To date, studies show conflicting results for PM2.5 exposures based on mode of commuting, and few studies compare multiple modes of transportation simultaneously along a common route, making inter-modal comparisons difficult. In this study, we examined breathing zone PM2.5 exposures for six different modes of commuting  simultaneously on a single 2.7 km (1.68 mile) arterial urban route in Salt Lake City, Utah (USA) during peak “rush hour” times. Using previously published minute ventilation rates, we estimated the inhaled dose and exposure rate for each mode of commuting. Mean PM2.5 concentrations ranged from 5.20 μg/m3 for driving with windows closed to 15.21 μg/m3 for driving with windows open. The estimated inhaled doses over the 2.7 km route were 6.83 μg for walking, 2.78 μg for cycling, 1.28 μg for light-rail train, 1.24 μg for driving with windows open, 1.23 μg for bus, and 0.32 μg for driving with windows closed. Similarly, the exposure rates were highest for cycling (18.0 μg/hr) and walking (16.8 μg/hr), and lowest for driving with windows closed (3.7 μg/hr). Our findings support previous studies showing that active commuters receive a greater PM2.5 dose and have higher rates of exposure than commuters using automobiles or public transportation. Our findings also support previous studies showing that driving with windows closed is protective against traffic-related PM2.5 exposure.Traffic-related air pollution in urban areas contributes significantly to commuters’ daily PM However, to date there are relatively few studies on PM2.5 exposures by type of commuting in the United States. Some studies show higher PM2.5 exposures for cyclists [2.5 compared to automobile or public transit [2.5 exposure by transportation mode.One strategy to reduce daily PMcyclists  and walkcyclists , 12 compcyclists  or lowercyclists , 15. Com transit , 17. The transit . Thus, a2.5 exposures is difficult for a number of reasons. Zuurbier et al. (2010) note that in most previous studies, particulate exposures were not measured simultaneously by mode of commuting [2.5 measures based on differences in sampling equipment [Understanding inter-modal differences in commuters’ PMommuting . This isommuting , 20. Somet al. 20 note thaommuting , 21, 22.quipment , making quipment . There iet al. 20 note thaquipment , or onlyquipment . Anotherquipment .2.5 for six different commuting types  on a single prescribed arterial urban route during peak “rush hour” times. To control for variation in background PM2.5 we used identical instruments for both personal and ambient monitoring. Finally, we estimated the PM2.5 inhaled dose and rate of exposure for each commuting mode based on previously published minute ventilation rates [In this study, we examined personal exposures to PMon rates , 25, 26.2  [Salt Lake City (SLC) is the capital and largest municipality in Utah, with a population of 192,672 according to the 2015 United States census . Populatple/mi2) . Air pol2.5 concentrations on 15 study personnel using TSI SidePak AM510 personal exposure monitors  as they travelled the 2.7 km (1.68 mi) route via walking, biking, riding the bus, riding the light rail system, driving with windows open, and driving with windows closed. Study personnel were university faculty (n = 3) and students (n = 12), all of which were in good physical health. For both automobiles, the internal circulation system was turned on during all study periods. All study personnel were non-smokers. Data collection occurred from Aug 8th–Aug 11th, 2016. Air pollution monitoring was conducted from approximately 8:00–10:00 AM, and again from 4:00–6:00 PM during peak commuting times. Samples were collected simultaneously for all modes of transportation beginning at the same time. At times, study workers were shifted to less efficient modes in order to provide enough overall sampling time per mode. However, individual data collection events occurred more expeditiously for some modes of transportation (e.g. light rail and driving) than for others. For the purposes of this study, a data collection event was defined as the study worker initiating data logging on the SidePak, travelling the length of the route in one direction using their designated mode of transportation, and then discontinuing data logging. Once the data collection event was completed, the study worker crossed to the other side of the road and repeated this process. For study workers riding the light rail, SidePak data logging was initiated approximately 5 minutes before arrival of the train, and was stopped upon exiting the train. The worker then reset the SidePak and rode the next available train in the opposite direction. There were no busses that ran the entire length of the study roadway, so we were obliged to use a route that covered approximately 44% (1.19 km) of the entire study route on the north side of the road only , calibrated pump flow rates to 1.7 L/min  using a Defender 510 volumetric flow calibrator , zeroed the 670 nm laser using an in-line HEPA filter, and charged the batteries. All pre- and post-calibrations were within ±5% of the 1.7 L/min target flow rate. All of the SidePak AM510 instruments were calibrated by the manufacturer to ISO 12103–1 A1 test dust (Arizona road dust) prior to using them in this study.SidePak AM510s (n = 9) were set to record PMThe inlet to the SidePak AM510 was attached to the collar or shoulder of the study worker’s shirt so it was within the person’s breathing zone. For the purposes of this study, we defined the breathing zone as the sphere of air located within 25 cm (10 in) of the individual’s nostrils . To cont2.5 was measured with one stationary SidePak AM510 instrument placed on the south side of the road , which was expected due to the short, 1-second time interval used on the SidePaks. Thus, to compare means between the two vans, we performed a t-test taking into account serial correlation in the data. Our results showed a statistically significant difference between the two vans (p = 0.029). Means and standard errors were 5.2 μg/m3 (SE = 0.17) and 4.9 μg/m3 (SE = 0.13) for the 2016 and 2013 models, respectively. Although the difference between the two vans was statistically significant, we believe the 0.3 μg/m3 difference was not necessarily clinically significant from an overall exposure standpoint. JMP version 12.0  was used for this analysis.Two university-owned Toyota Sienna mini-vans, 2013 and 2016 models, were used in this study for monitoring breathing zone exposures for driving with windows open and driving with windows closed. For driving with windows closed, van air conditioning was used with the air recirculating in the cabin (not drawing outside air into the vehicle). Both vehicles used the same model of cabin air filter (Toyota part number 87139-YZZ20), which had an atmospheric dust efficiency rating of 0.3 ~ 0.5 μm: 20±10% at 240m2.5 [Relative humidity (RH) greater than 50% can cause hygroscopic enlargement of particles less than 2.5 μm, resulting in the SidePak overestimating the concentration of PM2.5 . Ambient3 with a zero stability of ±0.001 mg/m3 over 24 hours. In order to account for this known limit, readings below 0.001 mg/m3 were recoded as 0.0007 mg/m3, which is the detection limit divided by the square root of two. This allowed us to use readings where the SidePak read 0 mg/m3 due to the monitor’s inability to detect particle counts that low (the particle count is never truly zero), and proceed with analysis at those time points. We then calculated the ratio of each commuting type to the background levels recorded at each second. The ratios were averaged and reported along with standard errors for overall particulate exposures by commuting type and by commuting type and time of day (morning or afternoon rush hour). Statistical differences between groups were calculated using linear regression within R: A Language and Environment for Statistical Computing (v 3.3.2) [We cleaned the data by matching each data point of commuter  with the particulate levels measured by the stationary monitor by date and second. The detection limit of the SidePak recorded by TSI is 0.001mg/mv 3.3.2) .2.5 per type of commuting, we used the averages of minute ventilation rates based off of different studies. Car transportation, with both windows opened and closed, had a minute ventilation rate of 11.8 L/min [2.5 per trip) was then calculated by multiplying the mean PM2.5 concentration for each commuting type by the minute ventilation rate (L/min) and then by multiplying by the conversion factor 1m3/1000 L and by the average number of minutes per trip [To calculate the inhaled dose of PM.8 L/min . Light r.8 L/min . Cycling.8 L/min . Cycling.8 L/min . The inhper trip . Exposur2.5 concentrations for six different commuting types and one stationary monitor in downtown Salt Lake City, Utah. We collected between 4524 (bus) and 39,725  data points for each type of commuting. The disparity in the numbers of points is due to inherent differences in commuting. The bus comes infrequently, whereas walkers were able to record values continuously. It should also be noted that due to a technical malfunction, data for driving with windows closed was only recorded on 8/8/16 and 8/11/16. While this malfunction was unfortunate, the particulate matter for driving windows closed was very consistent, with a standard deviation in the raw data of only 4.6 ug/m3. Overall standard deviations for the other modes of transportation varied between 9.9 ug/m3 (bike) and 17.2 ug/m3 (light rail). The standard deviations for driving with windows open, bus and walking were 11.4 ug/m3, 11.6 ug/m3 and 12.6 ug/m3, respectively. It is therefore unlikely that the comparative results were strongly affected by the omission of driving windows closed on the other days.We recorded real-time PM2.5 experienced by the different commuting types was between 5.21 ug/m3 (driving with windows closed) and 15.21 ug/m3 (driving with windows open). Mean exposures were 12.49 ug/m3 for light rail, 12.21 ug/m3 for walking, 12.62 ug/m3 for biking and 13.03 ug/m3 for taking the bus. While these averages are of interest, they may be differentially influenced by background pollution levels. Therefore, we focus the reporting of results below on the ratio of personal exposure by commuting type to background levels at each recorded second.Using the clean data matched by second to the ambient monitoring station, the average concentration of PM2.5 by commuting type and background monitor are shown in The number of data points (seconds), means, medians, variances and standard errors of the ratios of PMThe linear regression analysis showed that all commuting types were statistically significantly different from the reference type, driving windows closed (p < 0.001). Heteroscedasticity was not present in the residuals as determined by the Breush-Pagan test.2.5 exposure rates that were 4.8 and 4.5 times higher, respectively, than driving with windows closed. This was due largely to the elevated minute ventilation rates associated with active commuting and the lower PM2.5 concentration inside cars with windows closed. Exposure rates for driving with windows open, light rail, and bus were 2.9, 2.6, and 2.7 times higher, respectively, than driving with windows closed.As shown in 2.5 concentrations were highest for driving with windows open and bus travel, and lowest for driving with windows closed. However, other factors such as travel time, mode of commuting, and minute ventilation rates can significantly influence the rate of exposure and overall inhaled dose of particulate matter [2.5 for active commuters to be 2–7 times greater than for automobile and public transit commuters over comparable routes [2.5 lung deposition than car commuters, largely due to increased respiration rates [2.5 for cyclists and walkers in our study were approximately 9 and 21 times greater, respectively, than driving with windows closed over the same 2.7 km route.In this study, we examined fine particulate matter exposure for six different commuting types simultaneously on a single prescribed urban route during “rush hour” times. Breathing zone PMe matter , 16. Cone routes , 16. Simon rates . By comp3); however, inhaled dose (μg) and exposure rate (μg/hr) may be better measures for understanding health outcomes associated with commuting. Several previous studies show no difference or lower PM2.5 concentrations for cyclists and walkers than for automobile commuters [2.5, PM10, and soot concentrations were among automobile and bus commuters, but the highest exposure rates were found among high-traffic cyclists [2.5 exposures during cycling occurred at a rate of 100.4 μg/hr, compared to 51.9 μg/hr for diesel car [2.5 exposure [2.5 contribution attributable to commuting for cyclists and walkers may be 4–5 times higher than among automobile users. There is currently little data on the long-term health effects of increased air pollution exposure among active commuters, warranting additional research in this area [To date, most studies compare exposures across modes of transportation in terms of breathing zone concentrations  to result in the lowest exposure rates. Closing the vehicle windows and turning on the air recirculation system creates a mostly-closed, protective bubble around the automobile’s occupants. However, Knibbs, de Dear, and Atkinson (2009) described that even vehicles with closed windows experience some air flow through “leaks” . A varie2.5 concentrations compared to driving with the car windows closed. In their study, the car’s air conditioner was turned off, and the ventilation system was set to a “moderate” level [2.5 concentrations, respectively, compared to driving with windows closed. In our study, the car’s air conditioning system was used with the air recirculating on in the cabin (not drawing outside air into the vehicle). However, other studies show in-car exposures can be higher than for cyclists and walkers [Our findings support Briggs et al. (2008) who showed that walking exposed commuters to 2.2 times higher mean PMe” level . Our res walkers , 17. In  walkers , car win2.5 concentrations in busses in our study were due to self-pollution via leaks and air exchange between the cabin and outdoor air when the bus doors were opened during stops or from windows being opened.A combination of leaks when windows are closed and infiltration of outdoor air when windows and doors are open likely explain the high level of air pollution in busses. Sabin et al. (2005) describe that busses, especially older ones, can self-pollute (i.e. particles enter the bus even though the windows are closed), and passenger exchange can exacerbate this effect . Hammond2.5 levels among all transportation modes with the exception of driving with windows closed. The most pronounced differences were seen for driving with windows open and walking, which had more than 10 times greater average variation than driving with windows closed. The high variation in exposure for driving with windows open may be explained by greater PM2.5 concentrations entering the vehicle while idling at stoplights and lower concentrations experienced while driving. The variability experienced by walkers is likely due to microenvironments along the walking path [2.5 measures also fluctuated significantly by day and time of day. For instance, the highest average variation in exposure for all modes (except light rail) occurred on the morning of August 11. Similarly, the lowest average variation for all modes (except bus) occurred in the afternoon on August 8 . The SidePak AM510 instruments measured PM2.5 concentrations indirectly using 90° light scattering based on ISO 12103–1 A1 calibration test dust. Thus, actual ambient PM2.5 concentrations may have differed from our readings due to differences in particle size, shape, refractive index, and chemical composition; however, we assume that any measurement errors were systematic and roughly proportionate among all modes of commuting. Finally, although we did re-zero the SidePak instruments each night, we did not check for zero drift immediately before or after sampling. It is possible that temperature differences between the laboratory and sampling site location caused the SidePaks to read higher in the afternoon as temperatures increased.This study was limited to examining commuter data on one roadway in a single metropolitan city during summer, and therefore may not be generalizable to other locations or seasons. We were unable to account for the large variability in PM2.5 exposure in adults and children may depend on its chemical makeup [2.5 constituents vary by mode of commuting. This may provide additional valuable information to help direct future public health policy recommendations specific to commuting. This study was also limited in that we did not measure minute ventilation rates, but used values reported in previous studies. This may have resulted in under or over-estimation of actual inhaled dose and exposure rates due to differences in the fitness levels of study participants, speed of travel, weather conditions, and grade (flat vs sloped terrain) between this and other studies. In some settings, active transportation users choose different routes than those using motorized transportation. This study fixed route choice in order to make comparisons between transportation modes. Strengths of this study include the use of an identical background monitor at the sampling location, measurement of six commuting modes simultaneously on a common route, and controlling for dander released from clothing by using the same over-shirt for all data collection personnel.Studies suggest the health effects associated with PMl makeup –47. Fondl makeup , suggest2.5 doses and exposure rates. The higher exposure doses and rates estimated in our study were largely a function of increased respiration rate and time of travel for active commuters. Furthermore, driving with windows closed with the car’s internal air circulation system turned on was protective, resulting in the lowest PM2.5 breathing zone concentration, inhaled dose, and exposure rate. Future research should evaluate differences in fine particle constituents based on mode of commuting, including in congested conditions. These comparisons should also include relevant biological measurements. Furthermore, we recommend future studies to evaluate public awareness of how transportation choices influence commuters’ daily exposures.In conclusion, our data suggests that active commuting , when compared with other modes of transportation over comparable distances and routes, results in significantly higher PM


Eptesicus serotinus) maternity colonies located in the North-East of France. This species is currently considered as the main EBLV-1 reservoir. Multievent capture-recapture models were used to determine the factors influencing bat rabies transmission as this method accounts for imperfect detection and uncertainty in disease states. Considering the period of study, analyses revealed that survival and recapture probabilities were not affected by the serological status of individuals, confirming the capacity of bats to be exposed to lyssaviruses without dying. Five bats have been found with EBLV-1 RNA in the saliva at the start of the study, suggesting they were caught during virus excretion period. Among these bats, one was interestingly recaptured one year later and harbored a seropositive status. Along the survey, some others bats have been observed to both seroconvert  and serorevert . Peak of seroprevalence reached 34% and 70% in site A and B respectively. On one of the 2 sites, global decrease of seroprevalence was observed all along the study period nuanced by oscillation intervals of approximately 2–3 years supporting the oscillation infection dynamics hypothesized during a previous EBLV-1 study in a Myotis myotis colony. Seroprevalence were affected by significantly higher seroprevalence in summer than in spring. The maximum time observed between successive positive serological statuses of a bat demonstrated the potential persistence of neutralizing antibodies for at least 4 years. At last, EBLV-1 serological status transitions have been shown driven by age category with higher seroreversion frequencies in adults than in juvenile. Juveniles and female adults seemed indeed acting as distinct drivers of the rabies virus dynamics, hypothesis have been addressed but their exact role in the EBLV-1 transmission still need to be specified.This study describes two longitudinal serological surveys of European Bat Lyssavirus type 1 (EBLV-1) antibodies in serotine bat ( Eptesicus serotinus) maternity colonies previously found exposed to European Bat Lyssavirus type 1 (EBLV-1) was assessed using capture-recapture methodology. The two roosting site were located in the North-East of France. Animals were trapped, banded, and blood samples were collected to study their status regarding EBLV-1 exposure. Using capture-recapture models, the authors found that seropositive status of bats did not affect the survival abilities of individuals. Seroprevalence of EBLV-1 antibodies within the study showed an oscillation interval of approximately 2–3 years and a higher evidence of contact with the virus in summer than in spring. The maximum duration observed between successive positive serological statuses in the bat population also demonstrated a survival for at least 4 years after the exposition. This study confirms the ability of bats to survive despite circulation of lyssaviruses within the colony. Bats could indeed provide a valuable key to improving human health, currently facing numerous zoonotic epidemic issues.A multi-annual survey of two serotine bat ( Chiroptera is the second largest order of mammals after Rodentia. They have a worldwide geographical range, with the exception of Antarctica and are represented by more than 1 100 different species , 3 and iPteropus medius) in Sri Lanka, a new candidate in the formal classification of lyssaviruses . The transition probability from seropositive to seronegative was 0.99 [0.02–0.99] (a boundary estimate that was difficult to interpret) and 0.21 [0.09–0.40] from seronegative to seropositive. The probability of judging a positive result as positive was 1 while the probability of judging a negative result as negative was 0.81 [0.75–0.86].On site B, the best-ranked model indicated that the survival probability of female bats was 0.78 [0.73–0.83]. The transition probability from seropositive to seronegative was 0.89 [0.53–0.98] and 0.25 [0.06–0.61] from seronegative to seropositive for adult female bats and 0.15 [0.07–0.27] and 0.05 [0.02–0.12] for juveniles female bats respectively.2011 = 0.26 [0.19–0.37]; OR2012 = 0.65 [0.47–0.90]; OR2013 = 0.40 [0.29–0.55]) and a lower frequency of seropositive cases in spring than in summer (ORspring = 0.46 [0.35–0.60]).The evolution of corrected bat EBLV-1 seroprevalence on both sites A and B are presented in 2013 = 2.40 [1.61–3.60]).On site B, seroprevalence peaked at 70% [59.9–78.9] in spring 2013 then progressively decreased to reach 21.6% [11.5–34.9] in May 2015. The logistic regression did not detect any significant differences between seasons, but the frequency of seropositive cases in 2013 was higher than in 2012  affected by RABV in the United States . EEBLV-1 antibody carriage in serotine bats was not correlated with mortality probability. In both site A and B, peak-seroprevalences (34 and 70% respectively) were detected one or two years after the first detection of EBLV-1 positive carcasses. While we detected oscillation seroprevalences in time, at annual level, seroprevalences were found higher in summer compare to spring, suggesting that rearing period could increase virus circulation. We pointed out differences of serological statuses between adult female and juveniles and the need for further assessment. A better understanding of this mechanism, whether of ecological, biological and/or immunological origin, is indeed a real challenge and of great interest as elucidating zoonotic virus persistence in bats concomitant to unaffected survival could help to solve human health challenges.S1 Table(DOCX)Click here for additional data file.


