Complement C3d enables protective immunity capable of distinguishing spontaneously transformed from non-transformed cells.

Immune-surveillance depends in part on the recognition of peptide variants by T cell antigen receptors. Given that both normal B cells and malignant B cells accumulate mutations we chose a murine model of multiple myeloma to test conditions to induce cell-mediated immunity targeting malignant plasma cell (PC) clones but sparing of normal PCs. Revealing a novel function for intracellular C3d, we discovered that C3d engaged T cell responses against malignant plasma cells in the bone marrow of mice that had developed multiple myeloma spontaneously. Our results show that C3d internalized by cells augments immune surveillance by several mechanisms. In one, C3d induces a master transcription regulator, E2f1, to increase the expression of long non-coding (lnc) RNAs, to generate peptides for MHC-I presentation and increase MHC-I expression. In another, C3d increases expression of RNAs encoding ribosomal proteins linked to processing of defective ribosomal products (DRiPs) that arise from non-canonical translation and known to promote immunosurveillance. Cancer cells are uniquely susceptible to increased expression and presentation of mutant peptides given the extent of protein misfolding and accumulation of somatic mutations. Accordingly, although C3d can be internalized by any cell, C3d preferentially targets malignant clones by evoking specific T cell mediated immunity (CMI) and sparing most non-transformed polyclonal B cells and plasma cells with lower mutation loads. Malignant plasma cell deletion was blocked by cyclosporin or by CD8 depletion confirming that endogenous T cells mediated malignant clone clearance. Besides the potential for therapeutic application our results highlight how intracellular C3d modifies cellular metabolism to augment immune surveillance. One Sentence Summary: We show that intracellular soluble fragment 3d of complement (C3d) induces regression of spontaneous multiple myeloma in mice reducing tumor burden by 10 fold, after 8 weeks. C3d enables cell-mediated immunity to target multiple myeloma clones sparing non-transformed polyclonal B cells and plasma cells with lower mutation loads. We show that C3d increases the expression of ribosomal subunits associated with the translation of defective ribosomal products (DRiPs). C3d also decreases expression of protein arginine methyl transferase (PRMT) 5 which in turn relieves E2f1 repression increasing the expression of Lnc RNAs and derived peptides that evoke anti-tumor cellular immunity. The approach increases MHC-I expression by tumor cells and generates a CMI response that overcomes tumor immune-evasion strategies. Significance: Tumors are immunogenic in part because of somatic mutations that originate novel peptides that once presented on MHC engage cell-mediated immunity (CMI). However, in spite of the higher mutation load most tumors evade immunity. We discovered that a component of the complement system (C3d) overcomes tumor immune evasion by augmenting expression of ribosomal proteins and lncRNAs linked to the presentation of novel peptides by tumor cells. C3d induced CMI targets cancer cells sparing non transformed cells uncovering a novel function for complement in immune surveillance.

Alternative Direct Restorative Materials for Dental Amalgam: A Concise Review Based on an FDI Policy Statement.

Dental restorative procedures remain a cornerstone of dental practice, and for many decades, dental amalgam was the most frequently employed material. However, its use is declining, mainly driven by its poor aesthetics and by the development of tooth-coloured adhesive materials. Furthermore, the Minamata Convention agreed on a phase-down on the use of dental amalgam. This concise review is based on a FDI Policy Statement which provides guidance on the selection of direct restorative materials as alternatives to amalgam. The Policy Statement was informed by current literature, identified mainly from PubMed and the internet. Ultimately, dental, oral, and patient factors should be considered when choosing the best material for each individual case. Dental factors include the dentition, tooth type, and cavity class and extension; oral aspects comprise caries risk profiles and related risk factors; and patient-related aspects include systemic risks/medical conditions such as allergies towards certain materials as well as compliance. Special protective measures (eg, a no-touch technique, blue light protection) are required when handling resin-based materials, and copious water spray is recommended when adjusting or removing restorative materials. Cost and reimbursement policies may need to be considered when amalgam alternatives are used, and the material recommendation requires the informed consent of the patient. There is no single material which can replace amalgam in all applications; different materials are needed for different situations. The policy statement recommends using a patient-centred rather than purely a material-centred approach. Further research is needed to improve overall material properties, the clinical performance, the impact on the environment, and cost-effectiveness of all alternative materials.

