Seasonal variation of particle-induced oxidative potential of airborne particulate matter in Beijing.

An in vitro plasmid scission assay (PSA), the cell apoptosis assay, and ICP-MS were employed to study the oxidative potentials and trace element compositions of the airborne particulate matter (PM) in Beijing during a one year-long field campaign from June 2010 to June 2011. The cell damages induced by PM reveled by the cell apoptosis assay showed a similar variation pattern to the DNA damages obtained by PSA, verifying the feasibility of the PSA in analyzing the oxidative capacity of PM samples. The PSA experiments showed that the particle-induced DNA damage was highest in summer, followed by spring, winter and autumn in descending order. The percentages of the oxidative damages to plasmid DNA induced by the water-soluble fractions of PM under the particle doses from 10 to 250µg/ml were generally lower than 45%, with some values peaking at above 50%. The peak values were frequently present in late spring (i.e. April and May) and early summer (i.e. June) but they were scarcely observed in other seasons. These peak values were mostly associated with haze days or the days with low wind speed (less than 4m/s), indicating that the PM samples during haze had higher oxidative potential than those during non-haze periods. The oxidative potential induced by the water-soluble fraction of the PM displayed a significant positive correlation with the concentrations of the water-soluble elements Cd, Cs, Pb, Rb, Zn, Be and Bi, demonstrating that the particle-induced oxidative potentials were mainly sourced from these elements. The exposure risk represented by the mass concentration of these elements in unit volume of atmosphere was higher in summer and winter, and lower in autumn and spring. The haze day PM samples not only had higher level of oxidative potentials but also had higher concentrations of water-soluble elements.

Source Apportionment of Elemental Carbon in Beijing, China: Insights from Radiocarbon and Organic Marker Measurements.

Elemental carbon (EC) or black carbon (BC) in the atmosphere has a strong influence on both climate and human health. In this study, radiocarbon ((14)C) based source apportionment is used to distinguish between fossil fuel and biomass burning sources of EC isolated from aerosol filter samples collected in Beijing from June 2010 to May 2011. The (14)C results demonstrate that EC is consistently dominated by fossil-fuel combustion throughout the whole year with a mean contribution of 79% ± 6% (ranging from 70% to 91%), though EC has a higher mean and peak concentrations in the cold season. The seasonal molecular pattern of hopanes (i.e., a class of organic markers mainly emitted during the combustion of different fossil fuels) indicates that traffic-related emissions are the most important fossil source in the warm period and coal combustion emissions are significantly increased in the cold season. By combining (14)C based source apportionment results and picene (i.e., an organic marker for coal emissions) concentrations, relative contributions from coal (mainly from residential bituminous coal) and vehicle to EC in the cold period were estimated as 25 ± 4% and 50 ± 7%, respectively, whereas the coal combustion contribution was negligible or very small in the warm period.

Issues related to aircraft take-off plumes in a mesoscale photochemical model.

The physical and chemical characteristics of aircraft plumes at the take-off phase are simulated with the mesoscale CAMx model using the individual plume segment approach, in a highly resolved domain, covering the Athens International Airport. Emission indices during take-off measured at the Athens International Airport are incorporated. Model predictions are compared with in situ point and path-averaged observations (NO, NO2) downwind of the runway at the ground. The influence of modeling process, dispersion properties and background air composition on the chemical evolution of the aircraft plumes is examined. It is proven that the mixing properties mainly determine the plume dispersion. The initial plume properties become significant for the selection of the appropriate vertical resolution. Besides these factors, the background NOx and O3 concentration levels control NOx distribution and their conversion to nitrogen reservoir species.

Evaluation of a closed tunnel for field-scale measurements of nitrous oxide fluxes from an unfertilized grassland soil.