Schizophyllum commune BPSM01, Panusgiganteus BPSM27, Pleurotussp. BPSM34, Lentinussp. BPSM37, Pleurotusdjamor BPSM41 and Lentinula sp. BPSM45) were analysed for their nutritional , antioxidant potential. The present findings also suggested that the wild edible mushroom strains do not have only nutritional values but also can be used as an accessible source of natural antioxidants.The diversity of wild mushrooms was investigated from two protected forest areas in India and 231 mushroom specimens were morphologically identified. Among them, 76 isolates were screened for their antimicrobial potential against seven bacterial and fungal pathogens. Out of 76 isolates, 45 isolates which displayed significant antimicrobial activities were identified using ITS rRNA gene amplification and subsequently phylogenetically characterized using random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers. Sequencing of the ITS rRNA region classified the isolates into 16 genera belonging to 11 families. In total, 11 RAPD and 10 ISSR primers were selected to evaluate genetic diversity based on their banding profile produced. In total 337 RAPD and 312 ISSR bands were detected, among which percentage of polymorphism ranges from 34.2% to 78.8% and 38.6% to 92.4% by using RAPD and ISSR primers respectively. Unweighted Pair-Group Method with Arithmetic Mean (UPGMA) trees of selected two methods were structured similarly, grouping the 46 isolates into two clusters which clearly showed a significant genetic distance among the different strains of wild mushroom, with an similarity coefficient ranges from 0.58 to 1.00 and 0.59 to 1.00 with RAPD and ISSR analysis respectively. This reporthas highlighted both DTR and MNP forests provide a habitat for diverse macrofungal species, therefore having the potential to be used for the discovery of antimicrobials. The report has also demonstrated that both RAPD and ISSR could efficiently differentiate wild mushrooms and could thus be considered as efficient markers for surveying genetic diversity. Additionally, selected six wild edible mushroom strains ( Mushrooms are fleshy fungi and have become attractive as a source for the development of several extracellular enzymes with agricultural and biotechnological significance. Worldwide, they are well recognized as supplementary food due to their high nutritional values , 2 and mEven today, most of the tribal people in rural areas use wild mushrooms as their mainstay which sometimes leads to fatality due to the lack of knowledge in differentiating between edible and non-edible/poisonous mushrooms . MoreoveCurrently, amplification of the Internal Transcribed Spacers region of ribosomal DNA (ITS rDNA) has been proven as a powerful tool for the identification and phylogenetic analysis of mushrooms –18. In tPa” and sold them at local market for the betterment of their family. Hence, the present work was focused on the collection of wild fruiting bodies from two protected forest areas of Mizoram, Northeast India and was morphologically and molecularly identified by the amplification of the Internal Transcribed Spacer (ITS) region of ribosomal RNA (rRNA). Mycelial cultures were raised and screened for their antimicrobial activities. Furthermore, the identified mushrooms were genetically characterized by using RAPD and ISSR molecular markers. All the collected specimens were deposited in the Directorate of Mushroom Research, Solan, India. We hypothesized that the present study in general will help the local mushroom hunters to differentiate between edible and poisonous fruiting bodies. Overall the selected potential isolates can be exploited for the development of effective antimicrobials.Northeastern India is well known for its rich biodiversity and for its un-tapped bioresources which fallsunder Indo-Burma Mega biodiversity Hot Spot . Though,The following protected forest areas were investigated inthe present study:2 (Dampa Tiger Reserve (DTR) , which is one of the largest wildlife sanctuaries in Mizoram. It covers an area of ≈ 500 km2  and the 2 (Murlen Nation Park (MNP)  is located close to China Hills and covers an area of ≈ 100 km2 . Altitudin situ if possible or else the specimens were retained for later identification. The samples were collected in sterile paper bags and brought into the laboratory. Morphological parameters  were assessed from fresh samples as per Largent and Stuntz [Selected forests were visited for three consecutive years (2013–15) during rainy and winter sessions (June to December). Permission to collect wild mushrooms from the protected forests was obtained from the Chief Wildlife Warden, Environment and Forest Department, Government of Mizoram, India issued by Mr. LiankimaLailung, Conservator of Forest (WL), Mizoram, India. The wild mushrooms were identified d Stuntz . DigitalThe collected fruiting bodies were used to raise mycelial culture as per the modified method of Bolla et al. . BrieflyStaphylococcus aureus: MTCC-96; Escheridia coli: MTCC-739; Pseudomonas aeruginosa: MTCC-2453), three fungal pathogens  and a yeast pathogen . All the tested pathogens were obtained from the Microbial Type Culture Collection (MTCC), Chandigarh, India and were maintained at Molecular Microbiology and Systematics Laboratory, Department of Biotechnology, Mizoram University on specific media as per instructions.Antimicrobial activities were tested using three bacterial pathogens  of cell free extractof mushroom isolate and the plates were incubated at 37°C for 24 h. All experiments were carried out in triplicates and the zone of inhibition was measured in mm.Antibacterial activity was determined by modified agar well diffusion method . BrieflyTCCGTAGGTGAACCTGCGG-3’) and ITS4 (5’-TCCTCCGCTTATTGATATGC-3’) [2, 2.5mm dNTPs, 10 pmols of each primer, 50 ng of template DNA and 1 unit of Taq DNA polymerase. The PCR was carried out in the Applied Biosystems thermal cycler with an initial denaturation at 95°C for 5 min, followed by 30 cycles of denaturation at 95°C for 1 min, annealing at 57°C for 1 min, extension at 72°C for 2 min and a final extension at 72°C for 7 min. The amplified PCR product was separated on 1.5% agarose gel and visualized under Bio-Rad Gel Documentation System XR+ . The products were purified by using the Pure-link PCR Purification Kit (In-vitrogen) and sequenced commercially at Sci-Genom Labs Pvt. Ltd, Cochin, India.Total genomic DNA was extracted from the freshly grown mycelial mat as described by Cenis  with minATGC-3’) . PCR rea2, 0.5 mMdNTPs, 5 pmols of each primer, 1 unit of Taq DNA polymerase and 50 ng template DNA. Negative control reaction without DNA was included in each set. The PCR amplification was carried out with the following conditions: initial denaturation at 95°C for 5 min, followed by 35 cycles of denaturation at 94°C for 35s, annealing at 32°C for 45 s, extension at 72°C for 2 min and a final extension of 10 min at 72°C. The amplified PCR product was separated on 1.5% agarose gel using TAE buffer and examined under gel documentation system as described earlier. Ten ISSR primers were selected  Ltd,  were useselected  and ampl2, 2.0 μl of 2.5 mM dNTPs, 0.5 μl of 10 pmol ISSR primer,1 μl of DMSO (100%), 0.5 μl of BSA (10 mg/ml) and 2U Taq DNA Polymerase.The amplification conditions were as follows: initial denaturation at 94°C for 4 min followed by 35 cycles at 94°C for 50 s, at 45°C for 50 s, and at 72°C for 2 min with a final extension step at 72°C for 7 min. The amplified fragments were separated on 1.2% agarose gel using 1X TAE buffer and the banding patterns were examined under gel documentation system asmentioned above.The PCR reaction mixture was performed in 25 μl total volume, containing 50 ng of genomic DNA, 2.5 μl of 10X Taq Buffer with 15mM MgClSolanumkurzii(KC535794) as an out group. The robustness of the phylogenetic tree was tested by bootstrap analysis using 1,000 replicates using p-distance model [The obtained ITS sequences were compared with type strains retrieved from GenBank/NCBI database using BlastN search program and were aligned using Clustal W . Evolutice model . Trees wSYS version 2.2).Amplified fragments of RAPD and ISSR were scored as present (1), or absent (0), the fragments produced were numbered sequentially and entered into a binomial matrix . Only frSamples (1 g each) were dried for overnight in the oven maintained at 105°C and weighed, then placed at 575°C for 5 h. Then, the samples were weighed for final mass and difference between the mass at 105°C and 575°C was taken as ash content .Carbohydrate content was estimated by phenol sulphuric acid method with glucose as standard .Protein content was estimated by Bradford method using BSA as standard .The fat was extracted by Chloroform-methanol extraction and estimation method .The lyophilized powder (1.5 g) was extracted by stirring with 40 mL of methanol (25°C at 150 rpm) for 2 h and subsequently filtered through Whatman No. 4 paper. The residue was then extracted with 20 mL of methanol (25°C at 150 rpm) for 2 h. The combined methanolic extracts were evaporated at 40°C  to dryness and stored at 4°C until analysis .2CO3; finally 4.3 mL of distilled water was added to the solution. The reaction mixture was incubated for 60 min in the dark at room temperature and then the absorbance was measured at 725 nm using a microplate spectrophotometer UV-vis . Total phenolic content was expressed as mg of Gallic acid equivalents (GAE) per gram of dry sample (mg GAE/g). The measurements were done in triplicate.Total phenolic content was measured according to the method given by Singleton et al. . Briefly™ GO, Thermo Scientific, MA, USA) after incubation for 30 min in darkness at room temperature. The flavonoid content was calculated in mg Quercetin equivalents per g of dried sample (mg QAE/g).The flavonoids content was determined by the calorimetric method of aluminium chloride according to methodology previously described by Chang et al . In thisO+) was produced by reacting ABTS stock solution with 2.45 mM potassium persulfate  and allowing the mixture to stand in the dark at room temperature for 12–16 h before use. The ABTSO+solution was diluted with water to an absorbance of 0.70 (±0.02) at 734 nm. The reaction mixture consisted of 0.07 mL of extract and 3 mL of the ABTS radical. After incubation for 6 min, absorbance was determined in spectrophotometer at 734 nm. The antioxidant activity was calculated by using the following equation.ABTS radical scavenging activity was determined according to Re et al  with somcontrol = Absorbance of negative control at the moment of solution preparationAsample = Absorbance of sample after 45 minA50 values were calculated as the concentration of the sample (mg) required to scavenge 50% of the ABTS free radical. The IC50 is often used to express the amount of concentration of extracts need to scavenge 50%of the free radicals.The IC-1 and 64 scans per second were recorded [The selected wild mushroom samples were dried and mixed with FT-IR grade potassium bromide . The samples were grounded in agate pestle and mortar for obtaining pellets by hydraulic press. The absorbance Fourier transform infrared (FT-IR) spectra of the samples were recorded using JASCO FT-IR 6800 within the scanning range of 400–4000 cmrecorded .http://bioinformatics.psb.ugent.be/webtools/Venn/).The data were calculated in mean ± SD of the values found in triplicates from antimicrobial screening and analysed by one way of analysis of variance (ANOVA) to compare statistical significant differences (P<0.05) between antimicrobial activities of all isolates using Duncan’s multiple range test of SPSS software version 6.0. All the experiments were done in triplicates and the mean was taken. Relative abundance of mushroom isolates was compared between the selected natural forests by using Sigma Plot 12.0 version. Venn diagram was prepared online by using Bioinformatics and evolutionary genomics site .Lactarius, Trametes, Calocybe, Agaricus, Pleurotus, Schizophyllum, Cantharellus, Russula, Boletus, Termitomyces, Pholiota and Lentinus. Relative abundance of the wild mushrooms was compared between the two selected forests at the genus level (Boletus was the most abundant genus at DTR and MNP with 20.16 and 12.1% respectively. Genus Russula(10.48%) and Amanita (6.45%) were the most dominant genus in DTR as compared to MNP with 9.34% and 3.73% respectively. At the same time, genus Mycena and Trametes were dominant in MNP with 2.8% and 5.6% respectively. However, 19 genera were found in common among both the studied locations  and Ascomycota . Anatomical characteristics were also recorded of all collected fruiting bodies . Some ofus level  and the ocations . Moreovein vitro antagonistic activity against the selected fungal plant pathogens by dual culture plate assay and showed diverse antagonistic effects. Out of 76, 45 isolates showed positive growth inhibitory activity against atleast three fungal pathogens, with the percentage of inhibition ranging from 25% to 85% . Five isolates BPSM  exhibited positive antagonistic activity against all the tested pathogens. Among them, isolate BPSM30 indicated highest antifungal activity (78%) against F. proliferatum whereas isolate BPSM41 showed maximum antifungal activity (80%) against F. oxy f. sp. ciceri. The strongest antagonistic activity was found in BPSM33 isolate against F. oxysporum with 85.71% inhibition as compared to control.Out of 231, 76 isolates could be grown on artificial media and were screened for their antimicrobial potential. All isolates were tested for their % to 85% . All isoP. aeruginosa, S. aureus and E. coli. Out of 76 isolates, 37 isolates showed positive effects against two out of three pathogens and all isolates showed positive effect against P. aeruginosa except isolates BPSM17 and BPSM32. E.coliis the strongest pathogen against all the tested isolates except BPSM . The result showed that isolate BPSM01 displayed greatest inhibitory effect (9.63 mm) followed by BPSM48 (9.6 mm) and BPSM47 (9.56 mm) against P. aeruginosa. The result indicated that isolates BPSM01 and BPSM47 had the most significant effect against S. aureusgrowth with 9.6 mm and 9.46 mm respectively. Isolate BPSM03 exhibited broad spectrum antibacterial activity against E. coli(5.36 mm) and S. aureus (4.36 mm), therefore it can be considered as the most potent isolate for further studies  followed by Xylaria , Schizophyllum , Pleurotus , Marasmiellus , Microporus , Auricularia , Polyporus , Lentinus , Bjerkandera , Panus , Gymnopus , Pholiota , Fomitopsis , Hymenopellis  and Lentinula . The phylogenetic tree was constructed based on the neighbor-joining method with Kimura 2-parameter model in accordance with lowest BIC and highest AICc values using Mega 5.05. Transition/Transversion values were also estimated using Mega 5.05 (R = 1.75). ITS rRNA gene analysis classified the isolates into 16 genera and 11 families  with a bootstrap value of 75%  to maximum polymorphic of 78.8% (R5) was observed with an average of 56.8% polymorphism. Two primers showed more than 46% polymorphism. The dendrogram based on RAPD data was constructed by UPGMA analysis. The dendrogram grouped the 45 isolates into two clusters (A & B) with similarity coefficient ranges from 0.58 to 1.00. Cluster A was the largest cluster comprised 39 isolates and divided into two sub clusters A1 and A2. Cluster A1 consist of 25 isolates i.e.  similar .Trametes. Though, the dendrogram was similar with RAPD analysis, there was a slight difference from RAPD analysis. For example, Schizophyllum sp. BPSM  had the closest relationship with Pleurotus sp. BPSM  in the RAPD analysis, while it had similarly found in ISSR analysis where Schizophyllum sp. had the closest relationship with Pleurotus sp. Cluster B showed all the isolates belongs to Trametes in the RAPD analysis, while cluster B showed highly similar result in ISSR analysis  while the minimum percentage of polymorphic was observed using primer P2 (38.6%) with an average of 72.5%. Two primers showed more than 52% polymorphism. Dendrogram was constructed by UPGMA on the basis of the distance matrix by ISSR analysis. It divided the isolates into two clusters (A & B) with similarity coefficient ranges from 0.59 to 1.00. Cluster A was the largest cluster containing 42 isolates. Cluster A divided into two sub cluster A1 and A2. Cluster A1 consist of 34 isolates and Cluster A2 contain 8 isolates. Cluster B was composed of 3 isolates all belong to genus analysis .Pleurotusdjamor BPSM41  showed the highest level of ash content among the mushrooms studied. Three out of six mushrooms  showed almost the same level of proteins 30.3, 34.3 and 35.59 g/100 dw respectively. Carbohydrates content is highest in PanusgiganteusBPSM27 (80 g/100g dw), on the other hand the level of carbohydrates is extremely lower in the other samples ranging from 20–80 g/100g dw. Regarding fat contents, Lentinula sp. BPSM45 showed the highest fat content (6.26 g/100g dw) and Pleurotussp. BPSM34 revealed the lowest fat content (2.32 g/100g dw).Nutritional value and energetic contribution of the selected six wild mushrooms is presented in Bioactive properties of the selected six edible mushrooms were assessed by three different assays measuring total phenolic contents, total flavonoids content and scavenging activity of ABTS radical . As showS. commune BPSM01 exhibited the highest TPC value among the studied mushrooms (55.19 mg GAE/g) followed by Pleurotusdjamor(BPSM41) and Pleurotussp. (BPSM34) with 52.27 and 41.44 mg GAE/g, respectively. The lowest TPC value was recorded in Lentinussp. (BPSM37) with 12.76 mg GAE/g. Total flavonoid content (TFC) was quantified as milli-gram of Quercetin equivalent per gram of extract ranges from 0.373 to 0.744 mg of QAE/g. P. djamor BPSM41 showed the maximum TFC (0.744 mg QAE/g) which is followed by Lentinula sp. BPSM45 and Pleurotus sp. BPSM34 containing 0.652 mg QAE/g and 0.528 mg QAE/g, respectively. Panusgiganteus BPSM27 (0.373 mg QAE/g) and Lentinussp. BPSM37 (0.382 mg QAE/g) showed the lowest values of TFC among the mushrooms studied.50 values range from 3.25 to 11.43 mg/ml. The lower the IC50 value of mushroom extract, the higher was their free radical scavenging activity. Among the mushroom extracts we have studied, S. commune BPSM01 with an IC50 of 3.25 mg/ml and Pleurotusdjamor BPSM41 with an IC50 of 3.73 mg/ml indicates a significant antioxidant activity in the methanolic extract of these mushrooms.In this study for scavenging activity of ABTS radical, the IC-1 with functional group of ketone/aldehyde C = O stretch was greatly reduced in P. giganteus BPSM27 when compared to S. commune BPSM01, it shows that solubilization of the hemicellulose has taken place in P. giganteus BPSM27. The hemicellulose peak is totally absent in Pleurotus sp. BPSM34, Lentinussp. BPSM37 and Lentinula sp. BPSM45; it might be due to the complete solubilization of the hemicellulose part present in the samples. Reduction in lignin peak intensity of S. commune BPSM01 of wave number 2937 cm-1 with a functional group of C-H stretching was observed when compared to sample the rest of the studied mushrooms. Similarly reduction in peak intensity for lignin of wave number 1639 cm-1 and 1556 cm-1 was noticed in S. commune BPSM01 and Lentinussp. BPSM37 when compared to P. giganteus BPSM27, Pleurotussp. BPSM34, P. djamor BPSM41 and Lentinula sp. BPSM45. Functional groups representing cellulose and hemicellulose were observed at wave numbers 1200 cm-1 for P. djamor BPSM41 and Lentinula sp. BPSM45. Similarly for S. commune BPSM01, cellulose and hemicellulose peak with different functional group of glycosidic linkage was noticed at the wave number of 930 cm-1. Reduction in peak intensity for Lentinussp. BPSM37 and increase in P. djamor BPSM41 of wave number 1035 cm-1 with a functional group of C-O, C = C, and C-C-O stretching representing cellulose, hemicellulose and lignin was observed respectively, when compared to S. commune BPSM01, P. giganteus BPSM27, Pleurotussp. BPSM34 and Lentinussp. BPSM37.Mushroom samples were analysed in ATR-FT-IR  and different wave numbers, functional groups and their corresponding polymer of mushroom sample were presented in Agaricusbisporus[Volvariellabombycina [Pleurotussajor-caju [Pleurotuspulmonarius [Lycoperdonperlatum [Lentinus tuber-regium [Lentinussquarrosulus [Schizophyllumradiatumin the extracellular biosynthesis of silver nanoparticles and showed antimicrobial activity against gram positive and gram negative bacteria with anti-oxidative properties under submerged fermentation. Moreover, Ramberg et al. [Trameteshas immunomodulatory and anticancer effects.More than 200 different mushroom species have been identified from different parts of the world. Previous researchers have shown the importance of mushrooms for the production of novel secondary metabolites having antimicrobial potential , 49. In sbisporus, Volvariombycina , Pleurotjor-caju , Pleurotmonarius , Lycoperperlatum , Lentinur-regium  and Lentrrosulus . Recentlrrosulus  reportedg et al.  suggesteAmong the studied locations, it was revealed that the diversity of wild macrofungi was higher in DTR than in MNP. It is likely that the variation of wild macrofungi in different ecosystems is governed by different climatic conditions such as temperature, pH of soil, physiognomy, succession and geography. This finding was similarly reported by Angelini et al. , where sPleurotus pulmonarius (BPSM36) showing antibacterial activities against P. aeruginosa (9.4 mm) and S. aureus (7.7 mm). These findings were similarly reported by Chowdhury et al. [P. ostreatus has broad spectrum antibacterial activity. Isolate BPSM45 identified as Lentinula sp. showed antibacterial activity against all tested bacterial pathogens which was in concordance with the findings of Ishikawa et al. [Lentinula edodes has antibacterial action against B. cereus, S. aureus and E. coli. We found that all isolates were highly resistant against E. coli and C. albicans. Previous researchers have similarly reported that both the pathogens were found to be strongly effective against mushrooms [All the wild macrofungi tested were found to have various degrees of antimicrobial effects against gram positive and gram negative bacterial pathogens. Similarly, a previous study has also reported antimicrobial activity from the extract of mushrooms . In our . aureus .7 mm. Tha et al.  who showushrooms . Four stushrooms , Ramesh ushrooms , Chowdhuushrooms  who provin vitro antagonistic activity against three fungal pathogens and 45 isolates (19.48%) showed strong antimicrobial activity against at least two out of three tested pathogens. Interestingly, five isolates  found to inhibit all selected fungal pathogens, similar findings were reported by Jagadish et al. [Lentinula sp.Exhibited the highest percentage of inhibition (55.3%) against F. oxysporum, which is significantly higher than the finding reported by Osman et al. [F. oxysporumby Lentinula edodes LC202 and Lentinula edodes LC2141. Most of the selected isolates in the present study showed antagonistic potential against F. oxysporum f. sp. ciceri, a potent pathogen causing diseases in several crops. The potential isolates  have shown strong antifungal activity which has the potential to develop as a biocontrol agent. Park et al. [TCCGTAGGTGAACCTGCGG-3’) and ITS4 (5’-TCCTCCGCTTATTGATAGC-3’) primers to amplify the ITS region to identify isolates of medicinal mushroom Hericium erinaceum whereas Dentinger et al. [All the isolates were screened for their h et al.  and Udu-h et al.  who demon et al.  who showk et al.  has usedr et al.  have user et al. , and pror et al. , primarir et al. ,71 and sPleurotus sp. (BPSM10 and BPSM34) has been closely clustered together. This finding was similarly reported by Yin et al. [P. eryngii clustered together based on RAPD grouping. It proved that effectiveness of the molecular marker in the genetic diversity analysis of mushrooms at the genus level. The results obtained in the present study provided information regarding the genetic diversity of the strains by combined RAPD/ISSR analysis. In the present study, both RAPD/ISSR fingerprinting analysis clearly showed closely related Schizophyllum sp. strains and formed a mono-subclade at the coefficient of 90% and 95% respectively. Similarly Auriculariapolytricha  clustered together in both RAPD and ISSR technique, which further proved that the selected markers can be used to differentiate at the genus level. The RAPD technique has been successively used to differentiate among different strains of Agaricus bisporus [Ganoderma lucidum complex [Lentinula edodes[Agaricusbisporus[Trametes strains were clustered together and showed high levels of genetic diversity in ISSR markers, and been used in the identification and taxonomy for more than two decades [Lentinula edodes[Cordycepssinensis[Auriculariapolytricha[Agaricus bisporus[The analysis of genetic diversity is important to understand the genetic variation among the closely related organisms based on their genetic makeup. In RAPD analysis, both n et al.  and Ro en et al.  who repobisporus , Ganoder complex , Lentinula edodes; Agaricusbisporus. RAPD hasbisporus. Tramete decades . It is ala edodes, Cordycessinensis, Auriculolytricha and Agar bisporus.Lentinula sp. BPSM45 showed the highest ash content 21.406 g/100g. On the other hand, our study revealed