM2.5 exposures were approximately 1.5 and 2 times higher than U.S. EPA (15 μg/m3) and CalEPA (12 μg/m3) long-term standards , respectively . However, considering nonsedentary individuals only, average physical activity was 24% lower in high- than in low-walkability neighborhoods (410 vs. 543 min/week). This finding suggests that neighborhood type may have differing impacts on the number of people participating in physical activities, average physical activity among all individuals, and average physical activity among nonsedentary individuals.Average per capita physical activity was 50% higher in high- than in low-walkability neighborhoods ].The self-reported purpose of physical activity differs by neighborhood . For exa2.5 was smaller , but the difference between neighborhoods was slightly larger than for physical inactivity . O3 shows the reverse spatial pattern as PM2.5  but a smaller difference in mortality between neighborhoods . Attributable IHD mortality rates for NOx ].Sensitivity analysis 1: scaling method for minutes of physical activity. Results  indicate that our alternative assumptions reduce the variability in physical activity among neighborhoods. Specifically, the Monte Carlo simulation increases the share of nonsedentary individuals  but also yields reductions in estimated IHD mortality differences among neighborhoods. Our core conclusions are similar among the Monte Carlo simulations.Sensitivity analysis 2: air pollution model. Central tendencies varied by pollutant and model; however, trends in the core conclusions  were similar where it was possible to compare . In general, differences in estimated IHD mortality rates between high- and low-walkability neighborhoods were larger when using the alternate models; therefore, base-case results reported above may be conservative estimates  of air pollution spatial variability.Sensitivity analysis 3: physical activity dose–response. Our results did not change appreciably when using the linear dose–response curves .http://dx.doi.org/10.1289/ehp.1103806)]. Prior literature further explores socioeconomic aspects of this topic  within strata of age  and according to income and ethnicity [high income  and white vs. low income  and nonwhite]. The results reveal similar trends in risk differences between neighborhoods for each strata, suggesting that our results are robust to accounting for differences in income, ethnicity, and age. Details are in the Supplemental Material [pp. 11–14, Table S6 . This trade-off suggests that the net health impact of neighborhoods may depend in part on spatial patterns of air pollution.Our analysis summarizes between-neighborhood variations in two risk factors  using a time–activity travel diary for one region. We found risks were differential when stratified by neighborhood walkability. Specifically, when comparing estimated IHD mortality rates among neighborhoods, differences attributable to physical inactivity were modest and comparable to differences attributable to individual air pollutants. Because of spatial patterns associated with each pollutant, urban residents were often highly exposed to at least one but not all pollutants , affecting estimates of travel time . The SCAx, PM2.5, O3), and physical inactivity. Clearly, further analyses incorporating other risk factors  linked to the built environment are warranted. Interaction between physical activity and air pollution may vary on an even smaller scale than we have investigated in the present study . Future analyses could use age-specific risks of IHD mortality for air pollution and physical inactivity. Our analysis is descriptive  in nature; more research is needed to explore causality between urban form and health risks .Our work is motivated by the goal of understanding and designing clean, healthy, sustainable cities . Our invDespite these limitations, our results are relevant to health officials, sustainability scientists, and urban planners. To our knowledge, ours is the first analysis that directly compares health risks for both air pollution and physical inactivity among neighborhoods based on activity patterns for a random sample of residents in an urban area, and thus is the first to quantify relationships between urban form and the health impacts of physical activity and air pollution. We found that attributes of the built environment were associated with both air pollution exposure and physical inactivity. These results emphasize that to be health protective, neighborhoods designed to decrease risks from one factor must avoid unintentionally increasing risks from other factors.2.5 and NOx were greater in high- than in low-walkability neighborhoods, whereas estimated IHD mortality due to increased O3 was greater in low- than in high-walkability neighborhoods. In general, differences in estimated IHD mortality between neighborhoods were comparable for exposure to air pollutants and physical inactivity. Our results suggest complex within-urban spatial trade-offs in health risks associated with air pollution and physical inactivity. Efforts to design healthy neighborhoods should account for many factors, including air pollution and physical inactivity, and not address one concern at the expense of others.We compared the health impacts attributable to air pollution and physical inactivity among neighborhoods for one cohort . A larger proportion of our Southern California study population was classified as nonsedentary in high- versus low-walkability neighborhoods (25% vs. 13%). However, because only a small share of the total population was classified as physically active, we estimated only moderate differences in IHD mortality rates attributable to physical inactivity between neighborhood types. Spatial patterns of estimated attributable IHD mortality rates varied by pollutant: estimated mortality due to increased PM


Mice were orally treated with distilled water or black maca during 35 days starting 7 days after surgery. Memory and learning were assessed using the water Morris maze (from day 23–27) and the step-down avoidance test (days 34 and 35). At the end of each treatment, mice were sacrificed by decapitation and brains were dissected out for MDA, Ache and MAO determinations. Black maca (0.5 and 2.0 g/kg) increased step-down latency when compared to OVX control mice. Black maca decreased MDA and Ache levels in OVX mice; whereas, no differences were observed in MAO levels. Finally, black maca improved experimental memory impairment induced by ovariectomy, due in part, by its antioxidant and Ache inhibitory activities.The present study aims to test two different doses of aqueous extract of black maca on learning and memory in ovariectomized (OVX) mice and their relation with malonalehyde (MDA), acetylcholinesterase (Ache) and monoamine oxidase (MAO) brain levels. Female mice were divided into five groups: (i) naive (control), (ii) sham, (iii) OVX mice and OVX mice treated with (iv) 0.50 g kg Lepidium meyenii Walp., known as maca, grows over 4000 m altitude in the central Peruvian Andes. Previous studies were focused to demonstrate the traditional fertility-enhancing properties of the hypocotyls of maca [in vitro and in vivo [ of maca –3. In ad in vivo .Lepidium, including maca, belong to the cruciferous (Brassicaceae) family and it is possible that this plant may have effects on cognitive functions. In fact, many reports have claimed other non-traditional properties of maca including its capacity to reduce menstrual and menopausal symptoms in women, promote mental clarity, restore hormonal balance in women and improve memory [Epidemiological studies found a significant correlation between dietary intake of vegetables and improvement in cognitive function in elderly people . For inse memory –10. Preve memory . In addie memory .Ovariectomy is a well-known animal model to induce memory impairment in rodents . It was Considering that: (i) maca is used for its capacity to improve memory, (ii) maca has antioxidant activity, (iii) black maca shows the greatest effect on cognitive function, (iv) black maca showed acetylcholinesterase (Ache) inhibitory activity and no effect on monoamine oxidase (MAO) levels in male mice and (v) ovariectomy is an female animal model that induce memory impairment related to an increased oxidative stress and cholinergic and MAO dysfunction, the present study aims to evaluate the effect of black maca in ovariectomy-induced memory impairment in female mice and its effect on malonalehyde (MDA), Ache and MAO brain levels.ad libitum. All animal experiments were conducted in compliance with “Guide of the care and use of laboratory animals" [Female mice of 3-months old from the Kunming strain (28.29 ± 0.28 g) were used for the study. Mice were maintained at ambient temperature (23.0 ± 2.0°C) with a 12 : 12 h light/dark cycle in the animal house of the Institute of Medicinal Plant Development (IMPLAD). Mice were provided with laboratory chow and tap water animals" .