Occurrence and enological properties of two new non-conventional yeasts (Nakazawaea ishiwadae and Lodderomyces elongisporus) in wine fermentations.

The microbial diversity of wine alcoholic fermentation is not restricted to the presence and activity of Saccharomyces yeast strains. Some non-Saccharomyces species have been described as part of the fermentative microbiota, specially found in the initial steps of wine fermentations. These species may play roles from wine spoilage to flavor quality enhancement. From a large number of wine fermentations (429 wine samples), analyzed by ITS-amplicon sequencing to define their mycobiome, 2 non-conventional yeast species (Nakazawaea ishiwadae and Lodderomyces elongisporus) were detected, in a very limited number of samples but in significant levels of relative abundance. One strain of each species was isolated and their technological and enological potential have been characterized in this work. Compared with the Saccharomyces cerevisiae Viniferm Revelacion wine strain, the studied N. ishiwadae BMK17.1 and L. elongisporus BMK12.5 strains showed, as expected, a lower tolerance and growth fitness in high ethanol concentrations. However, N. ishiwadae BMK17.1 was able to grow also at 15% ethanol and L. elongisporus BMK12.5 at 10% reaching, in the latter case, slightly higher efficiency rates than S. cerevisiae at this level. Contrary to most non-Saccharomyces yeasts, these species were able to growth in presence of high doses of potassium-metabisulfite, reaching in both cases higher efficiency rates than S. cerevisiae. A notable affinity of L. elongisporus BMK12.5 for high pH values was clearly observed. Their fermentation kinetics and the final chemical-analytical characterization were studied in micro-fermentation assays, using synthetic grape must. L. elongisporus BMK12.5 was able to complete, in single inoculation, the sugar fermentation after 19 days, but, N. ishiwadae BMK17.1 left about 80 g/L sugars at this time. Co-inoculation assays (in a 1:100 proportion of S. cerevisiae:non-Saccharomyces strains) finished sugar consumption with similar kinetics than the S. cerevisiae single inoculation, in the case of L. elongisporus BMK12.5 co-inoculation, and with lower kinetics when using N. ishiwadae BMK17.1. A remarkable malic acid consumption and a low acetic acid production was associated with L. elongisporus BMK12.5 fermentations, together with a high production of 3-methyl-1-butanol and 2-phenylethanol, and the release of high amounts of proteins into the wines. N. ishiwadae BMK17.1, although unable to finish the fermentation itself, showed a high production of oligosaccharides and volatile compounds such as isobutanol or isobutyric acid. This work reports, for the first time, the occurrence and enological potential of two strains pertaining to the non-conventional yeast genera Lodderomyces and Nakazawaea.

A DNA Vaccine Protects Human Immune Cells against Zika Virus Infection in Humanized Mice.

A DNA vaccine encoding prM and E protein has been shown to induce protection against Zika virus (ZIKV) infection in mice and monkeys. However, its effectiveness in humans remains undefined. Moreover, identification of which immune cell types are specifically infected in humans is unclear. We show that human myeloid cells and B cells are primary targets of ZIKV in humanized mice. We also show that a DNA vaccine encoding full length prM and E protein protects humanized mice from ZIKV infection. Following administration of the DNA vaccine, humanized DRAG mice developed antibodies targeting ZIKV as measured by ELISA and neutralization assays. Moreover, following ZIKV challenge, vaccinated animals presented virtually no detectable virus in human cells and in serum, whereas unvaccinated animals displayed robust infection, as measured by qRT-PCR. Our results utilizing humanized mice show potential efficacy for a targeted DNA vaccine against ZIKV in humans.

Combination of IL-10 and IL-2 induces oligoclonal human CD4 T cell expansion during xenogeneic and allogeneic GVHD in humanized mice.

