Rationale Novel therapeutic vaccines for allergic asthma, based on House Dust Mite allergens, effective with fewer administration would be a very advantageous alternative over conventional allergy ...vaccines and would allow a greater extension of the immunotherapy approach, reducing the consumption of symptomatic medications. In Cuba and other Caribbean countries the species Dermatophagoides siboney is very relevant as a cause of respiratory allergy.The objective of this research was to perform the first evaluation in humans of the tolerability and safety of a D. siboney allergen vaccine, adjuvanted with Neisseria meningitidis B proteoliposome (PROLINEM-DS).
Site-specific recombination technologies are powerful new tools for the manipulation of genomic DNA in insects that can improve transgenesis strategies such as targeting transgene insertions, ...allowing transgene cassette exchange and DNA mobilization for transgene stabilization. However, understanding the fitness cost implications of these manipulations for transgenic strain applications is critical. In this study independent piggyBac-mediated attP target-sites marked with DsRed were created in several genomic positions in the Mexican fruit fly, Anastrepha ludens. Two of these strains, one having an autosomal (attP_F7) and the other a Y-linked (attP_2-M6y) integration, exhibited fitness parameters (dynamic demography and sexual competitiveness) similar to wild type flies. These strains were thus selected for targeted insertion using, for the first time in mexfly, the phiC31-integrase recombination system to insert an additional EGFP-marked transgene to determine its effect on host strain fitness. Fitness tests showed that the integration event in the int_2-M6y recombinant strain had no significant effect, while the int_F7 recombinant strain exhibited significantly lower fitness relative to the original attP_F7 target-site host strain. These results indicate that while targeted transgene integrations can be achieved without an additional fitness cost, at some genomic positions insertion of additional DNA into a previously integrated transgene can have a significant negative effect. Thus, for targeted transgene insertions fitness costs must be evaluated both previous to and subsequent to new site-specific insertions in the target-site strain.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Nasal provocation test (NPT) is indicated to confirm the diagnosis of allergic rhinitis to house dust mites. Methods An open, non-randomized, controlled clinical trial was carried out in 50 patients ...with allergic rhinitis sensitized to D. siboney mite and 50 non-allergicsubjects.
Rationale A nasal allergen provocation test (NAPT) is performed to confirm the diagnosis of allergic rhinitis to house dust mite, in the situation of discrepancy between the symptoms and the results ...of skin prick test (SPT) and/or serum specific immunoglobulin E. In Cuba, sensitization to house dust mites (Dermatophagoides pteronyssinus (Dp), Dermatophagoides siboney and Blomia tropicalis) is a major cause of allergic rhinitis. Conclusions Nasal allergen provocation test with the Dermatophagoides pteronyssinus is effective and safety by the diagnosis of allergic rhinitis to this mite.
We report on immobilizing H(2)/Zn-porphyrin-Zn-phthalocyanine conjugates onto single wall carbon nanotubes and by using the excellent stability of the resulting suspensions we were able to ...demonstrate for the first time the sequence of energy transfer-electron transfer in SWNT donor-acceptor conjugates.
The multisubunit cation/proton antiporter 3 family, also called Mrp, is widely distributed in all three phylogenetic domains (Eukarya, Bacteria, and Archaea). Investigations have focused on Mrp ...complexes from the domain Bacteria to the exclusion of Archaea, with a consensus emerging that all seven subunits are required for Na
/H
antiport activity. The MrpA subunit from the MrpABCDEFG Na
/H
antiporter complex of the archaeon Methanosarcina acetivorans was produced in antiporter-deficient Escherichia coli strains EP432 and KNabc and biochemically characterized to determine the role of MrpA in the complex. Both strains containing MrpA grew in the presence of up to 500 mM NaCl and pH values up to 11.0 with no added NaCl. Everted vesicles from the strains containing MrpA were able to generate a NADH-dependent pH gradient (ΔpH), which was abated by the addition of monovalent cations. The apparent K
values for Na
and Li
were similar and ranged from 31 to 63 mM, whereas activity was too low to determine the apparent K
for K
Optimum activity was obtained between pH 7.0 and 8.0. Homology molecular modeling identified two half-closed symmetry-related ion translocation channels that are linked, forming a continuous path from the cytoplasm to the periplasm, analogous to the NuoL subunit of complex I. Bioinformatics analyses revealed genes encoding homologs of MrpABCDEFG in metabolically diverse methane-producing species. Overall, the results advance the biochemical, evolutionary, and physiological understanding of Mrp complexes that extends to the domain Archaea IMPORTANCE: The work is the first reported characterization of an Mrp complex from the domain Archaea, specifically methanogens, for which Mrp is important for acetotrophic growth. The results show that the MrpA subunit is essential for antiport activity and, importantly, that not all seven subunits are required, which challenges current dogma for Mrp complexes from the domain Bacteria A mechanism is proposed in which an MrpAD subcomplex catalyzes Na
/H
antiport independent of an MrpBCEFG subcomplex, although the activity of the former is modulated by the latter. Properties of MrpA strengthen proposals that the Mrp complex is of ancient origin and that subunits were recruited to evolve the ancestral complex I. Finally, bioinformatics analyses indicate that Mrp complexes function in diverse methanogenic pathways.
