Oxidative stress is tightly regulated by cytochrome c phosphorylation and respirasome factors in mitochondria

Respiratory cytochrome c has been found to be phosphorylated at tyrosine 97 in the postischemic brain upon neuroprotective insulin treatment, but how such posttranslational modification affects mitochondrial metabolism is unclear. Here, we report the structural features and functional behavior of a phosphomimetic cytochrome c mutant, which was generated by site-specific incorporation at position 97 of p-carboxymethyl-l-phenylalanine using the evolved tRNA synthetase method. We found that the point mutation does not alter the overall folding and heme environment of cytochrome c, but significantly affects the entire oxidative phosphorylation process. In fact, the electron donation rate of the mutant heme protein to cytochrome c oxidase, or complex IV, within respiratory supercomplexes was higher than that of the wild-type species, in agreement with the observed decrease in reactive oxygen species production. Direct contact of cytochrome c with the respiratory supercomplex factor HIGD1A (hypoxia-inducible domain family member 1A) is reported here, with the mutant heme protein exhibiting a lower affinity than the wild-type species. Interestingly, phosphomimetic cytochrome c also exhibited a lower caspase-3 activation activity. Altogether, these findings yield a better understanding of the molecular basis for mitochondrial metabolism in acute diseases, such as brain ischemia, and thus could allow the use of phosphomimetic cytochrome c as a neuroprotector with therapeutic applications.España, Junta de Andalucía BIO-198España MINECO BFU2015-71017/BM

Nondestructive Assessment of Citrus Fruit Quality and Ripening by Visible–Near Infrared Reflectance Spectroscopy

As non-climacteric, citrus fruit are only harvested at their optimal edible ripening stage. The usual approach followed by producers and packinghouses to establish the internal quality and ripening of citrus fruit is to collect fruit sets throughout ripening and use them to determine the quality attributes (QA) by standard and, in many cases, destructive and time-consuming methods. However, due to the large variability within and between orchards, the number of measured fruits is seldom statistically representative of the batch, resulting in a fallible assessment of their internal QA (IQA) and a weak traceability in the citrus supply chain. Visible/near-infrared reflectance spectroscopy (Vis–NIRS) is a nondestructive method that addresses this problem, and has proved to predict many IQA of a wide number of fruit including citrus. Yet, its application on a daily basis is not straightforward, and there are still several questions to address by researchers in order to implement it routinely in the crop supply chain. This chapter reviews the application of Vis–NIRS in the assessment of the quality and ripening of citrus fruit, and makes a critical evaluation on the technique’s limiting issues that need further attention by researchers

CATÁLOGO DE LOS BRIÓFITOS DE LA PROVINCIA DE MURCIA (SURESTE DE ESPAÑA)

