Downregulation of HbFPS1 affects rubber biosynthesis of Hevea brasiliensis suffering from tapping panel dryness.

Tapping panel dryness (TPD) is a century-old problem that has plagued the natural rubber production of Hevea brasiliensis. TPD may result from self-protective mechanisms of H. brasiliensis in response to stresses such as excessive hormone stimulation and mechanical wounding (bark tapping). It has been hypothesized that TPD impairs rubber biosynthesis; however, the underlying mechanisms remain poorly understood. In the present study, we firstly verified that TPD-affected rubber trees exhibited lower rubber biosynthesis activity and greater rubber molecular weight compared to healthy rubber trees. We then demonstrated that HbFPS1, a key gene of rubber biosynthesis, and its expression products were downregulated in the latex of TPD-affected rubber trees, as revealed by transcriptome sequencing and iTRAQ-based proteome analysis. We further discovered that the farnesyl diphosphate synthase HbFPS1 could be recruited to small rubber particles by HbSRPP1 through protein-protein interactions to catalyze farnesyl diphosphate (FPP) synthesis and facilitate rubber biosynthesis initiation. FPP content in the latex of TPD-affected rubber trees was significantly decreased with the downregulation of HbFPS1, ultimately resulting in abnormal development of rubber particles, decreased rubber biosynthesis activity, and increased rubber molecular weight. Upstream regulator assays indicated that a novel regulator, MYB2-like, may be an important regulator of downregulation of HbFPS1 in the latex of TPD-affected rubber trees. Our findings not only provide new directions for studying the molecular events involved in rubber biosynthesis and TPD syndrome and contribute to rubber management strategies, but also broaden our knowledge of plant isoprenoid metabolism and its regulatory networks.

Stimuli-Responsive Crystalline Smart Materials: From Rational Design and Fabrication to Applications.

Stimuli-responsive smart materials that can undergo reversible chemical/physical changes under external stimuli such as mechanical stress, heat, light, gas, electricity, and pH, are currently attracting increasing attention in the fields of sensors, actuators, optoelectronic devices, information storage, medical applications, and so forth. The current smart materials mostly concentrate on polymers, carbon materials, crystalline liquids, and hydrogels, which have no or low structural order (i.e., the responsive groups/moieties are disorderly in the structures), inevitably introducing deficiencies such as a relatively low response speeds, energy transformation inefficiencies, and unclear structure-property relationships. Consequently, crystalline materials with well-defined and regular molecular arrays can offer a new opportunity to create novel smart materials with improved stimuli-responsive performance. Crystalline materials include framework materials (e.g., metal-organic frameworks, MOFs; covalent organic frameworks, COFs) and molecular crystals (e.g., organic molecules and molecular cages), which have obvious advantages as smart materials compared to amorphous materials. For example, responsive groups/moieties can be uniformly installed in the skeleton of the crystal materials to form ordered molecular arrays, making energy transfer between external-stimulus signals and responsive sites much faster and more efficiently. Besides that, the well-defined structures facilitate in situ characterization of their structural transformation at the molecular level by means of various techniques and high-tech equipment such as in situ spectra and single-crystal/powder X-ray diffraction, thus benefiting the investigation and understanding of the mechanism behind the stimuli-responsive behaviors and structure-property relationships. Nevertheless, some unsolved challenges remain for crystalline smart materials (CSMs), hampering the fabrication of smart material systems for practical applications. For instance, as the materials' crystallinity increases, their processability and mechanical properties usually decrease, unavoidably hindering their practical application. Moreover, crystalline smart materials mostly exist as micro/nanosized powders, which are difficult to make stimuli-responsive on the macroscale. Thus, developing strategies that can balance the materials' crystallinity and processability and establishing macroscale smart material systems are of great significance for practical applications.In this Account, we mainly summarize the recent research progress achieved by our groups, including (i) the rational design and fabrication of new stimuli-responsive crystalline smart materials, including molecular crystals and framework materials, and an in-depth investigation of their response mechanism and structure-property relationship and (ii) creating chemical/physical modification strategies to improve the processability and mechanical properties for crystalline materials and establishing macroscale smart systems for practical applications. Overall, this Account summarizes the state-of-the-art progress of stimuli-responsive crystalline smart materials and points out the existing challenges and future development directions in the field.

Metabolomics Analysis of the Effect of GAT-2 Deficiency on Th1 Cells in Mice.

