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Bacillus thuringiensis (Bt) has been successfully used commercially for more than 60 years for biocontrol of insect pests. Since 1996, transgenic plants expressing Bt crystal (Cry) proteins have been used commercially to provide protection against insects that predate on corn and cotton. More recently, Bt Cry proteins that target nematodes have been discovered. One of these, Cry14Ab, has been expressed in transgenic soybean plants and found to provide significant protection against the soybean cyst nematode, Heterodera glycines. However, to date there has been no description of high-level resistance to any Cry14A family protein in nematodes. Here, we describe forward genetic screens to identify such mutants using the nematode Caenorhabditis elegans. Although non-conditional screens failed to identify highly resistant C. elegans, a conditional (temperature-sensitive) genetic screen identified one mutant, bre-6(ye123) (for Bt protein resistant), highly resistant to both Cry14Aa and Cry14Ab. The mutant comes at a high fitness cost, showing significant delays in growth and development and reduced fecundity. bre-6(ye123) hermaphrodites are only weakly resistant to copper intoxication, indicating that the mutant is not highly resistant to all insults. Backcrossing-whole genome sequencing was used to identify the gene mutated in ye123 as the nuclear hormone receptor nhr-31. RNAi, DNA rescue, and CRISPR analyses confirm that resistance to Cry14Aa intoxication in bre-6(ye123) is due to mutation of nhr-31 and was renamed nhr-31(ye123). As predicted for a mutation in this gene, nhr-31(ye123) animals showed significantly reduced expression of most of the subunits of the C. elegans vacuolar ATPase (vATPase). Mutants in the vATPase subunits unc-32 and vha-7 also show resistance to Cry14Aa and/or Cry14Ab. These data demonstrate that nhr-31 and the vATPase play a significant role in the intoxication of C. elegans by Cry14A family proteins, that reduction in vATPase levels result in high resistance to Cry14A family proteins, and that such resistance comes at a high fitness cost. Based on the relative difficulty of finding resistant mutants and the fitness cost associated with the vATPase pathway, our data suggest that transgenic Cry14Ab plants may hold up well to resistance by nematode parasites., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Kim et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Leishmaniasis, a critical Neglected Tropical Disease caused by Leishmania protozoa, represents a significant global health risk, particularly in resource-limited regions. Conventional treatments are effective but suffer from serious limitations, such as toxicity, prolonged treatment courses, and rising drug resistance. Herein, we highlight the potential of inorganic nanomaterials as an innovative approach to enhance Leishmaniasis therapy, aligning with the One Health concept by considering these treatments' environmental, veterinary, and public health impacts. By leveraging the adjustable properties of these nanomaterials─including size, shape, and surface charge, tailored treatments for various diseases can be developed that are less harmful to the environment and nontarget species. We review recent advances in metal-, oxide-, and carbon-based nanomaterials for combating Leishmaniasis, examining their mechanisms of action and their dual use as standalone treatments or drug delivery systems. Our analysis highlights a promising yet underexplored frontier in employing these materials for more holistic and effective disease management.

Retroviruses and closely related LTR retrotransposons export full-length, unspliced genomic RNA (gRNA) for packaging into virions and to serve as the mRNA encoding GAG and POL polyproteins. Because gRNA often includes splice acceptor and donor sequences used to splice viral mRNAs, retroelements must overcome host mechanisms that retain intron-containing RNAs in the nucleus. Here we examine gRNA expression in Cer1, an LTR retrotransposon in C. elegans which somehow avoids silencing and is highly expressed in germ cells. Newly exported Cer1 gRNA associates rapidly with the Cer1 GAG protein, which has structural similarity with retroviral GAG proteins. gRNA export requires CERV (C. elegans regulator of viral expression), a novel protein encoded by a spliced Cer1 mRNA. CERV phosphorylation at S214 is essential for gRNA export, and phosphorylated CERV colocalizes with nuclear gRNA at presumptive sites of transcription. By electron microscopy, tagged CERV proteins surround clusters of distinct, linear fibrils that likely represent gRNA molecules. Single fibrils, or groups of aligned fibrils, also localize near nuclear pores. During the C. elegans self-fertile period, when hermaphrodites fertilize oocytes with their own sperm, CERV concentrates in two nuclear foci that are coincident with gRNA. However, as hermaphrodites cease self-fertilization, and can only produce cross-progeny, CERV undergoes a remarkable transition to form giant nuclear rods or cylinders that can be up to 5 microns in length. We propose a novel mechanism of rod formation, in which stage-specific changes in the nucleolus induce CERV to localize to the nucleolar periphery in flattened streaks of protein and gRNA; these streaks then roll up into cylinders. The rods are a widespread feature of Cer1 in wild strains of C. elegans, but their function is not known and might be limited to cross-progeny. We speculate that the adaptive strategy Cer1 uses for the identical self-progeny of a host hermaphrodite might differ for heterozygous cross-progeny sired by males. For example, mating introduces male chromosomes which can have different, or no, Cer1 elements., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Sun et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Argonaute/small RNA pathways and heterochromatin work together to propagate transgenerational gene silencing, but the mechanisms behind their interaction are not well understood. Here, we show that induction of heterochromatin silencing in C. elegans by RNAi or by artificially tethering pathway components to target RNA causes co-localization of target alleles in pachytene nuclei. Tethering the nuclear Argonaute WAGO-9/HRDE-1 induces heterochromatin formation and independently induces small RNA amplification. Consistent with this finding, HRDE-1, while predominantly nuclear, also localizes to peri-nuclear nuage domains, where amplification is thought to occur. Tethering a heterochromatin-silencing factor, NRDE-2, induces heterochromatin formation, which subsequently causes de novo synthesis of HRDE-1 guide RNAs. HRDE-1 then acts to further amplify small RNAs that load on downstream Argonautes. These findings suggest that HRDE-1 plays a dual role, acting upstream to initiate heterochromatin silencing and downstream to stimulate a new cycle of small RNA amplification, thus establishing a self-enforcing mechanism that propagates gene silencing to future generations., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Germline Argonautes direct transcriptome surveillance within perinuclear membraneless organelles called nuage. In C. elegans, a family of Vasa-related Germ Line Helicase (GLH) proteins localize in and promote the formation of nuage. Previous studies have implicated GLH proteins in inherited silencing, but direct roles in small-RNA production, Argonaute binding, or mRNA targeting have not been identified. Here we show that GLH proteins compete with each other to control Argonaute pathway specificity, bind directly to Argonaute target mRNAs, and promote the amplification of small RNAs required for transgenerational inheritance. We show that the ATPase cycle of GLH-1 regulates direct binding to the Argonaute WAGO-1, which engages amplified small RNAs. Our findings support a dynamic and direct role for GLH proteins in inherited silencing beyond their role as structural components of nuage., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Nuclease-directed genome editing is a powerful tool for investigating physiology and has great promise as a therapeutic approach to correct mutations that cause disease. In its most precise form, genome editing can use cellular homology-directed repair (HDR) pathways to insert information from an exogenously supplied DNA-repair template (donor) directly into a targeted genomic location. Unfortunately, particularly for long insertions, toxicity and delivery considerations associated with repair template DNA can limit HDR efficacy. Here, we explore chemical modifications to both double-stranded and single-stranded DNA-repair templates. We describe 5'-terminal modifications, including in its simplest form the incorporation of triethylene glycol (TEG) moieties, that consistently increase the frequency of precision editing in the germlines of three animal models ( Caenorhabditis elegans , zebrafish, mice) and in cultured human cells., Competing Interests: KG, GD, HG, ES, JW Co-inventor on patent applications related to this work (Application number: 16/384, 612 ), ZC, PK, YY, JG, PX, XZ, AO, MS, FI, PL, AE, NL, JR No competing interests declared, AM Co-inventor on patent application related to this work (Application number: 16/384, 612 ), CM The authors (K.S.G, A.M, G.A.D, H.G, J.K.W, E.J.S and C.C.M) have a patent application pending related to the findings described (Application number: 16/384, 612 ). Craig C. Mello is a co-founder and Scientific Advisory Board member of CRISPR Therapeutics, and Erik J. Sontheimer is a cofounder and Scientific Advisory Board member of Intellia herapeutics., (© 2021, Ghanta et al.)