Microhardness and Gloss of a New CAD/CAM Hybrid Material Following Accelerated Aging and Coffee Immersion.

PURPOSE: To evaluate and compare the effect of accelerated aging and coffee immersion on the microhardness and gloss of a new computer-aided design and computer-aided manufacturing (CAD/CAM) hybrid material (Crystal Ultra) to those of contemporary restorative materials. MATERIALS AND METHODS: A total of 160 specimens (12 × 14 × 1 mm ± 0.05 mm) were obtained from IPS e.max (IPS), VITA Enamic (VE), Crystal Ultra (CU), Lava Ultimate (LU), and CeraSmart (CS) high-translucency CAD/CAM blocks. The Vickers microhardness and gloss of the specimens were determined after thermocycling and coffee immersion. Data analysis was performed using SPSS (α = .05). RESULTS: IPS and CS specimens exhibited the highest (572.66 ± 11.30) and lowest (61.92 ± 3.91) microhardness, respectively. The highest gloss was observed with IPS specimens (3.31 ± 0.32), and LU specimens showed the lowest gloss (2.33 ± 0.06). A significant difference in gloss was observed between the materials at all measurement intervals (P < .01), except at T0 (P = .43). IPS specimens showed no significant changes in either group at any measurement interval. CONCLUSIONS: The microhardness and gloss of the new CU material were comparable to those of the tested contemporary hybrid restorative materials. Glass-ceramic showed superior hardness and gloss compared to hybrid restorative materials. Accelerated aging with thermocycling and staining significantly affected the microhardness and gloss of all tested CAD/CAM materials.

Evidence-based clinical practice guideline on restorative treatments for caries lesions: A report from the American Dental Association.

BACKGROUND: An expert panel convened by the American Dental Association (ADA) Council on Scientific Affairs together with the ADA Science and Research Institute's program for Clinical and Translational Research conducted a systematic review and developed recommendations for the treatment of moderate and advanced cavitated caries lesions in patients with vital, nonendodontically treated primary and permanent teeth. TYPES OF STUDIES REVIEWED: The authors searched for systematic reviews comparing carious tissue removal (CTR) approaches in Ovid MEDLINE, Embase, Cochrane Database of Systematic Reviews, and Trip Medical Database. The authors also conducted a systematic search for randomized controlled trials comparing direct restorative materials in Ovid MEDLINE, Embase, Cochrane Central Register of Controlled Trials, ClinicalTrials.gov, and the World Health Organization International Clinical Trials Registry Platform. The authors used the Grading of Recommendations Assessment, Development, and Evaluation approach to assess the certainty of the evidence and formulate recommendations. RESULTS: The panel formulated 16 recommendations and good practice statements: 4 on CTR approaches specific to lesion depth and 12 on direct restorative materials specific to tooth location and surfaces involved. The panel conditionally recommended for the use of conservative CTR approaches, especially for advanced lesions. Although the panel conditionally recommended for the use of all direct restorative materials, they prioritized some materials over the use of others for certain clinical scenarios. PRACTICAL IMPLICATIONS: The evidence suggests that more conservative CTR approaches may decrease the risk of adverse effects. All included direct restorative materials may be effective in treating moderate and advanced caries lesions on vital, nonendodontically treated primary and permanent teeth.

Effect of a modified adhesive system with encapsulated arginine and calcium carbonate on dentin permeability.