−1 of pentobarbital (ip). Bilateral ovariectomies were performed using a dorsolateral approach. Ovaries and surrounding fat tissue were removed and the incision was closed by suturing the muscles and skin. Similar surgical procedures were carried out for the sham operated animals except that the ovaries were not removed. Experiments were performed one week (7 days) after they were ovariectomized.Naive mice were anesthetized with 50 mg kgThe dried hypocotyls of black maca were obtained from Carhuamayo, Junin at 4000 m altitude. Irma Fernandez, a botanist of the Department of Pharmaceutical Sciences, Universidad Peruana Cayetano Heredia, authenticated the identity of the plant (voucher number IFV1885). The aqueous extract was prepared according to the traditional method. In brief, the pulverized dried hypocotyls were placed in a container with water and boiled. The preparation will be left standing to cool, filtered, freezed (−70°C) and lyophilized (Lyophilizer freeze Mobile12). One gram of dried black maca hypocotyls produced 0.46 g of lyophilized black maca.−1) extract of black maca or vehicle (distilled water) for 35 days between 9:00 h and 12:00 h. Animals were divided in the following groups (n = 10): naive, sham, OVX control groups and two groups of OVX mice treated with two doses aqueous extract of black maca (0.5 and 2.0 g kg−1). No differences between groups regarding body weight were observed at the beginning of the experiment (data not shown).In all experiments, a feeding needle N°18  for oral administration was used to administer aqueous  to reach the platform were recorded during these trials.A trial began by placing the animal in the water facing the wall of the pool at one of the starting points. If the animal failed to escape on the platform within 120 s, it was gently placed there by the experimenter and allowed to stay for 15 s. The inter-trial interval was 5–10 min. Three escape trails were given to all mice per day for four consecutive days (days 23–26 of each treatment). The escape latency(s), swim distance and average speed (cm sTwenty four (day 27) hours after the last training trial in the escape acquisition test, mice were submitted to the probe trial in which the platform was removed. In the 60-s probe trial, the time in the target quadrant  and the number of target crossings (number of crossings over the former location of the platform) were obtained as a measure for spatial memory.3) whose floor was made of parallel bronze bars. The left end of the grid was occupied by a 4 cm diameter, 5 cm high wooden platform. The behaviour of mice was recorded in a personal computer using an automated tracking system  coupledThe step-down avoidance test was performed 6 days after the Morris water maze as previously described . Before At the end of the experiments, mice were sacrificed by decapitation and the brains were dissected out. The brains were homogenized in 10.0 ml of saline (NaCl 0.9%), centrifuged at 3500 g for 15 min and the supernatant obtained for biochemical determinations . The determination of MDA, Ache and MAO levels were performed using commercial kits . All samples were run in a same assay to avoid between-assay variation. Protein concentrations were determined by the Lowry method  using boP-value in the ANOVA test was significant, post hoc Scheffé test was run when indicated. When variances were not homogeneous , the Kruskal-Wallis test was used to assess differences between groups. If the result was statistically significant, differences between pair of medians were assessed using the Mann-Whitney U-test. A value of P < .05 was considered to be statistically significant.Data were analyzed using the statistical package STATA 8.0 for personal computer . Data are presented as mean ± SEM. Homogeneity of variances was assessed using a Bartlett test. Variables with homogeneous variances (body weights and those corresponded to water Morris maze) were analyzed by analysis of variance (ANOVA). If the F4,44 = 1.80, P = NS). At the end of the study, differences in body weight gain were observed between groups . OVX mice showed a higher body weight increase than mice in naive  and sham  groups. Black maca did not alter the effect of ovariectomy on body weight . In fact, the increase in body weight in OVX mice treated with black maca was higher than naive and sham groups (P < .05).No differences among groups were observed in body weight at the beginning of the experiments . When OVX control group was compared against naive and sham groups, OVX mice showed longer swimming distance than naive and sham groups during days 2 , 3  and 4 . No differences between mice in naive and sham groups were observed (P > .05). From day 2 to 4, mice treated with 0.5 and 2.0 g kg−1 of black maca showed a reduction in swimming distance than OVX mice in control group reaching similar values to those of naive and sham groups   . F4,44 = 0.12, P > .05). Ovariectomy significantly increased the time to reach the fixed platform with respect to naive and sham mice in days 2 , 3  and 4 . Black maca reduced the escape latency when compare to the OVX control group during day 2 , 3  and 4  (All groups showed no significant changes in escape latency during the first day (P < .05) .F4,44 = 1.87, P = NS).Average speed was not affected by any treatment  and lower number of crossings  than mice in naive and sham groups. Both doses of black maca prevented the decrease in the time spent in the target quadrant  and the number of crossings of the previous location of the platform  . OVX mice treated with both doses of black maca showed higher values of step-down latency than OVX mice treated with vehicle (P < .05)  . P < .05). No differences were observed between naive and sham groups (P > .05). OVX mice receiving two different doses of black maca showed lower MDA and Ache brain levels than OVX mice (P < .05). MDA and Ache levels in black maca groups were similar than values in control groups (naïve and sham). OVX mice treated with 0.5 g kg−1 of black maca showed higher Ache values when compare with naive group (P < .05). No differences in MDA and Ache levels were observed in OVX mice treated with black maca (P > .05) (P > .05). MDA and Ache brain levels were increased in OVX mice when compare with naive and sham groups (P > .05) . In addir = −0.48, P = .002; r = −0.36, P = .021, resp.) and number of crossings  in the spatial probe of the water Morris maze. No correlations were observed between MDA and Ache brain levels and latency  and number of errors  in the step-down test.MDA, Ache and MAO values for each group were log-transformed for correlation analyses to determine the relation between these variables and those from the memory tests. Negative correlations were observed between MDA and Ache values and total time in the target quadrant  . For insR,3S)-1-methyltetrahydro-β-carboline-3-carboxylic acid, a molecule which is reported to be present in maca, may affect the central nervous system negatively [β-carbolines arises from a Pietet-Splenger condensation between l-tryptophan and aldehydes and this reaction is temperature- and pH-dependent [β-carbolines in the aqueous extract. Data from this study and that from others [As observed previously , resultsgatively . Tetrahyependent . The latm others , 12 suggThe neuro-protective effects of plants containing polyphenolic compounds such as quercetin  and anthin vivo [In the present study, 7 days after ovariectomy no differences in body weight were observed between groups as previously reported . An incrin vivo , 52.Finally, these results demonstrated that black maca can enhance learning and memory in OVX mice and this effect might be related, at least in part, to its ability to reduce LPO and Ache in OVX mice.National Nature Science Foundation of China, NSFC in 2004 (30472016/C03020701); International Technologic Collaboration Project, grant no. 2006DFA21740; Peruvian National Council of Sciences, Technology and Innovation (CONCYTEC) through the grant PROCOM 2005.