In animals, Argonaute small-RNA pathways scan germline transcripts to silence self-replicating genetic elements. However, little is known about how endogenous gene expression is recognized and licensed. Here, we show that the presence of introns and, by inference, the process of mRNA splicing prevents default Argonaute-mediated silencing in the C. elegans germline. The silencing of intronless genes is initiated independently of the piRNA pathway but nevertheless engages multiple components of the downstream amplification and maintenance mechanisms that mediate transgenerational silencing, including both nuclear and cytoplasmic members of the worm-specific Argonaute gene family (WAGOs). Small RNAs amplified from intronless mRNAs can trans-silence cognate intron-containing genes. Interestingly, a second, small RNA-independent cis-acting mode of silencing also acts on intronless mRNAs. Our findings suggest that cues put in place during mRNA splicing license germline gene expression and provide evidence for a splicing-dependent and dsRNA- and piRNA-independent mechanism that can program Argonaute silencing., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

In Caenorhabditis elegans, targeted genome editing techniques are now routinely used to generate germline edits. The remarkable ease of C. elegans germline editing is attributed to the syncytial nature of the pachytene ovary which is easily accessed by microinjection. This protocol describes the step-by-step details and troubleshooting tips for the entire CRISPR-Cas genome editing procedure, including gRNA design and microinjection of ribonucleoprotein complexes, followed by screening and genotyping in C. elegans , to help accessing this powerful genetic animal system. For complete details on the use and execution of this protocol, please refer to Ghanta and Mello (2020)., Competing Interests: Some of the findings described here are part of the patent applications filed by the University of Massachusetts Medical School with K.S.G and C.C.M. as co-inventors. C.C.M. is a scientific founder and advisory board member of CRISPR therapeutics., (© 2021.)

Germlines shape and balance heredity, integrating and regulating information from both parental and foreign sources. Insights into how germlines handle information have come from the study of factors that specify or maintain the germline fate. In early Caenorhabditis elegans embryos, the CCCH zinc finger protein PIE-1 localizes to the germline where it prevents somatic differentiation programs. Here, we show that PIE-1 also functions in the meiotic ovary where it becomes SUMOylated and engages the small ubiquitin-like modifier (SUMO)-conjugating machinery. Using whole-SUMO-proteome mass spectrometry, we identify HDAC SUMOylation as a target of PIE-1. Our analyses of genetic interactions between pie-1 and SUMO pathway mutants suggest that PIE-1 engages the SUMO machinery both to preserve the germline fate in the embryo and to promote Argonaute-mediated surveillance in the adult germline., Competing Interests: HK, YD, SL, MZ, WT, DC, MD, CM No competing interests declared, (© 2021, Kim et al.)

Eukaryotic cells use guided search to coordinately control dispersed genetic elements. Argonaute proteins and their small RNA cofactors engage nascent RNAs and chromatin-associated proteins to direct transcriptional silencing. The small ubiquitin-like modifier (SUMO) has been shown to promote the formation and maintenance of silent chromatin (called heterochromatin) in yeast, plants, and animals. Here, we show that Argonaute-directed transcriptional silencing in Caenorhabditis elegans requires SUMOylation of the type 1 histone deacetylase HDA-1. Our findings suggest how SUMOylation promotes the association of HDAC1 with chromatin remodeling factors and with a nuclear Argonaute to initiate de novo heterochromatin silencing., Competing Interests: HK, YD, GZ, YY, DC, MD, CM No competing interests declared, (© 2021, Kim et al.)

In several species, Piwi/piRNA genome silencing defects cause immediate sterility that correlates with transposon expression and transposon-induced genomic instability. In C. elegans, mutations in the Piwi-related gene (prg-1) and other piRNA deficient mutants cause a transgenerational decline in fertility over a period of several generations. Here we show that the sterility of late generation piRNA mutants correlates poorly with increases in DNA damage signaling. Instead, sterile individuals consistently exhibit altered perinuclear germ granules. We show that disruption of germ granules does not activate transposon expression but induces multiple phenotypes found in sterile prg-1 pathway mutants. Furthermore, loss of the germ granule component pgl-1 enhances prg-1 mutant infertility. Environmental restoration of germ granule function for sterile pgl-1 mutants restores their fertility. We propose that Piwi mutant sterility is a reproductive arrest phenotype that is characterized by perturbed germ granule structure and is phenocopied by germ granule dysfunction, independent of genomic instability.