To modify an adhesive system with halloysite clay nanotubes (HNTs) containing arginine and calcium carbonate and to evaluate their cytocompatibility, viscosity and efficacy in reducing dentin permeability. HNTs containing arginine and calcium carbonate were incorporated into the primer and adhesive of a three-step adhesive system (SBMP), and their viscosity was measured. Discs (n = 4/group) were prepared: SBMP (control), HNT-PR (modified primer), HNT-ADH (modified adhesive) and HNT-PR + ADH (modified primer and adhesive) were evaluated regarding cell death and viability. Dentin discs were prepared and randomly assigned into the following treatments (n = 10): NC (no treatment), SBMP, HNT-PR, HNT-ADH, HNT-PR + ADH and COL (Colgate® Sensitive Pro-relief™ prophylaxis paste). After, they were submitted to an erosive-abrasive cycling. Dentin permeability (hydraulic conductance) was evaluated at baseline, 24 h after treatment and after cycling. Both the modified primer and adhesive showed significantly higher viscosity than their controls. Group HNT-PR resulted in significantly higher cytotoxicity when compared to SBMP and HNT-PR + ADH groups. Group HNT-ADH resulted in the highest cell viability compared to all other groups. All groups showed significantly lower dentin permeability when compared to the NC group. Post-cycling, SBMP and HNT-ADH groups showed significantly lower permeability when compared to COL group. The addition of encapsulated arginine and calcium carbonate did not affect the cytocompatibility of the materials nor their ability to reduce dentin permeability.

Release and MMP-9 Inhibition Assessment of Dental Adhesive Modified with EGCG-Encapsulated Halloysite Nanotubes.

Degradation of the collagen fibrils at the dentin-resin interface by the enzymatic activity of matrix metalloproteinases (MMPs) has been known to permit some dental restoration complications, such as microleakage, secondary caries, and, ultimately, restoration failures. This study aimed to evaluate a modified adhesive by adding an MMP inhibitor from green tea extract with and without nanotube encapsulation to sustain the drug release. Epigallocatechin-3-gallate (EGCG) and Halloysite nanotubes (HNTs) were prepared to produce three variant combinations of modified adhesive (EGCG, EGCG-encapsulated HNT, and EGCG-free HNT). The drug loading efficiency and EGCG release over time were evaluated using UV-vis spectrometry. MMP-mediated ß-casein (BCN) cleavage rate assays were used to determine the ability of the EGCG in eluates of the adhesive to inhibit MMP-9 activities. For up to 8 weeks, HNT encapsulation reduced release to a statistically significant level. MMP-mediated ß-casein cleavage rate assays showed a significant decrease for the EGCG groups compared to the non-EGCG adhesive groups. Furthermore, the use of HNT for EGCG encapsulation to modify a dental adhesive helped slow down the rate of EGCG release without impacting its MMP inhibitory capabilities, which may help to maintain the dentin-resin interface's integrity over the long term after dental restoration placement.

Metabolic reprogramming by immune-responsive gene 1 up-regulation improves donor heart preservation and function.

Preservation quality of donor hearts is a key determinant of transplant success. Preservation duration beyond 4 hours is associated with primary graft dysfunction (PGD). Given transport time constraints, geographical limitations exist for donor-recipient matching, leading to donor heart underutilization. Here, we showed that metabolic reprogramming through up-regulation of the enzyme immune response gene 1 (IRG1) and its product itaconate improved heart function after prolonged preservation. Irg1 transcript induction was achieved by adding the histone deacetylase (HDAC) inhibitor valproic acid (VPA) to a histidine-tryptophan-ketoglutarate solution used for donor heart preservation. VPA increased acetylated H3K27 occupancy at the IRG1 enhancer and IRG1 transcript expression in human donor hearts. IRG1 converts aconitate to itaconate, which has both anti-inflammatory and antioxidant properties. Accordingly, our studies showed that Irg1 transcript up-regulation by VPA treatment increased nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2) in mice, which was accompanied by increased antioxidant protein expression [hemeoxygenase 1 (HO1) and superoxide dismutase 1 (SOD1)]. Deletion of Irg1 in mice (Irg1-/-) negated the antioxidant and cardioprotective effects of VPA. Consistent with itaconate's ability to inhibit succinate dehydrogenase, VPA treatment of human hearts increased itaconate availability and reduced succinate accumulation during preservation. VPA similarly increased IRG1 expression in pig donor hearts and improved its function in an ex vivo cardiac perfusion system both at the clinical 4-hour preservation threshold and at 10 hours. These results suggest that augmentation of cardioprotective immune-metabolomic pathways may be a promising therapeutic strategy for improving donor heart function in transplantation.

Direct materials for restoring caries lesions: Systematic review and meta-analysis-a report of the American Dental Association Council on Scientific Affairs.