The classic neurotransmitter γ-aminobutyric acid (GABA) has been shown to shape the activation and function of immune cells. There are four high-affinity GABA transporters (GATs, including GAT-1, GAT-2, GAT-3, and GAT-4) responsible for the transmembrane transport of GABA in mice. To explore the effect of GAT-2 on type 1 helper T (Th1) cells, naïve CD4+ T cells were isolated from splenocytes of GAT-2 knockout (KO) and wild-type (WT) mice and cultured for Th1 cell differentiation, and then, metabolomics analysis of Th1 cells was performed via gas chromatography coupled to time-of-flight mass spectrometry added with multivariate analyses. Based on the variable importance projection value > 1 and P < 0.05, a total of nine differentially expressed metabolites (DEMs) were identified between WT and KO. Then, DEMs were mapped to the KEGG database, and five metabolic pathways were significantly enriched, including the cysteine and methionine metabolism, the riboflavin metabolism, the purine metabolism, the glycerolipid metabolism, and the glycerophospholipid metabolism. Collectively, our metabolomics analysis revealed that deficiency of GAT-2 influenced the metabolomics profile of Th1 cells, which will provide insights into T cell response to GAT-2 deficiency in mice. Data are available via MetaboLights with identifier MTBLS3358.

Changes in the Microbiome of the Inner Surface of Clear Aligners After Different Usage Periods.

Clear aligners are removable orthodontic appliances that cover the tooth surface. The microbial composition and pH of the inner surface of aligners directly affect the enamel health. In this study, eight subjects who used the same type of clear aligners were instructed to brush their teeth normally and to not clean their aligners until sampling. Saliva and the contents of the inner surface of the aligners (liquid and plaque) were collected at 0 h (T0), 4 h (T4), 8 h (T8), 12 h (T12), and 24 h (T24) after usage, and pH values and microbial compositions were measured. The microbial composition was analyzed with 16S rRNA gene sequencing, and changes were assessed based on operational taxonomic unit abundance. The pH, alpha diversity values, and abundance of specific microbes on the inner surface of the aligners gradually decreased from T0 to T24 (P < 0.05). An insignificant increase in microbial community beta diversity was observed from T0 to T24. Principal component analysis revealed that the microbial composition at T0 was different from at T12 and T24. The relative abundances of phylum Firmicutes (P < 0.01), orders Lactobacillales and Bacteroidales (P < 0.05), and genus Streptococcus and species Streptococcus infantis increased significantly, while those of genera Actinomyces and Rothia and species Rothia aeria decreased significantly at T24 (P < 0.05). These findings reveal that uncleaned aligners might lead to enamel damage, especially after continuous usage for 12 h. Thus, clear aligners should be cleaned after 12 h of usage or at least within 24 h of usage.

Impact of Hyperbaric Oxygen on the Healing of Teeth Extraction Sockets and Alveolar Ridge Preservation.

OBJECTIVES: The purpose of this study was to investigate the role of hyperbaric oxygen (HBO) in the healing of teeth extraction sockets and in alveolar ridge preservation. This may provide an experimental basis for the widespread application of HBO in oral implantation. METHODS: A total of 32 beagle dogs were included in the study and randomly divided equally between an HBO group treated with hyperbaric oxygen (100% O2, 2.4 atm, 90 min/day, 5 times/week, 6 weeks) and a normobaric oxygen (NBO) group treated with normal air in the same chamber. The lateral incisors of the maxillary and mandible of each dog were extracted, and the right upper and lower incisor extraction sockets (A2C2) were allowed to heal naturally, while left upper and lower incisor sockets (B2D2) received implants of a commercial bone substitute. At 4 and 8 weeks after surgery, clinical observation, cone-beam computerized tomography (CBCT), histomorphology observation, and expression levels of vascular endothelial growth factor (VEGF) and bone morphogenetic protein 2 (BMP-2) were analyzed to evaluate new bone formation, mineralization, and reconstruction. RESULTS: After 4 and 8 weeks, bone width and lip contour of the extraction socket in the NBO group were significantly reduced and collapsed in comparison with the HBO group. CBCT showed that the difference in vertical height between the alveolar crest of the labial tongue and palatal side of the extraction sockets was smaller in the HBO than NBO group. There was a significant difference in new bone formation (P < 0.05) and bone mineral density (P < 0.05) between the HBO and NBO groups, and the HBO group showed significantly greater new bone and bone reconstruction based on histology. Furthermore, the expression levels of VEGF and BMP-2 were higher in the HBO group. CONCLUSION: HBO reduced bone resorption and promoted early bone formation, bone mineralization, and reconstruction in the extraction sockets. HBO greatly reduced the healing time of the extraction sockets and promoted alveolar ridge preservation, thus showing promise for the clinic.