Emissions of the major greenhouse gas NO from soils are characterized by huge spatial variability. An upscaling based on conventional small-scale chamber measurements is thus questionable and may involve a considerable amount of uncertainty. In this feasibility study, we evaluated the applicability of a large, closed tunnel for field-scale measurements of NO fluxes from an unfertilized grassland soil. The tunnel, coupled to an open-path Fourier transform infrared spectrometer, covered 500 m. During a 2-yr campaign, concurrent closed-chamber measurements (area of 0.045 m) were performed at the tunnel plot. The tunnel system enabled high-density and precise NO concentration measurements under dry, stable, nocturnal atmospheric conditions, but higher wind speeds and rain limited its application. To calculate an unbiased, predeployment NO flux from the increase of NO concentrations during tunnel deployment, we propose a novel approach based on inverse modeling (IMQ0). We show that IMQ0 is appropriate for the specific non-steady state tunnel setup. Compared with conventional models, which were developed for gas flux calculation from concentration gradients measured in vented closed chambers, IMQ0 is most accurate. Whereas NO fluxes obtained from the tunnel measurements were generally small and at a typical background level, the chamber measurements revealed high spatial and temporal variability of NO emissions, including slight NO uptake and precipitation-triggered emission peaks. The cumulative NO fluxes of both methods differed by one order of magnitude and were smaller for the tunnel measurements. We argue that the chambers were occasionally susceptible to detection of hotspots and hot moments of NO emission. However, these emissions were evidently not representative for the field scale. Compared with available greenhouse gas measurement techniques, we conclude that the tunnel may serve as a gap-filling method between small-scale chamber and ecosystem-level micrometeorological techniques, particularly during stable nocturnal conditions.

Effect of maternally supplied n-3 and n-6 oils on the fatty acid composition and mononuclear immune cell distribution of lymphatic tissue from the gastrointestinal tract of suckling piglets.

Fatty acids are essential for immune cell function. Maternal dietary fatty acid supply influences body fat composition of their offspring. As a first step to study immunonutritional interactions at an early age of pigs, four sows were fed a diet containing sunflower oil or oil from seal blubber during pregnancy and lactation. Corresponding piglets were sacrificed at three consecutive time points in the suckling period and their mesenteric lymph nodes and spleen were analysed by gas chromatography for levels of fatty acid. At the same time mononuclear cells of these organs and of the intestinal lymphoid tissue from the jejunum were isolated and subpopulations characterised by flow cytometry. Levels of fatty acids from the lymphatic organs of the piglets were significantly influenced by the maternal diet. The concentration of the fatty acids 20:5n-3, 22:5n-3 and 22:6n-3 were higher in the spleen and mesenteric lymph node of piglets suckled to sows of the test diet. Additionally, suckling time affected the levels of some long chain polyunsaturated fatty acids. Dietary effects were seen on some subpopulations including CD4-CD8alpha+ lymphocytes of the mesenteric lymph nodes and CD4+CD8alpha+ lymphocytes of the lamina propria, which were higher in the group fed seal blubber oil. The levels of CD21+ B-cells were higher in the group fed sunflower oil. The results indicate that the maternal diet and suckling time affect the fatty acid status of the investigated lymphatic tissues of piglets, but may have minor effects on the investigated lymphocyte subpopulations.

Application of direct thermal desorption gas chromatography time-of-flight mass spectrometry for determination of nonpolar organics in low-volume samples from ambient particulate matter and personal samplers.

Direct thermal desorption and in-situ derivatization thermal desorption methods in conjunction with gas chromatography time-of-flight mass spectrometry have been characterized and evaluated for analysis of trace components from filters loaded with ambient particulate matter (PM). The limits of quantification were in the range of 7-24 pg for n-alkanes, 20 pg for hopanes, and 4-22 pg for polycyclic aromatic hydrocarbons (PAH). The limit of quantification was defined as the minimum amount of substance that conforms to the minimum distinguishable signal plus 9 times the standard deviation of this background signal from PM-loaded filters. The method has been successfully applied to low-volume samples from ambient PM collected with stationary and personal samplers. Stationary samples were collected in winter 2008 and 2010 in Augsburg, Germany. Sample aliquots of 0.2-0.3 m³ from stationary sampling were analyzed. High diurnal variation in concentration and source contribution was found especially during periods with low wind speed and low mixing layer height. High contributions of solid fuel combustion (wood and coal) were found in evening and nighttime samples, leading to peak PAH concentrations at midnight more than 10 times higher than at noon. Finally, the method was applied to samples collected by means of a personal sampler, i.e. a micro aethalometer, in Xi'an, China. Quantitative data on n-alkanes, hopanes, and PAH were obtained from sample volumes of 17 and 24 l. The impact of different sources such as vehicular and biogenic emissions could be distinguished.

