Ribosomal oxygenases are structurally conserved from prokaryotes to humans

2-Oxoglutarate (2OG)-dependent oxygenases have important roles in the regulation of gene expression via demethylation of N-methylated chromatin components1,2 and in the hydroxylation of transcription factors3 and splicing factor proteins4. Recently, 2OG-dependent oxygenases that catalyse hydroxylation of transfer RNA5,6,7 and ribosomal proteins8 have been shown to be important in translation relating to cellular growth, TH17-cell differentiation and translational accuracy9,10,11,12. The finding that ribosomal oxygenases (ROXs) occur in organisms ranging from prokaryotes to humans8 raises questions as to their structural and evolutionary relationships. In Escherichia coli, YcfD catalyses arginine hydroxylation in the ribosomal protein L16; in humans, MYC-induced nuclear antigen (MINA53; also known as MINA) and nucleolar protein 66 (NO66) catalyse histidine hydroxylation in the ribosomal proteins RPL27A and RPL8, respectively. The functional assignments of ROXs open therapeutic possibilities via either ROX inhibition or targeting of differentially modified ribosomes. Despite differences in the residue and protein selectivities of prokaryotic and eukaryotic ROXs, comparison of the crystal structures of E. coli YcfD and Rhodothermus marinus YcfD with those of human MINA53 and NO66 reveals highly conserved folds and novel dimerization modes defining a new structural subfamily of 2OG-dependent oxygenases. ROX structures with and without their substrates support their functional assignments as hydroxylases but not demethylases, and reveal how the subfamily has evolved to catalyse the hydroxylation of different residue side chains of ribosomal proteins. Comparison of ROX crystal structures with those of other JmjC-domain-containing hydroxylases, including the hypoxia-inducible factor asparaginyl hydroxylase FIH and histone Nε-methyl lysine demethylases, identifies branch points in 2OG-dependent oxygenase evolution and distinguishes between JmjC-containing hydroxylases and demethylases catalysing modifications of translational and transcriptional machinery. The structures reveal that new protein hydroxylation activities can evolve by changing the coordination position from which the iron-bound substrate-oxidizing species reacts. This coordination flexibility has probably contributed to the evolution of the wide range of reactions catalysed by oxygenases

Scans for signatures of selection in Russian cattle breed genomes reveal new candidate genes for environmental adaptation and acclimation

Domestication and selective breeding has resulted in over 1000 extant cattle breeds. Many of these breeds do not excel in important traits but are adapted to local environments. These adaptations are a valuable source of genetic material for efforts to improve commercial breeds. As a step toward this goal we identified candidate regions to be under selection in genomes of nine Russian native cattle breeds adapted to survive in harsh climates. After comparing our data to other breeds of European and Asian origins we found known and novel candidate genes that could potentially be related to domestication, economically important traits and environmental adaptations in cattle. The Russian cattle breed genomes contained regions under putative selection with genes that may be related to adaptations to harsh environments (e.g., AQP5, RAD50, and RETREG1). We found genomic signatures of selective sweeps near key genes related to economically important traits, such as the milk production (e.g., DGAT1, ABCG2), growth (e.g., XKR4), and reproduction (e.g., CSF2). Our data point to candidate genes which should be included in future studies attempting to identify genes to improve the extant breeds and facilitate generation of commercial breeds that fit better into the environments of Russia and other countries with similar climates

Role of hypoxia-inducible factors in epigenetic regulation via histone demethylases.