At postpartum, an occurrence of 72.0% for severe fatty liver along with 73.5% of subclinical ketosis (SCK) was found. The multiple regression model that best explained the association between milk production in the previous lactation (MYP) and TAG at first week postpartum was as follows: TAG, % = −11.2 + 3.16 (prepartum body condition) + 0.0009176 (MYP) . Logistic regression indicated that Holstein × Carora crossbred cows tended to have 27% higher relative risk than Holstein to experience SCK, whereas prepartum liver TAG greater than 3% tended to be associated with a higher relative risk for SCK compared to cows with TAG ≤3%.The present on-farm research evaluated the occurrence of fatty liver syndrome and its predisposing risk factors for multiparous dairy cows from a commercial herd in Venezuela. Liver biopsy samples were collected at 35 days (d) prepartum (Holstein,  Fatty liver (hepatic lipidosis) and ketosis  are metabolic alterations that occur in high producing dairy cows during the periparturient period. Although distinctly different, they are both part of a generalized lipid mobilization syndrome named fatty liver syndrome (FLS), severe fatty liver, or fat cow syndrome. Therefore, FLS is characterized by a high hepatic triacylglycerols infiltration and the subsequent ketosis development associated with retained fetal membranes and metritis at early lactation –3. Dry matter intake reductions along with high energy demand for milk production are the main risk factors for FLS . Fatty lAnother factor that could predispose to the development of FLS would be the length of dry period as stated by McCormack  but, at n = 14 and Holstein × Carora crossbred, n = 17), 19 , and 20  cows were actually sampled at prepartum, 1st week, and 5th week postpartum, respectively. Both during the dry period and after calving, cows were loosed housed. All cows were fed from 30 to 35 days before calving date with a total mixed ration , located in Quibor, Lara State, Venezuela. This is a dry forest premountain area, which has an average temperature of 25.4°C, a relative humidity of 69%, and an average annual rainfall of 485.5 mm. Forty multiparous (2 to 7 lactations) pregnant cows were randomly selected from a group of 60 to be sampled by liver biopsy at 35 days (d) prepartum as well as 1 to 7 d, and 28 to 35 d postpartum. However, due to logistic problems only a total of 31  were calculated from proximal composition using the summative equations proposed by NRC [During the study, each ration was sampled twice for chemical analysis: moisture, ash, ether extract (EE) and crude protein (CP) (the Kjeldahl method) , neutrald by NRC .Body condition scores (BCS) were measured prepartum and during the first, third, and fifth weeks postpartum, using the scale of 1 to 5 points with a range of 0.25 units . The weiβ-hydroxybutyrate in a semiquantitative measurement scale. Positive cows were those that have a color scale corresponding to milk β-hydroxybutyrate concentrations ≥200 μmol/L whereas negative cows were those with no reaction to the test or those whose color scale corresponded to milk β-hydroxybutyrate concentrations ≤100 μmol/L [The detection of subclinical ketosis was achieved in a total of 34 cows in the first, third, and fifth weeks postpartum by the reaction of a sample of fresh milk with sodium nitroprusiato  to detect levels of 0 μmol/L . Information on production records was collected from the farm, retrospectively. Total milk yield, both in the current and the previous lactations, was adjusted to 305 days. The length of dry period (number of days during dry period) was calculated for each cow. Therefore, the following performance parameters were available: prepartum body condition (PBC), body condition at calving (BCC), milk production in the previous lactation (MYP), dry period length (DPL), and number of lactations (NLAC). Also information was collected on the events of mastitis, metritis, rebreeding, foot problems, and deaths. Associations between milk yield in previous lactation (MYP), PBC, and liver TAG or glycogen concentrations were examined by Pearson's correlation using PROC CORR in SAS . The assP ≤ 0.15) in the statistical score analysis type 3 GEE . At the end, the following independent variables remained in the model: breed  and liver TAG concentration at prepartum . The relative risks were calculated instead of odds ratios by using the ESTIMATE option. The relative risks compare the likelihood of success or failure in each group at the level of the independent variable under investigation. By using PROC GLM of SAS [P = 0.05 to P = 0.15. Factors affecting SCK were analyzed using logistic regression by PROC GENMOND of SAS  under thM of SAS , an analn = 31) was 2.9% . At the first week postpartum (n = 19), the average was 6.1% , while in the fifth week postpartum (n = 20), the average liver TAG concentration reached 6.9% . The mean concentration of liver glycogen (fresh basis) five weeks before calving (n = 31) was 3.6% . For the first week postpartum (n = 19), the average reached 1.2% . Finally, at the fifth week postpartum (n = 15), the average liver glycogen concentration was 2.7% . All liver TAG and glycogen concentrations had a normal distribution . The average concentration of liver TAG (fresh basis) five weeks before calving  , which mIn retrospect, there were no differences in prepartum body weight between cows with or without fatty liver . HoweverP = 0.11) (P > 0.15) was found between PBC and liver TAG at first week postpartum. Obese cows have an increased adipose tissue lipolysis and a decreased food intake during the peripartum, which predisposes to the development of fatty liver [In the present study, severe postpartum fatty liver was present in Holstein cows associated with a higher prepartum body condition (PBC) as compared to Holstein × Carora cows  . Gerloffβ-hydroxybutyrate were considered positive, Rosendo et al. [n = 6; Holstein × Carora, n = 3). The incidence of SCK in the first nine weeks of lactation varies from 9 to 34% as reviewed by Rosendo et al. [For recording the incidence of SCK (new cases), ketotest results above 200 uM of milk o et al. . Among ao et al. . The higo et al. . The relationship between SCK, breed, and prepartum liver TAG levels was analyzed by logistic regression . CrossbrThe analysis showed that fatty liver  was present in those cows that produced more milk  or with liver glycogen concentration at the first week postpartum (P > 0.15). But on the other hand, MYC was negatively associated with liver glycogen concentration at the fifth week postpartum . At the present study, no associations were found between milk production during the current lactation (MYC) and liver TAG concentrations during the first and fifth weeks postpartum  and tended to be negatively associated with the concentration of liver GLY at the fifth week postpartum . None associations were found (P > 0.15) between MYP and liver GLY concentration at prepartum or at first week postpartum, as well as between MYP and liver TAG concentration at prepartum. The multiple regression model that best explained the association between MYP and the liver TAG concentration at first week postpartum was as follows: liver TAG concentration = −11.2 + 3.16 (PBC) + 0.0009176 (MYP) . The milk production in the previous lactation (MYP) tended to be positively associated with liver TAG concentration during both the first and fifth weeks postpartum . Others have found similar results (r = −0.72 and −0.39) [The present study found a negative correlation between liver TAG and liver GLY concentrations at the first week postpartum (d −0.39) , 37.P > 0.15)  and large length  did not affect the liver concentrations of TAG or GLY ( > 0.15) . Moreove > 0.15) . Some auThe present on-farm research revealed a high occurrence of prepartum mild fatty liver, accompanied by a high occurrence of severe fatty liver, up to 72%, and an extremely high incidence of subclinical ketosis during the postpartum period. There was a high percentage of prepartum obesity even though the level of energy for the prepartum ration was below that recommended by the NRC. Fatty liver syndrome developed in those cows with a greater potential to produce milk but associations between lactation performance and fatty liver are still unclear. Crossbred cows, Holstein × Carora, tended to have a higher relative risk (27% more than Holstein cows) to experience subclinical ketosis. Therefore, the study suggests breed differences in the susceptibility to both fatty liver and SCK that deserve further study. Based on TAG values greater than 3%, postpartum fatty liver syndrome might be predicted from 35 days before calving.


