In vitro and in vivo evaluation of iRoot BP Plus as a coronal sealing material for regenerative endodontic procedures.

OBJECTIVES: To investigate in vitro effects of a nanoparticle bioceramic material, iRoot BP Plus, on stem cells from apical papilla (SCAP) and in vivo capacity to induce pulp-dentin complex formation. MATERIALS AND METHODS: The sealing ability of iRoot BP Plus was measured via scanning electron microscopy (SEM). SCAP were isolated and treated in vitro by iRoot BP Plus conditioned medium, with mineral trioxide aggregate (MTA) conditioned medium and regular medium used as controls, respectively. Cell proliferation was assessed by BrdU labeling and MTT assay and cell migration was evaluated with wound healing and transwell assays. Osteo/odontogenic potential was evaluated by Alizarin red S staining and qPCR. Pulp-dentin complex formation in vivo was assessed by a tooth slice subcutaneous implantation model. RESULTS: iRoot BP Plus was more tightly bonded with the dentin. There was no difference in SCAP proliferation between iRoot BP Plus and control groups (P > 0.05). iRoot BP Plus had a greater capacity to elevated cell migration (P < 0.05) and osteo/odontogenic marker expression and mineralization nodule formation of SCAP compared with MTA groups (P < 0.05). Furthermore, the new continuous dentine layer and pulp-like tissue was observed in the iRoot BP Plus group in vivo. CONCLUSIONS: iRoot BP Plus showed excellent sealing ability, promoted the migration and osteo/odontogenesis of SCAP and induced pulp-dentin complex formation without affecting the cell proliferation, which indicated iRoot BP Plus was a promising coronal sealing material in REPs. CLINICAL RELEVANCE: The coronal sealing materials play crucial roles for the outcomes of REPs. This study showed that iRoot BP Plus has good coronal sealing and promote pulp-dentin complex formation compared with MTA, providing experimental evidences for the clinical application of iRoot BP Plus as a promising coronal seal material in REPs.

Key process for reducing grains arsenic by applying sulfate varies with irrigation mode: Dual effects of microbe-mediated arsenic transformation and iron plaque.

Sulfate affects the transformation of arsenic (As) in soil and its absorption by plant roots. However, the influence of sulfate and irrigation interactions on the mobility of As in the soil-rice system remains poorly understood. To address this gap, we conducted a pot experiment with varying sulfate levels and irrigation modes to examine their effects on rice As translocation, soil As forms, iron plaque formation, and microorganisms involved in As transformation. The addition of exogenous sulfate significantly reduced grain As levels by a maximum of 60.1%, 46.7%, and 70.5% under flooding (F), flooding-moist alternate (FM), moist (M) conditions, respectively. However, the changes in soil available As did not fully correspond to grains As content. Soil available As was only reduced by sulfate under the FM treatment, which limited grains As accumulation under this condition. The reduction in grains As content under F and M conditions was mainly attributed to sulfate-induced increases in soil pH, which in turn inhibited As translocation and promoted iron plaque formation. Additionally, both irrigation mode and sulfate fertilization independently or interactively influenced the abundance of Sulfuritalea, Koribacter, Geobacter, and Sulfuriferula, thereby affecting the As forms in soil through the Fe/S redox process. Specifically, under F and FM conditions, SO42--S inhibited Geobacter but stimulated Fe-oxidizing bacteria, possibly resulting in increased As bound to Fe/Mn oxides (As-F3). Under M condition, SO42--S levels regulated As adsorption and release through the participation of Fe/S cycle bacteria, specifically influencing the adsorbed As fraction (As-F2). Therefore, the addition of SO42--S hindered As translocation to grains by promoting As sequestration in the iron plaque and facilitating microbe-mediated As immobilization through the Fe/S cycle, which was dependent on soil moisture. These results can be used as a guide for sulfur fertilizer application under different soil moisture with the goal of minimizing rice grain As.

Effects of feeding on different parts of Ailanthus altissima on the intestinal microbiota of Eucryptorrhynchus scrobiculatus and Eucryptorrhynchus brandti (Coleoptera: Curculionidae).

