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Purpose: To report 2-year outcomes of trans-epithelial accelerated corneal collagen cross-linking (TE-ACXL) procedure in the treatment of progressive keratoconus patients. Patients and Methods: Twenty-four eyes from 24 patients who underwent TE-ACXL (6mW/[cm.sup.2] for 15 minutes) were included in this retrospective interventional study. Best-corrected visual acuity (BCVA), keratometry values, thinnest corneal thickness (PachyMin) and topometric indexes were analysed preoperatively and at 6-month, 12-month, 18-month and 24-month postoperative. Progression was assessed by increase [greater than or equal to]1.00D in maximum keratometry (Kmax); increase [greater than or equal to]1.00D in corneal astigmatism; decrease [greater than or equal to]2% in PachyMin; increase [greater than or equal to]0.42 in D-index. Results: There were no complications during or after TE-ACXL. No significant differences ([DELTA]) were observed between baseline and 12-month or 24-month postoperative: [DELTA]BCVA (-0.08 [+ or -] 0.25, p=0.190; -0.04 [+ or -] 0.17, p = 0.588), [DELTA]Kmax (-0.08 [+ or -] 1.32, p=0.792; -1.04 [+ or -] 1.89, p=0.135), [DELTA]Astigmatism (-0.15 [+ or -] 0.89, p=0.485; -0.24 [+ or -] 1.38, p=0.609), [DELTA]PachyMin (-0.56 [+ or -] 15.70, p=0.882; 0.56 [+ or -] 18.74, p=0.931), [DELTA]Index Surface Variation ([DELTA]ISV) (-2.11 [+ or -] 10.27, p=0.395; -4.67 [+ or -] 17.32, [DELTA]=0.442), [DELTA]Index Vertical Asymmetry ([DELTA]IVA) (-0.05 [+ or -] 0.17, p=0.208; -0.08 [+ or -] 0.26, p=0.397), [DELTA]Index Height Decentration ([DELTA]IHD) (0.00 [+ or -] 0.02, p=0.368; -0.01 [+ or -] 0.04, p=0.484), [DELTA]KI (0.00 [+ or -] 0.05, p=0.851; 0.01 [+ or -] 0.06, p=0.877) and [DELTA]D-index (0.15 [+ or -] 1.14, p=0.512; 0.06 [+ or -] 1.36, p=0.892). Eleven to 33% of patients had disease progression at 24-month postoperative according to the parameters used to determine progression. Conclusion: Although some patients maintain disease progression, TE-ACXL seems to be a safe and effective treatment for keratoconus over the 2-year follow-up period. Studies with longer follow-up periods and larger patient cohorts are recommended. Keywords: cornea, keratoconus, disease progression, transepithelial, cross-linking

Abstract Phytoremediation is an ecofriendly technique to clean heavy metals from contaminated soil by the use of high biomass producing plant species. Chelators can help to improve this biological technique by increasing metal solubility. Therefore, a pot experiment was conducted to determine the effect of the chelators EDTA and citric acid (CA) in phytoremediation of Ni contaminated soil by using Brassica napus (canola). Two cultivars of B. napus, Con-II (tolerant) and Oscar (sensitive), were selected after screening and exposed to NiSO4 at 30 ppm at the time of sowing. CA (10 mM) and EDTA (1.5 mM) were applied either alone or in combination with each other after two weeks of Ni treatments. Different parameters like morpho-physiological and biochemical data were recorded after 15 days of chelate application. The results highlighted the successful use of chelating agents (CA and EDTA) not only to ameliorate Ni stress but also to enhance Ni accumulation which is prerequisite for phytoremediation. The basal application of 10 mMCA and 1.5 mM EDTA concentration proved to be effective for the growth of plants. The combination of chelating agents failed to show any synergistic effects. Resumo A fitorremediação é uma técnica ecologicamente correta para limpar metais pesados de solo contaminado pelo uso de espécies vegetais produtoras de alta biomassa. Os quelantes podem ajudar a melhorar esta técnica biológica aumentando a solubilidade do metal. Para tanto, foi realizado um experimento em vaso para determinar o efeito dos quelantes EDTA e ácido cítrico (AC) na fitorremediação de solo contaminado com Ni, utilizando Brassica napus (canola). Duas cultivares de B. napus, Con-II (tolerante) e Oscar (sensível) foram selecionadas após triagem e expostas a NiSO4 a 30 ppm no momento da semeadura. CA (10 mM) e EDTA (1,5 mM) foram aplicados sozinhos ou em combinação um com o outro após duas semanas de tratamentos com Ni. Diferentes parâmetros como dados morfofisiológicos e bioquímicos foram registrados após 15 dias de aplicação de quelato. Os resultados destacaram o uso bem-sucedido de agentes quelantes (CA e EDTA) não apenas para melhorar o estresse de Ni, mas também para aumentar o acúmulo de Ni, um pré-requisito para a fitorremediação. A aplicação basal da concentração de 10 mMCA e 1,5 mM de EDTA mostrou-se eficaz para o crescimento das plantas. A combinação de agentes quelantes não mostrou quaisquer efeitos sinérgicos.

