Bax induces cytochrome c release by multiple mechanisms in mitochondria from MCF7 cells

Bax, a pro-apoptotic member of the Bcl-2 family of proteins has the ability to form transmembrane pores large enough to allow cytochrome c (Cyt c) release, as well as to activate the mitochondrial permeability transition pore (mPTP); however, no differential study has been conducted to clarify which one of these mechanisms predominates over the other in the same system. In the present study, we treated isolated mitochondria from MCF7 cells with recombinant protein Bax and tested the efficacy of the mPTP inhibitor cyclosporin A (CsA) and of the Bax channel blocker (Bcb) to inhibit cytochrome c release. We also, induced apoptosis in MCF7 cell cultures with TNF-α plus cycloheximide to determine the effect of such compounds in apoptosis induction via mPTP or Bax oligomerization. Cytochrome c release was totally prevented by CsA and partially by Bcb when apoptosis was induced with recombinant Bax in isolated mitochondria from MCF7 cells. CsA increased the number of living cells in cell culture, as compared with the effect of Bax channel blocker. These results indicate that mPTP activation is the predominant pathway for Bax-induced cytochrome c release from MCF7 mitochondria and for apoptosis induction in the whole cell.     

Abatement of odorant compounds in one- and two-phase biotrickling filters under steady and transient conditions

The removal of hydrophobic volatile organic compounds (VOCs) still remains the main restriction in the biological treatment of odorous emissions due to mass transfer limitations. The addition of a non-aqueous phase to conventional biotrickling filters (BTF) may overcome this limitation by enhancing VOCs transport from the gas to the microorganisms. This study compared the long-term and transient performance of a one- (1P) and two-liquid phase (2P; with silicone oil as non-aqueous phase) BTFs for the removal of four VOCs (butanone, toluene, alpha-pinene, and hexane) at empty bed residence times (EBRT) ranging from 47 to 6 s. Removal efficiencies (RE) >96 % were obtained for butanone, toluene, and alpha-pinene in both bioreactors regardless of the EBRT, while higher hexane REs were recorded in the 2P-BTF (81–92 %) compared to the 1P-BTF (60–97 %). The two-phase system always showed a more consistent performance, being able to better withstand step VOC concentration increases and starvation periods, although it was more affected by liquid recycling shutdowns due to a reduced VOC mass transfer. The analysis of the microbial communities showed a high biodiversity and richness despite the low C source spectrum and high community evenness and richness. In this context, the presence of silicone oil mediated the development of a highly different phylogenetic composition of the communities.     

Síndrome de la bolsa de orina púrpura en dos pacientes institucionalizados

Purple urine bag syndrome (PUBS) is an uncommon but particularly striking phenomenon characterised by a chemical reaction involving the urine, plastic and certain enzymes from some sulphatase- and phosphatase-producing bacteria, including Proteus mirabilis, Escherichia coli and Morganella morganii, amongst others. Following this reaction, the catheter and the bag may be stained red, blue or purple. This phenomenon tends to occur in patients with multiple pathology and with urinary catheters, as part of a urinary tract infection. We describe two clinical cases of PUBS in institutionalised patients with permanent urinary catheters.     

Evaluation of the ratio of ω6 : ω3 fatty acids and vitamin e levels in the diet on the reproductive performance of cockerels

Three hundred and twenty 30-week old White Leghorn cockerels were housed in individual cages and distributed in a completely randomized factorial design of 5?×?3, with five oil sources (sunflower, soybean, canola, linseed and fish/soybean) and three levels of antioxidant (30, 200 and 400?mg of vitamin E/kg). The aim of this study was to evaluate the effect of the ratio of ω6?:?ω3 fatty acids by the inclusion of different oil sources and of dietary supplementation with vitamin E on the reproductive performance of cockerels. The use of the fish/soybean combination determined the lowest total antioxidant status of the semen. However, the addition of vitamin E to the fish/soybean-oil-based diet resulted in a linear increase in semen volume, motility and sperm vigour in the 38th week and again in the 52nd week for motility and for sperm vigour and fertility rate in the periods from 50?–?53 and 41?–?53 weeks of age. The use of canola oil in the diet resulted in the highest fertility rate during 50?–?53 and 41?–?53 weeks of life. Animals receiving the soybean oil based diet showed the smallest fertility rate in the range from 50?–?53 weeks of age and concomitantly the highest level of cholesterol in the spermatozoa in the range from 47?–?51 weeks. An interaction between the vitamin E level and soybean oil was verified by the linear increase in motility and sperm vigour at 38 weeks of age. Later, the contrary was shown by a linear reduction in fertility in the periods from 44?–?46, 47?–?49 and 41?–?53 weeks of age. Cockerels that had been fed on the sunflower-oil-based diet showed the highest content of saturated fatty acids in the spermatozoa from 48?–?51 weeks. An interaction effect was observed between the vitamin E level and sunflower oil shown by a linear increase in the content of saturated fatty acids in the spermatozoa and a linear reduction in the C18?:?1ω9, C18?:?2ω6 and PUFA (C18?:?2ω6?+?C20?:?4ω6) contents in the spermatozoa at 48?–?51 weeks and in sperm volume at 52 weeks of age.     