Eukaryotic cells regulate 5'-triphosphorylated RNAs (ppp-RNAs) to promote cellular functions and prevent recognition by antiviral RNA sensors. For example, RNA capping enzymes possess triphosphatase domains that remove the γ phosphates of ppp-RNAs during RNA capping. Members of the closely related PIR-1 (phosphatase that interacts with RNA and ribonucleoprotein particle 1) family of RNA polyphosphatases remove both the β and γ phosphates from ppp-RNAs. Here, we show that C. elegans PIR-1 dephosphorylates ppp-RNAs made by cellular RNA-dependent RNA polymerases (RdRPs) and is required for the maturation of 26G-RNAs, Dicer-dependent small RNAs that regulate thousands of genes during spermatogenesis and embryogenesis. PIR-1 also regulates the CSR-1 22G-RNA pathway and has critical functions in both somatic and germline development. Our findings suggest that PIR-1 modulates both Dicer-dependent and Dicer-independent Argonaute pathways and provide insight into how cells and viruses use a conserved RNA phosphatase to regulate and respond to ppp-RNA species., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2020 Elsevier Inc. All rights reserved.)

CRISPR genome editing has revolutionized genetics in many organisms. In the nematode Caenorhabditis elegans , one injection into each of the two gonad arms of an adult hermaphrodite exposes hundreds of meiotic germ cells to editing mixtures, permitting the recovery of multiple indels or small precision edits from each successfully injected animal. Unfortunately, particularly for long insertions, editing efficiencies can vary widely, necessitating multiple injections, and often requiring coselection strategies. Here, we show that melting double-stranded DNA (dsDNA) donor molecules prior to injection increases the frequency of precise homology-directed repair (HDR) by several fold for longer edits. We describe troubleshooting strategies that enable consistently high editing efficiencies resulting, for example, in up to 100 independent GFP knock-ins from a single injected animal. These efficiencies make C. elegans by far the easiest metazoan to genome edit, removing barriers to the use and adoption of this facile system as a model for understanding animal biology., (Copyright © 2020 Ghanta and Mello.)

GCNA proteins are expressed across eukarya in pluripotent cells and have conserved functions in fertility. GCNA homologs Spartan (DVC-1) and Wss1 resolve DNA-protein crosslinks (DPCs), including Topoisomerase-DNA adducts, during DNA replication. Here, we show that GCNA mutants in mouse and C. elegans display defects in genome maintenance including DNA damage, aberrant chromosome condensation, and crossover defects in mouse spermatocytes and spontaneous genomic rearrangements in C. elegans. We show that GCNA and topoisomerase II (TOP2) physically interact in both mice and worms and colocalize on condensed chromosomes during mitosis in C. elegans embryos. Moreover, C. elegans gcna-1 mutants are hypersensitive to TOP2 poison. Together, our findings support a model in which GCNA provides genome maintenance functions in the germline and may do so, in part, by promoting the resolution of TOP2 DPCs., (Copyright © 2019 Elsevier Inc. All rights reserved.)

The purpose of this in vitro study was to measure the vertical, positive-horizontal, and negative-horizontal misfit (VM, PHM, and NHM, respectively) of the zirconia three-element prosthetic framework, fabricated using different methods, and compare them with conventional fabrication methods (lost-wax casting). Furthermore, this study aimed to evaluate the influence of the misfit values on the biomechanical behavior of the 3-unit fixed prosthetic frameworks using three-dimensional finite element analysis (3D-FEA). Forty frameworks (n = 10) were fabricated as follow: G1, Cerec Bluecam; G2, iTero; G3, 3Series; and G4, conventional method. The samples were randomized to measure marginal misfit using a high-precision three-dimensional (3D)-optical microscope. The results were submitted to analysis of variance (ANOVA), with the significance level set at 5%. The mean VM values of each group were used in creating the models by 3D-FEA with the misfit found in optical microscopy. The programs used were the InVesalius, Rhinoceros, SolidWorks, FEMAP and NEiNastran. The von Mises map was plotted for each model. The G4 showed the lowest mean VM value (16.73 μm), followed by G3 (20.71 μm), G2 (21.01 μm), and G1 (41.77 μm) (p < 0.001). G2 was more accurate than G1 (p < 0.05) and similar to G3 (p = 0.319). For PHM, G4 was the most accurate and did not present overextended values. With regard to NHM, the computer-aided design and computer-aided manufacturing (CAD-CAM) systems were more accurate (-61.91 μm) than G4 (-95.36 μm) (p = 0.014). In biomechanical analysis, stress concentration caused by oblique loading is greater than caused by axial loading. In axial loading, G4 was the most favorable while G1 was the least favorable, biomechanically, in oblique loading, similar stress patterns were observed in all the models. The prosthetic screw was the most overloaded structure, but the material did not influence the stress distribution. The misfit prostheses showed a greater degree of stress than the controls (without misfit). The manufacturing method influenced the marginal misfit of the frameworks, with the conventional method being the most accurate and the Cerec Bluecam System (closed system) the least accurate. Biomechanically, fitting prostheses were more favorable than misfit prostheses., (Copyright © 2019. Published by Elsevier B.V.)

Purpose: To compare the marginal vertical misfit between implant-supported frameworks fabricated using CAD/CAM systems and the conventional technique (lost-wax casting)., Materials and Methods: This review was performed according to PRISMA criteria and registered on PROSPERO (CRD42017055685). An electronic search was performed independently by two examiners in the MEDLINE (Pubmed), Embase, Web of Science, and Cochrane Library databases to find studies published up to April 2018., Results: The database search yielded 507 references. After removing duplicate references, 384 studies remained. Eleven in vitro studies were selected according to the eligibility criteria (inter-reader κ = 0.88). Nine different CAD/CAM systems were used to fabricate 172 frameworks of different materials, including zirconia, monolithic lithium disilicate, and metallic alloys. Subgroup analyses were performed for different types and retention systems of the frameworks. In the general analysis, marginal misfit observed with the CAD/CAM systems was lower than with the conventional method (P = .003), as was observed in the subgroup analysis for single-unit frameworks (P < .00001). For fixed (P = .89), cemented (P = .60), and screwed (P = .18) frameworks, no significant difference was observed between the evaluated techniques., Conclusion: The CAD/CAM systems showed improved marginal fit over the conventional lost-wax casting technique for fabricating single-unit frameworks; however, in the subgroup analyses, no difference was observed for the fixed implant-supported type or for the retention systems evaluated.

CRISPR-based genome editing using ribonucleoprotein complexes and synthetic single-stranded oligodeoxynucleotide (ssODN) donors can be highly effective. However, reproducibility can vary, and precise, targeted integration of longer constructs-such as green fluorescent protein tags remains challenging in many systems. Here, we describe a streamlined and optimized editing protocol for the nematode Caenorhabditis elegans We demonstrate its efficacy, flexibility, and cost-effectiveness by affinity-tagging 14 Argonaute proteins in C. elegans using ssODN donors. In addition, we describe a novel PCR-based, partially single-stranded, "hybrid" donor design that yields high efficiency editing with large (kilobase-scale) constructs. We use these hybrid donors to introduce fluorescent protein tags into multiple loci, achieving editing efficiencies that approach those previously obtained only with much shorter ssODN donors. The principals and strategies described here are likely to translate to other systems, and should allow researchers to reproducibly and efficiently obtain both long and short precision genome edits., (Copyright © 2018 G. A. Dokshin et al.)