However, quantitative data are sparse concerning 131I-related risk of these common diseases.Background: Thyroid dysfunction after exposure to low or moderate doses of radioactive iodine-131 (131I exposure during childhood (≤ 18 years) due to fallout from the Chernobyl accident.Objective: Our goal was to assess the prevalence of thyroid dysfunction in association with 131I dose estimates in a Belarusian cohort of 10,827 individuals screened for various thyroid diseases.Methods: We conducted a cross-sectional analysis of hypothyroidism, hyperthyroidism, autoimmune thyroiditis (AIT), serum concentrations of thyroid-stimulating hormone (TSH), and autoantibodies to thyroperoxidase (ATPO) in relation to measurement-based 131I thyroid dose was 0.54 (0.23) Gy . We found significant positive associations of 131I dose with hypothyroidism  and serum TSH concentration. The excess odds ratio per 1 Gy for hypothyroidism was 0.34  and varied significantly by age at exposure and at examination, presence of goiter, and urban/rural residency. We found no evidence of positive associations with antibody-positive hypothyroidism, hyperthyroidism, AIT, or elevated ATPO.Results: Mean age at exposure (± SD) was 8.2 ± 5.0 years. Mean (median) estimated 131I dose and hypothyroidism in the Belarusian cohort is consistent with that previously reported for a Ukrainian cohort and strengthens evidence of the effect of environmental 131I exposure during childhood on hypothyroidism, but not other thyroid outcomes.Conclusions: The association between  Overall, 1,760 petabecquerel (PBq = 1015 Bq) of iodine-131 (131I) and 85 PBq of 137Cs were released into the environment [United Nations Scientific Committee on the Effects of Atomic Radiation (UNSCEAR) 2010].The most severe accident in the history of nuclear industry occurred on 26 April 1986, at the Chernobyl nuclear power plant located in Ukraine, about 10 km south of the border with Belarus. The radioactive fallout contained short-lived, mainly radioiodines, and long-lived radionuclides such as cesium-137  are more common than thyroid cancer and could result in substantial morbidity among those exposed. Some earlier studies reported an increase in thyroid-stimulating hormone (TSH) levels and the prevalence of juvenile hypothyroidism among children exposed to 131I following Chernobyl  and prevalence of subclinical hypothyroidism ; noneligible age (n = 114); inadequate dose estimates (n = 104); self-reported history of any thyroid disease before screening examination including, for example, nodular or diffuse goiter, thyroiditis, hypo- and hyperthyroidism (n = 542), or benign thyroid surgery (n = 58) or thyroid hormone intake (n = 168); lack of TSH measurements (n = 59) or TSH measured using AxSYM method  (n = 12); or lack of thyroid volume measurement (n = 66). The final sample for analysis was 10,827.The study was reviewed and approved by the institutional review boards of the participating organizations in Belarus and the United States, and all study participants or their guardians (for subjects who were ≤ 16 years of age at screening) signed informed consent.Screening procedure. Individuals were screened in one of the two fixed centers in Minsk or Gomel or by mobile teams at local medical facilities. The screening procedure included thyroid palpation and ultrasonographic examination by an ultrasonographer, independent thyroid palpation by an endocrinologist, collection of sociodemographic information and medical history data, collection of blood sample and spot urine sample, and dosimetry interview. In the case of thyroid nodule or focal lesion with the largest diameter ≥ 10 mm detected by either palpation or ultrasonography, thyroid nodule 5–10 mm with ultrasound evidence of malignancy , or diffusely abnormal thyroid structure accompanied by unexplained cervical lymphadenopathy, the study subject was referred for fine needle aspiration biopsy , and autoantibodies to thyroglobulin (ATG) were measured in serum samples with LUMitest immunochemiluminescence assays  using a Berthold 953 luminometer . TSH, ATPO, and ATG concentrations were measured in each study subject with a sufficient serum sample, while FT4 was measured only in those with TSH levels outside the reference range. For a portion of the cohort, TSH  and FT4 (n = 311) were measured with IMx immunochemiluminescence assays (Abbott Laboratories) using a Berthold 953 luminometer. All assays were conducted according to the manufacturer’s instruction.Urinary iodine concentrations (micrograms per liter) were measured photometrically using the Sandell–Kolthoff reaction as modified by Diagnostic criteria. After evaluating the range of TSH values in a reference sample from our cohort, we set reference limits for serum TSH concentration between 0.3 and 4.0 mIU/L. We defined hypothyroidism as a serum TSH concentration > 4 mIU/L, the upper limit of the reference range. Participants with serum TSH concentration > 4 mIU/L and FT4 < 10 pmol/L were considered to be cases of overt hypothyroidism. Hyperthyroidism was defined as serum TSH concentration < 0.3 mIU/L, the lower limit of the reference range, and participants with FT4 > 25 pmol/L were considered to be cases of overt hyperthyroidism. Elevated levels of ATPO (ATPO-positivity) and ATG (ATG-positivity) were defined as ATPO > 60 U/mL and ATG > 60 U/mL, respectively, consistent with BRAHMS recommendation. Participants with serum TSH concentration > 4 mIU/L and ATPO > 60 U/mL were considered to be cases of antibody-positive hypothyroidism. Autoimmune thyroiditis (AIT) was defined based on a combination of laboratory , ultrasound (hypoechoic gland with heterogeneous/granular structure) and palpatory (firm gland) findings, as described elsewhere  and computed corresponding 95% CIs based on logistic regression analysis using the GMBO module of Epicure statistical software , depended on a vector of covariates x that described the background (in the absence of 131I exposure) prevalence, and the estimated 131I thyroid dose d. For modeling prevalence of a specific thyroid outcome , we did not exclude participants with other thyroid outcomes from the analysis.software . We assuThe background adjustment factors were outcome-specific and included sex, age at examination , oblast (an administrative subdivision similar to a state or province) of residency at examination, rural or urban residency at examination, self-reported current cigarette smoking, self-reported current vitamin consumption, self-reported history of any thyroid disease in parents or siblings, year and season of examination, level of urinary iodine , presence of diffuse goiter based on thyroid palpation, ATPO and ATG concentrations . For each studied outcome, we retained in the final model those background factors whose inclusion into the model significantly improved the model fit, based on the likelihood ratio test comparing models with and without the covariate included at the 0.05 alpha level. We also retained those factors previously associated with the outcome of interest in non-irradiated populations.0(x), and a dose–response function, h(d). We fitted a simple linear dose–response model, γ = γ0(x) × (1 + βd), where β is the excess odds ratio (EOR), the parameter that measures the increase in EOR per unit increase in dose (EOR/Gy). We evaluated departure from linearity by fitting a linear-quadratic γ = γ0(x) × (1 + βd + θ2d) dose–response model. We also evaluated a categorical dose–response model using these categories—< 0.1–0.249, 0.25–0.49, 0.5–0.99, 1.0–2.49, 2.50–4.99, 5.00–9.99, ≥ 10.0 Gy—to assure a reasonably proportional increment in 131I dose. In addition to analysis of hypothyroidism on dichotomous scale (TSH ≤ 4.0 vs. TSH > 4.0 mIU/L), we analyzed continuous TSH concentrations using generalized linear regression models, where the mean of ln(TSH + 1) was described as a linear function of the same background adjustment factors used in the logistic model and 131I dose. We fitted statistical models over the entire dose range and performed sensitivity analyses excluding subjects with high 131I doses of ≥ 10 Gy and ≥ 5 Gy. We tested for dose–response trends by modeling exposure both as a continuous variable and as an ordinal categorical variable coded using integer values (0–7).Under a multiplicative OR model, γ can be written as a product of background