Expanding the scope of N → S acyl transfer in native peptide sequences

Understanding the factors that influence N → S acyl transfer in native peptide sequences, and discovery of new reagents that facilitate it, will be key to expanding its scope and applicability. Here, through a study of short model peptides in thioester formation and cyclisation reactions, we demonstrate that a wider variety of Xaa-Cys motifs than originally envisaged are capable of undergoing efficient N → S acyl transfer. We present data for the relative rates of thioester formation and cyclisation for a representative set of amino acids, and show how this expanded scope can be applied to the production of the natural protease inhibitor Sunflower Trypsin Inhibitor-1 (SFTI-1)

Encyclopedia of psychopharmacology

Definition of microiontophoresis and related methods as found in the Encyclopedia of Psychopharmacology.peer-reviewe

An eclipsing binary distance to the Large Magellanic Cloud accurate to 2 per cent

In the era of precision cosmology it is essential to determine the Hubble Constant with an accuracy of 3% or better. Currently, its uncertainty is dominated by the uncertainty in the distance to the Large Magellanic Cloud (LMC) which as the second nearest galaxy serves as the best anchor point of the cosmic distance scale. Observations of eclipsing binaries offer a unique opportunity to precisely and accurately measure stellar parameters and distances. The eclipsing binary method was previously applied to the LMC but the accuracy of the distance results was hampered by the need to model the bright, early-type systems used in these studies. Here, we present distance determinations to eight long-period, late- type eclipsing systems in the LMC composed of cool giant stars. For such systems we can accurately measure both the linear and angular sizes of their components and avoid the most important problems related to the hot early-type systems. Our LMC distance derived from these systems is demonstrably accurate to 2.2 % (49.97 +/- 0.19 (statistical) +/- 1.11 (systematic) kpc) providing a firm base for a 3 % determination of the Hubble Constant, with prospects for improvement to 2 % in the future.Comment: 34 pages, 5 figures, 13 tables, published in the Nature, a part of our data comes from new unpublished OGLE-IV photometric dat

On form factors in N=4 sym

In this paper we study the form factors for the half-BPS operators $\mathcal{O}^{(n)}_I$ and the $\mathcal{N}=4$ stress tensor supermultiplet current $W^{AB}$ up to the second order of perturbation theory and for the Konishi operator $\mathcal{K}$ at first order of perturbation theory in $\mathcal{N}=4$ SYM theory at weak coupling. For all the objects we observe the exponentiation of the IR divergences with two anomalous dimensions: the cusp anomalous dimension and the collinear anomalous dimension. For the IR finite parts we obtain a similar situation as for the gluon scattering amplitudes, namely, apart from the case of $W^{AB}$ and $\mathcal{K}$ the finite part has some remainder function which we calculate up to the second order. It involves the generalized Goncharov polylogarithms of several variables. All the answers are expressed through the integrals related to the dual conformal invariant ones which might be a signal of integrable structure standing behind the form factors.Comment: 35 pages, 7 figures, LATEX2

Lentiviral Engineered Fibroblasts Expressing Codon Optimized COL7A1 Restore Anchoring Fibrils in RDEB

Cells therapies, engineered to secrete replacement proteins, are being developed to ameliorate otherwise debilitating diseases. Recessive dystrophic epidermolysis bullosa (RDEB) is caused by defects of type VII collagen (C7), a protein essential for anchoring fibril formation at the dermal-epidermal junction (DEJ). Whilst allogeneic fibroblasts injected directly into the dermis can mediate transient disease modulation, autologous gene-modified fibroblasts should evade immunological rejection and support sustained delivery of C7 at the DEJ. We demonstrate the feasibility of such an approach using a therapeutic grade, self-inactivating-lentiviral vector, encoding codon optimized COL7A1, to transduce RDEB fibroblasts under conditions suitable for clinical application. Expression and secretion of C7 was confirmed, with transduced cells exhibiting supra-normal levels of protein expression and ex vivo migration of fibroblasts was restored in functional assays. Gene modified RDEB fibroblasts also deposited C7 at the DEJ of human RDEB skin xenografts placed on NOD-scid IL2Rgamma(null) recipients, with reconstruction of human epidermal structure and regeneration of anchoring fibrils at the DEJ. Fibroblast mediated restoration of protein and structural defects in this RDEB model strongly supports proposed therapeutic applications in man