H2O2-responsive polymeric micelles with a benzil moiety for efficient DOX delivery and AIE imaging.

Nano drug delivery is a promising domain in biomedical theranostics and has aroused more and more attention in recent years. We report here an amphiphilic polymer TPG1, bearing a H2O2-sensitive benzil and an AIE fluorophore tetraphenylethene (TPE) unit, which is able to self-assemble into spherical nanosized micelles in aqueous solution. Doxorubicin (DOX) can be encapsulated into TPG1 micelles efficiently with the loading capability of up to 59% by weight. The benzil moiety could be cleaved via the Baeyer-Villiger type reaction in the presence of H2O2, leading to the decomposition of TPG1 micelles and release of DOX. In vitro studies indicated that DOX-loaded TPG1 micelles can be internalized by cancer cells, followed by unloading encapsulated DOX under the stimulation of H2O2. The drug release process can be monitored by the AIE fluorescence from the degradation products containing a TPE moiety. MTT assays against HeLa and HepG2 cancer cells demonstrated that DOX-loaded micelles showed good anticancer efficacy. The polymer TPG1 and the corresponding decomposed products showed great biocompatibility. Our data suggest that TPG1 has the potential to be employed for the controlled drug delivery system.

Impact of Hyperbaric Oxygen on Tissue Healing around Dental Implants in Beagles.

BACKGROUND The impact of hyperbaric oxygen (HBO) on the healing of soft tissues around dental implants was studied in a beagle model. MATERIAL AND METHODS Beagle dogs were randomized to receive implants, followed by postoperative HBO therapy or not (n=10 per group). On postoperative days 3, 7, and 14, tissue specimens were paraffin-embedded and analyzed by hematoxylin-eosin and Masson staining, as well as immunohistochemistry against CD31. RESULTS Scores for inflammation pathology based on hematoxylin-eosin staining and mean optical density of collagen fibers were significantly different between the HBO and control groups on postoperative days 3 and 7 (P<0.05), but not on day 14. Mean optical density due to anti-CD31 staining was significantly higher in the HBO group on postoperative days 3, 7, and 14 (P<0.05). CONCLUSIONS These results suggest that HBO may promote early osteogenesis and soft tissue healing after implantation.

Effects of Structural Changes in Subchondral Bone on Articular Cartilage in a Beagle Dog Model.

OBJECTIVE: Using MR T2-mapping and histopathologic score for articular cartilage to evaluate the effect of structural changes in subchondral bone on articular cartilage. METHODS: Twenty-four male Beagle dogs were randomly divided into a subchondral bone defect group (n = 12) and a bone cement group (n = 12). Models of subchondral bone defectin the medial tibial plateau and subchondral bone filled with bone cement were constructed. In all dogs, the left knee joint was used as the experimental sideand the right knee as the sham side. The T2 value for articular cartilage at the medial tibial plateau was measured at postoperative weeks 4, 8, 16, and 24. The articular cartilage specimens were stained with hematoxylin and eosin, and evaluated using the Mankin score. RESULTS: There was a statistically significant difference (P < 0.05) in Mankin score between the bone defect group and the cement group at postoperative weeks 16 and 24. There was a statistically significant difference in the T2 values between the bone defect group and its sham group (P < 0.05) from week 8, and between the cement group and its sham group (P < 0.05) from week 16. There was significant difference in T2 values between the two experimental groups at postoperative week 24 (P < 0.01). The T2 value for articular cartilage was positively correlated with the Mankin score (ρ = 0.758, P < 0.01). CONCLUSION: Structural changes in subchondral bone can lead to degeneration of the adjacent articular cartilage. Defects in subchondral bone cause more severe degeneration of cartilage than subchondral bone filled with cement. The T2 value for articular cartilage increases with the extent of degeneration. MR T2-mapping images and the T2 value for articular cartilage can indicate earlycartilage degeneration.

In vitro and in vivo CT imaging using bismuth sulfide modified with a highly biocompatible Pluronic F127.

