Expression and Immunoreactivities of Hepatitis E Virus Genotype 3 Open Reading Frame-2 (ORF-2) Recombinant Proteins Expressed in Insect Cells

Hepatitis E virus (HEV) is a fecal-orally transmitted virus that is endemic in many geographical areas with poor sanitary conditions and inadequate water supplies. In Europe, a low-endemic area, an increased number of autochthonous sporadic human cases of patients infected with HEV strains of genotype 3, have been reported lately. The relatively high prevalence of HEV genotype 3 infections in European pigs has raised concerns about a potential zoonotic transmission to humans. Determination of HEV seroprevalence in pigs would help to clarify its incidence and possible zoonotic implications. To this purpose, we have expressed and partially characterized swine genotype 3 HEV open reading frame-2 proteins upon infection of Sf21 insect cells with recombinant baculoviruses. The use of the expressed proteins as diagnostic antigens for the detection of antibodies to HEV has been further assayed with human and swine sera.     

Morphological diversity of Struvite crystals produced by Myxococcus Coralloides and Myxococcus Xanthus

We studied the morphological diversity of struvite crystals produced by Myxococcus coralloides and Myxococcus xanthus in different culture conditions. We discussed the similarities of these crystals with the struvite morphology studied previously following the theory of the periodic bond chains.     

Graviola attenuates DMBA-induced breast cancer possibly through augmenting apoptosis and antioxidant pathway and downregulating estrogen receptors

Breast cancer is a global public health problem where it is the second most prevalent cancer. Historical cancer treatment with graviola has been reported. This study aimed to investigate the protective effects of graviola on 7,12-dimethylbenz[a]anthracene (DMBA)–induced rat breast cancer. Fifty female Wistar rats were allocated into four groups: control group (gastro-gavaged by sesame oil), DMBA-treated group (gastro-gavaged a single dose of DMBA [50 mg/kg body mass, diluted in 1 ml sesame oil]) at the age 57 days, DMBA+G37-treated group (gastro-gavaged a single dose of DMBA [50 mg/kg body mass, diluted in 1 ml sesame oil]) at the age of 57 days plus graviola (200 mg/kg body mass) two times weekly (p.o.) at the age of 37 days till the end of the experiment, and DMBA+G57-treated group (received a single dose of DMBA [50 mg/kg body mass, diluted in 1 ml sesame oil]) plus graviola (200 mg/kg body mass) two times weekly at the age of 57 days until the end of the experiment. After the 30-week experimental period, blood samples were collected. Then, animals were sacrificed to determine the apoptotic indices, antioxidant status, and mammary gland tumor marker (CA 15-3). The DMBA upregulated the expression of one of the main anti-apoptotic genes: B-cell lymphoma protein 2 (BCL2) and estrogen receptor alpha (ER-α) gene. Moreover, it significantly increased breast lipid peroxidation and serum CA 15-3 but decreased breast antioxidant enzymatic activities (glutathione peroxidase, glutathione S-transferase, catalase, and superoxide dismutase). Nevertheless, administration of DMBA and graviola especially DMBA+G37 induced apoptosis through at least 1.5-fold in gene expression levels of pro-apoptotic genes: BCL2-associated X protein (BAX), tumor suppressor gene (P53), and cysteinyl-aspartic acid-protease-3 (caspase-3). A critical role of P53 in the regulation of the BCL2 and BAX has been reported. These proteins can determine if the cell undergoes apoptosis or cancels the process. Once the BAX gene activates caspase-3, there is no irreversible way toward cell death. Also, graviola ameliorated the DMBA effects on antioxidant enzymatic activities and tumor marker CA 15-3. This study concludes that graviola ameliorated DMBA-induced breast cancer potentially through upregulating apoptotic genes, downregulating the ER-α gene, increasing antioxidants, and decreasing lipid peroxidation levels.

     

Human exposure to polybrominated diphenyl ethers (PBDE), as evidenced by data from a duplicate diet study, indoor air, house dust, and biomonitoring in Germany

Polybrominated diphenyl ethers (PBDE) are used as flame retardants in a wide variety of products. As part of the Integrated Exposure Assessment Survey (INES), this study aimed to characterize the exposure of an adult German population using duplicate diet samples, which were collected daily over seven consecutive days, and indoor air and house dust measurements. Our study population consisted of 27 female and 23 male healthy subjects, aged 14–60 years, all of whom resided in 34 homes in southern Bavaria. In these 34 residences the air was sampled using glass fiber filters and polyurethane foams and the dust was collected from used vacuum cleaner bags.The median (95th percentile) daily dietary intake of six Tetra- to HeptaBDE congeners was 1.2 ng/kg b.w. (3.3 ng/kg b.w.) or 67.8 ng/day (208 ng/day) (calculated from the 7-day median values of each study subject). Concentrations in indoor air and dust (cumulative Tri- to DecaBDE congener readings) ranged from 8.2 to 477 pg/m³ (median: 37.8 pg/m³) and 36.6 to 1580 ng/g (median: 386 ng/g), respectively. For some congeners, we identified a significant correlation between air and dust levels.The median (95th percentile) blood concentration of total Tetra- to HexaBDE congener readings was 5.6 (13.2) ng/g lipid. No significant sex differences were observed, but higher blood concentrations were found in younger participants. Using a simplified toxicokinetic model to predict the body burden from exposure doses led to results that were of the same order of magnitude as the measured blood concentrations.Based on these measurements and given our exposure assumptions, we estimated for the total tetra- to heptabrominated congener count an average (high) comprehensive total daily intake of 1.2 ng/kg b.w. (2.5 ng/kg b.w.). Overall, our results suggest that dietary exposure is the dominant intake pathway at least in our study population, responsible for 97% (average intake) and 95% (high intake) of the total intake of an adult population.     