Clinical-pathological study on β-APP, IL-1β, GFAP, NFL, Spectrin II, 8OHdG, TUNEL, miR-21, miR-16, miR-92 expressions to verify DAI-diagnosis, grade and prognosis

Traumatic brain injury (TBI) is one of the most important death and disability cause, involving substantial costs, also in economic terms, when considering the young age of the involved subject. Aim of this paper is to report a series of patients treated at our institutions, to verify neurological results at six months or survival; in fatal cases we searched for βAPP, GFAP, IL-1β, NFL, Spectrin II, TUNEL and miR-21, miR-16, and miR-92 expressions in brain samples, to verify DAI diagnosis and grade as strong predictor of survival and inflammatory response. Concentrations of 8OHdG as measurement of oxidative stress was performed. Immunoreaction of β-APP, IL-1β, GFAP, NFL, Spectrin II and 8OHdG were significantly increased in the TBI group with respect to control group subjects. Cell apoptosis, measured by TUNEL assay, were significantly higher in the study group than control cases. Results indicated that miR-21, miR-92 and miR-16 have a high predictive power in discriminating trauma brain cases from controls and could represent promising biomarkers as strong predictor of survival, and for the diagnosis of postmortem traumatic brain injury

Helical Chirality: a Link between Local Interactions and Global Topology in DNA

DNA supercoiling plays a major role in many cellular functions. The global DNA conformation is however intimately linked to local DNA-DNA interactions influencing both the physical properties and the biological functions of the supercoiled molecule. Juxtaposition of DNA double helices in ubiquitous crossover arrangements participates in multiple functions such as recombination, gene regulation and DNA packaging. However, little is currently known about how the structure and stability of direct DNA-DNA interactions influence the topological state of DNA. Here, a crystallographic analysis shows that due to the intrinsic helical chirality of DNA, crossovers of opposite handedness exhibit markedly different geometries. While right-handed crossovers are self-fitted by sequence-specific groove-backbone interaction and bridging Mg2+ sites, left-handed crossovers are juxtaposed by groove-groove interaction. Our previous calculations have shown that the different geometries result in differential stabilisation in solution, in the presence of divalent cations. The present study reveals that the various topological states of the cell are associated with different inter-segmental interactions. While the unstable left-handed crossovers are exclusively formed in negatively supercoiled DNA, stable right-handed crossovers constitute the local signature of an unusual topological state in the cell, such as the positively supercoiled or relaxed DNA. These findings not only provide a simple mechanism for locally sensing the DNA topology but also lead to the prediction that, due to their different tertiary intra-molecular interactions, supercoiled molecules of opposite signs must display markedly different physical properties. Sticky inter-segmental interactions in positively supercoiled or relaxed DNA are expected to greatly slow down the slithering dynamics of DNA. We therefore suggest that the intrinsic helical chirality of DNA may have oriented the early evolutionary choices for DNA topology

Smallholder farmers' adaptation to climate change and determinants of their adaptation decisions in the Central Rift Valley of Ethiopia