IL-10 is a crucial anti-inflammatory cytokine which can also exert a seemingly divergent immunostimulatory effects under certain conditions. We found high levels of the cytokine in a xenogeneic GVHD model where NOD-scid IL2rγcnull (NSG) mice were transplanted with human PBMCs in presence of IL-2. Presence of exogenous IL-10 altered the kinetics of IL-2 induced human T cell reconstitution in vivo, showing an initial delay, followed by rapid expansion. Further, compared to IL-2 alone, treatment with IL-2 in combination with IL-10 increased survival in most animals and completely protected ∼20% of mice from GVHD. Additionally, IL-2 induced expansion of both CD4+ and CD8+ xenoreactive T cells whereas a combination of IL-2 and IL-10 resulted in selective expansion of CD4+ T cells only. TCR Vß repertoire analysis of CD4+ T cells showed that in contrast to IL-2 alone, simultaneous presence of both cytokines drastically reduced the Vß repertoire of the expanded CD4+ T cells. Highly restricted Vß usage was also observed when the cytokine combination was tested in an allogeneic GVHD model where NOD-scid IL2rγcnull mice expressing HLA-DR4 (NSG-DR4) were transplanted with purified CD4+ T cells from HLA-DR4 negative donors. Taken together, our results demonstrate that IL-10 can profoundly modulate the subset composition and repertoire of responding T cells during GVHD.

Gene therapy with plasmids encoding IFN-ß or IFN-α14 confers long-term resistance to HIV-1 in humanized mice.

Because endogenous interferon type I (IFN-I) produced by HIV-1 infection might complicate the analysis of therapeutically administered IFN-I, we tested different humanized mouse models for induction of IFN-I during HIV-1 infection. While HIV-1 induced high levels of IFN-α in BLT mice, IFN-I was undetectable following infection in the Hu-PBL mouse model, in which only T cells expand. We therefore tested the effect of treatment with Pegylated IFN-2 (pegasys), in Hu-PBL mice. Pegasys prevented CD4 T cell depletion and reduced the viral load for 10 days, but the effect waned thereafter. We next expressed IFN-I subsets (IFN-α2, -α6, -α8, -α14, and -ß) in Hu-PBL mice by hydrodynamic injection of plasmids encoding them and 2 days later infected the mice with HIV-1. CD4 T cell depletion was prevented in all subtypes of IFN-I-expressing mice by day 10. However, at day 40 post-infection, protection was seen in IFN-ß- and IFN-α14-expressing mice, but not the others. The viral load followed an inverse pattern and was highest in control mice and lowest in IFN-ß- and IFN-α14-expressing mice until day 40 after infection. These results show that gene therapy with plasmids encoding IFN-ß and -α14, but not the commonly used -α2, confers long-term suppression of HIV-1 replication.

A novel curcumin-like dienone induces apoptosis in triple-negative breast cancer cells.

PURPOSE: According to the World Health Organization (WHO), breast cancer is the most common cancer affecting women worldwide. In the USA ~12.3 % of all women are expected to be diagnosed with various types of breast cancer, exhibiting varying degrees of therapeutic response rates. Therefore, the identification of novel anti-breast cancer drugs is of paramount importance. METHODS: The 1,5-diaryl-3-oxo-1,4-pentadienyl pharmacophore was incorporated into a number of cytotoxins. Three of the resulting dienones, 2a, 2b and 2c, were tested for their anti-neoplastic potencies in a variety of human breast cancer-derived cell lines, including the triple negative MDA-MB-231 cell line and its metastatic variant, using a live-cell bio-imaging method. Special emphasis was put on dienone 2c, since its anti-cancer activity and its mode of inflicting cell death have so far not been reported. RESULTS: We found that all three dienones exhibited potent cytotoxicities towards the breast cancer-derived cell lines tested, whereas significantly lower toxicities were observed towards the non-cancerous human breast cell line MCF-10A. The dienones 2b and 2c exhibited the greatest selective cytotoxicity at submicromolar concentration levels. We found that these two dienones induced phosphatidylserine externalization in MDA-MB-231 cells in a concentration-dependent manner, suggesting that their cytotoxic effect might be mediated by apoptosis. This possibility was confirmed by our observation that the dienone 2c can induce mitochondrial depolarization, caspase-3 activation, cell cycle disruption and DNA fragmentation in MDA-MB-231 cells. CONCLUSION: Our findings indicate that dienone 2c uses the mitochondrial/intrinsic pathway to inflict apoptosis in triple negative MDA-MB-231 breast cancer-derived cells. This observation warrants further assessment of dienone 2c as a potential anti-breast cancer drug.