Eucryptorrhynchus brandti and Eucryptorrhynchus scrobiculatus (Coleoptera: Curculionidae) are two monophagous weevil pests that feed on Ailanthus altissima (Mill.) Swingle but differ in their diet niche. In the field, adults of E. brandti prefer to feed on the trunk of A. altissima, whereas adults of E. scrobiculatus prefer to feed on the tender parts. We conducted Illumina sequencing of 16S rRNA to examine changes in bacterial diversity in the adults of these two weevil species after they fed on different parts of A. altissima (trunk, 2-3-year-old branches, annual branches, and petioles). Proteobacteria, Tenericutes, and Firmicutes were the dominant phyla in E. brandti (relative abundance was 50.64, 41.56, and 5.63%, respectively) and E. scrobiculatus (relative abundance was 78.63, 11.91, and 7.41%, respectively). At the genus level, Spiroplasma, endosymbionts2, Unclassified Enterobacteriaceae, and Lactococcus were dominant in E. brandti, and Unclassified Enterobacteriaceae, Wolbachia and Spiroplasma, and endosymbionts2 were dominant in E. scrobiculatus. Linear discriminant analysis effect size analysis revealed microbial biomarkers in the different treatment group of adults of both weevil species. Adults of E. brandti may require the trunk, and adults of E. scrobiculatus may require the petioles and annual branches to maintain the high diversity of their gut microbes. The results of this study indicate that feeding on different parts of A. altissima affects the composition and function of the microbes of E. brandti and the microbial composition of E. scrobiculatus. Variation in the abundance of Wolbachia and Spiroplasma in E. brandti and E. scrobiculatus is associated with dietary niche changes, and this might explain the evolution of reproductive isolation between these two sibling weevil species.

[Effects of Iron Intensity-regulated Root Microbial Community Structure and Function on Cadmium Accumulation in Rice].

Exploring the effects of exogenous iron (Fe) on cadmium (Cd) in rice is of great significance for ensuring food security. The accumulation of Cd and the changes in the microbial community structure in rice roots under three Fe concentrations (5, 50, and 500 µmol·L-1 EDTA-Na2Fe) were studied through a hydroponic experiment. The results showed that the increase in the environmental Fe concentration promoted the formation of iron plaque on the rice roots, and both Fe-deficiency and Fe-sufficiency would enhance the adsorption and fixation of Cd on the root surface. Compared with that of normal Fe levels (50 µmol·L-1), Fe deficiency increased Cd accumulation in rice roots and shoots by 49.76% and 15.68%, respectively. Although Fe sufficiency also increased Cd accumulation in the roots by 18.39%, the Cd concentration in shoots was significantly reduced by 35.95% compared with that of the normal Fe. 16S rRNA high-throughput sequencing was used to determine the root microbial community structure, and through PCA, LEfSe, and RDA analysis, it was found that compared with normal Fe, an Fe-deficient environment reduced the abundance and uniformity of root microbes. Proteobacteria and Bacteroidetes at the phylum level were the dominant flora, Fe deficiency inhibited the increase in the relative abundance of Bacteroidetes, and high-concentration Fe reduced the relative abundance of Proteobacteria. At the genus level, the relative abundance of functional microorganisms Ensifer, Rhodopila, Bdellovibrio, and Dyella were different under different Fe environments, which may have affected the absorption and accumulation of Cd by rice by affecting the formation of Fe plaque on the root and other biochemical processes. In addition, the effect of an Fe-deficient environment on microbial functions was higher than that of the Fe sufficient environment. This study investigated the changes in the rice root microbial community structure and the ability of rice to absorb and transport Cd under different Fe environments, which provided a theoretical basis and an important reference for the inhibition of Fe on Cd accumulation in rice in Cd-polluted paddy soil in southern China.

cGAS-STING pathway: post-translational modifications and functions in sterile inflammatory diseases.

Cytoplasmic microbial and host aberrant DNAs act as danger signals and trigger host immune responses. Upon recognition, the cytosolic DNA sensor cyclic GMP-AMP synthase (cGAS) catalyzes the production of a second messenger 2'3'-cGAMP, which activates endoplasmic reticulum (ER)-associated stimulator of interferon (IFN) genes (STING) and ultimately leads to the induction of type I IFNs and inflammatory genes that collectively initiate host immune defense against microbial invasion. Inappropriate activation or suppression of this signaling pathway has been implicated in the development of some autoimmune diseases, sterile inflammation, and cancers. In this review, we describe how the activity of cGAS and STING is regulated by host post-translational modifications and summarize the recent advances of cell-specific cGAS-STING activation and its association in sterile inflammatory diseases. We also discuss key outstanding questions in the field, including how our knowledge of cGAS-STING pathway could be translated into clinical applications.

The Lysosomal Rag-Ragulator Complex Licenses RIPK1 and Caspase-8-mediated Pyroptosis by Yersinia.