Background: The agricultural sector remains the main source of livelihoods for rural communities in Ethiopia, but faces the challenge of changing climate. This study investigated how smallholder farmers perceive climate change, what adaptation strategies they practice, and factors that influence their adaptation decisions. Both primary and secondary data were used for the study, and a multinomial logit model was employed to identify the factors that shape smallholder farmers’ adaptation strategies. Results: The results show that 90% of farmers have already perceived climate variability, and 85% made attempts to adapt using practices like crop diversification, planting date adjustment, soil and water conservation and management, increasing the intensity of input use, integrating crop with livestock, and tree planting. The econometric model indicated that education, family size, gender, age, livestock ownership, farming experience, frequency of contact with extension agents, farm size, access to market, access to climate information and income were the key factors determining farmers’ choice of adaptation practice. Conclusion: In the Central Rift Valley of Ethiopia, climate change is a pressing problem, which is beyond the capacity of smallholders to respond to autonomously. Farmers’ capacity to choose effective adaptation options is influenced by household demography, as well as positively by farm size, income, access to markets, access to climate information and extension, and livestock production. This implies the need to support the indigenous adaptation strategies of the smallholder farmers with a wide range of institutional, policy, and technology support; some of it targeted on smaller, poorer or female-headed households. Moreover, creating opportunities for non-farm income sources is important as this helps farmers to engage in those activities that are less sensitive to climate change. Furthermore, providing climate change information, extension services, and creating access to markets are crucial

Common Variants at 10 Genomic Loci Influence Hemoglobin A(1C) Levels via Glycemic and Nonglycemic Pathways

OBJECTIVE Glycated hemoglobin (HbA1c), used to monitor and diagnose diabetes, is influenced by average glycemia over a 2- to 3-month period. Genetic factors affecting expression, turnover, and abnormal glycation of hemoglobin could also be associated with increased levels of HbA1c. We aimed to identify such genetic factors and investigate the extent to which they influence diabetes classification based on HbA1c levels. RESEARCH DESIGN AND METHODS We studied associations with HbA1c in up to 46,368 nondiabetic adults of European descent from 23 genome-wide association studies (GWAS) and 8 cohorts with de novo genotyped single nucleotide polymorphisms (SNPs). We combined studies using inverse-variance meta-analysis and tested mediation by glycemia using conditional analyses. We estimated the global effect of HbA1c loci using a multilocus risk score, and used net reclassification to estimate genetic effects on diabetes screening. RESULTS Ten loci reached genome-wide significant association with HbA1c, including six new loci near FN3K (lead SNP/P value, rs1046896/P = 1.6 × 10−26), HFE (rs1800562/P = 2.6 × 10−20), TMPRSS6 (rs855791/P = 2.7 × 10−14), ANK1 (rs4737009/P = 6.1 × 10−12), SPTA1 (rs2779116/P = 2.8 × 10−9) and ATP11A/TUBGCP3 (rs7998202/P = 5.2 × 10−9), and four known HbA1c loci: HK1 (rs16926246/P = 3.1 × 10−54), MTNR1B (rs1387153/P = 4.0 × 10−11), GCK (rs1799884/P = 1.5 × 10−20) and G6PC2/ABCB11 (rs552976/P = 8.2 × 10−18). We show that associations with HbA1c are partly a function of hyperglycemia associated with 3 of the 10 loci (GCK, G6PC2 and MTNR1B). The seven nonglycemic loci accounted for a 0.19 (% HbA1c) difference between the extreme 10% tails of the risk score, and would reclassify ∼2% of a general white population screened for diabetes with HbA1c. CONCLUSIONS GWAS identified 10 genetic loci reproducibly associated with HbA1c. Six are novel and seven map to loci where rarer variants cause hereditary anemias and iron storage disorders. Common variants at these loci likely influence HbA1c levels via erythrocyte biology, and confer a small but detectable reclassification of diabetes diagnosis by HbA1c

The Distances of the Magellanic Clouds

The present status of our knowledge of the distances to the Magellanic Clouds is evaluated from a post-Hipparcos perspective. After a brief summary of the effects of structure, reddening, age and metallicity, the primary distance indicators for the Large Magellanic Cloud are reviewed: The SN 1987A ring, Cepheids, RR Lyraes, Mira variables, and Eclipsing Binaries. Distances derived via these methods are weighted and combined to produce final "best" estimates for the Magellanic Clouds distance moduli.Comment: Invited review article to appear in ``Post Hipparcos Cosmic Candles'', F. Caputo & A. Heck (Eds.), Kluwer Academic Publ., Dordrecht, in pres