A check-list of the bryophytes of Murcia province is provided. Bryophyte diversity in Murcia consists of 200 taxa (74 genera) of mosses and 32 taxa (20 genera) of liverworts. Pseudocrossidium obtusulum is reported as new to the Iberian Peninsula. A total of 30 taxa are new record from Murcia: Acaulon casasianum, Astomum levieri, Bryoerythrophyllum recurvirostrum, Crossidium laevipilum, Entosthodon convexus, Grimmia dissimulata, Gymnostomum lanceolatum, Hedwigia ciliata, Orthotrichum affine, O. pumilum, O. scanicum, O. schimperi, O. striatum, O. tenellum, O. tortidontium, O. vittii, Philonotis fontana, Pleuridium acuminatum, Pohlia cruda, Pterygoneurum subsessile, Rhynchostegiella tenella, Schistidium crassipilum, Schistidium singarense, Syntrichia virescens, Tortula canescens, T. mucronifolia, Weissia brachycarpa, W. condensa var. armata, Athalamia hyalina, and Jungermannia atrovirens. Thirty eight taxa (33 mosses and 5 liverworts) are excluded from the checklist or represent doubtful reports for Murcia.Se realiza un catalogo de los briofitos de la provincia de Murcia. La diversidad briofitica de Murcia se estima en 200 taxones (74 generos) de musgos y 32 taxones de hepaticas (20 generos). Pseudocrossidium obtusulum se cita por primera vez en la Peninsula Iberica. Un total de 30 taxones son novedades para la provincia de Murcia: Acaulon casasianum, Astomum levieri, Bryoerythrophyllum recurvirostrum, Crossidium laevipilum, Entosthodon convexus, Grimmia dissimulata, Gymnostomum lanceolatum, Hedwigia ciliata, Orthotrichum affine, O. pumilum, O. scanicum, O. schimperi, O. striatum, O. tenellum, O. tortidontium, O. vittii, Philonotis fontana, Pleuridium acuminatum, Pohlia cruda, Pterygoneurum subsessile, Rhynchostegiella tenella, Schistidium crassipilum, Schistidium singarense, Syntrichia virescens, Tortula canescens, T. mucronifolia, Weissia brachycarpa, W. condensa var. armata, Athalamia hyalina y Jungermannia atrovirens. Treinta y ocho taxones (33 musgos y 5 hepaticas) se han excluido del catalogo de la provincia o se citan como dudosos

Transcriptional profiling of fetal hypothalamic TRH neurons

RIGHTS : This article is licensed under the BioMed Central licence at http://www.biomedcentral.com/about/license which is similar to the 'Creative Commons Attribution Licence'. In brief you may : copy, distribute, and display the work; make derivative works; or make commercial use of the work - under the following conditions: the original author must be given credit; for any reuse or distribution, it must be made clear to others what the license terms of this work are.AbstractBackgroundDuring murine hypothalamic development, different neuroendocrine cell phenotypes are generated in overlapping periods; this suggests that cell-type specific developmental programs operate to achieve complete maturation. A balance between programs that include cell proliferation, cell cycle withdrawal as well as epigenetic regulation of gene expression characterizes neurogenesis. Thyrotropin releasing hormone (TRH) is a peptide that regulates energy homeostasis and autonomic responses. To better understand the molecular mechanisms underlying TRH neuron development, we performed a genome wide study of its transcriptome during fetal hypothalamic development. ResultsIn primary cultures, TRH cells constitute 2% of the total fetal hypothalamic cell population. To purify these cells, we took advantage of the fact that the segment spanning -774 to +84 bp of the Trh gene regulatory region confers specific expression of the green fluorescent protein (GFP) in the TRH cells. Transfected TRH cells were purified by fluorescence activated cell sorting, various cell preparations pooled, and their transcriptome compared to that of GFP- hypothalamic cells. TRH cells undergoing the terminal phase of differentiation, expressed genes implicated in protein biosynthesis, intracellular signaling and transcriptional control. Among the transcription-associated transcripts, we identified the transcription factors Klf4, Klf10 and Atf3, which were previously uncharacterized within the hypothalamus. ConclusionTo our knowledge, this is one of the first reports identifying transcripts with a potentially important role during the development of a specific hypothalamic neuronal phenotype. This genome-scale study forms a rational foundation for identifying genes that might participate in the development and function of hypothalamic TRH neurons.Published versio

Sustainable strategies based on glycine–betaine analogue ionic liquids for the recovery of monoclonal antibodies from cell culture supernatants