BACKGROUND: The goal of restoring caries lesions is to protect the pulp, prevent progression of the disease process, and restore the form and function of the tooth. The purpose of this systematic review was to determine the effect of different direct restorative materials for treating cavitated caries lesions on anterior and posterior primary and permanent teeth. TYPE OF STUDIES REVIEWED: The authors included parallel and split-mouth randomized controlled trials comparing the effectiveness of direct restorative materials commercially available in the United States placed in vital, nonendodontically treated primary and permanent teeth. Pairs of reviewers independently conducted study selection, data extraction, and assessments of risk of bias and certainty of the evidence using the Grading of Recommendations Assessment, Development and Evaluation approach. The authors conducted pair-wise meta-analyses to summarize the evidence and calculated measures of association and their 95% CIs. RESULTS: Thirty-eight randomized controlled trials were eligible for analysis, which included data on Class I and Class II restorations on primary teeth and Class I, Class II, Class III, Class V, and root surface restorations on permanent teeth. Included studies assessed the effect of amalgam, resin composite, compomer, conventional glass ionomer cement, resin-modified glass isomer cement, and preformed metal crowns. Moderate to very low certainty evidence suggested varying levels of effectiveness across restorative materials. CONCLUSIONS AND PRACTICAL IMPLICATIONS: Owing to a relatively low event rate across various outcomes indicating restoration failure, there was limited evidence to support important differences between direct restorative materials used in practice.

Bioactivity of Dental Restorative Materials: FDI Policy Statement.

The term bioactivity is being increasingly used in medicine and dentistry. Due to its positive connotation, it is frequently utilised for advertising dental restorative materials. However, there is confusion about what the term means, and concerns have been raised about its potential overuse. Therefore, FDI decided to publish a Policy Statement about the bioactivity of dental restorative materials to clarify the term and provide some caveats for its use in advertising. Background information for this Policy Statement was taken from the current literature, mainly from the PubMed database and the internet. Bioactive restorative materials should have beneficial/desired effects. These effects should be local, intended, and nontoxic and should not interfere with a material's principal purpose, namely dental tissue replacement. Three mechanisms for the bioactivity of such materials have been identified: purely biological, mixed biological/chemical, or strictly chemical. Therefore, when the term bioactivity is used in an advertisement or in a description of a dental restorative material, scientific evidence (in vitro or in situ, and preferably in clinical studies) should be provided describing the mechanism of action, the duration of the effect (especially for materials releasing antibacterial substances), and the lack of significant adverse biological side effects (including the development and spread of antimicrobial resistance). Finally, it should be documented that the prime purpose, for instance, to be used to rebuild the form and function of lost tooth substance or lost teeth, is not impaired, as demonstrated by data from in vitro and clinical studies. The use of the term bioactive dental restorative material in material advertisement/information should be restricted to materials that fulfil all the requirements as described in the FDI Policy Statement.

Xenotransplantation in transition.

The application of xenotransplantation of porcine organs and tissues for treatment of disease, sought for more than a century, might soon be realized. Until now, the immune response of recipients against xenogeneic organs and tissues posed the main obstacle to clinical application. However, decades of research into this immune response and identification of other molecular barriers together with advances in genetic engineering and cloning of large animals and immune therapeutics coalesced to support prolonged survival and function of porcine organ grafts in nonhuman primates. This experimental progress in turn sparks consideration of clinical trials. The papers in this special section provide authoritative views concerning the immune hurdles that still limit and potentially still preclude clinical application of xenotransplantation. Xenoreactive antibodies elicited in T cell-dependent B cell-responses constitute the most important hurdle and control of these responses impels use of intense regimens of immunosuppression. These antibodies pose a danger to xenografts and potentially compromise subsequent allografts. However, new insights into the specificity of these antibodies, the pathways and kinetics of production and genetic determinants of pathogenicity offer novel opportunities for intervention. Likewise, the rapid ability to propose and test new strategies in nonhuman primate models hastens needed advances. However further progress will depend on development and validation of laboratory methods for identification and assay of pathogenic immune responses and evaluation of the response to therapy.

TNFRSF13B in B cell responses to organ transplantation.