Eukaryotic chromatin is subject to multiple posttranslational histone modifications such as acetylation, methylation, phosphorylation, and ubiquitination. These various covalent modifications have been proposed to constitute a "histone code," playing important roles in the establishment of global chromatin environments, transcription, DNA repair, and DNA replication. Among these modifications, histone methylation specifies regulatory marks that delineate transcriptionally active and inactive chromatin. These histone methyl marks were considered irreversible; however, recent identification of site-specific histone demethylases demonstrates that histone methylation is dynamically regulated, which may allow cells to rapidly change chromatin conformation to adapt to environmental stresses or intrinsic stimuli. Of major interest is the observation that these histone demethylase enzymes, which are in the Jumonji gene family, require oxygen to function and, in some cases, are induced by hypoxia in an HIFalpha-dependent manner. This provides a new mechanism for regulation of the response to hypoxia

Roles of Salivary Components in Streptococcus mutans Colonization in a New Animal Model Using NOD/SCID.e2f1−/− Mice

Streptococcus mutans plays an important role in biofilm formation on the tooth surface and is the primary causative agent of dental caries. The binding of S. mutans to the salivary pellicle is of considerable etiologic significance and is important in biofilm development. Recently, we produced NOD/SCID.e2f1−/− mice that show hyposalivation, lower salivary antibody, and an extended life span compared to the parent strain: NOD.e2f1−/−. In this study we used NOD/SCID.e2f1−/− 4 or 6 mice to determine the roles of several salivary components in S. mutans colonization in vivo. S. mutans colonization in NOD/SCID.e2f1−/− mice was significantly increased when mice were pre-treated with human saliva or commercial salivary components. Interestingly, pre-treatment with secretory IgA (sIgA) at physiological concentrations promoted significant colonization of S. mutans compared with sIgA at higher concentrations, or with human saliva or other components. Our data suggest the principal effects of specific sIgA on S. mutans occur during S. mutans colonization, where the appropriate concentration of specific sIgA may serve as an anti-microbial agent, agglutinin, or an adherence receptor to surface antigens. Further, specific sIgA supported biofilm formation when the mice were supplied 1% sucrose water and a non-sucrose diet. The data suggests that there are multiple effects exerted by sIgA in S. mutans colonization, with synergistic effects evident under the condition of sIgA and limited nutrients on colonization in NOD/SCID.e2f1−/− mice. This is a new animal model that can be used to assess prevention methods for dental biofilm-dependent diseases such as dental caries

Optoelectronic characterization of CuInGa(S)2 thin films grown by spray pyrolysis for photovoltaic application