Purpose: The objective of this systematic review with meta-analysis was to evaluate surface treatment in yttria-stabilized tetragonal zirconia polycrystal ceramic (Y-TZP) on the shear bond strength (SBS) values between Y-TZP (core ceramic) and veneer ceramic, compared to untreated specimens., Materials and Methods: This review was registered at PROSPERO platform under the number CRD42016036493. The systematic review of the extracted publications was performed to compare the effect of surface treatment on SBS between Y-TZP ceramic and veneering ceramic. A comprehensive review of the literature from the earliest available dates through January 2017 was performed in the PubMed/Medline, Embase, Scopus, and Cochrane Library databases., Results: A total of 15 studies were identified for the inclusion of data, with only in vitro studies. A random-effect model found statistically significant differences between untreated and treated surfaces of Y-TZP (p < 0.00001; MD: 2.84; 95% CI: 2.19 to 3.49). In another analysis, a random-effect model found statistically significant differences between the groups that only performed the associations of treatments and control group (p < 0.00001; MD: 3.19; 95% CI: 2.11 to 4.28)., Conclusions: Surface treatment in Y-TZP improved the values of SBS between the Y-TZP and veneer ceramic. The associations between two or more treatments also showed positive effect on the bond strength due the cumulative effect of the treatments., (© 2017 by the American College of Prosthodontists.)

The aim of this is was evaluate the vertical and horizontal marginal adaptation of 3-unit fixed partial denture frameworks fabricated using different techniques and CAD/CAM systems. A total of 40 framework specimens were fabricated and divided into four groups as follows: lost-wax casting (G1); lost-wax casting with welding (G2); extraoral optical scanning of models (3S/DWOS) (G3); intraoral optical scanning (Cerec Bluecam/Sirona) (G4). A reference model was used to simulate a fixed partial denture with three elements (with a central pontic). The frameworks of G1 and G2 were cast in nickel-chromium (NiCr) alloy, whereas those of G3 and G4 were milled in zirconia. In all groups, vertical and horizontal marginal adaptation (over-contour and under-contour) was evaluated using a three-dimensional optical microscope (Quick Scope, Mitutoyo). The results showed higher vertical marginal misfit in G1 than in the other groups (p<0.001). Regarding horizontal marginal misfit, higher over-contour values occurred in G3 than in the other groups (p<0.001). G3 did not show under-contour at all, whereas the other groups did not differ from each other in this regard (p>0.05). Within the limitations of this study, it can be concluded that lost-wax casting with welding is a viable alternative to the use of CAD/CAM systems to fabricate frameworks of three-unit FPDs, since the techniques yielded similar vertical misfit values. Extra-oral CAD/CAM systems showed the highest horizontal misfit (over-contour) compared to other groups.

Animal cells have a remarkable capacity to adopt durable and heritable gene expression programs or epigenetic states that define the physical properties and diversity of somatic cell types. The maintenance of epigenetic programs depends on poorly understood pathways that prevent gain or loss of inherited signals. In the germline, epigenetic factors are enriched in liquid-like perinuclear condensates called nuage. Here, we identify the deeply conserved helicase-domain protein, ZNFX-1, as an epigenetic regulator and component of nuage that interacts with Argonaute systems to balance epigenetic inheritance. Our findings suggest that ZNFX-1 promotes the 3' recruitment of machinery that propagates the small RNA epigenetic signal and thus counteracts a tendency for Argonaute targeting to shift 5' along the mRNA. These functional insights support the idea that recently identified subdomains of nuage, including ZNFX-1 granules or "Z-granules," may define spatial and temporal zones of molecular activity during epigenetic regulation., (Copyright © 2018 Elsevier Inc. All rights reserved.)

In metazoans, Piwi-related Argonaute proteins engage piRNAs (Piwi-interacting small RNAs) to defend the genome against invasive nucleic acids, such as transposable elements. Yet many organisms-including worms and humans-express thousands of piRNAs that do not target transposons, suggesting that piRNA function extends beyond genome defense. Here, we show that the X chromosome-derived piRNA 21ux-1 downregulates XOL-1 (XO Lethal), a master regulator of X chromosome dosage compensation and sex determination in Caenorhabditis elegans. Mutations in 21ux-1 and several Piwi-pathway components sensitize hermaphrodites to dosage compensation and sex determination defects. We show that the piRNA pathway also targets xol-1 in C. briggsae, a nematode species related to C. elegans. Our findings reveal physiologically important piRNA-mRNA interactions, raising the possibility that piRNAs function broadly to ensure robust gene expression and germline development., (Copyright © 2018. Published by Elsevier Inc.)

Protein-coding genes undergo a wide array of regulatory interactions with factors that engage non-coding regions. Open reading frames (ORFs), in contrast, are thought to be constrained by coding function, precluding a major role in gene regulation. Here, we explore Piwi-interacting (pi)RNA-mediated transgene silencing in C. elegans and show that marked differences in the sensitivity to piRNA silencing map to the endogenous sequences within transgene ORFs. Artificially increasing piRNA targeting within the ORF of a resistant transgene can lead to a partial yet stable reduction in expression, revealing that piRNAs not only silence but can also "tune" gene expression. Our findings support a model that involves a temporal element to mRNA regulation by germline Argonautes, likely prior to translation, and suggest that piRNAs afford incremental control of germline mRNA expression by targeting the body of the mRNA, including the coding region., (Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.)