Single assay for simultaneous detection and differential identification of human and avian influenza virus types, subtypes, and emergent variants.

For more than four decades the cause of most type A influenza virus infections of humans has been attributed to only two viral subtypes, A/H1N1 or A/H3N2. In contrast, avian and other vertebrate species are a reservoir of type A influenza virus genome diversity, hosting strains representing at least 120 of 144 combinations of 16 viral hemagglutinin and 9 viral neuraminidase subtypes. Viral genome segment reassortments and mutations emerging within this reservoir may spawn new influenza virus strains as imminent epidemic or pandemic threats to human health and poultry production. Traditional methods to detect and differentiate influenza virus subtypes are either time-consuming and labor-intensive (culture-based) or remarkably insensitive (antibody-based). Molecular diagnostic assays based upon reverse transcriptase-polymerase chain reaction (RT-PCR) have short assay cycle time, and high analytical sensitivity and specificity. However, none of these diagnostic tests determine viral gene nucleotide sequences to distinguish strains and variants of a detected pathogen from one specimen to the next. Decision-quality, strain- and variant-specific pathogen gene sequence information may be critical for public health, infection control, surveillance, epidemiology, or medical/veterinary treatment planning. The Resequencing Pathogen Microarray (RPM-Flu) is a robust, highly multiplexed and target gene sequencing-based alternative to both traditional culture- or biomarker-based diagnostic tests. RPM-Flu is a single, simultaneous differential diagnostic assay for all subtype combinations of type A influenza viruses and for 30 other viral and bacterial pathogens that may cause influenza-like illness. These other pathogen targets of RPM-Flu may co-infect and compound the morbidity and/or mortality of patients with influenza. The informative specificity of a single RPM-Flu test represents specimen-specific viral gene sequences as determinants of virus type, A/HN subtype, virulence, host-range, and resistance to antiviral agents.

Microbiota and SCFA in lean and overweight healthy subjects.

Obesity has recently been linked to the composition of human microbiota and the production of short chain fatty acids (SCFAs). However, these findings rely on experimental studies carried out using rather small and defined groups of volunteers or model animals. Our aim was to evaluate differences within the human intestinal microbiota and fecal SCFA concentration of lean and obese subjects. A total of 98 subjects volunteered to take part in this study. The BMI in kg/m(2) of 30 volunteers was within the lean range, 35 were overweight and 33 were obese. The fecal microbiota was characterized by real-time PCR analyses. With the primers used herein we were able to cover 82.3% (interquartile range of 68.3-91.4%) of the total microbiota detectable with a universal primer. In addition, the concentration of SCFA was evaluated. The total amount of SCFA was higher in the obese subject group (P = 0.024) than in the lean subject group. The proportion of individual SCFA changed in favor of propionate in overweight (P = 0.019) and obese subjects (P = 0.028). The most abundant bacterial groups in faeces of lean and obese subjects belonged to the phyla Firmicutes and Bacteroidetes. The ratio of Firmicutes to Bacteroidetes changed in favor of the Bacteroidetes in overweight (P = 0.001) and obese subjects (P = 0.005). Our results are in line with previous reports suggesting that SCFA metabolism might play a considerable role in obesity. However, our results contradict previous reports with regard to the contribution of various bacterial groups to the development of obesity and this issue remains controversial.

The lectin-like domain of tumor necrosis factor-alpha improves alveolar fluid balance in injured isolated rabbit lungs.