[EN] Copper-indium gallium disulfide (CIGS) is a good absorber for photovoltaic application. Thin films of CIGS were prepared by spray pyrolysis on glass substrates in the ambient atmosphere. The films were characterized by different techniques, such as structural, morphological, optical and electrical properties of CIGS films were analyzed by X-ray diffraction (XRD), scanning electron microscopy (SEM), atomic force microscopy (AFM), spectrophotometer and Hall effect, respectively. After optimization, the deposited films structure, grain size, and crystallinity became more important with an increase of annealing time at 370 degrees C for 20 min. Transmission electron microscopy (TEM) analysis shows that the interface sheets are well crystallized and the inter planer distance are 0.25 nm, 0.28 nm, and 0.36 nm. The atomic force microscopy (AFM) observation shows that the grain size and roughness can be tolerated by optimizing the annealing time. The strong absorbance and low transmittance were observed for the prepared films with a suitable energy bandgap about 1.46 eV. The Hall effect measurement system examined that CIGS films exhibited optimal electrical properties, resistivity, carrier mobility, and carrier concentration which were determined to be 4.22 x 10(6) omega cm, 6.18 x 10(2) cm(2) V-1 S-1 and 4.22 x 10(6) cm(-3), respectively. The optoelectronic properties of CIGS material recommended being used for the photovoltaic application.Prof. Bouchaib HARTITI, The Senior Associate at ICTP, is very grateful to ICTP for permanent support. Prof. Mohamed Ebn Touhami, Director of the University Center for Analysis, Expertise, Transfer of Technology and Incubation, Kenitra, Morocco, is very grateful to CUA2TI for financial support. Thanks to Doctor Diogo M.F. Santos for the supervision of Amal Bouich's work during her research in CeFEMA research center. The authors also thank researchers from CeFEMA (IST-ULisboa, Portugal) and CUA2TI (FS-Kenitra Morocco) for their help.Bouich, A.; Hartiti, B.; Ullah, S.; Ullah, H.; Ebn Touhami, M.; Santos, DMF.; Marí, B. (2019). Optoelectronic characterization of CuInGa(S)2 thin films grown by spray pyrolysis for photovoltaic application. Applied Physics A. 125(8):1-9. https://doi.org/10.1007/s00339-019-2874-4S191258T. Feurer, P. Reinhard, E. Avancini, B. Bissig, J. Löckinger, P. Fuchs, S. Buecheler, Progress in thin film CIGS photovoltaics–Research and development, manufacturing, and applications. Prog. Photovolt. Res. Appl. 25(7), 645–667 (2017)A. Zegadi, M.A. Slifkin, M. Djamin, A.E. Hill, R.D. Tomlinson, A photoacoustic study of CuInxGa1− xSe2 alloys. Phys. Status Solidi (A) 133(2), 533–540 (1992)T.H. Sajeesh, A.R. Warrier, C.S. Kartha, K.P. Vijayakumar, Optimization of parameters of chemical spray pyrolysis technique to get n and p-type layers of SnS. Thin Solid Films 518(15), 4370–4374 (2010)J. Liu, D. Zhuang, H. Luan, M. Cao, M. Xie, X. Li, Preparation of Cu (In, Ga) Se2 thin film by sputtering from Cu (In, Ga) Se2 quaternary target. Progr. Nat. Sci. Mater. Int. 23(2), 133–138 (2013)M.I. Hossain, Fabrication and characterization of CIGS solar cells with In2 S3 buffer layer deposited by PVD technique. Chalcogenide Lett. 9(5), 185–191 (2012)M.A. Mughal, R. Engelken, R. Sharma, Progress in indium (III) sulfide (In2S3) buffer layer deposition techniques for CIS, CIGS, and CdTe-based thin film solar cells. Sol. Energy 120, 131–146 (2015)M. Powalla, M. Cemernjak, J. Eberhardt, F. Kessler, R. Kniese, H.D. Mohring, B. Dimmler, Large-area CIGS modules: Pilot line production and new developments. Sol. Energy Mater Sol. Cells 90(18–19), 3158–3164 (2006)M.E. Calixto, P.J. Sebastian, R.N. Bhattacharya, R. Noufi, Compositional and optoelectronic properties of CIS and CIGS thin films formed by electrodeposition. Sol. Energy Mater. Sol. Cells 59(1–2), 75–84 (1999)S. Jung, S. Ahn, J.H. Yun, J. Gwak, D. Kim, K. Yoon, Effects of Ga contents on properties of CIGS thin films and solar cells fabricated by co-evaporation technique. Curr. Appl. Phys. 10(4), 990–996 (2010)S. R. Ovshinsky, X. Deng, R. Young, U.S. Patent No. 5,231,047. Washington, DC: U.S. Patent and Trademark Office (1993).M. Kaelin, D. Rudmann, A.N. Tiwari, Low cost processing of CIGS thin film solar cells. Sol. Energy 77(6), 749–756 (2004)Fangdan Jiang, Jiayou Feng, Effect of temperature on selenization process of metallic Cu–In alloy precursors. Thin Solid Films 515(4), 1950–1955 (2006)S. Shirakata, Y. Kannaka, H. Hasegawa, T. Kariya, S. Isomura, Properties of Cu (In, Ga) Se2 thin films prepared by chemical spray pyrolysis. Jpn. J. Appl. Phys. 38(9R), 4997 (1999)Y.K. Kumar, G.S. Babu, P.U. Bhaskar, V.S. Raja, Effect of starting-solution pH on the growth of Cu2ZnSnS4 thin films deposited by spray pyrolysis. Phys. Status Solidi (A) 206(7), 1525–1530 (2009)M. Ajili, M. Castagné, N.K. Turki, Characteristics of CuIn1− xGaxS2 thin films synthesized by chemical spray pyrolysis. J. Lumin. 150, 1–7 (2014)B.J. Babu, S. Velumani, A. Kassiba, R. Asomoza, J.A. Chavez-Carvayar, J. Yi, Deposition and characterization of graded Cu (In1-xGax) Se2 thin films by spray pyrolysis. Mater. Chem. Phys. 162, 59–68 (2015)S.F. Varol, G. Babür, G. Çankaya, U. Kölemen, Synthesis of sol–gel derived nano-crystalline ZnO thin films as TCO window layer: effect of sol aging and boron. RSC Adv. 4(100), 56645–56653 (2014)J.A. Frantz, R.Y. Bekele, V.Q. Nguyen, J.S. Sanghera, A. Bruce, S.V. Frolov, I.D. Aggarwal, Cu (In, Ga) Se2 thin films and devices sputtered from a single target without additional selenization. Thin Solid Films 519(22), 7763–7765 (2011)C. Calderón, G. Gordillo, P. Bartolo-Pérez, F. Mesa, Effect of the deposition conditions on the optical, morphological and compositional properties of CuIn1− xGaxSe2 thin films prepared by a multistage process. 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Optik 183, 137–147 (2019