the highest energetic contribution of Panusgiganteus BPSM27 among the selected mushrooms, due to the highest content of carbohydrates and proteins with low fat content. The level of antioxidant activity in mushrooms is largely varied from one species to another and the solvent used for extraction. This could be due to their capability to extract bioactive substances [Among the nutritional contribution of the selected six mushrooms we have studied, bstances . Apart fbstances ,85,86.S. commune 25.03 mg GAE/g where as the TPC of S. commune BPSM01 was 55.19 mg GAE/g which is the highest TPC in this study. Flavonoids were not detected in P. Ostreatusin the findings of Matilla et al [Phenolic group is the major antioxidant compound found in mushrooms, . There ala et al  and Chowla et al . It is rS. commune BPSM01 with an IC50 of 3.25 mg/ml which is the most significant antioxidant activity in the methanolic extract of these mushrooms.The DPPH and ABTS assays are known for their reproducibility; ease of application and low cost  The ABTSMicroporus sp. and Schizophyllum commune respectively. Further testing of these strains for the discovery of novel and efficient antimicrobials should be considered. Microporus sp. strain BPSM 33 showed the highest antagonistic potential against the phytopathogen F. oxysporum with 85.71% of inhibition and could be used for the development of biocontrol agent against F. oxysporum. Furthermore, the genetic variation among the potential strains showing antimicrobial potential was considered by using RAPD and ISSR molecular markers. We reported that the selected markers can be used for assessing genetic diversity of wild mushrooms, considering both marker systems gave similar results in the present study. We also conclude that the wild edible mushroom strains does not have only nutritional values but also can be used as an accessible source of natural antioxidants.The macrofungal diversity from two protected forest areas in India was undertaken and their antimicrobial potential was investigated. For the first time 231 wild macrofungi were collected and documented from the selected locations. The significant antifungal and antibacterial activity was found in


We aimed to describe the trends and associated factors of hypertension among residents aged ≥15 yr in Guangzhou, China.Three standardized cross-sectional health surveys were conducted in 2004, 2009 and 2013 using a multi-stage cluster sampling method, and a total of 69128 qualified participants were included in the study. The data were obtained through physical health examination and questionnaire survey.P<0.001) from 42.8 to 47.5 yr, and the average body mass index slightly increased (P<0.001) from 22.4 to 23.0. Logistic regression analysis shows that higher age, male, higher body mass index, smoking and drinking alcohol were potential risk factors for hypertension.The age-standardised prevalence of hypertension increased from 12.5% to 16.0% between 2004 and 2009 and declined from 16.0% to 14.0% between 2009 and 2013, and crude prevalence respectively was 14.6%, 19.1% and 18.8% in 2004, 2009 and 2013. The proportion of optimal blood pressure dropped from 51.1% to 33.2%, high-normal blood pressure increased from 20.1% to 28.9%, grade 1 hypertension and grade 2 or 3 hypertension increased from 11.5% to 13.6% and 3.9% to 5.8% between 2004 and 2013. The average age was significantly increased (Both crude and age-standardized prevalence of hypertension were initially increased, but subsequently decreased in Guangzhou during 2004–2013. The optimal blood pressure population decreased significantly while the high-normal blood pressure population increased substantially during the survey period. Hypertension is the main cause of mortality in China as well as worldwide because of its high frequency and major risk of cardiovascular disease ,2. The h2), normal weight (18.5–23.9 kg/m2), overweight (24.0–27.9 kg/m2) or obese (≥28 kg/m2).In this study, hypertension was defined an average systolic BP (SBP) ≥140 mmHg, and/or an average diastolic BP (DBP) ≥90 mmHg, or currently taking antihypertensive drugs (self-reported). Definitions and classifications of BP levels (mmHg) in our analysis were based on the 2007 Guidelines for the Management of Arterial Hypertension  to 167 million Yuan (2014), with an average growth rate of 9.7% over the last decade . With thThe Community Health Diagnosis of Guangzhou is a program to assess health status of community residents in Guangzhou, which carried out by Guangzhou Center for Disease Control and Prevention every five years. A representative sample of permanent residents (aged 15 yr or older) in Guangzhou City was recruited. Three standardized cross-sectional health surveys were conducted in 2004, 2009 and 2013.The sample size of each survey was decided based on the Community Health Diagnosis Technical Manuals, recommended by the Department of Maternal and Child Health Care and Community Health of the Ministry of Health in China. Namely, using a family as the basic unit, 800 households were randomly sampled from community of more than 50000 people (two neighbourhoods nearby were combined if the population size of the neighbourhood was less than 50000), and single individuals were categorized as a family, a total of 9600 households. All family members were interviewed in this survey, including infants, children, adolescents, pregnant people and aged people. Participants between 15 to 80 yr old were selected except pregnant women. The sample size of each survey was 27299, 23467 and 18362 participants in 2004, 2009 and 2013. We selected this age range because BP is more susceptible to social economic factors than other age groups. The response rates for survey 2004, survey 2009 and survey 2013 were 93.1%, 98.87%, and 90.5%, respectively.Informed consent was taken from the participants and the study was approved by Ethics Committee of the university.A five-stage cluster random sampling plan was used to recruit participants in 2004, 2009 and 2013, respectively. In the first stage, one community health service center from each district was randomly selected as a sample point. Therefore, 12 locations were sampled and used for all three surveys. In the second stage, two neighborhoods (urban) or townships  were randomly sampled from each district. If a community health service center had jurisdiction over only one neighbourhood, then the neighbourhood was the sample point. The third stage of sampling was conducted in each selected neighbourhood or township. Four residential committees/villages were randomly selected from each neighbourhood or township and eight residential committees/villages were directly selected from the neighbourhood when the community health service center’s jurisdiction was only one neighbourhood. In the fourth stage, 100 households were randomly sampled from each residential committee/village. A residential committee with less than 100 households was merged with another committee. In the fifth stage, all family members who live in Guangzhou for at least 6 months were selected. Samplings from first to third stage were completed by the Guangzhou Center for Disease Control and Prevention (CDC). Sampling in the fourth and fifth stage was completed by each sample point according to the principle of random sampling and was implemented with the Guangzhou CDC’s approval .All participants were interviewed, completed a questionnaire and provided with a health examination at the central survey sites or households. Guangzhou CDC was responsible for developing the questionnaire. Our study chose part of the questionnaire, including demographic characteristics , personal health history (common chronic disease) and lifestyle (drinking and smoking habits). If the respondents could not finish the questionnaire during the investigation process , then a person knowledgeable background could reply on their behalf. Health examinations were conducted by physicians received specific training for the health assessments. Height, weight, and BP were collected in the health examination. Height and weight were measured before breakfast in the morning while wearing light clothes and without shoes. The participants were told to avoid alcohol, coffee, tea and exercise before the BP measurement. The BP measurement was taken twice for the right arm while participants were in a sitting position, and each measurement was separated by a 5-min interval. If the difference between the two readings were greater than 6 mmHg, then a third measure was taken.The average two or three readings (if necessary) were used for analysis. To ensure the reliability of survey data, systematic training was provided for investigators to ensure understanding of the study objective, meaning and methods. Investigation forms were checked by staff for errors or omissions after the questionnaire recovery, so that phone calls could be made to supplement the information. The replacement rate was not allowed to exceed 10%.F test. Age-standardised prevalence was calculated based on China's sixth census data in 2010 using the age groups 20–34 yr, 35–49 yr, 50–64 yr and 65–80 yr. Multinomial logistic regression was used to estimate potential risk factors for hypertension, including age, sex, nationality, BMI, smoking, drinking alcohol from the three surveys. Statistical analyses were conducted used SPSS 21.0 . Two-tailed P values <0.05 were considered statistically significant. The 95% confidence intervals were calculated and presented.Epidata (ver. 3.0) was used to establish database. All data were double inputted and checked. The frequency differences between research variables and trends were compared using the chi-squared test or trend of chi-square test. BP and age were compared using the P<0.001) while those aged 50–64 yr and 65–80 yr showed an increasing trend (P<0.001) between 2004 and 2013. Young people aged 15–34 yr decreased from 36.1% in 2004 to 24.2% in 2013, and people over age 50 yr increased from 33.1% to 47.0% during the same time. The mean ages in 2003, 2009 and 2013 were 42.8, 43.6 and 47.5 yr, respectively, which was a statistically significant increase (P<0.001). People with a BMI ≥24 or 28 have increased from 19.4% to 27.3% and 4.6% to 10.7% in 2004 to 2013 (P<0.001). The population also had a higher BP, due to an SBP increase from 117.9 to 123.2 mmHg and a DBP increase from 75.1 to 76.4 mmHg during 2004–2013 (P<0.001).P<0.001), but those aged 50–80 yr increased from 2004–2009 and declined from 2009–2013. We can find a similar law after gender stratification in both age-standardized and crude rates. The hypertension prevalence increases with residents’ age, with a peak in 65–80 yr old.Age was significantly associated with greater odds of hypertension in 2004, 2009 and 2013, the danger of suffering hypertension rises with the increasing of age .2004=2.01, 95%CI: 1.76 to 2.29; OR2009=2.08, 95%CI: 1.87 to 2.32; OR2013=2.05, 95%CI:1.86 to 2.26) and ≥28  was significantly associated with greater odds of hypertension compared with those BMI ranged from 18.5 to 23.9. People ever smoked  was significantly associated with greater odds of hypertension in 2013 compared with those never smoked.Male  was significantly associated with greater odds of hypertension in 2009. People whose BMI ≥24  was significantly associated with greater odds of hypertension compared with those never drank alcohol in both 2004, 2009 and 2013.People ever drank alcohol , and a slight decline in 2009–2013 16.0% vs 14.0%), which was more obvious in individuals aged 50 yr or above. Studies of different magnitudes and different research groups demonstrated varying prevalence  in central China but a rising trend (P<0.0001) in other regions  model, recommended for further study, can be used to distinguish the separate age effect regarding the hypertension prevalence in continuous cross-section data. Furthermore, there are many factors affecting hypertension prevalence such as dietary habits, lifestyle, public resources, economics and health care , and thiHypertension prevalence of community residents in Guangzhou has increased between 2004 and 2009, with a moderated decline between 2009 and 2013. Age, BMI, sex, smoking and drinking alcohol were the influential factors for hypertension. With the urban and economic development of Guangzhou, hypertension prevalence will still increase. Public health strategies should be still focused on the aging population and hypertensive patients in Guangzhou. Meanwhile, we should also keep one eye on high-normal blood pressure population and to prevent them from turning into hypertensive patients.Ethical issues  have been completely observed by the authors.