Myxococcus xanthus'' killed cells as inducers of struvite crystallization. Its possible role in the biomineralization processes

We studied the possibility of struvite formation using killed cells of Myxococcus xanthus. Cells were killed by heat, UV light and sonication. In all cases, we show that struvite crystallization occurs and we propose that the dead cells or cell debris can act by themselves as heterogeneous crystallization nuclei. We also show that the slime, produced by Myxococcus in a large quantities, is not involved in this process.     

A new approach to graphical and numerical analysis of links between plant chemotaxonomy and secondary metabolism from HPLC data smoothed by a simplex mixture design

Summary. HPLC analysis of secondary metabolites represents an efficient tool for the studying of plant chemical diversity under different aspects: chemotaxonomy, metabolomics, adaptative responses to ecological factors, etc. Statistical analyses of HPLC databases, e.g. correlation analysis between HPLC peaks, can reliably provide information on the similarity/dissimilarity degrees between the chemical compounds. The similarities, corresponding to positive correlations, can be interpreted in terms of analogies between chemical structures, synchronic metabolisms or co-evolution of two compounds under certain environment conditions, etc. . In terms of metabolism, positive correlations can translate precursor-product relationships between compounds; negative correlations can be indicative of competitive processes between two compounds for a common precursor(s), enzyme(s) or substrate(s). Furthermore, the correlation analysis under a metabolic aspect can help to understand the biochemical origins of an observed polymorphism in a plant species. With the aim of showing this, we present a new approach based on a simplex mixture design, Scheffé matrix, which provides a correlation network making it possible to graphically visualise and to numerically model the metabolic trends between HPLC peaks. The principle of the approach consisted in mixing individual HPLC profiles representative of different phenotypes, then from a complete mixture set, a series of average profiles were calculated to provide a new database with a small variability. Several iterations of the mixture design provided a smoothed final database from which the relationships between the secondary metabolites were graphically and numerically analysed. These relationships were scale-dependent, namely either deterministic or systematic: the first consisted of a monotonic global trend covering the whole variation field of each metabolites’ pair; the second consisted of repetitive monotonic variations which gradually attenuated or intensified along a global trend. This new metabolomic approach was illustrated from 404 individual plants of Astragalus caprinus (Leguminoseae), belonging to four chemical phenotypes (chemotypes) on the basis of flavonoids analysed in their leaves. After smoothing, the relationships between flavonoids were numerically fitted using linear or polynomial models; therefore the co-response coefficients were easily interpreted in terms of metabolic affinities or competitions between flavonoids which would be responsible of the observed chemical polymorphism (the four chemotypes). The statistical validation of the approach was carried out by comparing Pearson correlations to Spearman correlations calculated from the smoothed and the crude HPLC database, respectively. Moreover, the signs of the smoothed relationships were finely supported by analogies and differences between the chemical structures of flavonoids, leading to fluent interpretation in relation to the pathway architecture.     

Physicochemical mechanisms controlling the periodic release of flammable gases from hanford tanks