The ADAM (a disintegrin and metalloprotease) protein family uniquely exhibits both catalytic and adhesive properties. In the well-defined process of ectodomain shedding, ADAMs transform latent, cell-bound substrates into soluble, biologically active derivatives to regulate a spectrum of normal and pathological processes. In contrast, the integrin ligand properties of ADAMs are not fully understood. Emerging models posit that ADAM-integrin interactions regulate shedding activity by localizing or sequestering the ADAM sheddase. Interestingly, 8 of the 21 human ADAMs are predicted to be catalytically inactive. Unlike their catalytically active counterparts, integrin recognition of these \"dead\" enzymes has not been largely reported. The present study delineates the integrin ligand properties of a group of non-catalytic ADAMs. Here we report that human ADAM11, ADAM23, and ADAM29 selectively support integrin [alpha]4-dependent cell adhesion. This is the first demonstration that the disintegrin-like domains of multiple catalytically inactive ADAMs are ligands for a select subset of integrin receptors that also recognize catalytically active ADAMs.

Porcine reproductive and respiratory syndrome (PRRS) caused by PRRS virus (PRRSV) is one of the most severe swine diseases that affects almost all swine-breeding countries. Nonstructural protein 2 (NSP2) is one of the most important viral proteins in the PRRSV life cycle. Our previous study showed that PRRSV NSP2 could induce the formation of aggresomes. In this study we explored the effects of aggresome formation on cells and found that NSP2 could induce autophagy, which depended on aggresome formation to activate aggrephagy. The transmembrane and tail domains of NSP2 contributed to aggrephagy and the cellular protein 14-3-3[epsilon] played an important role in NSP2-induced autophagy by binding the tail domain of NSP2. These findings provide information on the function of the C-terminal domain of NSP2, which will help uncover the function of NSP2 during PRRSV infection. Keywords: PRRSV, NSP2, Aggrephagy, 14-3-3[epsilon], Tail domain of NSP2

Background Sequencing technology and assembly algorithms have matured to the point that high-quality de novo assembly is possible for large, repetitive genomes. Current assemblies traverse transposable elements (TEs) and provide an opportunity for comprehensive annotation of TEs. Numerous methods exist for annotation of each class of TEs, but their relative performances have not been systematically compared. Moreover, a comprehensive pipeline is needed to produce a non-redundant library of TEs for species lacking this resource to generate whole-genome TE annotations. Results We benchmark existing programs based on a carefully curated library of rice TEs. We evaluate the performance of methods annotating long terminal repeat (LTR) retrotransposons, terminal inverted repeat (TIR) transposons, short TIR transposons known as miniature inverted transposable elements (MITEs), and Helitrons. Performance metrics include sensitivity, specificity, accuracy, precision, FDR, and F 1 . Using the most robust programs, we create a comprehensive pipeline called Extensive de-novo TE Annotator (EDTA) that produces a filtered non-redundant TE library for annotation of structurally intact and fragmented elements. EDTA also deconvolutes nested TE insertions frequently found in highly repetitive genomic regions. Using other model species with curated TE libraries (maize and Drosophila), EDTA is shown to be robust across both plant and animal species. Conclusions The benchmarking results and pipeline developed here will greatly facilitate TE annotation in eukaryotic genomes. These annotations will promote a much more in-depth understanding of the diversity and evolution of TEs at both intra- and inter-species levels. EDTA is open-source and freely available: https://github.com/oushujun/EDTA .

