De novo Light Harvesting Complexes as Model System to study Chromophor protein Interactions in the Native Membrane

The presented thesis has focused on the interactions between protein and pigments in photosynthetic membrane proteins, and the significance of these interactions in membrane protein assembly. The thesis has been divided into 3 Chapters, two are focused on the interactions between (bacterio)chlorophyll and proteins, and one is focused on the interactions, between carotenoid and proteins. In order to explore these interactions model proteins have been designed based on the peripheral antenna of Rhodobacter sphaeroides. In the model LH2 complexes, portions of the transmembrane helices, in particular, at the pigment binding sites, are replaced simplified alternating by alanine-leucine stretches. In the model sequence context, the effects of particular amino acids are amplified, and thus allow for convenient identification of potentially critical interaction motifs. This approach is employed to study the factors that contribute to pigment binding and pigment-protein assembly. To confirm the significance of thus identified motifs, they are subsequently also examined in the WT sequence context. In Chapter 3, it is shown that the residue at position -4 of the beta-subunit has a critical structural role for the proper organisation of the excitonically coupled BChl dimer in the antenna complex. In WT LH2, the residue at this position makes an H-bond to the C131 keto carbonyl group of one of the dimeric BChl molecules. The potential importance of such a H-bonding motif at the BChl/protein interface is demonstrated by use of the model LH2 in which the H-bond drives the folding and assembly of this transmembrane BChl-protein. The structural role of this residue at the BChl/protein interface is further demonstrated by the linear correlation between the LH2 spectral tuning and the residue-BChl contact. In Chapter 4, the aspect of diastereotopic ligation to the central Mg of BChl is explored, in particular, the consequences of BChl-ligation for folding and assembly of BChl-proteins. The analysis of H-bonding patterns in Chl-binding photosystem I and II showed that H-bonding at the (B)Chl-protein interface is structurally distinct depending on the ligation type. In essence, the C131 keto groups of (B)Chl ligated in the beta-position, contrary to those ligated in the betaposition, are frequently employed to associate Chl-helix units and thus involved in tertiary interactions. Disruption of such H-bonding interactions by site directed mutagenesis significantly altered the structural stability and assembly of the LH2 complex in the membrane. These findings suggest that H-bonding to -ligated bacteriochlorophyll is a key structural motif for the correct assembly of (bacterio)chlorophyll proteins. In Chapter 5, it is shown by mutational analysis of the carotenoid binding pocket of native and model LH2 complexes that the aromatic residues, in particular phenylalanine, are a key factor for carotenoid binding. The phenylalanine not only contributes to the stable Car binding but also lock the Car into a particular molecular configuration. The importance of aromatic residues in Car binding is further supported by statistical analyses of the plant photosystems which show that phenylalanine residues are frequently in the close vicinity of Car moelcules. This study provides, to the best of our knowledge, the first experimental evidence for the central role of aromatic residues in carotenoid binding and functional specification

Las fiestas populares en España. Siglos XVI-XVIII

Comunicación presentada en el Congreso "Il tempo libero. Economia e società. Secc. XIII-XVIII" celebrado en Prato (Italia) del 18 al 23 de abril de 1994.Publicad

Cannabidiol and Other Cannabinoids in Demyelinating Diseases

A growing body of preclinical evidence indicates that certain cannabinoids, including cannabidiol (CBD) and synthetic derivatives, may play a role in the myelinating processes and are promising small molecules to be developed as drug candidates for management of demyelinating diseases such as multiple sclerosis (MS), stroke and traumatic brain injury (TBI), which are three of the most prevalent demyelinating disorders. Thanks to the properties described for CBD and its interesting profile in humans, both the phytocannabinoid and derivatives could be considered as potential candidates for clinical use. In this review we will summarize current advances in the use of CBD and other cannabinoids as future potential treatments. While new research is accelerating the process for the generation of novel drug candidates and identification of druggable targets, the collaboration of key players such as basic researchers, clinicians and pharmaceutical companies is required to bring novel therapies to the patients

Detrimental Effect of Cannabidiol on the Early Onset of Diabetic Nephropathy in Male Mice