When dialysis access stenoses are dilated by noncoated balloons, respective primary and secondary patencies hardly reach 50 per cent and 85 per cent at one year. This study determines the primary and secondary patency rates at 6 and 12 months for venous stenoses treated by paclitaxel-coated balloon (PCB) angioplasty in native hemodialysis accesses.From 2012 to 2014, 70 venous stenoses in 41 patients benefited from PCB angioplasties. The patients’ mean age was 62.5 ± 13.8 years’ standard deviation (SD) with 75 per cent male gender, 31.7 per cent diabetes, and 65.9 per cent arterial hypertension. There were 58.5 per cent forearm fistulas and 41.5 per cent arm fistulas. Primary and secondary patency rates were prospectively established by using the Kaplan-Meier technique and tested by using the log-rank test.The primary patency rates ± SD were 81.4 ± 4.6 per cent and 60 ± 5.9 per cent at 6 and 12 months, respectively. The secondary patency rates ± SD reached 94.3 ± 2.8 per cent at 6 months and 91.4 ± 3.3 per cent at 12 months.This study shows an improvement of the primary and secondary patency rates at 6 and 12 months when venous stenoses of native dialysis fistulas are treated by PCB. The indications for PCB, however, remain to be established by larger randomized studies. As a consequence of the constant aging of the populations, the incidence of end-stage renal disease is steadily rising . When re23456The main concerns regarding native shunt failures are the anastomotic and venous stenoses that lower the shunts’ flow, which may lead to thrombosis, a complication that seriously impairs the long-term durability of dialysis accesses . Pathoge457The technique of the endovascular treatment of the venous strictures  has been extensively described. In short, plain balloons generally allow procedural success, high-pressure balloons help in difficult cases, and the utility of the more expensive cutting balloons remains unproven 89. That 865The purpose of our study was to determine the primary and secondary patencies at 6 and 12 months of the venous stenoses treated by a PCB.We conducted a nonrandomized prospective single-centre study to determine the primary and secondary patencies at 6 and 12 months of native arteriovenous fistulas venous stenoses treated by PCB percutaneous angioplasty. The study protocol was approved by our institutional review board, and written informed consent was obtained. The study began in February 2012 with the first PCB venous angioplasty and ended in November 2014, 12 months after the 70th PCB dilation. Patients less than 18 years old were excluded.Forty-one patients with a mean age ± SD of 62.5 ± 13.8 years (range 30–83 years), including 31 (75.6%) men and 10 (24.4%) women, benefited from dilations. Thirteen of the 41 (31.7%) patients were treated for diabetes, whereas 27 (65.9%) received antihypertensive drugs.Forearm fistulas were 24/41 (58.5%) and arm fistulas were 17/41 (41.5%), with 30/41 (73.2%) shunts on the left side. The mean age of the shunts was 67 months (range 10 days to 288 months), and 29/70 (41.4%) of the stenoses had had a previous treatment that was a balloon dilation in 25/29 (86%) of the cases.Seventy venous stenoses in these 41 shunts benefited from PCB angioplasties. Fifteen of 70 21.4%) stenoses were discovered via symptoms of shunt dysfunction, as listed in Table 71.4% stenAll dilation procedures were performed with patients under continuous electrocardiography and pulse oximetry monitoring; conscious sedation was achieved by an anesthesiologist for fragile or demanding patients; otherwise, 1 or 2 mg of midazolam were slowly administered intravenously before the balloon inflations.A 5F introducer sheath was inserted in the vein, and fistulography was performed, with or without a tourniquet, to assess the site, the severity and the length of the stricture, and the diameter of the adjacent healthy vein. A 0.035-inch straight, hydrophilic-coated, Terumo guidewire  was used for calibration purposes before angiographic measurements. The venous stenoses’  mean length and their mean distance to anastomosis ± SD were 17.2 ± 12 mm and 13.4 ± 12.2 cm. The mean severity of stenoses ± SD was 67.6 ± 9.4 per cent. When required by the size of the PCB, the 5F sheath was exchanged for a 6F one (mandatory for PCB with a diameter of 7 mm).Heparin was administered intravenously in doses up to 3,000 IU according to the patient’s weight. The stenosis was then crossed with the guidewire, using a 5F vertebral catheter  as a torque device. In very complex, anfractuous strictures, thinner guidewires were used. The guidewire was then possibly exchanged for an 0.018 metallic guidewire , and the PCB , with appropriate diameter (5–7 mm) and length (40–60 mm), was then inflated for one minute at prescribed pressure. If a waist persisted on the PCB, overinflation to 18 ATM was applied. A pressure-inflation device  was used for balloon-inflation monitoring in all cases Figures .When the dilation was incomplete, the stenosis being unbreakable or the diameter of the adjacent healthy vein being > 7 mm, we resorted, depending on the case, to cutting balloons  (five cases) or to using larger high-pressure balloons  of 8 or 9 mm in diameter (seven cases).For each patient, an echo-Doppler was performed 10 days after the procedure to check fistula flows in order to collect basis values (no overflow was noticed). Then, even without any symptom of dysfunction of the shunt, the patients benefited from echo-Doppler controls at 3, 6, and 12 months. If, in the 12-month period, the fistula showed any sign of significant stenotic resurgence, be it in clinical or echo-Doppler terms, the shunt was dilated again. Stenoses at another site than the previously dilated one were treated by conventional, high-pressure, or cutting balloons, and restenoses at the same site benefited from PCBs. The former were excluded from the secondary patency rates, whereas the latter were included.Primary patency for a dilated stenosis was defined as uninterrupted patency of the treated site after PCB dilation, and secondary patency was defined as patency of the treated stenosis following endovascular reintervention with a PCB at the same site. We did not resort to the recommended standard of assisted primary patency since we wanted to compare our results to the main studies, using noncoated balloons, which mostly mentioned primary and secondary patencies . We consPrimary and secondary patency rates for venous stenoses after PCB dilation were established by using the Kaplan-Meier technique and tested by using the log-rank test. Predictors of dilated stenosis patency, such as the length of the stenosis or the age of the fistula (cf. infra), were determined by using a Cox proportional hazards regression model with clinical and anatomic variables. Variables identified in the univariate Cox model with P < 0.2 were included in the multiple-regression model by using a stepwise regression. A two-sided P value of less than 0.05 was considered to indicate a statistically significant difference. Statistics calculation was carried out by the Luxembourg Institute of Health with the SAS version 9.4 software.An initial success rate  of 58/70 (82.9%) due to PCB waist or insufficient diameter of the PCB rose to 100 percent with additional procedures. The mean duration ± SD of a dilation was 19.1 ± 8.3 min. During the procedures, five cases of venous extravasation occurred at the site of dilation and were easily controlled by balloon inflation at 2 ATM and/or external compression. Spasms and elastic recoils occurred, but none needed to be treated. One case of infection of the puncture site responded well to broad-spectrum antibiotic therapy.During follow-up, three shunts were abandoned because early (one to four months) and unexpected extensive thrombosis occurred, and complex endovascular treatments were considered too heavy due to the patients’ poor clinical status (very old age and/or severe cardiac deficiency); these patients were dialysed via central catheters. The three fistulas comprised five PCB-dilated stenoses that were included in the data. Three patients died, unrelated to the procedures, and were censored.At 6 and 12 months, respectively, primary patency rates ± SD for the dilated venous stenoses were 81.4 ± 4.6 per cent and 60 ± 5.9 per cent. For the arm fistulas, primary patency rates were 80 ± 7.3 per cent and 53.3 ± 9.1 per cent at 6 and 12 months, whereas, for the forearm fistulas, they were 82.5 ± 6 per cent and 65 ± 7.5 per cent (Table The only significant factor negatively impacting venous primary patency was the length of the stenosis (hazard ratio (HR) per centimetre = 1.038, P = 0.008): the longer the stenosis, the shorter the primary patency Table .For all the dilated venous stenoses, secondary patency rates ± SD, at 6 and 12 months, were 94.3 ± 2.8 per cent and 91.4 ± 3.3 per cent. For the arm fistulas, they were 90 ± 5.5 per cent and 90 ± 5.5 per cent, whereas, for the forearm fistulas, they were 97.5 ± 4.2 per cent and 92.5 ± 4.2 per cent  and the higher stenosis severity (P = 0.037) Table >.In this study, we determined the primary and secondary patencies at 6 and 12 months, that is, 84.1 per cent, 60 per cent, 93.4 per cent, and 91.4 per cent, respectively, of native arteriovenous fistulas stenoses that were dilated by PCB. The patency rates we observed compare favourably to other series using noncoated balloons, even though our echo-Doppler surveillance program, which allows early detection of the stenoses, has possibly lowered the patency percentages. For example, one publication presented a primary patency with a conventional balloon at 12 months reaching 48.5 per cent, whereas, in another article, also using plain balloons, primary patency at 12 months was 41.4 per cent 4. HoweverThe greater length of the stenosis was the only negative predictor of primary patency, whereas the higher severity of the stricture and the younger age of the patient were the sole unfavourable predictors of secondary patency. As far as secondary patency significant predictors calculation is concerned, the small number of restenoses (22) may have biased the results. Classical predictors such as gender, diabetes, or fistula age did not statistically modify primary or secondary patencies. This was also noted in one study . HyperteThe good efficacy of the PCB results in the inhibition of neointimal hyperplasia by paclitaxel, a mitotic inhibitor. The problem is that engineering has a cost: a PCB is much more expensive (up to three times the price in Luxembourg) than a conventional or a high-pressure balloon, and it may, quite logically, be inflated only once. Moreover, some additional dilation procedures must be performed with high-pressure balloons, cutting balloons, or balloons exceeding the diameter of 7 mm .Should PCBs only be used in selective cases of long venous stenosis? Should they be kept for restenosis? As early unexpected thrombosis occurred in three shunts after PCB dilation, should short-term anticoagulation be considered? Since we recognize that our series is small, implying possible bias, and particularly for the secondary patency section, concerning 22 restenoses only, further randomized, multicentric studies, with larger cohorts of patients are needed to refine the indications for PCBs.PCBs appear to improve the primary and secondary patency rates after the treatment of venous stenoses in native hemodialysis fistulas. The main problem remains in their much higher cost, possibly indicating that they should be indicated for selected cases. Further larger and randomized studies must be launched to define the best indications of this expensive device more precisely.