The nature of magnetoelectric coupling in Pb(Zr,Ti)O3 -Pb(Fe,Ta)O3.

The coupling between magnetization and polarization in a room temperature multiferroic (Pb(Zr,Ti)O3 -Pb(Fe,Ta)O3 ) is explored by monitoring the changes in capacitance that occur when a magnetic field is applied in each of three orthogonal directions. Magnetocapacitance effects, consistent with P(2) M(2) coupling, are strongest when fields are applied in the plane of the single crystal sheet investigated.

The gender of cell lines matters when screening for novel anti-cancer drugs.

Current reports indicated that the gender origin of cells is important in all facets of experimental biology. To explore this matter using an anticancer high throughput screening platform, seven male- and seven female-derived human cell lines, six from cancer patients in each group, were exposed to 81 novel cytotoxins. In this screen, the findings revealed that 79 out of 81 of the compounds consistently inflicted higher levels of toxicity towards male derived cells, emphasizing that there is indeed a gender-related difference in cell sensitivity to these anti-neoplastic agents. This gender-related drug sensitivity and toxicity explored at the molecular and cellular level emerged from a drug discovery enterprise.

Novel 3,5-bis(arylidene)-4-oxo-1-piperidinyl dimers: structure-activity relationships and potent antileukemic and antilymphoma cytotoxicity.

Novel clusters of 3,5-bis(benzylidene)-4-oxo-1-piperidinyl dimers 3-5 were evaluated against human Molt4/C8 and CEM T-lymphocytes and human HeLa cervix adenocarcinoma cells as well as murine L1210 leukemia neoplasms. Several of these compounds demonstrated IC50 values in the submicromolar and low micromolar range and compounds possessing 4-fluoro, 4-chloro and 3,4,5-trimethoxy substituents in the series 3 and 4 were identified as potent molecules. A heat map revealed the very high cytotoxic potencies of representative compounds against a number of additional leukemic and lymphoma cell lines and displayed greater toxicity to these cells than nonmalignant MCF10A and Hs-27 neoplasms. These dienones are more refractory to breast and prostate cancers. The evaluation of representative compounds in series 3-5 against a panel of human cancer cell lines revealed them to be potent cytotoxins with average IC50 values ranging from 0.05 to 8.51 µM. In particular, the most potent compound 4g demonstrated over 382-fold and 590-fold greater average cytotoxic potencies in this screen than the reference drugs, melphalan and 5-fluorouracil, respectively. A mode of action investigation of two representative compounds 3f and 4f indicated that they induce apoptosis which is due, at least in part, to the activation of caspase-3 and depolarization of the mitochondrial membrane potential.

Años de vida potencial perdidos: su utilidad en el análisis de la mortalidad en México / Years of potential life lost: advantages for mortality analysis in Mexico

La evolución de la epidemiología de la mortalidad en los países en desarrollo requiere del uso de indicadores complementarios o alternativos a las tasas específicas para el análisis de la mortalidad por causa. Este trabajo presenta las principales causas de mortalidad en México durante 1983 por sexo, a través del indicador años de vida potencial perdidos. Se utilizaron y discutieron diversas alternativas de construcción del indicador. La aplicación del indicador a los datos del país demostró utilidad para evaluar el impacto de la mortalidad por enfermedades infecciosas y causas violentas. Asimismo, las diferencias en magnitud entre los sexos y los diferentes intervalos de edad permitieron identificar la fracción de los años de vida potencial perdidos atribuible a grupos de población determinados. Se considera que el indicador provee información valiosa para el análisis epidemiológico de la mortalidad por causas