prevalence odds of a specific thyroid outcome, denoted as γJ categories of different factors such as sex, age at exposure and examination, current smoking status, urban versus rural residency, oblast of residence at first screening examination, family history of thyroid disease, ATPO level, presence of goiter, and urinary iodine levels. We compared two nested models with and without an interaction term between dose and factor under investigation using likelihood ratio test with J – 1 degrees of freedom (df). A significant p-value indicated that the association between radiation and the prevalence of the outcome was not homogeneous across levels of the factor of interest.To evaluate factors interacting with dose, we allowed a linear dose–response trend β to vary within We estimated attributable risk at a given dose level as the ratio of excess cases to observed cases expressed as percentage. Excess cases were estimated as the difference between the numbers of observed and expected (in the absence of radiation exposure) cases.p < 0.05 to be statistically significant.We estimated the statistical significance of model parameters, test of trend, and comparison in goodness of fit between models using likelihood ratio chi-square tests with df equal to the difference in number of parameters between the models being compared. All tests were two-sided, and we considered Characteristics of study participants and prevalence of functional thyroid outcomes. The main characteristics of the study cohort  are summarized in 131I thyroid dose was 0.54 (0.23) Gy, ranging from 0.001 to 26.6 Gy.n = 319 including 18 cases of overt hypothyroidism); 1.26% for hyperthyroidism (n = 137 including 13 cases of overt hyperthyroidism); 5.74% for elevated ATPO (n = 622); and 0.80% for AIT (n = 87).The prevalence of functional thyroid outcomes in the study cohort was as follows: 2.95% for hypothyroidism (131I dose and thyroid outcomesAssociations between . Estimated ORs by categories of 131I thyroid dose and EORs/Gy based on a simple linear model adjusted for background factors are shown in http://dx.doi.org/10.1289/ehp.1205783). We found a nonmonotonic but significant positive trend with increasing 131I dose for hypothyroidism (p < 0.001) and a nonmonotonic nonsignificant positive trend for AIT (p = 0.07), but did not find evidence of positive trends for the other outcomes. Although the EOR/Gy for elevated ATPO levels was –0.07 , this finding was driven by a small number of cases exposed to high doses of 131I (n = 2), and the overall trend was nonsignificant (p = 0.30). The estimated EOR/Gy for hypothyroidism was 0.34  based on a linear dose–response model. However, over the entire dose range the linear-quadratic model fit the data for hypothyroidism significantly better than a simple linear model , there was no significant difference in fit between the two models (p = 0.69 for linear-quadratic vs. linear model comparison). The estimate of the EOR/Gy at a dose range < 10 Gy was 0.21 , somewhat lower than the linear estimate of 0.34 based on the entire dose range, yet statistically significant. The estimate of the EOR/Gy at a dose range < 5 Gy was 0.11 . Based on a linear model, 36 of 319 hypothyroidism cases (11.3%) in the study could be attributed to 131I thyroid exposure.131I thyroid dose over the entire dose range that was best described by the linear-quadratic function (p < 0.001 for linear-quadratic vs. linear model comparison) (data not shown). The estimated coefficient for an increase in TSH concentration per 1 Gy based on the linear model was 0.03 . In contrast to the analysis of TSH on a dichotomous scale, continuous TSH analysis showed a significant positive dose–response with 131I not only in the range of thyroid doses up to 10 Gy, but also in the range of doses up to 5 Gy (p for linear trend = 0.004). No evidence of better fit of linear-quadratic compared to a simple linear dose–response model was found at 131I thyroid doses < 5 Gy (p for two model comparison = 0.85). The estimated coefficient for an increase in TSH concentration per 1 Gy based on the linear model in the dose range up to 5 Gy was 0.02 . After hypothyroid participants were excluded , the estimated coefficient for TSH in association with a 1-Gy increase in exposure across the entire dose range was 0.02 .Analysis of TSH levels on a continuous scale using generalized linear models provided similar results to those based on the dichotomous definition of hypothyroidism: There was a significant increase in TSH concentration with increasing 131I.Analyses of AIT and other outcomes using nonlinear dose–response models or a limited dose range did not provide evidence of significant positive associations with 131I dose and hypothyroidismModification of associations between . Estimates of the linear dose–response slope for prevalent hypothyroidism by selected characteristics are summarized in p = 0.60), cigarette smoking (p = 0.76), urinary iodine level , family history of thyroid disease , or oblast of residence at first screening examination . We found a significant interaction of 131I dose with both age at exposure (p = 0.04) and age at examination (p = 0.03), with EOR/Gy decreasing with increasing age at exposure or at examination. The highest EORs/Gy were found in individuals youngest at the time of the accident (< 5 years) or at the time of screening (< 20 years). We observed a significantly higher EOR/Gy for hypothyroidism in rural compared with urban residents (p = 0.02) and among subjects without diffuse goiter compared with subjects with diffuse goiter (p = 0.02). ATPO-negative individuals had higher radiation-related risk of hypothyroidism  than ATPO-positive individuals  but the difference was of borderline statistical significance (p = 0.06). Assessment of interactions between 131I dose and the above factors on a continuous TSH scale provided consistent results in terms of statistical significance and direction except for smoking (data not shown); the increase in TSH concentration per 1 Gy in nonsmokers  compared with smokers  was statistically significantly lower (p = 0.04).131I thyroid doses and prevalent, predominantly subclinical hypothyroidism in about 11,000 residents of Belarus exposed to radioactive fallout from the Chernobyl accident at ≤ 18 years of age. Over the entire dose range, a linear-quadratic dose–response model fit the data best, with the quadratic component attributed largely to a few individuals with doses ≥ 10 Gy. We also found an association with TSH levels on a continuous scale. The dose–response data for TSH were best described by a linear-quadratic function over the entire dose range, but a significant linear dose response was found in the range of doses up to 5 Gy, suggesting that levels of 131I an order of magnitude below the typical doses (≥ 50 Gy) used to treat thyroid disease  (We found a significant association between low to moderate disease)  can indudisease) , the dos131I thyroid doses. Findings from other populations exposed to 131I, such as Marshall Islanders , for possible effect of calendar time. We did not find significant differences in associations between radiation and prevalence of any of the outcomes by year of examination (data not shown).In addition to variation by age, the association between radiation and prevalence of hypothyroidism also varied according to rural or urban residence and diagnosis of goiter. It is unclear what factors might be responsible for stronger associations among rural compared with urban residents and in nongoitrous compared with goitrous individuals, but differences in the past intake of dietary iodine could be important. We did not find significant variation of radiation–hypothyroidism association according to urinary iodine excretion levels, but these reflect current levels of iodine intake and are subject to high within-individual variability. Because individuals were screened over a relatively long period, we adjusted all our p for homogeneity = 0.06). Although in nonexposed populations autoimmune thyroid diseases or elevated thyroid antibody levels have been associated with higher risk of progression to overt hypothyroidism   compared with ATPO-positive individuals (ATPO > 60 U/mL)  . Hence,  fallout . In atom fallout , a conve fallout , althoug fallout . More pr131I dose and hyperthyroidism or AIT was found in