The study of a ternary system formed by two ionic liquids (ILs) (N-butyl-pyridinium chloride (bpyCl) and 3-butyl-1-methylimidazolium chloride (bmimCl)) and water has been carried out using UV–vis ...absorption measurements. The results were employed to design non-linear mathematical models based on artificial neural networks. The models provided accurate results for the estimation of the concentration of bpyCl and bmimCl (the best model offered the following mean prediction errors (MPEs): MPEbpyCl=3.81% and MPEbmimCl=5.78%). Different methodologies have been considered to describe the system, which has led to discussions regarding the relevance of number, type, and quality of independent variables used in the models. This study allowed improving the estimations achieved by different models and decreasing the error attained seven- and twelve-fold for the concentration estimation of bpyCl and bmimCl, respectively, from the worst to the best model.
Wastewater treatment plants produce solid and semi-solid sludge, which treatment minimises secondary environmental pollution because of wastewater treatment and obtaining new bioproducts. For this ...reason, in this paper, the co-pyrolysis of biogenic biomasses recovered from a biological reactor with immobilised fungal and bacterial biomass and a tertiary reactor with
Chlorella
sp. used for dye-contaminated wastewater treatment was carried out. Biogenic biomasses mixed with pine bark allowed the production and characterisation of two types of biochar. The raw material and biochar were on the “in vitro” germination of
Lolium
sp. seeds, followed by adsorption studies for malachite green (MG) dye using the raw material and the biochar. Results showed that using 60 mg L
−1
of a cationic coagulant at pH 6.5 allowed for the recovery of more than 90% of the microalgae after 50 min of processing. Two biochar resulted: BC
300
, at pH 5.08 ± 0.08 and BC
500
, at pH 6.78 ± 0.01. The raw material and both biochars were co-inoculated with growth-promoting bacteria; their viabilities ranged from 1.7 × 10
6
± 1.0 × 10
1
to 7.5 × 10
8
± 6.0 × 10
2
CFU g
−1
for total heterotrophic, nitrogen-fixing and phosphate-solubilising bacteria. Re-use tests on
Lolium
sp. seed germination showed that with the post-coagulation effluent, the germination was 100%, while with the biochar, with and without beneficial bacteria, the germination was 98 and 99%, respectively. Finally, BC
500
adsorbed the highest percentage of malachite green at pH 4.0, obtaining
q
ecal values of 0.5249 mg g
−1
(
R
2
: 0.9875) with the pseudo-second-order model.
Mitochondrial aldehyde dehydrogenase (ALDH2) has been proposed as a key enzyme in cardioprotection during ischemia–reperfusion processes. This proposal led to the search for activators of ALDH2 with ...the aim to develop cardioprotective drugs. Alda‐1 was the first activator of ALDH2 identified and its cardioprotective effect has been extensively proven in vivo; however, the mechanism of activation is not fully understood. A crystallographic study showed that Alda‐1 binds to the entrance of the aldehyde‐binding site; therefore, Alda‐1 should in essence be an inhibitor. In the present study, kinetic experiments were performed to characterize the effect of Alda‐1 on the properties of ALDH2 (kinetic parameters, determination of the rate‐limiting step, reactivity of the catalytic cysteine) and on the kinetic mechanism (type of kinetics, sequence of substrates entering, and products release). The results showed that Alda‐1 dramatically modifies the properties of ALDH2, the Km for NAD+ decreased by 2.4‐fold, and the catalytic efficiency increased 4.4‐fold; however, the Km for the aldehyde increased 8.6‐fold, thus, diminishing the catalytic efficiency. The alterations in these parameters resulted in a complex behavior, where Alda‐1 acts as inhibitor at low concentrations of aldehyde and as an activator at high concentrations. Additionally, the binding of Alda‐1 to ALDH2 made the deacylation less limiting and diminished the pKa of the catalytic cysteine. Finally, NADH inhibition patterns indicated that Alda‐1 induced a change in the sequence of substrates entry and products release, in agreement with the proposal of both substrates entering ALDH2 by the NAD+ entrance site.
Activator of human ALDH2 Alda‐1 protects cell integrity from diverse stress conditions; however, the activation mechanism is not fully understood. Kinetic characterization showed that binding of Alda‐1 to ALDH2 activates the catalytic cysteine reducing its pKa, which induces the switching of order of substrates binding and products release, additionally, Alda‐1 increases the rate of the limiting step of the reaction.
Abstract
Ethanol accumulation during fermentation contributes to the toxic effects in Saccharomyces cerevisiae, impairing its viability and fermentative capabilities. The iron–sulfur (Fe–S) cluster ...biogenesis is encoded by the ISC genes. Reactive oxygen species (ROS) generation is associated with iron release from Fe–S-containing enzymes. We evaluated ethanol toxicity, ROS generation, antioxidant response and mitochondrial integrity in S. cerevisiaeISC mutants. These mutants showed an impaired tolerance to ethanol. ROS generation increased substantially when ethanol accumulated at toxic concentrations under the fermentation process. At the cellular and mitochondrial levels, ROS were increased in yeast treated with ethanol and increased to a higher level in the ssq1∆, isa1∆, iba57∆ and grx5∆ mutants – hydrogen peroxide and superoxide were the main molecules detected. Additionally, ethanol treatment decreased GSH/GSSG ratio and increased catalase activity in the ISC mutants. Examination of cytochrome c integrity indicated that mitochondrial apoptosis was triggered following ethanol treatment. The findings indicate that the mechanism of ethanol toxicity occurs via ROS generation dependent on ISC assembly system functionality. In addition, mutations in the ISC genes in S. cerevisiae contribute to the increase in ROS concentration at the mitochondrial and cellular level, leading to depletion of the antioxidant responses and finally to mitochondrial apoptosis.
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