Anti-inflammatory and antidiabetogenic properties have been ascribed to cannabidiol (CBD). CBD-based medicinal drugs have been approved for over a lustrum, and a boom in the commercialization of CBD products started in parallel. Herein, we explored the efficacy of CBD in streptozotocin (STZ)-induced diabetic mice to prevent diabetic nephropathy at onset. Eight-to-ten-week-old C57BL6J male mice were treated daily intraperitoneally with 10 mg/kg of CBD or vehicle for 14 days. After 8 days of treatment, mice were challenged with STZ or vehicle (healthy-control). At the end of the study, non-fasting blood glucose (FBG) level was 276 ± 42 mg/dL in vehicle-STZ-treated compared to 147 ± 9 mg/dL (p ≤ 0.01) in healthy-control mice. FBG was 114 ± 8 mg/dL in vehicle-STZ-treated compared to 89 ± 4 mg/dL in healthy-control mice (p ≤ 0.05). CBD treatment did not prevent STZ-induced hyperglycemia, and non-FBG and FBG levels were 341 ± 40 and 133 ± 26 mg/dL, respectively. Additionally, treatment with CBD did not avert STZ-induced glucose intolerance or pancreatic beta cell mass loss compared to vehicle-STZ-treated mice. Anatomopathological examination showed that kidneys from vehicle-STZ-treated mice had a 35% increase of glomerular size compared to healthy-control mice (p ≤ 0.001) and presented lesions with a 43% increase in fibrosis and T cell infiltration (p ≤ 0.001). Although treatment with CBD prevented glomerular hypertrophy and reduced T cell infiltration, it significantly worsened overall renal damage (p ≤ 0.05 compared to vehicle-STZ mice), leading to a more severe renal dysfunction than STZ alone. In conclusion, we showed that CBD could be detrimental for patients with type 1 diabetes, particularly those undergoing complications such as diabetic nephropathy

EHP-101 alleviates angiotensin II-induced fibrosis and inflammation in mice

Some cannabinoids showed anti-inflammatory and antifibrotic activities. EHP-101 is an oral lipidic formulation of the novel non-psychotropic cannabidiol aminoquinone VCE-004.8, which showed antifibrotic activity in murine models of systemic sclerosis induced by bleomycin. We herein examined the effect of EHP-101 on cardiac and other organ fibrosis in a mouse model induced by Angiotensin II. VCE-004.8 inhibited TGFβ- and Ang II-induced myofibroblast differentiation in cardiac fibroblasts detected by α-SMA expression. VCE-004.8 also inhibited Ang II-induced ERK 1 + 2 phosphorylation, NFAT activation and mRNA expression of IL1β, IL6, Col1A2 and CCL2 in cardiac fibroblasts. Mice infused with Ang II resulted in collagen accumulation in left ventricle, aortic, dermal, renal and pulmonary tissues; oral administration of EHP-101, Ajulemic acid and Losartan improved these phenotypes. In myocardial tissue, Ang II induced infiltration of T cells and macrophages together with the accumulation of collagen and Tenascin C; those were all reduced by either EHP-101 or Losartan treatment. Cardiac tissue RNA-Seq analyses revealed a similar transcriptomic signature for both treatments for inflammatory and fibrotic pathways. However, the gene set enrichment analysis comparing data from EHP-101 vs Losartan showed specific hallmarks modified only by EHP-101. Specifically, EHP-101 inhibited the expression of genes such as CDK1, TOP2A and MKi67 that are regulated to the E2 factor family of transcription factors. This study suggests that the oral administration of EHP-101 prevents and inhibits cardiac inflammation and fibrosis. Furthermore, EHP-101 inhibits renal, pulmonary and dermal fibrosis. EHP-101 could offer new opportunities in the treatment of cardiac fibrosis and other fibrotic diseases

The adipokine lipocalin-2 in the context of the osteoarthritic osteochondral junction