Assessing the Fecal Microbiota: An Optimized Ion Torrent 16S rRNA Gene-Based Analysis Protocol

Assessing the distribution of 16S rRNA gene sequences within a biological sample represents the current state-of-the-art for determination of human gut microbiota composition. Advances in dissecting the microbial biodiversity of this ecosystem have very much been dependent on the development of novel high-throughput DNA sequencing technologies, like the Ion Torrent. However, the precise representation of this bacterial community may be affected by the protocols used for DNA extraction as well as by the PCR primers employed in the amplification reaction. Here, we describe an optimized protocol for 16S rRNA gene-based profiling of the fecal microbiota.     

Novel Pancreatic Endocrine Maturation Pathways Identified by Genomic Profiling and Causal Reasoning

We have used a previously unavailable model of pancreatic development, derived in vitro from human embryonic stem cells, to capture a time-course of gene, miRNA and histone modification levels in pancreatic endocrine cells. We investigated whether it is possible to better understand, and hence control, the biological pathways leading to pancreatic endocrine formation by analysing this information and combining it with the available scientific literature to generate models using a casual reasoning approach. We show that the embryonic stem cell differentiation protocol is highly reproducible in producing endocrine precursor cells and generates cells that recapitulate many aspects of human embryonic pancreas development, including maturation into functional endocrine cells when transplanted into recipient animals. The availability of whole genome gene and miRNA expression data from the early stages of human pancreatic development will be of great benefit to those in the fields of developmental biology and diabetes research. Our causal reasoning algorithm suggested the involvement of novel gene networks, such as NEUROG3/E2F1/KDM5B and SOCS3/STAT3/IL-6, in endocrine cell development We experimentally investigated the role of the top-ranked prediction by showing that addition of exogenous IL-6 could affect the expression of the endocrine progenitor genes NEUROG3 and NKX2.2.     

Increased Learning and Brain Long-Term Potentiation in Aged Mice Lacking DNA Polymerase μ

A definitive consequence of the aging process is the progressive deterioration of higher cognitive functions. Defects in DNA repair mechanisms mostly result in accelerated aging and reduced brain function. DNA polymerase µ is a novel accessory partner for the non-homologous end-joining DNA repair pathway for double-strand breaks, and its deficiency causes reduced DNA repair. Using associative learning and long-term potentiation experiments, we demonstrate that Polµ^−/− mice, however, maintain the ability to learn at ages when wild-type mice do not. Expression and biochemical analyses suggest that brain aging is delayed in Polµ^−/− mice, being associated with a reduced error-prone DNA oxidative repair activity and a more efficient mitochondrial function. This is the first example in which the genetic ablation of a DNA-repair function results in a substantially better maintenance of learning abilities, together with fewer signs of brain aging, in old mice.     

Comparative proteomic analysis of growth hormone secretagogue A233 treatment of murine macrophage cells J774A.2 indicates it has a role in antiviral innate response

BackgroundGrowth hormone secretagogues (GHS), among other factors, regulate the release of GH. The biological activity of the secretagogue peptide A233 as a promoter of growth and innate immunity in teleost fish has previously been demonstrated, but its role in the immune system of mammals is not well understood.MethodsThe effect of the peptide was investigated in J774A.2 macrophage cells using a comparative proteomics approach after 6 and 12 h of peptide stimulation.ResultsThe functional analysis of differentially modulated proteins showed that A233 peptide treatment appears to promote activation and ROS-dependent cytotoxic functions in macrophages and enhanced expression of antiviral protein complexes such as MAVS. In accordance with this hypothesis, we found that A233 treatment enhanced superoxide anion production and the IFN-γ level in J774A.2 cells and mouse splenocytes, respectively, and reduced viral load in a dengue virus mouse model of infection.ConclusionsThe growth hormone secretagogue A233 peptide promotes activation of ROS-dependent cytotoxic functions and exerts immunomodulatory effects that enable an antiviral state in a dengue virus mouse model.General SignificanceThe increase of IFN-γ level and the differential modulation of antiviral proteins by the A233 peptide suggest that the molecule could activate an innate immune response with a possible further impact in the treatment of acute and chronic diseases.