Antibodies directed against organ transplants are thought to pose the most vexing hurdle to enduring function and survival of the transplants, particularly organ xenotransplants, and accordingly basic and clinical investigation has focused on elucidating the specificity and pathogenicity of graft-specific antibodies. While much has been learned about these matters, far less is known about the B cells producing graft-specific antibodies and why these antibodies appear to injure some grafts but not others. With the goal of addressing those questions, we have investigated the properties of tumor necrosis factor receptor super family-13B (TNFRSF13B), which regulates various aspects of B cell responses. A full understanding of the functions of TNFRSF13B however is hindered by extreme polymorphism and by diversity of interactions of the protein. Nevertheless, TNFRSF13B variants have been found to exert distinct impact on natural and elicited antibody responses and host defense and mutations of TNFRSF13B have been found to influence the propensity for development of antibody-mediated rejection of organ transplants. Because B cell responses potentially limit application of xenotransplantation, understanding how TNFRSF13B diversity and TNFRSF13B variants govern immunity in xenotransplantation could inspire development of novel therapeutics that could in turn accelerate clinical implementation of xenotransplantation.

Accommodation in allogeneic and xenogeneic organ transplantation: Prevalence, impact, and implications for monitoring and for therapeutics.

Accommodation refers to acquired resistance of organs or tissues to immune or inflammatory reactions that might otherwise cause severe injury or rejection. As first observed in ABO-incompatible kidney transplants and heterotopic cardiac xenografts, accommodation was identified when organ transplants continued to function despite the presence of anti-graft antibodies and/or other reactants in the blood of recipients. Recent evidence suggests many and perhaps most organ transplants have accommodation, as most recipients mount B cell responses specific for the graft. Wide interest in the impact of graft-specific antibodies on the outcomes of transplants prompts questions about which mechanisms confer protection against such antibodies, how accommodation might be detected and whether and how rejection could be superimposed on accommodation. Xenotransplantation offers a unique opportunity to address these questions because immune responses to xenografts are easily detected and the pathogenic impact of immune responses is so severe. Xenotransplantation also provides a compelling need to apply these and other insights to decrease the intensity and toxicity of immunosuppression that otherwise could limit clinical application.

Microhardness and gloss of a new CAD-CAM hybrid material following accelerated aging and coffee immersion.

PURPOSE: To evaluate and compare the effect of accelerated aging and coffee immersion on the microhardness and gloss of a new CAD/CAM hybrid material (Crystal Ultra) to those of contemporary restorative materials. MATERIALS AND METHODS: A total of 160 specimens (12 x 14 x 1 mm ± 0.05 mm) were obtained from (IPS e.max [IPS], VITA Enamic [VE], Crystal Ultra [CU], Lava Ultimate [LU], and CeraSmart [CS]) high-translucency CAD/CAM blocks. The Vickers microhardness and gloss of the specimens were determined following thermocycling and coffee immersion. Data analysis was performed using SPSS (α = .05). RESULTS: IPS and CS specimens exhibited the highest (572.66 ± 11.30) and lowest (61.92 ± 3.91) microhardness, respectively. The highest gloss was observed with IPS specimens (3.31 ± 0.32), and LU specimens showed the lowest gloss (2.33 ± 0.06). A significant difference in gloss was observed between the materials at all measurement intervals (P < .01), except at T0 (P = .43). IPS specimens showed no significant changes in either group at any measurement interval. CONCLUSION: The microhardness and gloss of the new CU material were comparable to those of the tested contemporary hybrid restorative materials. Glass-ceramic showed superior hardness and gloss compared to hybrid restorative materials. Accelerated aging by thermocycling and staining significantly affected the microhardness and gloss of all the tested CAD/CAM materials.

A non-human primate model of acute liver failure suitable for testing liver support systems.