the Ukrainian cohort  were excluded.In our cohort screened at ages ranging from 11 to 33 years , hypothyroidism  was diagnosed in 319 subjects . We estimated that 36 cases (11.3%) could be attributed to There is controversy in the literature about the appropriate upper TSH reference limit . Calls hThe clinical consequences of moderately elevated TSH levels are the subject of continuing discussion in the context of risks versus benefits related to treatment of subclinical hypothyroidism . It has Although our study had significant strengths, there were some limitations. Among the 16,213 eligible and traced subjects, 11,970 (73.8%) underwent the first screening examination. However, self-selection bias related to dose seems unlikely because study participants and study personnel were unaware of subjects’ exposure level. We do not believe that exclusion from the study of 542 individuals with self-reported thyroid diseases diagnosed before the first screening examination biased the dose–response estimates because the EORs/Gy including and excluding these individuals were not meaningfully different for any outcome (data not shown). For example, for hypothyroidism the EORs/Gy were 0.50  and 0.34 , respectively. Because diagnostic criteria and laboratory assays used to diagnose thyroid diseases preceding screening examination could vary across medical institutions and differ from our outcome definitions, we considered it appropriate to exclude the 542 subjects from the analysis. Although we used two assays to measure TSH levels, there were no differences between the groups measured by IMx and LUMI assay in overall TSH distribution or in the assays reference limits established based on evaluation of the reference sample from our cohort (data not shown). Moreover, we found no difference in the estimates of EOR/Gy for hypothyroidism in the groups measured by different TSH assays (data not shown).At this stage we did not take into account the impact of uncertainties in dose estimates because work on assessment of thyroid dose uncertainties in the cohort is ongoing. Typically, radiation doses are estimated with a combination of Berkson and classical type measurement errors . Ignorin131I thyroid dose from the Chernobyl accident and prevalence of hypothyroidism (defined as serum TSH > 4.0 mIU/L) or serum TSH concentration among subjects exposed at ≤ 18 years of age. The excess OR per 1 Gy for hypothyroidism was 0.34 . Associations between radiation and prevalent hypothyroidism were stronger among younger individuals and those with ATPO levels ≤ 60 U/mL. No significant positive associations were observed between radioiodine exposure and antibody-positive hypothyroidism, thyroid autoimmunity, or hyperthyroidism. Further analysis incorporating data from subsequent screening cycles is required to assess temporal trends (transitory or persistent) and shed more light on the natural history of radiation-related effects on thyroid function.We found significant positive associations between estimated (606 KB) PDFClick here for additional data file.


Background. Oxidative stress is enhanced in metabolic syndrome (MetS) and believed to contribute to accelerated atherosclerosis. Weight loss is associated with lowered oxidative stress. Methods. We performed a cross-sectional study in 92 consecutive patients with metabolic syndrome and 80 without. A dietary intervention with moderately low-calorie diet  was carried out in 53 of metabolic syndrome patients. Oxidative stress, assessed by sNOX2-dp and urinary 8-iso-PGF2α, and antioxidant status, assessed by serum levels of vitamin E and adiponectin, were measured before and after 6 months. Results. Serum vitamin E/cholesterol ratio was significantly lower in metabolic syndrome compared to controls (P < 0.001) and decreased by increasing the number of metabolic syndrome components (P < 0.001). After six months, 23 and 30 patients showed >5% (group A) or <5% (group B) weight loss, respectively. Urinary 8-iso-PGF2α (−39.0%), serum sNOX2-dp (−22.2%), adiponectin (+125%), and vitamin E/cholesterol ratio (+129.8%) significantly changed only in A group. Changes in body weight and in serum adiponectin were independent predictors of vitamin E/cholesterol ratio variation. Conclusion. Our findings show that in metabolic syndrome moderate weight loss is associated with multiple health benefits including not only oxidative stress reduction but also enhancement of antioxidant status. The metabolic syndrome (MetS) has emerged as an important cluster of metabolic and nonmetabolic risk factors for atherosclerotic disease associated with insulin resistance , 2. MetSLifestyle modifications in the form of moderate caloric restriction, moderate increase in physical activity, and changes in dietary composition towards a less atherogenic, low-saturated fat, and prudent diet are the most common modalities for managing patients with MetS. Current guidelines on the management of the individual components of MetS emphasize that moderate weight loss should be a first-line therapy .Oxidative stress is believed to play a major role in vascular disease, as it is implicated in the initiation and progression of atherosclerosis; it determines inflammation of the artery wall, which ultimately leads to macrophage infiltration and anatomical and functional changes of endothelial wall.Oxidative stress is a typical feature of MetS and obesity –11 as shWeight loss has been associated with reduced oxidative stress and NOX2 activation which lead to an improvement of endothelial dysfunction . Vitamin E is a fat-soluble antioxidant that inhibits lipid peroxidation . Serum cBlood antioxidants, including vitamin E, are consistently lower in obese than nonobese adults and progressively lower with increases in BMI –19. HoweAim of the present study was twofold. First, to evaluate serum levels of vitamin E in patients with and without MetS. Second, to assess changes of vitamin E and markers of systemic oxidative stress in MS patients adhering to a moderately calorie-restricted, Mediterranean-type diet during a 6-month study period.The cross-sectional study vas performed in 172 consecutive outpatients, aged 40–80 years, referred to our metabolic outpatient clinic at the Department of Internal Medicine of the University Hospital in Rome. MetS was diagnosed in 92 patients following the recently harmonized criteria by major scientific organizations based on the concomitant presence of at least three of the following five clinical features: waist circumference  ≥102 cm in men and ≥88 cm in women, fasting blood glucose ≥100 mg/dL, triglycerides ≥150 mg/dL, HDL cholesterol <40 mg/dL in men and <50 mg/dL in women, and arterial systolic/diastolic blood pressure ≥130/≥85 mmHg .Written consent was obtained from enrolled subjects and the study was conformed to the ethical guidelines of the Declaration of Helsinki; the ethical committee of our Institution approved the study.Participants were instructed to consume a moderately calorie-restricted diet during the 6-month study period. They followed a balanced low-calorie , low-fat, high-carbohydrate diet  under supervision of the study nutritionist. The goal was to achieve a weight loss of approximately 5% of initial body weigh over six months. Patients were asked to increase the consumption of fruit, vegetables, low-fat dairy products, poultry, fish, and whole grain products, to limit the intake of red meat and of high-fat dairy, to have only moderate amounts of sugars and foods containing added sugars, and to have olive oil as the only seasoning and cooking fat. They received sample menus, recipes, and instructional nutrition labels in order to achieve a better healthy food selection and a daily negative energy balance. The diet was distributed in 5 meals, with 2 breaks during the day. At baseline and after the diet period a weekly food frequency questionnaire was administered. All foods were self-selected and prepared by the subjects and no supplements were prescribed. Energy requirements were calculated at baseline by estimating basal metabolism and resting energy expenditure by the Harris-Benedict equation. Subjects were asked to maintain the same level of physical activity throughout the study period.After overnight fasting and supine rest for at least 10 minutes, blood was withdrawn from an antecubital vein. Blood samples