piRNAs (Piwi-interacting small RNAs) engage Piwi Argonautes to silence transposons and promote fertility in animal germlines. Genetic and computational studies have suggested that C. elegans piRNAs tolerate mismatched pairing and in principle could target every transcript. Here we employ in vivo cross-linking to identify transcriptome-wide interactions between piRNAs and target RNAs. We show that piRNAs engage all germline mRNAs and that piRNA binding follows microRNA-like pairing rules. Targeting correlates better with binding energy than with piRNA abundance, suggesting that piRNA concentration does not limit targeting. In mRNAs silenced by piRNAs, secondary small RNAs accumulate at the center and ends of piRNA binding sites. In germline-expressed mRNAs, however, targeting by the CSR-1 Argonaute correlates with reduced piRNA binding density and suppression of piRNA-associated secondary small RNAs. Our findings reveal physiologically important and nuanced regulation of individual piRNA targets and provide evidence for a comprehensive post-transcriptional regulatory step in germline gene expression., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The aim of this systematic review and meta-analysis was to compare the survival rate of the implants and the peri-implant tissue changes associated with implants inserted in fresh extraction sockets and those inserted in healed sockets. This review has been registered at PROSPERO under the number CRD42016043309. A systematic search was conducted by two reviewers independently in the databases PubMed/MEDLINE, Embase, and the Cochrane Library using different search terms; articles published until November 2016 were searched for. The searches identified 30 eligible studies. A total of 3,049 implants were installed in a total of 1,435 patients with a mean age of 46.68 years and a minimum of 6 months of follow-up. The survival rate of delayed implants (98.38%) was significantly greater than immediate implants (95.21%) (p=.001). For the marginal bone loss (p=.32), implant stability quotients values (p=.44), and pocket probing depth (p=.94) there was no significant difference between the analysed groups. The immediate implants placed in fresh sockets should be performed with caution because of the significantly lower survival rates than delayed implants inserted in healed sockets., (Copyright © 2017 International Association of Oral and Maxillofacial Surgeons. Published by Elsevier Ltd. All rights reserved.)

Purpose: The aim of this study was to use three-dimensional finite element analysis to analyze the stress distribution transferred by single implant-supported prostheses placed in the anterior maxilla using different connections (external hexagon, internal hexagon, or Morse taper), inclinations of the load (0, 30, or 60 degrees), and surgical techniques for placement (monocortical/conventional, bicortical, or bicortical with nasal floor elevation)., Materials and Methods: Nine models representing a bone block of this region were simulated by computer-aided design software (InVesalius, Rhinoceros, SolidWorks). Each model received one implant, which supported a cemented metalloceramic crown. Using FEMAP software, finite elements were discretized while simulating a 178-N load at 0, 30, and 60 degrees relative to the long axis of the implant. The problem was solved in NEi Nastran software, and postprocessing was performed in FEMAP. Von Mises stress and maximum principal stress maps were made., Results: The von Mises stress analysis revealed that stress increased with increasing inclination of the load, from 0 to 30 to 60 degrees. Morse taper implants showed less stress concentration around the cervical and apical areas of the implant. The bicortical technique, associated or not with nasal floor elevation, contributed to decreasing the stress concentration in the apical area of the implant. Maximum principal stress analysis showed that the increase in inclination was proportional to the increase in stress on the bone tissue in the cervical area. Lower stress concentrations in the cortical bone were obtained with Morse taper implants and the bicortical technique compared with other connections and surgical techniques, respectively., Conclusion: Increasing the inclination of the applied force relative to the long axis of the implant tended to overload the peri-implant bone tissue and the internal structure of the implants. The Morse taper connection and bicortical techniques seemed to be more favorable than other connections or techniques, respectively, for restoring the anterior maxilla.

The recent discovery of the positive-sense single-stranded RNA (ssRNA) Orsay virus (OV) as a natural pathogen of the nematode Caenorhabditis elegans has stimulated interest in exploring virus-nematode interactions. However, OV infection is restricted to a small number of intestinal cells, even in nematodes defective in their antiviral RNA interference (RNAi) response, and is neither lethal nor vertically transmitted. Using a fluorescent reporter strain of the negative-sense ssRNA vesicular stomatitis virus (VSV), we show that microinjection of VSV particles leads to a dose-dependent, muscle tissue-tropic, lethal infection in C. elegans. Furthermore, we find nematodes deficient for components of the antiviral RNAi pathway, such as Dicer-related helicase 1 (DRH-1), to display hypersusceptibility to VSV infection as evidenced by elevated infection rates, virus replication in multiple tissue types, and earlier mortality. Strikingly, infection of oocytes and embryos could also be observed in drh-1 mutants. Our results suggest that the antiviral RNAi response not only inhibits vertical VSV transmission but also promotes transgenerational inheritance of antiviral immunity. Our study introduces a new, in vivo virus-host model system for exploring arbovirus pathogenesis and provides the first evidence for vertical pathogen transmission in C. elegans., (Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Objective: The purpose of this systematic review was to evaluate the use of mini implants to retain complete overdentures in terms of survival rates of mini implants, marginal bone loss, satisfaction, and quality of life., Data: This report followed the PRISMA Statement and PICO question. This review has been registered at PROSPERO under the number CRD42016036141., Source: Two independent reviewers performed a comprehensive search of studies published until September 2016 and listed in the PubMed/MEDLINE, Embase, and The Cochrane Library databases. The focused question was: is the use of mini implants feasible for prosthodontic rehabilitation with complete overdentures?, Results: The 24 studies selected for review evaluated 1273 patients whose mean age was 65.93 years; these patients had received 2494 mini implants and 386 standard implants for retaining overdenture prosthesis. The mean follow-up time was 2.48 years (range: 1-7 years). There was a higher survival rate of mini implants (92.32%). More frequent failures for maxillary (31.71%) compared with mandibular arches (4.89%). The majority of studies revealed marginal bone loss values similar to those of standard implants (<1.5mm). All studies verified an increase in satisfaction and quality of life after rehabilitation treatment with mini dental implants., Conclusion: The present systematic review indicates that the use of mini implants for retaining overdenture prosthesis is considered an alternative treatment when standard treatment is not possible, since it presents high survival rates, acceptable marginal bone loss, and improvements in variables related to satisfaction and quality of life., Clinical Significance: Based on the results of this study, the use of a minimum 4 and 6 mini implants can be considered a satisfactory treatment option for rehabilitation of the mandibular and maxillary arches respectively with a complete overdenture., (Copyright © 2016. Published by Elsevier Ltd.)

The advent of sexual reproduction and the evolution of a dedicated germline in multicellular organisms are critical landmarks in eukaryotic evolution. We report an ancient family of GCNA (germ cell nuclear antigen) proteins that arose in the earliest eukaryotes, and feature a rapidly evolving intrinsically disordered region (IDR). Phylogenetic analysis reveals that GCNA proteins emerged before the major eukaryotic lineages diverged; GCNA predates the origin of a dedicated germline by a billion years. Gcna gene expression is enriched in reproductive cells across eukarya - either just prior to or during meiosis in single-celled eukaryotes, and in stem cells and germ cells of diverse multicellular animals. Studies of Gcna- mutant C. elegans and mice indicate that GCNA has functioned in reproduction for at least 600 million years. Homology to IDR-containing proteins implicated in DNA damage repair suggests that GCNA proteins may protect the genomic integrity of cells carrying a heritable genome., Competing Interests: The authors declare that no competing interests exist.

