Localised laccase activity modulates distribution of lignin polymers in gymnosperm compression wood.

The woody stems of coniferous gymnosperms produce specialised compression wood to adjust the stem growth orientation in response to gravitropic stimulation. During this process, tracheids develop a compression-wood-specific S2 L cell wall layer with lignins highly enriched with p-hydroxyphenyl (H)-type units derived from H-type monolignol, whereas lignins produced in the cell walls of normal wood tracheids are exclusively composed of guaiacyl (G)-type units from G-type monolignol with a trace amount of H-type units. We show that laccases, a class of lignin polymerisation enzymes, play a crucial role in the spatially organised polymerisation of H-type and G-type monolignols during compression wood formation in Japanese cypress (Chamaecyparis obtusa). We performed a series of chemical-probe-aided imaging analysis on C. obtusa compression wood cell walls, together with gene expression, protein localisation and enzymatic assays of C. obtusa laccases. Our data indicated that CoLac1 and CoLac3 with differential oxidation activities towards H-type and G-type monolignols were precisely localised to distinct cell wall layers in which H-type and G-type lignin units were preferentially produced during the development of compression wood tracheids. We propose that, not only the spatial localisation of laccases, but also their biochemical characteristics dictate the spatial patterning of lignin polymerisation in gymnosperm compression wood.

Comparative multi-omics analysis reveals diverse latex-based defense strategies against pests among latex-producing organs of the fig tree (Ficus carica).

MAIN CONCLUSION: Latexes in immature fruit, young petioles and lignified trunks of fig trees protect the plant using toxic proteins and metabolites in various organ-dependent ways. Latexes from plants contain high amounts of toxic proteins and metabolites, which attack microbes and herbivores after exudation at pest-induced wound sites. The protein and metabolite constituents of latexes are highly variable, depending on the plant species and organ. To determine the diversity of latex-based defense strategies in fig tree (Ficus carica) organs, we conducted comparative proteomic, transcriptomic and metabolomic analyses on latexes isolated from immature fruit, young petioles and lignified trunks of F. carica after constructing a unigene sequence library using RNA-seq data. Trypsin inhibitors were the most abundant proteins in petiole latex, while cysteine proteases ("ficins") were the most abundant in immature fruit and trunk latexes. Galloylglycerol, a possible defense-related metabolite, appeared to be highly accumulated in all three latexes. The expression levels of pathogenesis-related proteins were highest in the latex of trunk, suggesting that this latex had adapted a defensive role against microbe attacks. Although young petioles and immature fruit are both unlignified soft organs, and potential food for herbivorous insects, unigenes for the sesquiterpenoid pathway, which likely produces defense-associated volatiles, and the phenylpropanoid pathway, which produces toxic furanocoumarins, were expressed less in immature fruit latex. This difference may indicate that while petioles and fruit protect the plant from attack by herbivores, the fruit must also attract insect pollinators at younger stages and animals after ripening. We also suggest possible candidate transcription factors and signal transduction proteins that are involved in the differential expression of the unigenes.

Orthodontic skeletal anchorage using a palatal external plate.

We have developed the Anchor-Lock external plate system, which is fitted on the palate for the purpose of orthodontic skeletal anchorage. The aim of this study was to introduce the Anchor-Lock and assess its success rate. The Anchor-Lock is composed of titanium screws of 2·0-mm diameter and a titanium plate of 1·0-mm thickness. The external plate is rigidly interlocked with the heads of the screws, which are implanted trans-mucosally into palatal bone. Three types of Anchor-Lock are available. These were applied to 137 orthodontic patients (104 females and 33 males) aged 10-54 years. Two types of plate were used, a straight-shaped plate applied to the hard palate and to the anterior palate and a double-Y-shaped plate applied after tooth-borne rapid maxillary expansion. Success rate of the Anchor-Lock was 92·0% overall. No significant difference in success rate was found by age or sex of patients. Type or screw length of the Anchor-Lock did not affect success rate significantly. Success rate was significantly increased by the use of the surgical stent. The Anchor-Lock was effectively applied to distalize and/or intrude the upper molars. The Anchor-Lock system appears suitable for clinical use as an alternative to conventional screw- and plate-type orthodontic implants.

Limiting silicon supply alters lignin content and structures of sorghum seedling cell walls.