Probe bismuth sulfide modified with Pluronic F127 (Bi2S3-PF127), which has high biocompatibility and dispersibility, is synthesized using triblock copolymer Pluronic F127 to modify hydrophobic Bi2S3 nanoparticles that are prepared by a hot injection method. TEM results show that most of the probe has a length of about 14.85 ± 1.70 nm and a breadth of about 4.79 ± 0.63 nm. After injected into the tail vein of a mouse, the probe has obvious CT contrast enhancement capability from x-ray CT imaging results. Meanwhile, the probe's in vivo toxicity is also studied. It is found that hematoxylin and eosin stains of major organs have no change. A biochemical analysis (alanine aminotransferase and aspartate aminotransferase) prove the probe has no adverse effects. The results of a blood analysis (white blood cell count, red blood cell count, hemoglobin, and platelet count) are also normal. The biological distribution of Bi by ICP-AES shows that most of nanoparticles are cleaned out after injection 48 h, and the circulation half-life of the probe is 5.0 h, suggesting that Bi2S3-PF127 has a long circulation and indicating that the Bi2S3-PF127 probe has good biocompatibility and safety.

Biodegradable PBAT microplastics adversely affect pakchoi (Brassica chinensis L.) growth and the rhizosphere ecology: Focusing on rhizosphere microbial community composition, element metabolic potential, and root exudates.

Biodegradable plastics (BPs) have gained increased attention as a promising solution to plastics pollution problem. However, BPs often exhibited limited in situ biodegradation in the soil environment, so they may also release microplastics (MPs) into soils just like conventional non-degradable plastics. Therefore, it is necessary to evaluate the impacts of biodegradable MPs (BMPs) on soil ecosystem. Here, we explored the effects of biodegradable poly(butylene adipate-co-terephthalate) (PBAT) MPs and conventional polyethylene (PE) MPs on soil-plant (pakchoi) system at three doses (0.02 %, 0.2 %, and 2 %, w/w). Results showed that PBAT MPs reduced plant growth in a dose-dependent pattern, while PE MPs exhibited no significant phytotoxicity. High-dose PBAT MPs negatively affected the rhizosphere soil nutrient availability, e.g., decreased available phosphorus and available potassium. Metagenomics analysis revealed that PBAT MPs caused more serious interference with the rhizosphere microbial community composition and function than PE MPs. In particular, compared with PE MPs, PBAT MPs induced greater changes in functional potential of carbon, nitrogen, phosphorus, and sulfur cycles, which may lead to alterations in soil biogeochemical processes and ecological functions. Moreover, untargeted metabolomics showed that PBAT MPs and PE MPs differentially affect plant root exudates. Mantel tests, correlation analysis, and partial least squares path model analysis showed that changes in plant growth and root exudates were significantly correlated with soil properties and rhizosphere microbiome driven by the MPs-rhizosphere interactions. This work improves our knowledge of how biodegradable and conventional non-degradable MPs affect plant growth and the rhizosphere ecology, highlighting that BMPs might pose greater threat to soil ecosystems than non-degradable MPs.

Mechanistic investigation of seeded growth in triblock copolymer stabilized gold nanoparticles.

We report the seeded synthesis of gold nanoparticles (GNPs) via the reduction of HAuCl4 by (L31 and F68) triblock copolymer (TBP) mixtures. In the present study, we focused on [TBP]/[Au(III)] ratios of 1-5 (≈1 mM HAuCl4) and seed sizes ~20 nm. Under these conditions, the GNP growth rate is dominated by both the TBP and seed concentrations. With seeding, the final GNP size distributions are bimodal. Increasing the seed concentration (up to ~0.1 nM) decreases the mean particle sizes 10-fold, from ~1000 to 100 nm. The particles in the bimodal distribution are formed by the competitive direct growth in solution and the aggregative growth on the seeds. By monitoring kinetics of GNP growth, we propose that (1) the surface of the GNP seeds embedded in the TBP cavities form catalytic centers for GNP growth and (2) large GNPs are formed by the aggregation of GNP seeds in an autocatalytic growth process.

Preparation of doxycycline-imprinted magnetic microspheres by inverse-emulsion suspension polymerization for magnetic dispersion extraction of tetracyclines from milk samples.