Corrigendum: The adipokine lipocalin-2 in the context of the osteoarthritic osteochondral junction. Scientific Reports, 6, 30666. https://doi.org/10.1038/srep30666Obesity and osteoarthritis (OA) form a vicious circle in which obesity contributes to cartilage destruction in OA, and OA-associated sedentary behaviour promotes weight gain. Lipocalin-2 (LCN2), a novel adipokine with catabolic activities in OA joints, contributes to the obesity and OA pathologies and is associated with other OA risk factors. LCN2 is highly induced in osteoblasts in the absence of mechanical loading, but its role in osteoblast metabolism is unclear. Therefore, because osteochondral junctions play a major role in OA development, we investigated the expression and role of LCN2 in osteoblasts and chondrocytes in the OA osteochondral junction environment. Our results showed that LCN2 expression in human osteoblasts and chondrocytes decreased throughout osteoblast differentiation and was induced by catabolic and inflammatory factors; however, TGF-beta 1 and IGF-1 reversed this induction. LCN2 reduced osteoblast viability in the presence of iron and enhanced the activity of MMP-9 released by osteoblasts. Moreover, pre-stimulated human osteoblasts induced LCN2 expression in human chondrocytes, but the inverse was not observed. Thus, LCN2 is an important catabolic adipokine in osteoblast and chondrocyte metabolism that is regulated by differentiation, inflammation and catabolic and anabolic stimuli, and LCN2 expression in chondrocytes is regulated in a paracrine manner after osteoblast stimulation.The authors acknowledge Mr. Oliver Shaw for performing the English revision and the support of Dr. Esbrit's. The authors' research is supported by research grants from Fondo de Investigación Sanitaria funded by the Instituto de Salud Carlos III (PI12/00144, PI13/00570, CP15/00007, PI14/00016 and PIE13/00024). R.G. is funded by the Instituto de Salud Carlos III through a Miguel Servet programme. A.V. is the recipient of a fellowship from the Fundación Conchita Rábago. A.G.M. was funded by the Universidad Carlos III de Madrid (Spain). R.L. and O.G. were funded by the Instituto de Salud Carlos III. O.G. is a member of the RETICS Programme, RD12/0009/0008 Instituto de Salud Carlos III (ISCIII). The research is supported by research grant from FEDE

The Transcriptome of Streptococcus pneumoniae Induced by Local and Global Changes in Supercoiling

The bacterial chromosome is compacted in a manner optimal for DNA transactions to occur. The degree of compaction results from the level of DNA-supercoiling and the presence of nucleoid-binding proteins. DNA-supercoiling is homeostatically maintained by the opposing activities of relaxing DNA topoisomerases and negative supercoil-inducing DNA gyrase. DNA-supercoiling acts as a general cis regulator of transcription, which can be superimposed upon other types of more specific trans regulatory mechanism. Transcriptomic studies on the human pathogen Streptococcus pneumoniae, which has a relatively small genome (∼2 Mb) and few nucleoid-binding proteins, have been performed under conditions of local and global changes in supercoiling. The response to local changes induced by fluoroquinolone antibiotics, which target DNA gyrase subunit A and/or topoisomerase IV, involves an increase in oxygen radicals which reduces cell viability, while the induction of global supercoiling changes by novobiocin (a DNA gyrase subunit B inhibitor), or by seconeolitsine (a topoisomerase I inhibitor), has revealed the existence of topological domains that specifically respond to such changes. The control of DNA-supercoiling in S. pneumoniae occurs mainly via the regulation of topoisomerase gene transcription: relaxation triggers the up-regulation of gyrase and the down-regulation of topoisomerases I and IV, while hypernegative supercoiling down-regulates the expression of topoisomerase I. Relaxation affects 13% of the genome, with the majority of the genes affected located in 15 domains. Hypernegative supercoiling affects 10% of the genome, with one quarter of the genes affected located in 12 domains. However, all the above domains overlap, suggesting that the chromosome is organized into topological domains with fixed locations. Based on its response to relaxation, the pneumococcal chromosome can be said to be organized into five types of domain: up-regulated, down-regulated, position-conserved non-regulated, position-variable non-regulated, and AT-rich. The AT content is higher in the up-regulated than in the down-regulated domains. Genes within the different domains share structural and functional characteristics. It would seem that a topology-driven selection pressure has defined the chromosomal location of the metabolism, virulence and competence genes, which suggests the existence of topological rules that aim to improve bacterial fitness