Sorghum has been recognized as a promising energy crop. The composition and structure of lignin in the cell wall are important factors that affect the quality of plant biomass as a bioenergy feedstock. Silicon (Si) supply may affect the lignin content and structure, as both Si and lignin are possibly involved in plant mechanical strength. However, our understanding regarding the interaction between Si and lignin in sorghum is limited. Therefore, in this study, we analyzed the lignin in the cell walls of sorghum seedlings cultured hydroponically with or without Si supplementation. Limiting the Si supply significantly increased the thioglycolic acid lignin content and thioacidolysis-derived syringyl/guaiacyl monomer ratio. At least part of the modification may be attributable to the change in gene expression, as suggested by the upregulation of phenylpropanoid biosynthesis-related genes under -Si conditions. The cell walls of the -Si plants had a higher mechanical strength and calorific value than those of the +Si plants. These results provide some insights into the enhancement of the value of sorghum biomass as a feedstock for energy production by limiting Si uptake.

Nitrogen deficiency results in changes to cell wall composition of sorghum seedlings.

Sorghum [Sorghum bicolor (L.) Moench] has been gaining attention as a feedstock for biomass energy production. While it is obvious that nitrogen (N) supply significantly affects sorghum growth and biomass accumulation, our knowledge is still limited regarding the effect of N on the biomass quality of sorghum, such as the contents and structures of lignin and other cell wall components. Therefore, in this study, we investigated the effects of N supply on the structure and composition of sorghum cell walls. The cell walls of hydroponically cultured sorghum seedlings grown under sufficient or deficient N conditions were analyzed using chemical, two-dimensional nuclear magnetic resonance, gene expression, and immunohistochemical methods. We found that the level of N supply considerably affected the cell wall structure and composition of sorghum seedlings. Limitation of N led to a decrease in the syringyl/guaiacyl lignin unit ratio and an increase in the amount and alteration of tissue distribution of several hemicelluloses, including mixed linkage (1 → 3), (1 → 4)-ß-D-glucan, and arabinoxylan. At least some of these cell wall alterations could be associated with changes in gene expression. Nitrogen status is thus one of the factors affecting the cell wall properties of sorghum seedlings.

An enteric polymer mitigates the effects of gastric pH on oral absorption of poorly soluble weak acid drugs from supersaturable formulations: A case study with dantrolene.

This study demonstrated that an enteric polymer can mitigate the effects of gastric pH on the oral absorption of a poorly water-soluble weak acid drug, dantrolene (DNT). An amorphous solid dispersion (ASD) of DNT with hydroxypropyl methylcellulose (HPMC) acetate succinate (ASD-HPMCAS) was prepared as the enteric released ASD (ER-SF). ASD with HPMC (ASD-HPMC) and DNT sodium salt were also used as immediate-release supersaturable formulations (IR-SFs) with and without water-soluble polymer, respectively. In vivo study with rats and in vitro study with a dissolution/permeation (D/P) system were performed to evaluate oral DNT absorption from each formulation under normal and high gastric pH conditions in rats and humans, respectively. The oral absorption of DNT from both IR-SFs in rats with a high gastric pH was significantly higher than that in rats with a normal gastric pH. In contrast, ASD-HPMCAS attenuated the difference in oral absorption between normal and high gastric pH conditions with significant improvement of DNT absorption. In vivo results implied that an enteric polymer delayed the onset of dissolution until after gastric emptying. ASD-HPMCAS generated supersaturation in the small intestine irrespective of gastric conditions, which was supported bythe in vitrostudy using the D/P system. This study suggested that an enteric polymer is useful to mitigate the inter- and intra-individual differences in oral absorption of poorly water-soluble weak acid drugs.

Optimal loading conditions for controlled movement of anterior teeth in sliding mechanics.

OBJECTIVE: To determine optimal loading conditions such as height of retraction force on the power arm and its position on the archwire in sliding mechanics. MATERIALS AND METHODS: A 3D finite element method (FEM) was used to simulate en masse anterior teeth retraction in sliding mechanics. The degree of labiolingual tipping of the maxillary central incisor was calculated when the retraction force was applied to different heights of a power arm set mesial or distal to the canine. RESULTS: When the power arm was placed mesial to the canine, at the level of 0 mm (bracket slot level), uncontrolled lingual crown tipping of the incisor was observed and the anterior segment of the archwire was deformed downward. At a power arm height of 5.5 mm, bodily movement was produced and the archwire was less deformed. When the power arm height exceeded 5.5 mm, the anterior segment of the archwire was raised upward and lingual root tipping occurred. When the power arm was placed distal to the canine, lingual crown tipping was observed up to a level of 11.2 mm. CONCLUSIONS: Placement of the power arm of an archwire between the lateral incisor and canine enables orthodontists to maintain better control of the anterior teeth in sliding mechanics. Both the biomechanical principles associated with the tooth's center of resistance and the deformation of the archwire should be taken into consideration for predicting and planning orthodontic tooth movement.

Real-time orthognathic surgical simulation using a mandibular motion tracking system.