A magnetic dispersion extraction method was developed based on a molecularly imprinted magnetic microsphere (MIMM) for the selective clean-up and enrichment of tetracycline antibiotics from milk samples. The MIMMs were prepared by inverse-emulsion suspension polymerization, using doxycycline, trimethylolpropane trimethacrylate, acrylamide, methacrylic acid, and surface-modified Fe3 O4 as a template molecule, crosslinker, functional monomer, and magnetic component, respectively. Synthesis and extraction conditions were optimized for obtaining excellent affinity and high selectivity. The magnetism, covering amount, and selectivity of the magnetic microspheres were characterized by a vibrating sample magnetometer, thermogravimetric analysis, and a competitive recognition experiment. The MIMMs were applied to separate tetracycline antibiotics from milk samples by magnetic dispersion extraction, and an enrichment factor of 9.28 and a good sample clean-up were obtained. The average recoveries of four tetracycline antibiotics were obtained in the range of 74.5-93.8% with a precision of 1.2-5.2%. The LODs and LOQs of the proposed method were in the range of 7.4-19.4 and 24.7-64.7 µg/kg, respectively. The results indicated that magnetic dispersion extraction using MIMMs is a powerful tool for food-sample pretreatment with high selectivity and a simplified procedure.

Effect of different amounts of bamboo charcoal on properties of biodegradable bamboo charcoal/polylactic acid composites.

Biodegradable composites were prepared from polylactic acid (PLA) and bamboo charcoal (BC) by melt blending and hot pressing. The effects of BC addition on the mechanical properties, water absorption, DMA, TGA, DSC, and CONE of BC/PLA composites were investigated. The microscopic morphology of the composites was analyzed by SEM. The results showed that for BC addition of 40 wt%, the mechanical strength, thermal properties, and flame retardant properties of the composites were improved compared with those of PLA, with a 2.24 % increase in flexural strength and a 1535 % increase (500 °C) in TG mass retention rate. The crystallinity increased by 129.66 %, the peak loss factor decreased by 31.15 %, the time required for combustion was delayed by 168 s, the peak heat release rate decreased by 29.40 %, the carbon residue rate detected by cone calorimetry increased by 48.50 %, and the peak mass loss rate decreased by 48.82 %. The addition of BC enhanced the crystallization capacity of PLA, and improved the thermal properties and flame retardant properties of the prepared composites. The results showed that materials prepared with a BC content of 40 wt% exhibited the best overall performance.

Fretting stimulation enhances bone growth at the interface between hydroxyapatite coating and bone.

Biologically fixed arthroplasty is limited in its development by the long postoperative recovery time and the low quality of solidity of the fixed interface in the short postoperative period. Therefore, fretting stimulation is used to accelerate the combination between bone tissue and the biological fixation interface of artificial joint prostheses. The effects of different compression loads and tangential micro-motion amplitude on the growth rate of bone tissue and the firm quality of fixation interface were studied by using two kinds of micro-motion stimuli: compression and tangential micro-motion. The mechanism of micro-motion stimulation to promote bone growth at the fixation interface was revealed. The results of binding force detection of biological fixation interface and bone tissue section staining showed that the bone tissue and hydroxyapatite coating interface had the most tendency to produce new bone tissue under compression load of 4 N. In the tangential fretting environment, the tangential fretting amplitude of ± 40 µm and the normal load of 7.5 N were the most conducive to bone growth, making the combination of bone tissue and titanium alloy prosthesis coated with hydroxyapatite more firm. The study is important for accelerating the integration and shortening the rehabilitation time after artificial joint replacement.

In vitro biocompatiability and mechanical properties of bone adhesive tape composite based on poly(butyl fumarate)/poly(propylene fumarate)-diacrylate networks.