Cannabidiol induces antioxidant pathways in keratinocytes by targeting BACH1

Cannabidiol (CBD) is a major non-psychotropic phytocannabinoid that attracted a great attention for its therapeutic potential against different pathologies including skin diseases. However, although the efficacy in preclinical models and the clinical benefits of CBD in humans have been extensively demonstrated, the molecular mechanism(s) and targets responsible for these effects are as yet unknown. Herein we characterized at the molecular level the effects of CBD on primary human keratinocytes using a combination of RNA sequencing (RNA-Seq) and sequential window acquisition of all theoretical mass spectrometry (SWATH-MS). Functional analysis revealed that CBD regulated pathways involved in keratinocyte differentiation, skin development and epidermal cell differentiation among other processes. In addition, CBD induced the expression of several NRF2 target genes, with heme oxygenase 1 (HMOX1) being the gene and the protein most upregulated by CBD. CRISPR/Cas9-mediated genome editing, RNA interference and biochemical studies demonstrated that the induction of HMOX1 mediated by CBD, involved nuclear export and proteasomal degradation of the transcriptional repressor BACH1. Notably, we showed that the effect of BACH1 on HMOX1 expression in keratinocytes is independent of NRF2. In vivo studies showed that topical CBD increased the levels of HMOX1 and of the proliferation and wound-repair associated keratins 16 and 17 in the skin of mice. Altogether, our study identifies BACH1 as a molecular target for CBD in keratinocytes and sets the basis for the use of topical CBD for the treatment of different skin diseases including atopic dermatitis and keratin disorders.This work was supported by the Medical Research Institute of the University of Dundee, Cancer Research UK (C52419/A22869) (LV) and Tenovus Scotland (T18/07) (LC) and by grant SAF2017-87701-R (EM) from the Ministry of the Economy and Competition (MINECO) co-financed with the European Union FEDER funds. InnoHealth Group and Emerald Health Biotechnology also supported this work.Ye

Análisis de los valores de hipertensión arterial en pacientes que emplean sistemas personalizados de dosificación.

post-print311 K

MISIONES ACTUALES DE LAS UNIVERSIDADES PÚBLICAS: UNA PERSPECTIVA SOCIOLÓGICA

[EN: From the New Sociological Institutionalism point of view, we have seen what the missions that institutionally Spanish public universities assume in their strategic plans are. It has been applied the content analysis approach. We have found that indicators are more frequent to teaching activities, in second place appeared research activities and in third place were knowledge transfer activities. Not all universities assume the same grade of institutionalization for each mission. In fact, we have obtained a typology of universities split in two groups, one focused to teaching and another focused to research and knowledge transfer[ES: Este artículo analiza, desde la perspectiva del Nuevo Institucionalismo Sociológico, cuáles son las misiones que las universidades públicas españolas asumen de forma institucional en sus planes estratégicos. Para tal fin, se ha utilizado el análisis de contenido. En general, los resultados indican que las universidades muestran mayor énfasis en la misión ¿Docencia¿, seguida de la ¿Investigación¿, y en último lugar en la ¿Transferencia de conocimiento¿, si bien este patrón no es homogéneo en todas las universidades. Se han identificado dos tipologías de universidades en función del peso otorgado a cada misión: las más centradas en ¿Docencia¿ frente a las focalizadas en ¿Investigación y Transferencia de Conocimiento¿.Palomares Montero, D.; Garcia Aracil, A.; Castro-Martinez, E. (2012). MISIONES ACTUALES DE LAS UNIVERSIDADES PÚBLICAS: UNA PERSPECTIVA SOCIOLÓGICA. Arbor. 188(753):171-192. doi:10.3989/arbor.2012.753n1011S171192188753Colyvas, J. A., & Powell, W. W. (2006). Roads to Institutionalization: The Remaking of Boundaries between Public and Private Science. Research in Organizational Behavior, 27, 305-353. doi:10.1016/s0191-3085(06)27008-4Colyvas, J. A., & Powell, W. W. (2006). Roads to Institutionalization: The Remaking of Boundaries between Public and Private Science. Research in Organizational Behavior, 27, 305-353. doi:10.1016/s0191-3085(06)27008-4Laredo, P. (2007). Revisiting the Third Mission of Universities: Toward a Renewed Categorization of University Activities? Higher Education Policy, 20(4), 441-456. doi:10.1057/palgrave.hep.8300169Meyer, J. W. (1977). The Effects of Education as an Institution. American Journal of Sociology, 83(1), 55-77. doi:10.1086/226506Taylor, J., & De Lourdes Machado, M. (2006). Higher education leadership and management: From conflict to interdependence through strategic planning. Tertiary Education and Management, 12(2), 137-160. doi:10.1080/13583883.2006.996716