Host cells initiate cell death programs to limit pathogen infection. Inhibition of transforming growth factor-ß-activated kinase 1 (TAK1) by pathogenic Yersinia in macrophages triggers receptor-interacting serine/threonine-protein kinase 1 (RIPK1)-dependent caspase-8 cleavage of gasdermin D (GSDMD) and inflammatory cell death (pyroptosis). A genome-wide clustered regularly interspaced short palindromic repeats (CRISPR) screen to uncover mediators of caspase-8-dependent pyroptosis identified an unexpected role of the lysosomal FLCN-FNIP2-Rag-Ragulator supercomplex, which regulates metabolic signalling and the mechanistic target of rapamycin complex 1 (mTORC1). In response to Yersinia infection, FADD, RIPK1 and caspase-8 were recruited to Rag-Ragulator, causing RIPK1 phosphorylation and caspase-8 activation. Pyroptosis activation depended on Rag GTPase activity and lysosomal tethering of Rag-Ragulator, but not mTORC1. Thus, the lysosomal metabolic regulator Rag-Ragulator instructs the inflammatory response to Yersinia.

Driving Hierarchical Collagen Fiber Formation for Functional Tendon, Ligament, and Meniscus Replacement.

Hierarchical collagen fibers are the primary source of strength in musculoskeletal tendons, ligaments, and menisci. It has remained a challenge to develop these large fibers in engineered replacements or in vivo after injury. The objective of this study was to investigate the ability of restrained cell-seeded high density collagen gels to drive hierarchical fiber formation for multiple musculoskeletal tissues. We found boundary conditions applied to high density collagen gels were capable of driving tenocytes, ligament fibroblasts, and meniscal fibrochondrocytes to develop native-sized hierarchical collagen fibers 20-40 µm in diameter. The fibers organize similar to bovine juvenile collagen with native fibril banding patterns and hierarchical fiber bundles 50-350 µm in diameter by 6 weeks. Mirroring fiber organization, tensile properties of restrained samples improved significantly with time, reaching ~1 MPa. Additionally, tendon, ligament, and meniscal cells produced significantly different sized fibers, different degrees of crimp, and different GAG concentrations, which corresponded with respective juvenile tissue. To our knowledge, these are some of the largest, most organized fibers produced to date in vitro. Further, cells produced tissue specific hierarchical fibers, suggesting this system is a promising tool to better understand cellular regulation of fiber formation to better stimulate it in vivo after injury.

An optical relay approach to very low cost hybrid polymer-complementary metal-oxide semiconductor electrophoresis instrumentation.

Electrophoresis is an integral part of many molecular diagnostics protocols and an inexpensive implementation would greatly facilitate point-of-care (POC) applications. However, the high instrumentation cost presents a substantial barrier, much of it associated with fluorescence detection. The cost of such systems could be substantially reduced by placing the fluidic channel and photodiode directly above the detector in order to collect a larger portion of the fluorescent light. In future, this could be achieved through the integration and monolithic fabrication of photoresist microchannels on complementary metal-oxide semiconductor microelectronics (CMOS). However, the development of such a device is expensive due to high non-recurring engineering costs. To facilitate that development, we present a system that utilises an optical relay to integrate low-cost polymeric microfluidics with a CMOS chip that provides a photodiode, analog-digital conversion and a standard serial communication interface. This system embodies an intermediate level of microelectronic integration, and significantly decreases development costs. With a limit of detection of 1.3±0.4nM of fluorescently end-labeled deoxyribonucleic acid (DNA), it is suitable for diagnostic applications.

[Histological observation of microscrews anchorage implant-bone interface loaded with different orthodontic condition].

PURPOSE: To study the stability of the microscrew implant anchorage under different loading force and different timing and observe the integration of the implant to the bone. METHODS: Thirty-six titanium alloy microscrew implants were implanted into three dogs' maxilla and mandible. The implants were divided into two groups: one with immediate implant loading and the other with delayed implant loading. In the delayed loading group, the microscrew implants that were implanted into the right maxilla and mandible were unloaded temporarily. Four weeks later, these dogs, together with the immediate loading group, whose microscrew implants were implanted into the left maxilla and mandible, were loaded at the same time. The mesial screw was unloaded. Two distal screws were loaded with force: 1.96N on the maxilla, and 3.92N on the mandibular. After 12 weeks, the dogs were sacrificed. The specimen were fixed and decalcified. Subsequently, immunohistochemical stain for BMP-2 was employed. The specimen were assessed with image analysis system to analyze the average grey scale. The data were analyzed using Student's t test and one-way ANOVA with SPSS 10.0 software package. RESULTS: BMP-2 staining showed that there was significant difference between the immediate loading group(1.96N,3.92N), delayed loading group(1.96N,3.92N) and non-loading group at average gray value (P<0.05). But there was no significant difference between the immediate loading group and delayed loading group (P>0.05). CONCLUSIONS: Both the immediate and the delayed implant loading can strengthen the attachment and integration of the implant to the bone. The timing of the loading force has no significant impact on the stabilization of the microscrew implants.