Acute hepatic failure is associated with high morbidity and mortality for which the only definitive therapy is liver transplantation. Some fraction of those who undergo emergency transplantation have been shown to recover native liver function when transplanted with an auxiliary hepatic graft that leaves part of the native liver intact. Thus, transplantation could have been averted with the development and use of some form of hepatic support. The costs of developing and testing liver support systems could be dramatically reduced by the availability of a reliable large animal model of hepatic failure with a large therapeutic window that allows the assessment of efficacy and timing of intervention. Non-lethal forms of hepatic injury were examined in combination with liver-directed radiation in non-human primates (NHPs) to develop a model of acute hepatic failure that mimics the human condition. Porcine hepatocyte transplantation was then tested as a potential therapy for acute hepatic failure. After liver-directed radiation therapy, delivery of a non-lethal hepatic ischemia-reperfusion injury reliably and rapidly generated liver failure providing conditions that can enable pre-clinical testing of liver support or replacement therapies. Unfortunately, in preliminary studies, low hepatocyte engraftment and over-immune suppression interfered with the ability to assess the efficacy of transplanted porcine hepatocytes in the model. A model of acute liver failure in NHPs was created that recapitulates the pathophysiology and pathology of the clinical condition, does so with reasonably predictable kinetics, and results in 100% mortality. The model allowed preliminary testing of xenogeneic hepatocyte transplantation as a potential therapy.

Allogeneic and xenogeneic lymphoid reconstitution in a RAG2 -/- IL2RG y/- severe combined immunodeficient pig: A preclinical model for intrauterine hematopoietic transplantation.

Mice with severe combined immunodeficiency are commonly used as hosts of human cells. Size, longevity, and physiology, however, limit the extent to which immunodeficient mice can model human systems. To address these limitations, we generated RAG2 -/- IL2RG y/- immunodeficient pigs and demonstrate successful engraftment of SLA mismatched allogeneic D42 fetal liver cells, tagged with pH2B-eGFP, and human CD34+ hematopoietic stem cells after in utero cell transplantation. Following intrauterine injection at day 42-45 of gestation, fetuses were allowed to gestate to term and analyzed postnatally for the presence of pig (allogeneic) and human (xenogeneic) B cells, T-cells and NK cells in peripheral blood and other lymphoid tissues. Engraftment of allogeneic hematopoietic cells was detected based on co-expression of pH2B-eGFP and various markers of differentiation. Analysis of spleen revealed robust generation and engraftment of pH2B-eGFP mature B cells (and IgH recombination) and mature T-cells (and TCR-ß recombination), T helper (CD3+CD4+) and T cytotoxic (CD3+CD8+) cells. The thymus revealed engraftment of pH2B-eGFP double negative precursors (CD4-CD8-) as well as double positive (CD4+, CD8+) precursors and single positive T-cells. After intrauterine administration of human CD34+ hematopoietic stem cells, analysis of peripheral blood and lymphoid tissues revealed the presence of human T-cells (CD3+CD4+ and CD3+CD8+) but no detectable B cells or NK cells. The frequency of human CD45+ cells in the circulation decreased rapidly and were undetectable within 2 weeks of age. The frequency of human CD45+ cells in the spleen also decreased rapidly, becoming undetectable at 3 weeks. In contrast, human CD45+CD3+ T-cells comprised >70% of cells in the pig thymus at birth and persisted at the same frequency at 3 weeks. Most human CD3+ cells in the pig's thymus expressed CD4 or CD8, but few cells were double positive (CD4+ CD8+). In addition, human CD3+ cells in the pig thymus contained human T-cell excision circles (TREC), suggesting de novo development. Our data shows that the pig thymus provides a microenvironment conducive to engraftment, survival and development of human T-cells and provide evidence that the developing T-cell compartment can be populated to a significant extent by human cells in large animals.

Effect of post-irradiation polymerization on selected mechanical properties of six direct resins.

This study evaluated the post-irradiation mechanical property development of six resin composite-based restorative materials from the same manufacturer starting at 1 h post irradiation, followed by 24 h, 1 week, and 1 month after fabrication. Samples were stored in 0.2M phosphate buffered saline until testing. Flexural strength, flexural modulus, flexural toughness, modulus of resiliency, fracture toughness, and surface microhardness were performed at each time interval. Mean data was analyzed by Kruskal Wallis and Dunn's post hoc testing at a 95% level of confidence (α=0.05). Results were material specific but overall, all resin composite material mechanical properties were found to be immature at 1 h after polymerization as compared to that observed at 24 h. It may be prudent that clinicians advise patients, especially those receiving complex posterior composite restorations, to guard against overly stressing these restorations during the first 24 h.