We developed a new orthognathic surgical simulation system able to predict both occlusal correction and mandibular repositioning in three dimensions. This system uniquely integrates the real motion of the dental cast model with the virtual motion of the reconstructed cranio-facial model. The skeletal change of the mandibular osteotomy is simulated on the PC monitor while the occlusal change is confirmed by checking the cast model on the simulator. The simulation process is easily repeated and the operator can make several attempts to determine the final mandibular position. The occlusal relationship at the simulated mandibular posture is registered and the occlusal wafer splint, which ensures intermaxillary fixation, is fabricated on the simulator. This surgical simulation system appears to satisfy clinical demands well and is an important facilitator of communication between orthodontists and surgeons. Here, we outline the system and apply it to a demonstration case of orthognathic surgery.

Mixed-reality simulation for orthognathic surgery.

BACKGROUND: Mandibular motion tracking system (ManMoS) has been developed for orthognathic surgery. This article aimed to introduce the ManMoS and to examine the accuracy of this system. METHODS: Skeletal and dental models are reconstructed in a virtual space from the DICOM data of three-dimensional computed tomography (3D-CT) recording and the STL data of 3D scanning, respectively. The ManMoS uniquely integrates the virtual dento-skeletal model with the real motion of the dental cast mounted on the simulator, using the reference splint. Positional change of the dental cast is tracked by using the 3D motion tracking equipment and reflects on the jaw position of the virtual model in real time, generating the mixed-reality surgical simulation. ManMoS was applied for two clinical cases having a facial asymmetry. In order to assess the accuracy of the ManMoS, the positional change of the lower dental arch was compared between the virtual and real models. RESULTS: With the measurement data of the real lower dental cast as a reference, measurement error for the whole simulation system was less than 0.32 mm. In ManMoS, the skeletal and dental asymmetries were adequately diagnosed in three dimensions. Jaw repositioning was simulated with priority given to the skeletal correction rather than the occlusal correction. In two cases, facial asymmetry was successfully improved while a normal occlusal relationship was reconstructed. Positional change measured in the virtual model did not differ significantly from that in the real model. CONCLUSIONS: It was suggested that the accuracy of the ManMoS was good enough for a clinical use. This surgical simulation system appears to meet clinical demands well and is an important facilitator of communication between orthodontists and surgeons.

Involvement of HMGB1 and RAGE in IL-1ß-induced gingival inflammation.

OBJECTIVE: Extracellularly released high mobility group box 1 (HMGB1) protein behaves as a cytokine, promotes inflammation and participates in the pathogenesis of several disorders in peripheral organs. The role of HMGB1 and receptor for advanced glycation end products (RAGE) expressed in gingival inflammatory tissues was explored. METHODS: Real time PCR was applied to assay HMGB1 and RAGE mRNA expression in gingival epithelial and fibroblast cells induced by interleukin-1ß (IL-1ß). A highly selective inhibitor of inducible nitric oxide (iNOS) was employed. ELISA was done for measurement of HMGB1 concentrations in cell culture media of gingival epithelial and fibroblast cells. Immunohistochemistry was performed to analyse the expression and sub-cellular localization of HMGB1, together with RAGE, in specimens obtained from patients with chronic inflammation. RESULTS: A time-dependent response of HMGB1 and RAGE expression in gingival cells to IL-1ß induction was observed. IL-1ß promotes HMGB1 production in human gingival epithelial cells in a nitric oxide-dependent manner. HMGB1 and RAGE appeared highly expressed in gingival inflammatory tissues. CONCLUSION: These results demonstrate that HMGB1 and RAGE are abundantly expressed in gingiva and promptly released during gingival inflammation. We suggest a role for HMGB1/RAGE/iNOS signalling on inflamed gingival epithelial cells.

Wall-associated kinase 1 (WAK1) is crosslinked in endomembranes, and transport to the cell surface requires correct cell-wall synthesis.

The Arabidopsis thaliana wall-associated kinases (WAKs) bind to pectin with an extracellular domain and also contain a cytoplasmic protein kinase domain. WAKs are required for cell elongation and modulate sugar metabolism. This work shows that in leaf protoplasts a WAK1-GFP fusion protein accumulates in a cytoplasmic compartment that contains pectin. The WAK compartment contains markers for the Golgi, the site of pectin synthesis. The migration of WAK1-GFP to the cell surface is far slower than that of a cell surface receptor not associated with the cell wall, is influenced by the presence of fucose side chains on one or more unidentified molecules that might include pectin, and is dependent upon cellulose synthesis on the plasma membrane. WAK is crosslinked into a detergent-insoluble complex within the cytoplasmic compartment before it appears on the cell surface, and this is independent of fucose modification or cellulose synthesis. Thus, the assembly and crosslinking of WAKs may begin at an early stage within a cytoplasmic compartment rather than in the cell wall itself, and is coordinated with synthesis of surface cellulose.