The aim of this study was to evaluate the stress distribution on full-arch prostheses, considering axial implants and tilted implants with or without abutments. Two photoelastic models were made, being that in 1 model the implants were inserted perpendicular, whereas in the other model 2 implants were inserted perpendicular and 2 implants were inserted with tilting 30° to the distal area. The prosthetic situations were evaluated: A-axial implants and nonsegmented full-arch prosthesis; B-axial implants and segmented full-arch prosthesis; C-tilted distal implants and nonsegmented full-arch prosthesis; D-tilted distal implants and segmented full-arch prosthesis, and was applied an axial load of 100 N at 5 points. The results showed greater stress concentration for axial implants than tilted implants in anterior and posterior loading. There was no difference in stress distribution to the use of abutment in the tilted implants. Shorter cantilever with tilted implants improved the stress distribution; the use of abutment did not influence the stress distribution.

This systematic review evaluated the effect on bone formation and implant survival of combining platelet-rich plasma (PRP) with bone grafts in maxillary augmentation. A comprehensive review of articles listed in the PubMed/MEDLINE, Embase, and Cochrane Library databases covering the period January 2000 to January 2015 was performed. The meta-analysis was based on bone formation for which the mean difference (MD, in millimetres) was calculated. Implant survival was assessed as a dichotomous outcome and evaluated using the risk ratio (RR) with 95% confidence interval (CI). The search identified 3303 references. After inclusion and exclusion criteria were applied, 17 studies were selected for qualitative analysis and 13 for quantitative analysis. A total of 369 patients (mean age 51.67 years) and 621 maxillary sinus augmentations were evaluated. After the data analysis, additional analyses were performed of the implant stability quotient, marginal bone loss, and alveolar bone height measured by MD. The results showed no significant difference in implant stability (P=0.32, MD 1.00, 95% CI -0.98 to 2.98), marginal bone loss (P=0.31, MD 0.06, 95% CI -0.05 to 0.16), alveolar bone height (P=0.10, MD -0.72, 95% CI -1.59 to 0.14), implant survival (P=0.22, RR 1.95, 95% CI 0.67-5.69), or bone formation (P=0.81, MD -0.63, 95% CI -5.91 to 4.65). In conclusion, the meta-analysis indicates no influence of PRP with bone graft on bone formation and implant survival in maxillary sinus augmentation., (Copyright © 2016. Published by Elsevier Ltd.)

The aim of this study was to perform a systematic review and meta-analysis to evaluate the possible benefits of platform-switching (PSW) implants when compared to regular platform (RP) implants in the categories of bone preservation and longevity. This systematic review and meta-analysis was performed in accordance with the PRISMA statement, PICO question, and Jadad scale. The relative risk (RR) of failure and the mean difference for marginal bone loss were calculated considering a confidence interval (CI) of 95%. Heterogeneity and subgroup analyses were performed, and funnel plots drawn. Twenty-five studies (17 randomized controlled trials (RCTs) and eight prospective studies) involving 1098 patients and 2310 implants were analysed. The meta-analysis revealed a significant reduction in crestal bone loss for PSW implants compared with RP implants (-0.41mm, 95% CI -0.52 to -0.29, P<0.00001). However, there was no statistically significant difference in implant failure (RR 1.10, 95% CI 0.6-2.02, P=0.75). A reduction in bone loss with PSW implants was observed for the following subgroups: RCTs only, implants in the maxilla, and implants in the mandible. PSW implants presented lower bone resorption compared with RP implants. RCTs should be done to explain the possible biases., (Copyright © 2015 International Association of Oral and Maxillofacial Surgeons. Published by Elsevier Ltd. All rights reserved.)

Piwi-interacting RNAs (piRNAs) engage Piwi proteins to suppress transposons and are essential for fertility in diverse organisms. An interesting feature of piRNAs is that, while piRNA lengths are stereotypical within a species, they can differ widely between species. For example, piRNAs are mainly 29 and 30 nucleotides in humans, 24 to 30 nucleotides in D. melanogaster, and uniformly 21 nucleotides in C. elegans. However, how piRNA length is determined and whether length impacts function remains unknown. Here, we show that C. elegans deficient for PARN-1, a conserved RNase, accumulate untrimmed piRNAs with 3' extensions. Surprisingly, these longer piRNAs are stable and associate with the Piwi protein PRG-1 but fail to robustly recruit downstream silencing factors. Our findings identify PARN-1 as a key regulator of piRNA length in C. elegans and suggest that length is regulated to promote efficient transcriptome surveillance., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Several recent studies link parental environments to phenotypes in subsequent generations. In this work, we investigate the mechanism by which paternal diet affects offspring metabolism. Protein restriction in mice affects small RNA (sRNA) levels in mature sperm, with decreased let-7 levels and increased amounts of 5' fragments of glycine transfer RNAs (tRNAs). In testicular sperm, tRNA fragments are scarce but increase in abundance as sperm mature in the epididymis. Epididymosomes (vesicles that fuse with sperm during epididymal transit) carry RNA payloads matching those of mature sperm and can deliver RNAs to immature sperm in vitro. Functionally, tRNA-glycine-GCC fragments repress genes associated with the endogenous retroelement MERVL, in both embryonic stem cells and embryos. Our results shed light on sRNA biogenesis and its dietary regulation during posttesticular sperm maturation, and they also link tRNA fragments to regulation of endogenous retroelements active in the preimplantation embryo., (Copyright © 2016, American Association for the Advancement of Science.)

Influenza A virus (IAV) lacks the enzyme for adding 5' caps to its RNAs and snatches the 5' ends of host capped RNAs to prime transcription. Neither the preference of the host RNA sequences snatched nor the effect of cap-snatching on host processes is completely defined. Previous studies of influenza cap-snatching used poly(A)-selected RNAs from infected cells or relied on annotated host genes to define the snatched host RNAs, and thus lack details on many noncoding host RNAs including snRNAs, snoRNAs, and promoter-associated capped small (cs)RNAs, which are made by "paused" Pol II during transcription initiation. In this study, we used a nonbiased technique, CapSeq, to identify host and viral-capped RNAs including nonpolyadenylated RNAs in the same samples, and investigated the substrate-product correlation between the host RNAs and the viral RNAs. We demonstrated that noncoding host RNAs, particularly U1 and U2, are the preferred cap-snatching source over mRNAs or pre-mRNAs. We also found that csRNAs are highly snatched by IAV. Because the functions of csRNAs remain mostly unknown, especially in somatic cells, our finding reveals that csRNAs at least play roles in the process of IAV infection. Our findings support a model where nascent RNAs including csRNAs are the preferred targets for cap-snatching by IAV and raise questions about how IAV might use snatching preferences to modulate host-mRNA splicing and transcription., (© 2015 Gu et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.)