Chromosomal aberrations are relatively frequent pathologies in both humans and animals. Among them, translocations present a specific meiotic segregation pattern able to give a higher percentage of unbalanced gametes that can induce fertility problems. In this study, the meiotic segregation patterns of 1p, 1q and 18 Bubalus bubalis chromosomes were analyzed in both total sperm fraction and motile sperm fraction of a t(1p;18) carrier and a control bulls by triple-color FISH analysis with a pool of specific BAC probes. The frequencies of each total sperm fraction products in the carrier resulting from alternate, adjacent I, adjacent II and 3:1 segregation were 39%, 20%, 1% and 38%, respectively. On the other hand, the frequencies of each motile sperm fraction products in the carrier resulting from alternate, adjacent I, adjacent II and 3:1 segregation were 93%, 5%, 0% and 2%, respectively. The frequencies of normal sperms in the carrier were 27% and 69% in total sperm fraction and motile sperm fraction, respectively. The frequencies detected in motile sperm fraction were also validated by comparison with bull's progeny. To our knowledge, this is the first report on the meiotic segregation patterns in motile sperm fractions of B. bubalis bull carrying a chromosomal translocation. These data suggest that translocation has a very limited effect on aneuploidy in the gametes, and therefore, on the reproductive abilities of the bull.

Reducing Amyloid [beta] (A[beta]) in the brain is of fundamental importance for advancing the therapeutics for Alzheimer`s disease. The endogenous metallopeptidase neprilysin (NEP) has been identified as one of the key A[beta]-degrading enzymes. Delivery of NEP to the brain by utilizing the Brain Shuttle (BS) transport system offers a promising approach for clearing central A[beta]. We fused the extracellular catalytic domain of NEP to an active or inactive BS module. The two BS-NEP constructs were used to investigate the pharmacokinetic/pharmacodynamics relationships in the blood and the cerebrospinal fluid (CSF) in dose-response and multiple dosing. As previously shown, NEP was highly effective at degrading A[beta] in blood but not in the CSF compartment after systemic administration. In contrast, the NEP with an active BS module led to a significant CSF exposure of BS-NEP, followed by substantial A[beta] reduction in CSF and brain parenchyma. Our data show that a BS module against the transferrin receptor facilitates the transport of an A[beta] degrading enzyme across the blood-brain barriers to efficiently reduce A[beta] levels in both CSF and brain.

Biomolecular self-assembly is an emerging bottom-up approach for the synthesis of novel nanomaterials. DNA and viruses have both been used to create scaffolds but the former lacks chemical diversity and the latter lack spatial control. To date, the use of protein scaffolds to template materials on the nanoscale has focused on amyloidogenic proteins that are known to form fibrils or two-protein systems where a second protein acts as a cross-linker. We previously developed a unique approach for self-assembly of nanomaterials based on engineering [beta]-solenoid proteins (BSPs) to polymerize into micrometer-length fibrils. BSPs have highly regular geometries, tunable lengths, and flat surfaces that are amenable to engineering and functionalization. Here, we present a newly engineered BSP based on the antifreeze protein of the beetle Rhagium inquisitor (RiAFP-m9), which polymerizes into stable fibrils under benign conditions. Gold nanoparticles were used to functionalize the RiAFP-m9 fibrils as well as those assembled from the previously described SBAFP-m1 protein. Cysteines incorporated into the sequences provide site-specific gold attachment. Additionally, silver was deposited on the gold-labelled fibrils by electroless plating to create nanowires. These results bolster prospects for programable self-assembly of BSPs to create scaffolds for functional nanomaterials.

With the objective of assisting in the understanding of the chromosome evolution of Pentatomomorpha and in the quest to understand how the genome organizes/reorganizes for the chromosomal position of the 45S rDNA in this infraorder, we analyzed 15 species (it has being 12 never studied before by FISH) of Pentatomomorpha with the probe of 18S rDNA. The mapping of the 45S gene in the Coreidae family demonstrated that the species presented markings on the autosomes, with the exception of Acanthocephala parensis and Leptoglossus gonagra that showed markers on m-chromosomes. Most species of the Pentatomidae family showed marking in the autosomes, except for two species that had 45S rDNA on X sex chromosome (Odmalea sp. and Graphosoma lineatum) and two that showed marking on the X and Y sex chromosomes. Species of the Pyrrhocoridae family showed 18S rDNA markers in autosomes, X chromosome as well as in Neo X. The Largidae and Scutelleridae families were represented by only one species that showed marking on the X sex chromosome and on a pair of autosomes, respectively. Based on this, we characterized the arrangement of 45S DNAr in the chromosomes of 12 new species of Heteroptera and discussed the main evolutionary events related to the genomic reorganization of these species during the events of chromosome and karyotype evolution in Pentatomomorpha infraorder.