Hanford tanks contain more than 60 million gallons of high-level wastes produced by decades of extracting plutonium from irradiated uranium fuel. The wastes were concentrated to a thick slurry consistency by evaporation prior to storage to minimize space. The resulting concentrated waste properties introduced unanticipated, detrimental conditions affecting workers' and the public's health and safety and involving the release of potentially flammable gases. The released gases consist primarily of hydrogen, nitrous oxide, and ammonia. Dilution and sluicing were initially proposed to mitigate the flammable gas safety conditions. As a result of evaluations, the mechanisms and conditions that are thought to control the accumulation and spontaneous release of flammable gases were identified and confirmed. The technical rationale was established for developing operational approaches to mitigate the periodic generation of flammable gases in existing tanks and to avoid any reoccurrence of this serious safety problem during future waste management activities. The chemistry of the two highest risk tanks was examined to test the potential for reversing the conditions causing gas buildup and the consequences of sluicing without appropriate chemical conditioning. The identified mechanisms apply equally to the remaining flammable gas tanks at Hanford as well as to other waste tanks in the DOE complex, particularly those at Savannah River. Passive means of mitigating the flammable gas condition require less than 1:1 dilution, and sluicing wastes from tank 106-C can be accomplished without creating a flammable gas condition. Carbonate equilibria reactions and their effect on aluminum speciation are largely overlooked and provided the key for explaining the episodic release of flammable gases from tank wastes. The reaction of atmospheric carbon dioxide with a sodium hydroxide-rich waste solution produces carbonate precipitates. More importantly, this reaction lowers the pH of the waste and precipitates aluminum hydroxide as a gel. The wastes contain substantial amounts of complexing agents such as ethylene diamine tetraacidicacid (EDTA), hydroxy ethylene diamine triacidic acid (HEDTA), and their degradation products. These complexing agents stabilize the aluminum hydroxide gel together with chromium, manganese and iron hydroxides, and oxybydroxides under the resulting pH conditions. These complex species may coprecipitate and accumulate as a metastable layer in the middle and lower levels of th tank. The complexed aluminum hydroxide acts as a binding agent trapping other particulates in a microcrystalline mat. Microcrystalline particles such as sodium nitrite provide the structural strength for the mat. Once the gas accumulation below the gel layer achieves a critical buoyancy sufficient to rupture the microcrystalline mat, a gas release event occurs. The cycle of gas buildup and release continues each time the buoyancy of the trapped gas exceeds the hydrostatic pressure and the gels' plasticity modulus. Stokes Law predicts a particle settling rate in the tank of less than 50 days, well within the bistorical periodicity of GREs. Laboratory tests, forming the basis of a recent patent application, verify that large quantities of complexed aluminum hydroxide gel are produced by passing carbon dioxide through simulated waste solutions (Hobl, 1993) equivalent to those found in tank 101-SY. It was confirmed that a simple adjustment of pH witll redissolve the gel, thereby reducing viscosity and safely facilitating continuous flammable gas release. Additional experiments were undertaken to provide a basis for understanding the role of complexed aluminum hydroxides in the CO₂/NaOH/Al(OH)₃ (complexing agents)/NaAlO₂ system. This article examines a plausible mechanism for the periodic release of flammable gas and considerations for: (1) remediating existing flammable gas tanks through a combination of chemical treatment and mixer pumps; (2) diluting, combining, retrieving, and storing wastes; (3) preventing clogging of transfer lines; (4) sludge and soil washing; and (5) cribs, ponds, basins, and ground-water cleanup. This study provides a singificant breakthrough for tank waste management by explaining key mechanisms controlling episodic release of flammable gases. The breakthrough provides the bases for removing the tanks classified as flammable gas from the wathclist and has broad operational applications with a potential for billions of dollars in cost savings.     

MULTICRITERION ANALYSIS OF GROUNDWATER CONTAMINATION MANAGEMENT1

ABSTRACT: Multicriterion decision making (MCDM) techniques were used to analyze a groundwater contamination management problem from the viewpoint of conflicting multiple objectives. The groundwater management model was used to find a compromise strategy for trading off fresh water supply, containment of the waste, and total pumping cost in a hypothetical confined aquifer affected by previous waste disposal action. A groundwater flow model was used to formulate the hydraulic constraints. A linear system model was used to describe drawdown and velocity as functions of the decision variables which were pumping rates. The model determines the pumping location and rates. A modified c-constraint method was used to generate the set of nondominated solutions which were the alternative compromise strategies. Three different MCDM techniques, Compromise programming (CP), ELECTRA II and MCQA II, were used to select a “satisficing” alternative. Analysis of the results showed that, although these techniques follow different principles, the same preferred strategies were reached. Also, it was noticed that maintaining high groundwater velocities is expensive and difficult. In order to meet a two year target date, large amounts of water had to be pumped. Therefore, rapid restoration results in large pumping volumes and high costs.     

Removal of carbamazepine from urban wastewater by sulfate radical oxidation

The occurrence of bioactive trace pollutants such as pharmaceuticals in natural waters is an emerging issue. Numerous pharmaceuticals are not completely removed in conventional wastewater treatment plants. Advanced oxidation processes may represent an interesting alternative to completely mineralize organic trace pollutants. In this article, we show that sulfate radicals generated from peroxymonosulfate/Co^II are more efficient than hydroxyl radicals generated from the Fenton’s reagent (H₂O₂/Fe^II) for the degradation of the pharmaceutical compound, carbamazepine. The second-order rate constant for the reaction of SO₄ ^·− with carbamazepine is 1.92·10⁹ M⁻¹ s⁻¹. In laboratory grade water and in real urban wastewater, SO₄ ^·− yielded a faster degradation of carbamazepine compared to HO^· . Under strongly oxidizing conditions, a nearly complete mineralization of carbamazepine was achieved, while under mildly oxidizing conditions, several intermediates were identified by LC–MS. These results show for the first time in real urban wastewater that sulfate radicals are more selective than hydroxyl radicals for the oxidation of an organic pollutant and may represent an interesting alternative in advanced oxidation processes.