were taken into tubes containing EDTA and centrifuged at 3,000 rpm for 15 minutes to obtain plasma. Serum and plasma were divided into aliquots and stored at −80°C.Serum concentration of vitamin E was measured by HPLC using tocopheryl acetate as internal standard. Reagents included HPLC-grade ethanol, methanol and hexane , and tocopheryl acetate . A flow rate of 2.0 mL/min was used with a LC/233 Diode Array Detector  set at 0.02–0.1 attenuation. Levels were expressed as the ratio between serum alpha-tocopherol concentration (μmol/L) and serum total cholesterol concentration (mmol/L) [(mmol/L) , 20. Intα (8-iso-PGF2αUrinary 8-iso-prostaglandin F2) was measured by a previously described and validated enzyme immunoassay method [y method . Intra-aSerum levels of soluble NOX2-derived peptide (sNOX2-dp) were detected by ELISA method as previously described [escribed ; intraasAdiponectin serum levels were measured with a commercial immunoassay . Intra-assay and interassay coefficients of variation were 6% and 8%, respectively. t-test for unpaired data was used for the comparison of mean values. Group comparisons were performed by use of analysis of variance (ANOVA). Proportions and categorical variables were tested by the χ2-test and by the 2-tailed Fisher's exact method when appropriate. Multiple stepwise forward linear regression analyses were performed to identify the independent predictors of vitamin E/cholesterol ratio and vitamin E/cholesterol ratio changes after dietary intervention. To test the independent predictive role of vitamin E/cholesterol ratio for MetS, a logistic regression analysis was performed.Statistical analysis was performed using the SPSS statistical software version 13.0 for Windows . All variables were tested for normal distribution prior to analyses. Data are expressed as the mean ± SD for continuous variables. The correlation between variables was analysed with the Pearson test and the Cochran's linear trend test. Student's P values are two tailed; a P value of less than 0.05 was considered statistically significant.All t-test for independent groups, considering (i) expected difference in Vitamin E/cholesterol ratio to be detected between patients and controls = 0.6 μmol/mmol, (ii) standard deviations homogeneous between groups SD = 1.1 μmol/mmol, and (iii) type I error probability α = 0.05 and power 1-β = 0.90. This resulted in n = 71 per group, which was increased to n = 80. For the intervention study, as previously reported [n = 15 patients for group to participate in the intervention study.For the cross-sectional study, the number of MetS patients and controls was computed with respect to a two tailed Student's reported , we hypoμmol/mmol, P < 0.001).Clinical and biochemical characteristics of MetS patients and controls are reported in Ptrend < 0.001)  . P = 0.006), after controlling for confounding factors.As previous studies indicated that patients with vitamin E/cholesterol ratio <5 are at higher risk of vascular events , 24 we dPatients included in the interventional study had clinical characteristics similar to those of the entire population except for the exclusion of diabetes (data not shown). During dietary treatment some patients adhered more closely to the prescribed diet and, after six months, achieved much better weight loss than others. Therefore, for the purpose of the present analysis, participants were divided in two groups according to body weight loss higher or lower than 5%, respectively. A good compliance to dietary recommendations was found in 23 subjects (group A), who lost 7% of their initial weight over six months, while 30 subjects (group B) showed no significant weight changes (+0.1%) . BaselinMajor nutrient changes concerned dietary fat. In fact, in keeping with the study protocol, group A markedly reduced (−58.6%) dietary consumption of olive oil and seasoning fat, which led to an estimated decrease of vitamin E intake from 39.3 to 16.2 mg/day. Conversely, subjects belonging to group B showed no significant changes in dietary fat and vitamin E intake.α (−39.0%) and serum sNOX2-dp (−22.2%), was observed, together with an increase of serum adiponectin (+125%). In the same group, mean serum Vitamin E/cholesterol ratio increased significantly , while nonstatistically significant changes were observed in group B .Baseline and after six months characteristics of patients belonging to group A and group B are reported in r = −0.67; P < 0.01), BMI , waist circumference , total calories , fat calories , and intakes and changes of dietary consumption of cooking and seasoning oils . Moreover, a negative correlation was found with fasting blood glucose , HOMA-IR , total cholesterol , and triglycerides , while a positive correlation was observed with changes of serum adiponectin .At bivariate analysis performed in all subjects , a stronβ = − 0.37; t = − 2.7; P = 0.009) and in serum adiponectin  were the only independent predictors of Vitamin E/cholesterol ratio variation after controlling for age, gender, and changes of HOMA-IR, urinary 8-iso-PGF2α, and serum sNOX2-dp (adjusted R2 = 0.31). Conversely, weight loss  and changes of urinary 8-iso-PGF2α  and of serum Vitamin E/cholesterol ratio  were the independent predictors of adiponectin variation (adjusted R2 = 0.46).A multiple regression analysis showed that percent changes in body weight  and/or a decrease in antioxidant defence. Enhanced production of ROS is likely to occur via upregulation of NOX2, which, in fact, is overactivated in patients with MetS . Adults To the best of our knowledge, there are only two studies reporting vitamin E values in patients with MetS, with controversial results. The first  reports To further explore the relationship between vitamin E and MetS we investigated the effect of a six-month, low-calorie, dietary intervention on biomarkers of antioxidant status and oxidative stress in patients with the metabolic syndrome. In a previous paper referring to the same patients with MetS , we repoThe present study extends these findings demonstrating that moderate weight loss is associated not only to oxidative stress lowering, but also with an increase of antioxidant status. In fact, mean serum Vitamin E/cholesterol ratio raised by 130% in the group of patients achieving at least 5% reduction of body weight. The increase of vitamin E was coincident with the increase of serum adiponectin, another molecule with antioxidant property , 29. TogThis finding is of potentially clinical relevance as antioxidants have positive effects on metabolic and glycaemic control. Accordingly, an association was also observed between changes of Vitamin E/cholesterol ratio and changes of indexes of glycemic and lipid metabolism, which is in agreement with the hypothesis that oxidative stress may negatively influence glycaemic control by increasing insulin resistance , 31. FinIn our study we may exclude an increased dietary intake of alpha-tocopherol after weight loss. In fact, major dietary changes consisted in the reduction of olive oil and seasoning fat consumption, which lead to an estimated decrease of vitamin E intake from 39.3 to 16.2 mg/day, which is similar to the reported daily intake of healthy, nonobese Italians on a Mediterranean diet .We acknowledge that our study may have some limitations. Lack of randomization to two different diets could have biased the results. We decided, however, that randomizing to two diets, one with and another without calorie restriction, was unethical. An interesting alternative chance was provided by previous studies showing that in case of dietary restriction about 45% of subjects are poor responders to diet  and do nIn conclusion, our findings give further support to the concept that in MetS moderate weight loss is associated with multiple health benefits, some of which are related to an improvement in antioxidant status and a decrease of oxidative stress. Therefore, it is tempting to speculate that in this clinical setting the increase in antioxidant defences may not only reverse ROS generation (and atherosclerosis progression), but also improve insulin sensitivity and lipoprotein profile, thus increasing protection towards diabetes and cardiovascular disease.Finally, we confirm the importance of calorie restriction as the first-line intervention to reduce the cardiometabolic risk in patients with MetS.