OBJECTIVE: Identification of mechanisms that preserve optimal alveolar fluid balance during pulmonary edema is of great clinical importance. This study was performed to determine whether the lectin-like domain of tumor necrosis factor-alpha (designated TIP) can improve fluid balance in experimental lung injury by affecting alveolocapillary permeability and/or fluid clearance. DESIGN: Prospective, randomized laboratory investigation. SETTING: University-affiliated laboratory. SUBJECTS: Adult male rabbits. INTERVENTIONS: TIP, a scrambled peptide (scrTIP), dibutyryl cyclic adenosine monophosphate (db-cAMP), or saline was applied to isolated, ventilated, and buffer-perfused rabbit lungs by ultrasonic nebulization, after which hydrostatic edema or endo/exotoxin-induced lung injury was induced and edema formation was assessed. In studies evaluating the resolution of alveolar edema, 2.5 mL of excess fluid was deposited into the alveolar space of isolated lungs by nebulization in the absence or presence of TIP, scrTIP, amiloride, or ouabain or combinations thereof. MEASUREMENTS AND MAIN RESULTS: Microvascular permeability was largely increased during hydrostatic edema and endo/exotoxin-induced lung injury in saline-treated lungs, or lungs that received scrTIP, as assessed by capillary filtration coefficient (K(f,c)) and fluorescein isothiocyanate-labeled albumin flux across the alveolocapillary barrier. In contrast, TIP- or db-cAMP-treated lungs exhibited significantly lower vascular permeability upon hydrostatic challenge. Similarly, extravascular fluid accumulation, as assessed by fluid retention, wet weight to dry weight ratio, and epithelial lining fluid volume measurements, was largely inhibited by TIP or db-cAMP pretreatment. Furthermore, TIP increased sodium-potassium adenosine triphosphatase (Na,K-ATPase) activity 1.6-fold by promoting Na,K-ATPase exocytosis to the alveolar epithelial cell surface and increased amiloride-sensitive sodium uptake, resulting in a 2.2-fold increase in active Na+ transport, and hence improved clearance of excess fluid from the alveolar space. CONCLUSIONS: Aerosolized TIP improved alveolar fluid balance by both reducing vascular permeability and enhancing the absorption of excess alveolar fluid in experimental lung injury. These data may suggest a role for TIP as a potential therapeutic agent in pulmonary edema.

Phenotypic and functional aspects of the neonatal immune system as related to the maternal dietary fatty acid supply of sows.

The maternal-fetal transfer and subsequent uptake of sow milk enriched with n-6- or n-3-polyunsaturated fatty acids may not only influence neonatal body fat but may also have an impact on the immune function of newborn piglets. Sows were fed a diet containing sunflower oil as n-6-source or oil from seal blubber with long chain polyunsaturated n-3-fatty acids during pregnancy and lactation. Sow serum was investigated during pregnancy and serum and milk during lactation; piglet serum and liver were investigated in the suckling period until day 19. Piglet leukocyte subpopulations were characterised by flow cytometry and leukocyte proliferation was tested after stimulation with mitogens. No differences were noted in performance. The serum and milk fatty acid status of the sows was markedly influenced by the diet. Eicosapentaenoic acid (20:5n-3), 22:5n-3 and 22:6n-3 were higher (p < 0.001) in serum and liver of piglets delivered from sows fed the seal blubber oil. Piglets at birth had lower lymphocyte counts (p < 0.01) than piglets 19 days after birth. However, no influence of feeding the different oil sources was noted on lymphocyte phenotyping and leukocyte proliferation test. The results of the present study show that the maternal diet affected the fatty acid status of neonates, but much more in the sucking period. Immunological traits were not affected, probably as the mononuclear cell lineage is too immature around birth. Effects of PUFA n-3 might only be seen at a later time point or in the polymorphonuclear cell lineage as they were dominating right after birth.

Comparison of a xylanase and a complex of non starch polysaccharide-degrading enzymes with regard to performance and bacterial metabolism in weaned piglets.