Polyfumarate has been considered as injectable and biodegradable bone cement. However, its mechanical and degradation properties are particularly important. Therefore, the current study aimed to develop the properties by compositing poly (butyl fumarate)-based networks with hydroxyapatite nano-powders. In this regard, the poly (butyl fumarate) (PBF) matrix composite was compared with different components by evaluating their composition, mechanical properties, hydrophilicity, and biodegradability. Furthermore, their bioactivity in the phosphate-buffered saline (PBS) and, via applying mouse embryo osteoblast precursor cells (MC3T3-E1), their cell interaction, including adhesion, proliferation, and in vitro cytotoxicity assay, were assessed. The addition of hydroxyapatite improved the mechanical strength and modulus of PBF matrix composite. The composite reinforced with 3 wt% hydroxyapatite showed a higher lap-shear strength (1.68 MPa) and bonding strength (4.30 MPa), a maximum compression strength at fracture (95.18 MPa), modulus (925.29 MPa), and compression strength at yield (31.43 MPa), respectively. Also, hydrophilicity and in vitro degradation of the composite were enhanced in the presence of hydroxyapatite. In this condition, after a period of immersion (52 weeks) in PBS, the weight loss rate, and degradation rate of the composite increased. The composite proliferation, adhesion, and toxicity of MC3T3-E1 cells improved in comparison to the PBF matrix composite. Accordingly, controllable strength and degradation of the composite, along with its proven biocompatibility, make the composite a candidate for the treatment of comminuted fractures.

Development and characterization of a mouse floxed Bmp2 osteoblast cell line that retains osteoblast genotype and phenotype.

Bone morphogenetic protein 2 (Bmp2) is essential for osteoblast differentiation and osteogenesis. Generation of floxed Bmp2 osteoblast cell lines is a valuable tool for studying the effects of Bmp2 on osteoblast differentiation and its signaling pathways during skeletal metabolism. Due to relatively limited sources of primary osteoblasts, we have developed cell lines that serve as good surrogate models for the study of osteoblast cell differentiation and bone mineralization. In this study, we established and characterized immortalized mouse floxed Bmp2 osteoblast cell lines. Primary mouse floxed Bmp2 osteoblasts were transfected with pSV3-neo and clonally selected. These transfected cells were verified by PCR and immunohistochemistry. To determine the genotype and phenotype of the immortalized cells, cell morphology, proliferation, differentiation and mineralization were analyzed. Also, expression of osteoblast-related gene markers including Runx2, Osx, ATF4, Dlx3, bone sialoprotein, dentin matrix protein 1, osteonectin, osteocalcin and osteopontin were examined by quantitative RT-PCR and immunohistochemistry. These results showed that immortalized floxed Bmp2 osteoblasts had a higher proliferation rate but preserved their genotypic and phenotypic characteristics similar to the primary cells. Thus, we, for the first time, describe the development of immortalized mouse floxed Bmp2 osteoblast cell lines and present a useful model to study osteoblast biology mediated by BMP2 and its downstream signaling transduction pathways.

Phenotypic variation of fluoride responses between inbred strains of mice.

Excessive systemic exposure to fluoride (F) can lead to disturbances in bone homeostasis and dental enamel development. We have previously shown strain-specific responses to F in the development of dental fluorosis (DF) and in bone formation/mineralization. The current study was undertaken to further investigate F responsive variations in bone metabolism and to determine possible relationships with DF susceptibility. Seven-week-old male mice from FVB/NJ, C57BL/6J, C3H/HeJ, A/J, 129S1/SvImJ, AKR/J, DBA/2J, and BALB/cByJ inbred strains were exposed to NaF (0 or 50 ppm as F(-)) in drinking water for 60 days. Sera were collected for F, Ca, Mg, PO(4), iPTH, sRANKL, and ALP levels. Bone marrow cells were subjected to ex vivo cell culture for osteoclast potential and CFU colony assays (CFU-fibroblast, CFU-osteoblast, CFU-erythrocyte/granulocyte/macrophage/megakaryocyte, CFU-granulocyte/macrophage, CFU-macrophage, and CFU-granulocyte). Femurs and vertebrae were subjected to micro-CT analyses, biomechanical testing, and F, Mg, and Ca content assays. DF was evaluated using quantitative fluorescence and clinical criteria. Strain-specific responses to F were observed for DF, serum studies, ex vivo cell culture studies, and bone quality. Among the strains, there were no patterns or significant correlations between DF severity and the actions of F on bone homeostasis (serum studies, ex vivo assays, or bone quality parameters). The genetic background continues to play a role in the actions of F on tooth enamel development and bone homeostasis. F exposure led to variable phenotypic responses between strains involving dental enamel development and bone metabolism.

Fine mapping of dental fluorosis quantitative trait loci in mice.