Objective: Vitamin D deficiency is common in patients with ileal pouch-anal anastomosis (IPAA) for ulcerative colitis (UC). Whether vitamin D levels are further lowered in patients with concomitant IPAA and primary sclerosing cholangitis (PSC) is not known. The aim of this study was to evaluate the presence of PSC as a risk factor for vitamin D deficiency in patients with UC and IPAA.Methods: In this case control study, 74 patients with concurrent IPAA and PSC were included in the study group, and 79 patients with IPAA, but without PSC, served as controls. Forty-four variables were analyzed. Univariate analysis and multivariate analysis with stepwise logistic regression were performed.Results: A total 153 eligible patients were included, with 74 (48.4%) in the study group and 79 (51.6%) in the control group. Vitamin D level in the study group was 18.9 ± 1.4 ng/dL compared with 30.3 ± 1.7 ng/d in the control group (P = 0.011). Vitamin D deficiency (≤ 20 ng/dL) was present in 65 (42.5%) patients. PSC occurred in 49 (75.4%) of the 65 patients with vitamin D deficiency. In the multivariate analysis, only the presence of PSC  and vitamin D supplementation  remained associated with vitamin D deficiency.Conclusion: The presence of PSC was found to be an independent risk factor for vitamin D deficiency in UC patients with IPAA. These patients should be routinely screened and closely monitored for vitamin D deficiency. Metabolic disorders have been extensively reported in IPAA including anemia, iron deficiency, vitamin BPrimary sclerosing cholangitis (PSC) is the most common liver disorder associated with inflammatory bowel disease (IBD). It is more common in UC than in Crohn's disease (CD), with prevalence of 8% and 1%–3%, respectively , 5. The Vitamin D deficiency is also commonly seen in patients with IBD , with thVitamin D levels have not been studied in UC patients with IPAA and concomitant PSC. The presence of PSC, a cholestatic liver disorder, could potentially further burden vitamin D levels in patients with UC and IPAA, who have already been shown to have high rates of vitamin D deficiency. We hypothesized that patients with IPAA and PSC would a dual risk for vitamin D deficiency. The aim of this study was to evaluate if the presence of PSC is a risk factor for vitamin D deficiency in patients with UC and IPAA.The medical records of 1221 patients from the Pouchitis Registry at the Cleveland Clinic were reviewed. Of these, 153 eligible patients were identified, of whom 74 had PSC. Data were collected retrospectively from the years 2001–2014. This case control study was approved by the Cleveland Clinic Institutional Review Board.3 measurement after colectomy.Inclusion criteria were patients with (i) UC and with or without PSC and (ii) serum 25(OH)DExclusion criteria were (i) IPAA for familial adenomatous polyposis or other non-IBD conditions and (ii) patients with PSC who had been submitted to liver transplantation.After reviewing the medical records of 1221 patients from our Pouchitis Registry, 74 patients with PSC fulfilled the inclusion criteria (study group). From the remaining 1147 patients, 79 eligible age-matched controls (control group) were identified and included in the study.vs south). Definition of geographical location was as follows: a northern location was considered above the latitude of 37 degrees north, and a southern location was considered below the latitude of 37 degrees north. The presence or absence of PSC diagnosis was also evaluated.Twenty eight variables were analyzed. Variables known to impact vitamin D status were assessed. Demographic variables included age, sex, race, body mass index, smoking status and geographical location DMedication use was documented, including antibiotics, corticosteroids , and non-steroidal anti-inflammatory drugs (NSAID). Supplementation for calcium and vitamin D was also evaluated.3 were used to assess vitamin D status. Levels of 25(OH)D3 ≥ 30 ng/ml were considered normal. Low levels of vitamin D were considered when 25(OH)D3 was < 30 ng/ml. Low vitamin D levels were further subdivided into vitamin D insufficiency when 25(OH)D3 levels were between 21 and 29 ng/dl and vitamin D deficiency when 25(OH)D3 levels were ≤ 20 ng/dl.Blood levels of 25(OH)DThe primary outcome was to determine the frequency of vitamin D deficiency in PSC patients with UC and to evaluate the association of PSC as an independent risk factor for vitamin D deficiency in UC patients with IPAA.t test . Chi-square test  was used to compare categorical variables. Univariate analysis was used to identify potential risk factors for vitamin D deficiency.Statistical analysis was conducted using the SPSS software version 22 . Mean ± SD, or N (%) was used to present continuous variables. Continuous variables were compared using the Student P < 0.05) in the univariate analysis, along with known risk factors for vitamin D deficiency, were included in the multivariate logistic regression analysis. Stepwise approach was used for sifting through large numbers of potential independent variables.Factors identified as being significant (P= 0.150). There were 69 females (45.1%) in the overall population, with 27 (36.5%) in the PSC subgroup and 42 (53.2%) in the subgroup without PSC (P= 0.051).The mean vitamin D level in the overall population studied was 24.8 ± 1.5 ng/dL. The mean vitamin D level in patients with PSC compared with patients without PSC was 18.9 ± 1.4 ng/dL and 30.3 ± 1.7 ng/dL, respectively (P= 0.011). Vitamin D deficiency (levels ≤ 20 ng/dL) was present in 65 (42.5%) of the 153 patients, while 75.4% (49/65) of the patients with vitamin D deficiency had PSC.This study included 153 patients. Of these, 74 (48.4%) had UC, IPAA and PSC (study group), and 79 (51.6%) had UC and IPAA only. The mean age of the overall population studied was 51.4 ± 1.2 years. The mean age of patients with and without PSC was 48.8 ± 13.8 years and 52.8 ± 16.0 years, respectively (vs 73.4% (58/79), respectively (P < 0.003). The prevalence of vitamin D deficiency was 66.2% (49/74) vs 20.3% (16/79) P < 0.001 in patients with PSC and without PSC, respectively.When comparing the subgroup of patients with PSC to the patients without PSC, the prevalence of low levels of vitamin D was 91.9% (68/74) vs 23.8 ± 1.0 U/L, P < 0.001), alanine aminotransferase , bilirubin  and alkaline phosphatase  were found in PSC patients, as expected. The albumin levels were not different between the groups .In addition, higher levels of aspartate aminotransferase (75.7 ± 10.9 3 ≤ 20 ng/dl), and statistical analyses were performed to evaluate possible risk factors associated with vitamin D deficiency.This study was further subdivided in two separate samples based on the presence or absence of vitamin D deficiency (25(OH)D.In the univariate analysis, the following variables were associated with vitamin D deficiency: higher levels of total bilirubin, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase and creatinine; lower levels of calcium and transferrin; use of vitamin D supplementation, NSAID and corticosteroids and the presence of PSC as shown in P= 0.001) along with vitamin D supplementation  as shown in .In multivariable analysis, PSC was an independent risk factor for vitamin D deficiency in the multivariate analysis . In that study, vitamin D levels were assessed in 143 patients with PSC without mention of UC, of which 56 PSC patients received ursodeoxycholic acid (UDCA) and did not have end-stage liver disease, while 87 patients had advanced PSC and were being evaluated for liver transplantation. The prevalence of vitamin D deficiency in the UDCA treatment group, which consisted of patients without end-stage liver disease, was 14%, while 57% of the patients in the pre-transplantation group had vitamin D deficiency [There are limited data in the literature regarding vitamin D deficiency in patients with PSC. Jorgensen 3  by sunlight exposure. The vitamin D produced in the skin is transported to the liver bound to vitamin-D binding protein. In the liver, vitamin D undergoes the first hydroxylation to produce 25- hydroxy vitamin D3. Thus vitamin D metabolism is closely related to liver function. A second hydroxylation step then occurs in the kidney to form 1,25-hydroxyvitamin D3, which is the active vitamin D metabolite. Dietary intake of vitamin D3 is a smaller contribution to the total vitamin D pool, although this contribution may become important in geographic locations where the sunlight is less abundant, such as northern countries. Vitamin D acquired through the diet is absorbed mainly in the duodenum and jejunum and depends on bile-acids since it is a liposoluble vitamin [It is well known that the skin is the main source of vitamin D through conversion of 7– dehydrocholesterol to vitamin D vitamin , 19.Cholestatic diseases such as primary biliary cirrhosis and PSC are associated with vitamin D deficiency. The main mechanism of vitamin D deficiency was proposed to be impaired intestinal absorption of vitamin D due to insufficient intraluminal bile acid concentrations , 20. In 3 and increased catabolic removal of 25(OH)D3 [Vitamin D deficiency has also been shown to be common in patients with chronic liver disease in general with a prevalence ranging 46% to 75% , 22–24. 25(OH)D3 .vs 4.5 ± 0.3; P= 0.320), suggesting overall preserved liver synthetic function in the PSC patients studied. In addition, liver function tests in the PSC patients studied showed a cholestasis pattern, with aspartate aminotransferase of 75.7 ± 10.9, alananine aminotransferase of 75.9 ± 9.2, bilirubin of 2.4 ± 0.5 and alkaline phosphatase of 229.8 ± 25.3. This suggests that significant vitamin D deficiency in patients with IPAA and PSC occurs in the cholestatic phase of the disease before end-stage liver disease is established. This may be due to the fact that patients with UC and IPAA are already at risk for vitamin D deficiency due to adverse metabolic effects of the pouch construction, and dietary vitamin D sources may be significant for maintaining vitamin D levels. Cholestasis due to PSC may alter the thin balance that maintains vitamin D levels in these patients.Vitamin D deficiency can occur in early stages of PSC; however, the rates of vitamin D deficiency seem to be much higher when PSC reaches end-stage liver disease . AlthougThe association between the severity of the liver dysfunction and vitamin D deficiency was not assessed. Further prospective studies evaluating the association of vitamin D deficiency with the degree of liver dysfunction in PSC patients are needed to ascertain whether significant vitamin D deficiency already occurs in the cholestasis phase.P= 0.018). This can be explained by the fact that individuals with vitamin D deficiency are usually started on vitamin D supplementation. It may be interesting in the future to evaluate the exact dose and duration of vitamin D supplementation that could prevent vitamin D deficiency in these patients.In this study, vitamin D supplementation remained a risk factor for vitamin D deficiency in the multivariate analysis  were from northern locations.Although there were no differences in vitamin D levels between races, conclusions about race are difficult to ascertain in this study given the way this information is entered into our database, with patients being divided as Hispanics and non-Hispanics only. Because this is a retrospective study, it was not possible to ascertain patient compliance to vitamin D supplementation. It will also be interesting to compare different doses of vitamin D supplementation in patients with IPAA and PSC in the future to establish the exact dose of vitamin D supplementation that would prevent vitamin D deficiency in these patients.In conclusion, vitamin D deficiency was common in this patient population, suggesting the need for close evaluation of vitamin D levels. More studies are needed to clarify the actual mechanism by which PSC contributes to lower levels of vitamin D in IPAA patients.Conflict of interest statement: none declared.