Weaned piglets were fed a wheat based diet either non-supplemented or supplemented with a multi-enzyme preparation or a xylanase mono-enzyme preparation, respectively. Both enzyme preparations increased live weight gain nonsignificantly, but only animals of the xylanase group showed a trend (p = 0.076) for an improved feed conversion. Only precaecal digestibility of total amino acids was improved significantly when the mono-enzyme preparation was added. Improvements of digestibility of crude fat, crude protein and starch did not reach the significance level. Both enzyme preparations reduced jejunal viscosity, however viscosity in the colon was only reduced by the mono-enzyme preparation. Both enzymes significantly reduced Lactobacillus spp. cell numbers as well as bacterial metabolites in the stomach and showed similar nonsignificant modifications in jejunum contents except for acetate in the mono-enzyme group. Total jejunal bile acids were unchanged. Compared to the control, the ratio of the main conjugated to the main deconjugated bile acid was significantly higher in the mono-enzyme group. This study has shown that the mono- and multi-enzyme preparation can lead to improved performance in wheat based diets for piglets. Like in poultry, the main mode of action seems to be the reduction of small intestinal viscosity. However, the generation of fermentable carbohydrates by the multi-enzyme preparation may mask beneficial effects on performance due to the development of an active bile acid deconjugating microbiota in the small intestine.

Multipass open-path Fourier-transform infrared measurements for nonintrusive monitoring of gas turbine exhaust composition.

The detection limits for NO and NO2 in turbine exhausts by nonintrusive monitoring have to be improved. Multipass mode Fourier-transform infrared (FTIR) absorption spectrometry and use of a White mirror system were found from a sensitivity study with spectra simulations in the mid-infrared to be essential for the retrieval of NO2 abundances. A new White mirror system with a parallel infrared beam was developed and tested successfully with a commercial FTIR spectrometer in different turbine test beds. The minimum detection limits for a typical turbine plume of 50 cm in diameter are approximately 6 parts per million (ppm) for NO and 9 ppm for NO2 (as well 100 ppm for CO2 and 4 ppm for CO).

Effects of selenium deficiency on fatty acid metabolism in rats fed fish oil-enriched diets.

The hepatic fatty acid metabolism was investigated in rats stressed by selenium deficiency and enhanced fish oil intake. Changes in the composition of lipids, peroxides, and fatty acids were studied in the liver of rats fed either a Sedeficient (8 microg Se/kg) or a Se-adequate (300 microg Se/kg) diet, both rich in n-3 fatty acid-containing fish oil (100 g/kg diet) and vitamin E (146 mg alpha-tocopherol/kg diet). The two diets were identical except for their Se content. Se deficiency led to a decrease in hair coat density and quality as well as to changes in liver lipids, individual lipid fractions and phospholipid fatty acid composition of the liver. The low Se status did reduce total and reduced glutathione in the liver but did not affect the hepatic malondialdehyde level. In liver phospholipids (PL), Se deficiency significantly reduced levels of palmitic acid [16:0], fatty acids of the n-3 series such as DHA [22:6 n-3], and other long-chain polyunsaturates C-20-C-22, but increased n-6 fatty acids such as linoleic acid (LA) [18:2 n-6]. Thus, the conversion of LA to arachidonic acid was reduced and the ratio of n-6/n-3 fatty acids was increased. As in liver PL, an increase in the n-6/n-3 ratio was also observed in the mucosal total fatty acids of the small intestine. These results suggest that in rats with adequate vitamin E and enhanced fish oil intake, Se deficiency affects the lipid concentration and fatty acid composition in the liver. The changes may be related to the decreased levels of selenoenzymes with antioxidative functions. Possible effects of Se on absorption, storage and desaturation of fatty acids were also discussed.

Human papillomavirus type 16 L1 capsomeres induce L1-specific cytotoxic T lymphocytes and tumor regression in C57BL/6 mice.