Interval exercise versus continuous exercise in patients with moderate to severe chronic obstructive pulmonary disease – study protocol for a randomised controlled trial [ISRCTN11611768]

BACKGROUND: Physical exercise has become a cornerstone of management of chronic obstructive pulmonary disease (COPD) because it leads to clinically relevant improvements of exercise capacity and health-related quality of life (HRQL). Despite the scarcity of randomised trials directly comparing exercise protocols, current guidelines recommend high intensity continuous exercise for lower extremities as the probably most effective exercise modality. However, for patients admitted to inpatient respiratory rehabilitation programmes, it is often difficult to initiate such an exercise programme because they are severely limited by dyspnoea and leg fatigue and therefore unable to perform continuous exercise at higher intensities and for periods longer than 30 minutes. Interval exercise may be an attractive alternative for these COPD patients because it allows high intensity exercise with recovery periods. The aim of this study is to assess if interval exercise compared to high intensity continuous exercise is not of inferior effectiveness in terms of HRQL and exercise capacity improvements but associated with better exercise tolerance in patients with moderate to severe COPD at the beginning of a respiratory rehabilitation. METHODS/DESIGN: We will assign patients with moderately severe to severe COPD to either continuous exercise or interval exercise using a stratified randomisation. Patients will follow 12–15 exercise sessions during a comprehensive inpatient respiratory rehabilitation. Primary end point for effectiveness is HRQL as measured by the Chronic Respiratory Questionnaire (CRQ) two weeks after the end of rehabilitation and secondary endpoints include additional clinical outcomes such as functional exercise capacity, other HRQL measures, patients' experience of physical exercise as well as physiological measures of the effects of physical exercise such as cardiopulmonary exercise testing. Including expected drop-outs, we will need 52 patients per group to show differences corresponding to the minimal clinically important difference of the CRQ. Outcome assessors and investigators involved in data analysis will be blinded to group assignment until analyses have been carried out. DISCUSSION: Clinicians and the scientific community need evidence on the benefits and tolerance of exercise protocols available in clinical practice. The proposed trial will provide important and needed data on interval and continuous exercise for decision making in clinical practice