Background: Transcription start sites (TSSs) with pronounced and phased nucleosome arrays downstream and nucleosome-depleted regions upstream of TSSs are observed in various species., Results: We have characterized sequence variation and expression properties of this set of TSSs (which we call "Nucleocyclic TSSs") using germline and somatic cells of three medaka (Oryzias latipes) inbred isolates from different locations. We found nucleocyclic TSSs in medaka to be associated with higher gene expression and characterized by a clear boundary in sequence composition with potentially-nucleosome-destabilizing A/T-enrichment upstream (p < 10(-60)) and nucleosome- accommodating C/G-enrichment downstream (p < 10(-40)) that was highly conserved from an ancestor. A substantial genetic distance between the strains facilitated the in-depth analysis of patterns of fixed mutations, revealing a localization-specific equilibrium between the rates of distinct mutation categories that would serve to maintain the conserved sequence anisotropy around TSSs. Downstream of nucleocyclic TSSs, C to T, T to C, and other mutation rates on the sense strand increased around first nucleosome dyads and decreased around first linkers, which contrasted with genomewide mutational patterns around nucleosomes (p < 5 %). C to T rates are higher than G to A rates around nucleosome associated with germline nucleocyclic TSS sites (p < 5 %), potentially due to the asymmetric effect of transcription-coupled repair., Conclusions: Our results demonstrate an atypical evolutionary process surrounding nucleocyclic TSSs.

The aim of this study was to analyze the stress distribution of short implants supporting single unit or splinted crowns by the photo-elasticity method. Four photo-elastic models were produced: A (3.75×7 mm); B (3.75×7 mm, 3.75×7 mm and 3.75×7 mm); C (3.75×10 mm, 3.75×7 mm and 3.75×7 mm); D (3.75×13 mm, 3.75×7 mm and 3.75×7 mm). The prostheses were made with Ni-Cr alloy. A load of 100 N in the axial and oblique directions was applied, totaling 380 applications, individually capturing their images in each model. The data were randomized and analyzed qualitatively and quantitatively by 2 examiners. The oblique loading was significantly more damaging. The increase in length was favorable for stress distribution (p<0.05). The splinting was beneficial for the transmission of stresses mainly (p<0.05). The splinting of the crowns, as well as increasing the length of the first implant and axial loading was most beneficial in the stress distribution. Short splinted implants behaved better than single unit implants. Increasing of the length of the first implant significantly improved the stress distribution in all analyzed situations., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The regulation of mRNA translation is of fundamental importance in biological mechanisms ranging from embryonic axis specification to the formation of long-term memory. POS-1 is one of several CCCH zinc-finger RNA-binding proteins that regulate cell fate specification during C. elegans embryogenesis. Paradoxically, pos-1 mutants exhibit striking defects in endo-mesoderm development but have wild-type distributions of SKN-1, a key determinant of endo-mesoderm fates. RNAi screens for pos-1 suppressors identified genes encoding the cytoplasmic poly(A)-polymerase homolog GLD-2, the Bicaudal-C homolog GLD-3, and the protein NEG-1. We show that NEG-1 localizes in anterior nuclei, where it negatively regulates endo-mesoderm fates. In posterior cells, POS-1 binds the neg-1 3' UTR to oppose GLD-2 and GLD-3 activities that promote NEG-1 expression and cytoplasmic lengthening of the neg-1 mRNA poly(A) tail. Our findings uncover an intricate series of post-transcriptional regulatory interactions that, together, achieve precise spatial expression of endo-mesoderm fates in C. elegans embryos., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Background: Exercise therapy is an evidence-based intervention for the conservative management of knee osteoarthritis. It is hypothesized that exercise therapy could reduce the knee adduction moment. A systematic review was performed in order to verify the effects of exercise therapy on the knee adduction moment in individuals with knee osteoarthritis in studies that also assessed pain and physical function., Methods: A comprehensive electronic search was performed on MEDLINE, Cochrane CENTRAL, EMBASE, Google scholar and OpenGrey. Inclusion criteria were randomized controlled trials with control or sham groups as comparator assessing pain, physical function, muscle strength and knee adduction moment during walking at self-selected speed in individuals with knee osteoarthritis that underwent a structured exercise therapy rehabilitation program. Two independent reviewers extracted the data and assessed risk of bias. For each study, knee adduction moment, pain and physical function outcomes were extracted. For each outcome, mean differences and 95% confidence intervals were calculated. Due to clinical heterogeneity among exercise therapy protocols, a descriptive analysis was chosen., Findings: Three studies, comprising 233 participants, were included. None of the studies showed significant differences between strengthening and control/sham groups in knee adduction moment. In regards to pain and physical function, the three studies demonstrated significant improvement in pain and two of them showed increased physical function following exercise therapy compared to controls. Muscle strength and torque significantly improved in all the three trials favoring the intervention group., Interpretation: Clinical benefits from exercise therapy were not associated with changes in the knee adduction moment. The lack of knee adduction moment reduction indicates that exercise therapy may not be protective in knee osteoarthritis from a joint loading point of view. Alterations in neuromuscular control, not captured by the knee adduction moment measurement, may contribute to alter dynamic joint loading following exercise therapy. To conclude, mechanisms other than the reduction in knee adduction moment might explain the clinical benefits of exercise therapy on knee osteoarthritis., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Small interfering RNA (siRNA)-mediated RNA interference (RNAi) pathways are critical for the detection and inhibition of RNA virus replication in insects. Recent work has also implicated RNAi pathways in the establishment of persistent virus infections and in the control of DNA virus replication. Accumulating evidence suggests that diverse double-stranded RNAs produced by RNA and DNA viruses can trigger RNAi responses yet many viruses have evolved mechanisms to inhibit RNAi defenses. Therefore, an evolutionary arms race exists between host RNAi pathways and invading viral pathogens. Here we review recent advances in our knowledge of how insect RNAi pathways are elicited upon infection, the strategies used by viruses to counter these defenses, and discuss recent evidence implicating Piwi-interacting RNAs in antiviral defense.