We analyzed capsomeres of human papillomavirus type 16 (HPV16) consisting of the L1 major structural protein for their ability to trigger a cytotoxic T-cell (CTL) response. To this end, we immunized C57BL/6 mice and used the L1(165-173) peptide for ex vivo restimulation of splenocytes prior to analysis ((51)Cr release assay and enzyme-linked immunospot assay [ELISPOT]). This peptide was identified in this study as a D(b)-restricted naturally processed CTL epitope by HPV16 L1 sequence analysis, major histocompatibility complex class I binding, and (51)Cr release assays following immunization of C57BL/6 mice with HPV16 L1 virus-like particles (VLPs). HPV16 L1 capsomeres were obtained by purification of HPV16 L1 lacking 10 N-terminal amino acids after expression in Escherichia coli as a glutathione S-transferase fusion protein (GST-HPV16 L1 Delta N10). Sedimentation analysis revealed that the majority of the purified protein consisted of pentameric capsomeres, and assembled particles were not observed in minor contaminating higher-molecular-weight material. Subcutaneous (s.c.) as well as intranasal immunization of C57BL/6 mice with HPV16 L1 capsomeres triggered an L1-specific CTL response in a dose-dependent manner as measured by ELISPOT and (51)Cr release assay. Significant reduction of contaminating bacterial endotoxin (lipopolysaccharide) from the capsomere preparation did not diminish the immunogenicity. Antibody responses (serum and vaginal) were less robust under the experimental conditions employed. In addition, s.c. vaccination with HPV16 L1 capsomeres induced regression of established tumors expressing L1 determinants (C3 tumor cells). Our data demonstrate that capsomeres are potent inducers of CTL responses similar to completely assembled T=7 VLPs. This result is of potential relevance for the development of (combined prophylactic and therapeutic) HPV-specific vaccines, since capsomeres can be produced easily and also can be modified to incorporate heterologous sequences such as early HPV proteins.

The informatics response in disaster, terrorism, and war.

The United States currently faces several new, concurrent large-scale health crises as a result of terrorist activity. In particular, three major health issues have risen sharply in urgency and public consciousness--bioterrorism, the threat of widespread delivery of agents of illness; mass disasters, local events that produce large numbers of casualties and overwhelm the usual capacity of health care delivery systems; and the delivery of optimal health care to remote military field sites. Each of these health issues carries large demands for the collection, analysis, coordination, and distribution of health information. The authors present overviews of these areas and discuss ongoing work efforts of experts in each.

Crystal structure of oxidized flavodoxin, an essential protein in Helicobacter pylori.

The redox protein flavodoxin has been shown earlier to be reduced by the pyruvate-oxidoreductase (POR) enzyme complex of Helicobacter pylori, and also was proposed to be involved in the pathogenesis of gastric mucosa-associated lymphoid-tissue lymphoma (MALToma). Here, we report its X-ray structure, which is similar to flavodoxins of other bacteria and cyanobacteria. However, H. pylori flavodoxin has an alanine residue near the isoalloxazine ring of its cofactor flavin mononucleotide (FMN), while the other previously crystallized flavodoxins have a larger hydrophobic residue at this position. This creates a solute filled hole near the FMN cofactor of H. pylori flavodoxin. We also show that flavodoxin is essential for the survival of H. pylori, and conclude that its structure can be used as a starting point for the modeling of an inhibitor for the interaction between the POR-enzyme complex and flavodoxin.

Chimeric virus-like particles of the human papillomavirus type 16 (HVP 16) as a prophylactic and therapeutic vaccine

Infection by certain human papillomaviruses (HPV), most notably HPV types 16 and 18, is the mejor risk factor cervical cancer. Worldwide, this disease represents the second most frequent malignant tumor in women; thus, there is urgent need for efficient therapy and prevention. The natural history of cervical cancer and its precursors (cervical intraepithelial neoplasias), as well as animal experiments, strogly suggest that immune system controls both the primary infection (by neutralizing antibodies directed against the major structural protein L1) and the progression o the disease (via cytotoxic T cell specific for the viral oncoproteins expressed in transformed cells, e.g., E7). By the expression of an HPV 16 L1E7 fusion proteins, we have generated chimeric virus-like particles (CVLP). Immunization of mice with CVLPs induces neutralizing antibodies directed against L1 virus-like particles (devoid of the E7 portion) and E7-specific t cells as measured in vitro. Vaccinated animals are protected against tumor growth followings inoculation of syngeneic HPV 16-transformed cells. In addition, we observed a therapeutic effect of vaccination on pre-existing tumors. This data allowed us to concelude that CVLPs are suitable for prevention and therapy of HPV infection. A vaccine based on HPV 16 L1E7 CVLPs is currently under development