Sustained proliferation in cancer: mechanisms and novel therapeutic targets

Proliferation is an important part of cancer development and progression. This is manifest by altered expression and/or activity of cell cycle related proteins. Constitutive activation of many signal transduction pathways also stimulates cell growth. Early steps in tumor development are associated with a fibrogenic response and the development of a hypoxic environment which favors the survival and proliferation of cancer stem cells. Part of the survival strategy of cancer stem cells may manifested by alterations in cell metabolism. Once tumors appear, growth and metastasis may be supported by overproduction of appropriate hormones (in hormonally dependent cancers), by promoting angiogenesis, by undergoing epithelial to mesenchymal transition, by triggering autophagy, and by taking cues from surrounding stromal cells. A number of natural compounds (e.g., curcumin, resveratrol, indole-3-carbinol, brassinin, sulforaphane, epigallocatechin-3-gallate, genistein, ellagitannins, lycopene and quercetin) have been found to inhibit one or more pathways that contribute to proliferation (e.g., hypoxia inducible factor 1, nuclear factor kappa B, phosphoinositide 3 kinase/Akt, insulin-like growth factor receptor 1, Wnt, cell cycle associated proteins, as well as androgen and estrogen receptor signaling). These data, in combination with bioinformatics analyses, will be very important for identifying signaling pathways and molecular targets that may provide early diagnostic markers and/or critical targets for the development of new drugs or drug combinations that block tumor formation and progression

Prevalence of von Hippel-Lindau gene mutations in sporadic renal cell carcinoma: results from the Netherlands cohort study

BACKGROUND: Biallelic von Hippel-Lindau (VHL) gene defects, a rate-limiting event in the carcinogenesis, occur in approximately 75% of sporadic clear-cell Renal Cell Carcinoma (RCC). We studied the VHL mutation status in a large population-based case group. METHODS: Cases were identified within the Netherlands cohort study on diet and cancer, which includes 120,852 men and women. After 11.3 years of follow-up, 337 incident cases with histologically confirmed epithelial cancers were identified. DNA was isolated from paraffin material collected from 51 pathology laboratories and revised by one pathologist, leaving material from 235 cases. VHL mutational status was assessed by SSCP followed by direct sequencing, after testing SSCP as a screening tool in a subsample. RESULTS: The number of mutations was significantly higher for clear-cell RCC compared to other histological types. We observed 131 mutations in 114 out of 187 patients (61%) with clear-cell RCC. The majority of mutations were truncating mutations (47%). The mean tumor size was 72.7 mm for mutated tumors compared to 65.3 mm for wildtype tumors (p = 0.06). No statistically significant differences were observed for nuclear grade, TNM distribution or stage. In other histological types, we observed 8 mutations in 7 out of 48 patients (15%), 1 mutation in 1 of 6 oncocytoma, 3 mutations in 2 of 7 chromophobe RCC, 2 mutations in 2 of 30 papillary RCC, no mutations in 1 collecting duct carcinoma and 2 mutations in 2 of 4 unclassified RCC. CONCLUSION: VHL mutations were detected in 61% of sporadic clear-cell RCC. VHL mutated and wildtype clear-cell RCC did not differ with respect to most parameters

Potent and selective chemical probe of hypoxic signaling downstream of HIF-α hydroxylation via VHL inhibition

Chemical strategies to using small molecules to stimulate hypoxia inducible factors (HIFs) activity and trigger a hypoxic response under normoxic conditions, such as iron chelators and inhibitors of prolyl hydroxylase domain (PHD) enzymes, have broad-spectrum activities and off-target effects. Here we disclose VH298, a potent VHL inhibitor that stabilizes HIF-α and elicits a hypoxic response via a different mechanism, that is the blockade of the VHL:HIF-α protein-protein interaction downstream of HIF-α hydroxylation by PHD enzymes. We show that VH298 engages with high affinity and specificity with VHL as its only major cellular target, leading to selective on-target accumulation of hydroxylated HIF-α in a concentration- and time-dependent fashion in different cell lines, with subsequent upregulation of HIF-target genes at both mRNA and protein levels. VH298 represents a high-quality chemical probe of the HIF signalling cascade and an attractive starting point to the development of potential new therapeutics targeting hypoxia signalling