Approximately 75% of the human genome is transcribed, the majority of which does not encode protein. However, many noncoding RNAs (ncRNAs) are rapidly degraded after transcription, and relatively few have established functions, questioning the significance of this observation. Here we show that esBAF, a SWI/SNF family nucleosome remodeling factor, suppresses transcription of ncRNAs from ∼57,000 nucleosome-depleted regions (NDRs) throughout the genome of mouse embryonic stem cells (ESCs). We show that esBAF functions to both keep NDRs nucleosome-free and promote elevated nucleosome occupancy adjacent to NDRs. Reduction of adjacent nucleosome occupancy upon esBAF depletion is strongly correlated with ncRNA expression, suggesting that flanking nucleosomes form a barrier to pervasive transcription. Upon forcing nucleosome occupancy near two NDRs using a nucleosome-positioning sequence, we found that esBAF is no longer required to silence transcription. Therefore, esBAF's function to enforce nucleosome occupancy adjacent to NDRs, and not its function to maintain NDRs in a nucleosome-free state, is necessary for silencing transcription over ncDNA. Finally, we show that the ability of a strongly positioned nucleosome to repress ncRNA depends on its translational positioning. These data reveal a novel role for esBAF in suppressing pervasive transcription from open chromatin regions in ESCs., (© 2015 Hainer et al.; Published by Cold Spring Harbor Laboratory Press.)

Effective silencing by RNA-interference (RNAi) depends on mechanisms that amplify and propagate the silencing signal. In some organisms, small-interfering RNAs (siRNAs) are amplified from target mRNAs by RNA-dependent RNA polymerase (RdRP). Both RdRP recruitment and mRNA silencing require Argonaute proteins, which are generally thought to degrade RNAi targets by directly cleaving them. However, in C. elegans, the enzymatic activity of the primary Argonaute, RDE-1, is not required for silencing activity. We show that RDE-1 can instead recruit an endoribonuclease, RDE-8, to target RNA. RDE-8 can cleave RNA in vitro and is needed for the production of 3' uridylated fragments of target mRNA in vivo. We also find that RDE-8 promotes RdRP activity, thereby ensuring amplification of siRNAs. Together, our findings suggest a model in which RDE-8 cleaves target mRNAs to mediate silencing, while generating 3' uridylated mRNA fragments to serve as templates for the RdRP-directed amplification of the silencing signal., (Copyright © 2015 Elsevier Inc. All rights reserved.)

RNAi has become an essential tool in C. elegans research. This unit describes procedures for RNAi in C. elegans by microinjecting with dsRNA, feeding with bacteria expressing dsRNA, and soaking in dsRNA solution, as well as high-throughput methods for RNAi-based screens., (Copyright © 2015 John Wiley & Sons, Inc.)

Genome editing based on CRISPR (clustered regularly interspaced short palindromic repeats)-associated nuclease (Cas9) has been successfully applied in dozens of diverse plant and animal species, including the nematode Caenorhabditis elegans. The rapid life cycle and easy access to the ovary by micro-injection make C. elegans an ideal organism both for applying CRISPR-Cas9 genome editing technology and for optimizing genome-editing protocols. Here we report efficient and straightforward CRISPR-Cas9 genome-editing methods for C. elegans, including a Co-CRISPR strategy that facilitates detection of genome-editing events. We describe methods for detecting homologous recombination (HR) events, including direct screening methods as well as new selection/counterselection strategies. Our findings reveal a surprisingly high frequency of HR-mediated gene conversion, making it possible to rapidly and precisely edit the C. elegans genome both with and without the use of co-inserted marker genes., (Copyright © 2014 by the Genetics Society of America.)

Aim: This paper describes a case of a rehabilitation involving Computer Aided Design/Computer Aided Manufacturing (CAD-CAM) system in implant supported and dental supported prostheses using zirconia as framework., Background: The CAD-CAM technology has developed considerably over last few years, becoming a reality in dental practice. Among the widely used systems are the systems based on zirconia which demonstrate important physical and mechanical properties of high strength, adequate fracture toughness, biocompatibility and esthetics, and are indicated for unitary prosthetic restorations and posterior and anterior framework., Case Description: All the modeling was performed by using CAD-CAM system and prostheses were cemented using resin cement best suited for each situation., Conclusion: The rehabilitation of the maxillary arch using zirconia framework demonstrated satisfactory esthetic and functional results after a 12-month control and revealed no biological and technical complications., Clinical Significance: This article shows the important of use technology CAD/CAM in the manufacture of dental prosthesis and implant-supported.

Virus-host interactions drive a remarkable diversity of immune responses and countermeasures. We found that two RNA viruses with broad host ranges, vesicular stomatitis virus (VSV) and Sindbis virus (SINV), are completely restricted in their replication after entry into Lepidopteran cells. This restriction is overcome when cells are co-infected with vaccinia virus (VACV), a vertebrate DNA virus. Using RNAi screening, we show that Lepidopteran RNAi, Nuclear Factor-κB, and ubiquitin-proteasome pathways restrict RNA virus infection. Surprisingly, a highly conserved, uncharacterized VACV protein, A51R, can partially overcome this virus restriction. We show that A51R is also critical for VACV replication in vertebrate cells and for pathogenesis in mice. Interestingly, A51R colocalizes with, and stabilizes, host microtubules and also associates with ubiquitin. We show that A51R promotes viral protein stability, possibly by preventing ubiquitin-dependent targeting of viral proteins for destruction. Importantly, our studies reveal exciting new opportunities to study virus-host interactions in experimentally-tractable Lepidopteran systems.

Argonaute (AGO) proteins are key nuclease effectors of RNAi. Although purified AGOs can mediate a single round of target RNA cleavage in vitro, accessory factors are required for small interfering RNA (siRNA) loading and to achieve multiple-target turnover. To identify AGO cofactors, we immunoprecipitated the C. elegans AGO WAGO-1, which engages amplified small RNAs during RNAi. These studies identified a robust association between WAGO-1 and a conserved Vasa ATPase-related protein RDE-12. rde-12 mutants are deficient in RNAi, including viral suppression, and fail to produce amplified secondary siRNAs and certain endogenous siRNAs (endo-siRNAs). RDE-12 colocalizes with WAGO-1 in germline P granules and in cytoplasmic and perinuclear foci in somatic cells. These findings and our genetic studies suggest that RDE-12 is first recruited to target mRNA by upstream AGOs (RDE-1 and ERGO-1), where it promotes small RNA amplification and/or WAGO-1 loading. Downstream of these events, RDE-12 forms an RNase-resistant (target mRNA-independent) complex with WAGO-1 and may thus have additional functions in target mRNA surveillance and silencing., (Copyright © 2014 Elsevier Ltd. All rights reserved.)