In present study we have elucidated the role of 2758 A>G (rs696), in the recognition site of miR449a in the 3′ UTR of NFKB inhibitor alpha (NFKBIA) gene, in development of sporadic colorectal cancer. Colorectal cancer (CRC) is rated as second cause of cancer death. Genetic determinants are considered as driving forces in development of sporadic CRC. Single nucleotide polymorphisms (SNPs), are attributed as the main genetic factor in cancers susceptibility. MicroRNAs, are key players in post-translational gene regulation by binding to their specific recognition sequences located at 3' untranslated region (UTR) of mRNAs.  A case–control study using 143 CRC patients and 137 noncancerous counterparts were undertaken in order to determine rs696 genotypes using polymerase chain reaction– restriction fragment length polymorphism (PCR–RFLP) method. There was a significant difference for the genotype frequencies of rs696 between patients and controls. The frequencies of GG, AG, AA genotypes in the control group were 38.7, 45.3, and 16.1 %, respectively, and the genotype frequencies in case group were 19.6, 40.6, and 39.9 %, respectively. Our results suggest significant correlation between rs696 polymorphism and colorectal cancer risk Globally, more than one million people experience CRC every year, and it has become the third and second most common malignancy in men and women respectively , 2. DespMicroRNAs (miRNAs), small noncoding RNA molecules with poptosis , 17. miRpoptosis . Recent poptosis . NFKBIA 2758 A>G (rs696) variant located at recognition site of miR449a  method was used to determine rs696 genotypes. PCR was performed in a total 25µl volume containing, 2.5 µl 10x PCR buffer, 1.5 mM MgCl2, 200 μM dNTPs, 0.2μM each primer, 100 ng of genomic DNA, and 1 U of Taq DNA polymerase. Thermal cycling profile adjusted at 94◦ C for 5 min followed by 34 cycles of 94◦ C, 64◦ C and 72◦ C all for 30 seconds with a final extension step of 72◦ C for 5 min. Primer sequences, reaction conditions, and restriction enzyme used are given in HaeIII restriction endonuclease enzyme overnight at 37◦ C. Subsequently restriction enzyme was inactivated at 80◦ C for 20 minutes and then analyzed on 2% agarose gel. Fragment sizes of 535 and 410 bp were indicative of AG heterozygous genotype; a single 410 bp fragment displayed the presence of GG homozygous and one fragment of 535 bp indicated AA genotype . DNA was stored at -20genotype . To confStatistical analyses2) test were used to compare the differences of demographic characteristics distributions between CRC patients and the control group. Hardy–Weinberg equilibrium was tested among cases and controls using Pearson’s Chi square test. Genotype and allele frequency differences between cases and controls were evaluated using logistic regression analysis, with odds ratios (OR) and 95% confidence intervals (95% CI). P value < 0.05 was considered statistically significant. All data analyses were carried out using SPSS 22.0 statistical software. The t- test and the Chi-squared (χP = 0.10 and P = 0.34 respectively). However, we found a significant difference in physical activity (P = 0.007) between case and control groups. Also individuals in control group were more aspirin or NSAID users compared with sporadic CRC cases (P < 0.004). The case group had a higher proportion of male subjects (52.4% vs 39.4%). Our study population consisted of 143 CRC patients  and 137 CRC free individuals . The distributions of selected characteristics of the cases and controls are presented in  P < 0.001). In allele distribution analysis we found increased level of allele A in cases, compared with controls. This allele was associated with an increased risk of CRC . The distributions of genotype and allele frequency are shown in Genotyping data were in Hardy–Weinberg equilibrium for this SNP in the tested groups. There was a significant difference for the genotype frequencies of rs696 between patients and controls. The frequencies of GG, AG, AA genotypes in the control group were 38.7, 45.3, and 16.1 %, respectively, and the genotype frequencies in case group were 19.6, 40.6, and 39.9 %, respectively. This result demonstrated the significant association of AA genotype with risk of CRC in our study population . The analysis for correlation between NFKBIA genotypes and NSAIDs consumption demonstrated that in drug user patients, AA genotype was more frequent vs. AG, GG genotype . While in control group, the situation was reverse (38.9% vs. 80.6%). Data are shown in As mentioned above we also found that our case group consumed less aspirin or NSAID compared to control group  was more frequent in CRC patients. Also individuals possessing allele A were more frequently affected with CRC than those with G allele . et al. (et al. (In a study conducted in China AA genotype of rs696 polymorphism found to be correlated with elevated risk of nasopharyngeal carcinoma . Such aset al.  could no (et al.  indicateTo examine the interaction between the NFKBIA genotypes and NSAID use, we assessed the effect of NSAID use stratified by NFKBIA genotypes in drug user subgroup. The result showed that patients with AA genotype of this polymorphism appeared to have less benefit of NSAID use; it is plausible that NSAID use could affect by NFKBIA genotypes. Without doubt, agents such as NSAIDS that inhibit inflammatory events can reduce the risk of CRC. Based on the available reports 50% diminution in the incidence of colorectal cancer attributed to the regular aspirin users , 29. ButWe believe this work would further justify the role of NFKB1/NFKBIA axis in colon cancer susceptibility. Further evaluation of the role of rs696 in NFKBIA gene in other population with an expanded population size would help to reach a definite conclusion regarding the role of this polymorphism and may prove its utility as a CRC screening biomarker.


To evaluate changes in intraocular pressure (IOP) after clear corneal phacoemulsification (CCP) in normal patients.th day, lst, 2nd, 3rd month and 6 months after surgery.A prospective study including 273 normal patients selected for cataract extraction by CCP. Intraocular pressure was recorded on the 15For statistical analysis, Epi Info was used to determine the statistical significance of changes in IOP.P = 0.12), but significantly correlated with change in anterior chamber depth (ACD) (P = 0.002). The postoperative IOP was inversely related to preoperative ACD (P = 0.012). Age, sex and axial length were not significantly related to IOP reduction (P = 0.2–0.5)The mean age of 96 women and 177 men was 71 ± 12 years. The mean IOP before surgery was 14.18 ± 3.4 mmHg. Our patients showed a mean decrease in IOP of 2.25 mmHg (16%) compared to preoperative values. Change in IOP was not related to lens thickness (CCP was associated with a statistically significant reduction in IOP. The exact mechanism by which cataract surgery results in IOP reduction is unclear. CCP can be performed with the intent of achieving better IOP control. Cataract extraction surgery, independent of the technique used, induces variations in intraocular pressure (IOP). Although an elevation of the IOP in the early postoperative stage may be noted, many studies have reported a reduction in IOP.‐3 Some s4This was a prospective study including 273 normal patients selected for cataract extraction by phacoemulsification using a 3.2 mm clear corneal incision between June 2003 and January 2006. The study was approved by the Hassan II University Ethics Committee.A prospective analysis was performed using clinical charts focusing on patient age and sex, size of the capsulorhexis, and pre- and postoperative IOP. The axial length, lens thickness and anterior chamber depth (ACD) were measured during preoperative assessment three weeks before surgery using ultrasound A scan by contact technique. Central corneal thickness was not assessed. IOP was measured by Goldmann applanation tonometer by the same examiner preoperatively, on the 15th day, and subsequently one, two, three and six months after surgery. The mean change in IOP after surgery was calculated. The mean follow-up period was six months.Patients with history of ocular surgery, trauma, preoperative IOP greater than 21 mmHg, on ocular medication and who developed postoperative complication were excluded from the study.For statistical analysis, Epi Info was used to determine the statistical significance of changes in IOP. The statistical significance between the groups was estimated using Chi test. A valueAll surgeries were performed by the same surgeon.Most of the patients underwent surgery under peribulbar anesthesia using Lidocaine.Surgery involved a 3.2 mm superior clear corneal tunnel incision, injection of viscoelastic material into the anterior chamber, capsulorhexis of 5 mm, hydrodissection, in the bag phacoemulsification using phaco-chop technique, cortex aspiration, additional injection of viscoelastic material and insertion of foldable hydrophobic intraocular lens (IOL) in the capsular bag. The viscoelastic material was then removed. The corneal incision was closed by stromal hydration.Postoperatively, all patients were treated with topical combination of dexamethasone and Neomycin eyedrops during four weeks, and topical nonsteroidal antiinflammatory eyedrops four times daily for eight weeks.P = 0.012), after 1 month 11.98 ± 3.1 mmHg (P = 0.015), after two months 11.92 ± 2.4 mmHg (P = 0.005), after 3 months 11.84 ± 1.4 mmHg and after six months 11.82 ± 1.3 mmHg (P = 0.005).Two hundred and seventy three eyes of 273 patients were recruited for the study. The mean age of the 96 women and 177 men was 71 ± 12 years. The mean IOP before surgery was 14.18 ± 3.4 mmHg. Mean preoperative ACD was 2.96 mm and postoperative ACD was 4.09 mm. The mean lens thickness was 4.24 mm and axial length was 23 mm. A postoperative reduction of IOP was found as shown in The postoperative reduction in IOP in mmHg and in percentage is shown in Figures P = 0.011). Lens thickness was not significantly related to change in IOP (P: 0.12); however, it was significantly related to change in ACD (P: 0.002). The postoperative IOP was inversely related to preoperative ACD (P: 0.012). Age, sex, axial length were not significantly related to IOP reduction range of P: 0.2–0.5.The reduction of IOP measured after 15 days was 2.1 mmHg, after 1 month 2.26 mmHg, after three months 2.34 mmHg and after six months 2.36 mmHg. Our group had a mean decrease in IOP of 2.25 mmHg (15.77%) compared to preoperative values (Numerous studies have shown that cataract surgery by phacoemulsification with posterior chamber IOL induces a mid-and long-term lowering of IOP.‐10 Altho7IOP reduction has been shown to be more prominent after CCP than after phacoemulsification with sclerocorneal tunnel.14 Howeve20et al reported that the size of the capsulorhexis had an effect on the IOP after phacoemulsification; they showed that a capsulorhexis of 4 mm had a greater IOP lowering effect than a capsulorhexis of 6 mm.[The exact mechanism by which cataract surgery improves IOP is unclear. Many hypotheses have been presented in the literature, namely 1 of 6 mm. In our sStudies on patients with open-angle glaucoma have demonstrated a pressure lowering effect of CCP.1823 In e182In conclusion, cataract surgery by CCP induces reduction of IOP in normal patients. The pathogenic mechanisms are still unclear. CCP can be performed with the intent of achieving better IOP control.