Objectives. Mount Sakurajima in Japan is one of the most active volcanoes in the world. This work was conducted to examine the effect of volcanic ash on the chronic respiratory disease mortality in the vicinity of Mt. Sakurajima. Methods. The present work examined the standardized mortality ratios (SMRs) of respiratory diseases during the period 1968–2002 in Sakurajima town and Tarumizu city, where ashfall from the volcano recorded more than 10.000 g/m2/yr on average in the 1980s. Results. The SMR of lung cancer in the Sakurajima-Tarumizu area was 1.61 (95% CI = 1.44–1.78) for men and 1.67 (95% CI = 1.39–1.95) for women while it was nearly equal to one in Kanoya city, which neighbors Tarumizu city but located at the further position from Mt. Sakurajima, and therefore has much smaller amounts of ashfall. Sakurajima-Tarumizu area had elevated SMRs for COPDs and acute respiratory diseases while Kanoya did not. Conclusions. Cristobalite is the most likely cause of the increased deaths from those chronic respiratory diseases since smoking is unlikely to explain the increased mortality of respiratory diseases among women since the proportion of smokers in Japanese women is less than 20%, and SPM levels in the Sakurajima-Tarumizu area were not high. Further studies seem warranted. During 2, which was mainly noncrystalline [Ashes from Mt. Sakurajima contain approximately 60% of SiOstalline . Recentlstalline .Although many studies examined the health effects of Mt. Sakurajima's volcanic activity among the residents in its neighborhood , no evid Sakurajima town has the population of about 4.000 and is situated on the foot of Mt. Sakurajima. Tarumizu city has the population of about 20.000 and is located within 10 km from Mt. Sakurajima. Kanoya city, with the population of about 100.000, neighbors Tarumizu city but is located at the further position from Mt. Sakurajima and therefore has much smaller amounts of ashfall. None of those municipalities has serious air pollution from traffic exhaustion. In the present study, we examined mortality of respiratory diseases other than tuberculosis. One of them was malignant neoplasm of trachea, bronchus, and lung (ICD9th: 162). We simply called this disease entity “lung cancer” (ICD9th: 162) in the present study. Other important respiratory diseases excluding pulmonary tuberculosis are “diseases of respiratory system” (ICD9th: 460–519). This disease category is divided into the following groups in ICD9th: acute respiratory infections (ICD9th: 460–466), other diseases of upper respiratory infection (ICD9th: 470–478), pneumonia and influenza (ICD9th: 480–487), chronic obstructive pulmonary disease and allied conditions (ICD9th: 490–496), pneumoconiosis and other lung diseases due to external agents (ICD9: 500–508), and other diseases of respiratory system (ICD9th: 510–519). In this study, we combined “acute respiratory infections” and “pneumonia and influenza” (ICD9th: 480–487) and called this category as acute respiratory diseases . On the other hand, the disease categories of “other diseases of upper respiratory infection” (ICD9th: 470–478) and “other diseases of respiratory system” (ICD9th: 510–519) were not included in our study. We concluded that they were not important for the present study because the number of deaths from those diseases was small, and they were, in a sense, an ill-defined disease category. Needless to say, we were interested in “pneumoconiosis and other lung diseases due to external agents” (ICD9th: 500–508), which may be associated with volcanic activities. However, the publication by the local health authority does not present the number of deaths for this disease category.Standardized mortality ratios (SMRs) of deaths from lung cancer (ICD9th: 162), acute respiratory diseases , and COPDs (ICD9th: 490–496) in the Sakurajima-Tarumizu area (exposed area) and Kanoya city (control area) were calculated as a ratio between observed and expected numbers of deaths. The numbers of deaths from lung cancer (ICD9th: 162), acute respiratory diseases , and COPDs (ICD9th: 490–496) were obtained from the annual Vital Statistics Report published by the Kagoshima Prefectural Municipal Government. The expected numbers of deaths were obtained through multiplying age-gender-year-specific population numbers of Sakurajima-Tarumizu area or Kanoya city by corresponding gender-, age- (5-year category), and year-specific mortality rates for lung cancer (ICD9th: 162), acute respiratory diseases , and COPDs (ICD9th: 490–496) in Kagoshima Prefecture (population of about 1.6 million), which were obtained from the annual Vital Statistics Report published by the Kagoshima Prefectural Municipal Government. The 95% confidence intervals (95% CIs) of SMRs were obtained by the methods assuming poisson distribution [ The data on the amount of ashfall were obtained from the website of Kagoshima Prefectural Government. The SMR of lung cancer in Sakurajima-Tarumizu area was 1.61 (95% CI = 1.44–1.78) for men and 1.67 (95% CI = 1.39–1.95) for women, indicating that lung cancer mortality was significantly increased when compared to the entire Kagoshima Prefecture. Sakurajima-Tarumizu area had a significantly increased COPD mortalities  suggests long-term effects of Mt. Sakurajima, which has become active every 10–30 years over the last 100 years.Confounding of smoking is unlikely among women since the proportion of female smokers is less than 20% in Japan . The pro3, 0.025–0.035 mg/m3 and 0.027–0.036 mg/m3, respectively (Kagoshima Prefecture Government).Suspended particulate matter (SPM) is a possible factor that increased the mortality of lung cancer and COPDs in Sakurajima-Tarumizu area. However, Nishii et al. reported the lack of significant correlation between SPM concentrations and the amount of ashfall . At the −3 year increases lung cancer risk by 1.2-folds. In the Sakurajima-Tarumizu area, the cumulative amount of ashfall in the 1980s and 1990s was nearly 200,000 g m−2. Since the ash of Sakurajima contains approximately 7% of cristobalite, and its 10% is assumed to be respirable material (<4 um) based on the finding of Hillman [−3 year (=200.000 × 7% × 10%). The exposure to only 0.1% of 1400 g can increase lung cancer risk by 1.7-folds.On the basis of what was reported by Steenland et al. , we estiCristalline silica is known to cause silicosis or pneumoconiosis . HoweverThe present study has several limitations. One is the lack of information on silicosis. Therefore, whether lung cancer patients had silicosis or not is not known. In addition, the mortality of silicosis and related diseases cannot be examined because the information is not made public. However, the study of Wakisaka et al.  suggesteIn conclusion, cristobalite is the most likely cause of the increased deaths from those chronic respiratory diseases since smoking is unlikely to explain the increased mortality of respiratory diseases among women since the proportion of smokers in Japanese women is less than 20%, and SPM levels in the Sakurajima-Tarumizu area were not high. Further studies on the health effects of Mt. Sakurajima's volcanic activities seem warranted. Those studies should include the examination of indoor levels of crystalline silica.We selected Kanoya city as a control area since the ashfall was much less than Sakurajima-Tarumizu area while those two areas are closely located. Kanoya is slightly more urban than Sakurajima-Tarumizu. Therefore, we were afraid that the using Kanoya as a control may end up with underestimating the effects of volcanic activities on chronic respiratory disease mortality in Sakurajima-Tarumizu area. However, contrary to our expectation, Kanoya city had the mortality of respiratory disease mortality less than the entire Kagoshima, making the comparison more favorable for Sakurajima-Tarumizu area.Mt. Sakurajima in Japan is one of the most active volcanoes in the world. In the last century, the volcano became active every 10–30 years. Its volcanic ash contains up to 7 wt.% of cristobalite, which is known to be carcinogenic. However, the chronic health effects of Mt. Sakurajima's volcanic activities were not clear.This study showed that Mt. Sakurajima's volcanic activities increased the mortality of lung cancer and COPDs in the Sakurajima-Tarumizu area, which has the largest amount of ashfall from the volcano in its vicinity.Further studies should be conducted to confirm the association between the volcanic activities and chronic respiratory disease mortality. Those studies should include the examination of indoor levels of crystalline silica.