Genetic factors underlie the susceptibility and the resistance to dental fluorosis (DF). The A/J (DF susceptible) and 129P3/J (DF resistant) mouse strains have previously been used to detect quantitative trait loci (QTLs) associated with DF on chromosome (Chr) 2 and Chr 11. In the present study, increased marker density genotyping followed by interval mapping was performed to narrow the QTL intervals and improve the logarithm of the odds (to the base 10) (LOD) scores. Narrower intervals were obtained on Chr 2 where LOD ≥ 6.0 (57-84 cM or ≈ 51 Mb), LOD ≥ 7.0 (62-79 cM or ≈ 32 Mb), and LOD ≥ 8.0 (65-74 cM or ≈ 17 Mb); and on Chr 11 where LOD ≥ 6.0 (18-51 cM or ≈ 53 Mb), LOD ≥ 7.0 (28-48 cM or ≈ 34 Mb), and LOD ≥ 8.0 (31-45 cM or ≈ 22 Mb). Haplotype analysis between A/J and 129P3/J mice further reduced the QTL intervals. Accn1 was selected as a candidate gene based upon its location near the peak LOD score on Chr 11 and distant homology with the Caenorhabditis elegans fluoride-resistance gene, flr1. The severity of DF between Accn1(-/-) and wild-type mice was not significantly different. Hence, the loss of ACCN1 function does not modify DF severity in mice. Narrowing the DF QTL intervals will facilitate additional candidate gene selections and interrogation.

Multimode-ultrasound and microwave assisted natural ternary deep eutectic solvent sequential pretreatments for corn straw biomass deconstruction under mild conditions.

Mild and effective pretreatments are essential to deconstruct lignocellulosic biomass so as to reuse cellulose content for value-added products. In this study, sequential multimode-ultrasound and microwave with natural ternary deep eutectic solvent (NATDES) pretreatments were used to deconstruct corn straw and optimized factors such as NATDES, ultrasonic, and microwave parameters. Results indicated that the ultrasound-NATDES or microwave-NATDES pretreatment could remove 37.86% and 52.36% lignin, respectively. When using sequential multimode-ultrasound and microwave assisted NATDES pretreatment, the delignification efficiency increased to 61.50%, and the cellulose content increased from 34.70% to 76.08%. In addition, the delignification of sequential multimode-ultrasound and microwave assisted NATDES pretreatment (under the mild conditions of microwave heating at 60 °C and 60 min) increased to 57.39%, and the cellulose content increased to 59.98%, too. This highlighted the effect of the combined ultrasound and microwave technology. Finally, the microstructural changes of mercury intrusion porosimeters, scanning electron microscopy, thermogravimetric, X-ray diffraction and Fourier transform mid-infrared spectroscopy were conducted to confirm the effectiveness of this method to deconstruct corn straw. A mechanism of the deconstruction of corn straw biomass in NATDES with the assistance of the sequential multimode-ultrasound and microwave heating was proposed. This research could open a window for future use of biomass energy by deconstructing lignocellulosic biomasses using environmentally friendly pretreatment methods.

Immortalized mouse floxed Bmp2 dental papilla mesenchymal cell lines preserve odontoblastic phenotype and respond to BMP2.

Bone morphogenetic protein 2 (Bmp2) is essential for odontogensis and dentin mineralization. Generation of floxed Bmp2 dental mesenchymal cell lines is a valuable application for studying the effects of Bmp2 on dental mesenchymal cell differentiation and its signaling pathways during dentinogenesis. Limitation of the primary culture of dental mesenchymal cells has led to the development of cell lines that serve as good surrogate models for the study of dental mesenchymal cell differentiation into odontoblasts and mineralization. In this study, we established and characterized immortalized mouse floxed Bmp2 dental papilla mesenchymal cell lines, which were isolated from 1st mouse mandibular molars at postnatal day 1 and immortalized with pSV40 and clonally selected. These transfected cell lines were characterized by RT-PCR, immunohistochemistry, and analyzed for alkaline phosphatase activity and mineralization nodule formation. One of these immortalized cell lines, iBmp2-dp, displayed a higher proliferation rate, but retained the genotypic and phenotypic characteristics similar to primary cells as determined by expression of tooth-specific markers as well as demonstrated the ability to differentiate and form mineralized nodules. In addition, iBmp2-dp cells were inducible and responded to BMP2 stimulation. Thus, we for the first time described the establishment of an immortalized mouse floxed Bmp2 dental papilla mesenchyma cell line that might be used for studying the mechanisms of dental cell differentiation and dentin mineralization mediated by Bmp2 and other growth factor signaling pathways.