Monoclonal antibodies (mAbs) are of crucial interest for therapeutic purposes, particularly in vaccination and immunization, and in the treatment of life-threatening diseases. However, their downstream processing from the complex cell culture media in which they are produced still requires multiple steps, making mAbs extremely high-cost products. Therefore, the development of cost-effective, sustainable and biocompatible purification strategies for mAbs is in high demand to decrease the associated economic, environmental and health burdens. Herein, novel aqueous biphasic systems (ABS) composed of glycine–betaine analogue ionic liquids (AGB-ILs) and K2HPO4/KH2PO4 at pH 7.0, the respective three-phase partitioning (TPP) systems, and hybrid processes combined with ultrafiltration were investigated and compared in terms of performance as alternative strategies for the purification and recovery of anti-human interleukin-8 (anti-IL-8) mAbs, which are specific therapeutics in the treatment of inflammatory diseases, from Chinese Hamster Ovary (CHO) cell culture supernatants. With the studied ABS, mAbs preferentially partition to the IL-rich phase, with recovery yields up to 100% and purification factors up to 1.6. The best systems were optimized in what concerns the IL concentration, allowing to take advantage of IL-based three-phase partitioning approaches where a precipitate enriched in mAbs is obtained at the ABS interface, yielding 41.0% of IgG with a purification factor of 2.7 (purity of 60.9%). Hybrid processes combining the two previous techniques and an ultrafiltration step were finally applied, allowing the recovery of mAbs from the different fractions in an appropriate buffer solution for further biopharmaceutical formulations, while allowing the simultaneous IL removal and reuse. The best results were obtained with the hybrid process combining TPP and ultrafiltration, allowing to obtain mAbs with a purity higher than 60%. The recyclability of the IL was additionally demonstrated, revealing no losses in the purification and recovery performance of these systems for mAbs. The biological activity of anti-IL-8 mAbs is maintained after the several purification and recovery steps, indicating that the novel ABS, three-phase partitioning and hybrid processes comprising AGB-ILs are promising and sustainable strategies in mAbs downstream processing.publishe

Flora brio-pteridolítica de los bosques lauroides de Rhododendron ponticum L. del Parque Natural de los Alcornocales (Cádiz-Málaga, España)

RESUMEN. Flora brio-pteridolítica de los bosques lauroides de Rhododendron ponticum L. del Parque Natural de los Alcornocales (Cádiz-Málaga, España). Se ha estudiado parte de la diversidad brio- pteridofítica en el Parque Natural de los Alcornocales (Cádiz-Málaga; Sur de España). Nos hemos centrado en las comunidades caracterizadas por la presencia de Rhododendron ponticum, uno de los hábitats más representativos del Parque por su originalidad. Para ello hemos muestreado 23 localidades repartidas por todo el Parque y que varían por sus características climáticas. Se han identificado 171 especies de las que 111 son musgos, 2 antocerotas, 42 hepáticas y 17 helechos. Para cada una de las localidades (canutos) se aporta su diversidad total y las especies más representativas y exclusivas. Pretendemos contribuir con ello a una correcta identificación de zonas susceptibles de ser tenidas en cuenta para futuros planes de zonificación y conservación de este tipo de comunidades, únicas y frecuentemente olvidadas en los planteamientos conservacionistas a nivel regional y nacional.Palabras clave. Briófitos, pteridófitos, Rhododendron ponticum, bosque lauroides, Los Alcornocales, Andalucía, Península Ibérica.ABSTRACT. Bryo-pteridophytic flora in the lauroid forests (Rhododendron ponticum L.) of the Natural Park Los Alcornocales (Cadiz-Malaga, Spain). A great part of the bryo-pteridophytic diversity of the Natural Park of Los Alcornocales (Cadiz and Málaga provinces, Andalusia, south of Spain) has been studied. The study has been focused on the woody lauroid, relic communities of Rhododendron ponticum L., one of the more representative habitats of the Park because of its singularity. 23 localities extensively distributed in the Park have been inventoried, taking into account climactic variability. 171 species have been identified: 111 mosses, 2 anthoceros, 42 liverworts and 17 pteridophytes. Total species diversity and the most representative and/or exclusive species are given for each locality. We want to contribute to identifying areas to bear in mind for future conservation and land planning for this kind of communities, unique and frequently forgotten in territory politics at regional and national levels.Key words. Bryophytes, pteridophytes, Rhododendron ponticum, lauroid forests, Los Alcornocales, Andalusia, Iberian Peninsula