Accuracy Evaluation of a Three-Dimensional Model Generated from Patient-Specific Monocular Video Data for Maxillofacial Prosthetic Rehabilitation: A Pilot Study.

PURPOSE: To evaluate if the combination of a monoscopic photogrammetry technique and smartphone-recorded monocular video data could be appropriately applied to maxillofacial prosthesis fabrication. MATERIALS AND METHODS: Smartphone video and laser scanning data were recorded for five healthy volunteers (24.1 ± 0.7 years). Three-dimensional (3D) facial models were generated using photogrammetry software and a laser scanner. Smartphone-recorded video data were used to generate a photogrammetric 3D model. The videos were recorded at two resolutions: 1080 × 1920 (high resolution) and 720 × 1280 pixels (low resolution). The lengths of five nasal component parts (nose height, nasal dorsum length, nasal column length, nasal ala length, and nose breadth) were compared in the photogrammetric 3D models (as the test model) and the laser scanned 3D models (as the validation model) using reverse engineering software. RESULTS: There was a significant difference in the nasal dorsum length between the test model and the validation model (high resolution; 95% confidence interval, 2.05-5.07, Low resolution; confidence interval, 2.19-5.69). In contrast to the nasal dorsum length, there were no significant differences in nose height, nose breadth, nasal ala length, and nasal column length. CONCLUSION: Using smartphone-recorded video data and a photogrammetry technique may be a promising technique to apply in the maxillofacial prosthetic rehabilitation workflow.

Data on the effect of in vivo knockdown using artificial ErbB3 miRNA on Remak bundle structure.

Mature Schwann cells, the peripheral nervous system (PNS) glial cells, have two major roles for neuronal axons (Bunge, 1993) [1]. For large diameter axons, Schwann cells form myelin sheaths with multiple layers. For small diameter axons, they form Remak bundle composed only of single layer of the Schwann cell plasma membrane. In the PNS, ErbB3 forms a dimer with ErbB2 on the Schwann cell plasma membrane. ErbB3 plays a key role in myelination by myelinating Schwann cells, that is to say, its role in myelin thickness. Herein we provide the data regarding the effect of in vivo knockdown of ErbB3 on the thickness between an axon and a neighboring axon in Remak bundle, which is formed by non-myelinating Schwann cells. Since ErbB3 knockout mice are embryonically lethal, Schwann cell lineage-specific transgenic mice transcribing ErbB3 shRNA with an artificial miRNA backbone were generated and used in these experiments (Torii et al., 2014) [2].

Data on the effect of hypomyelinating leukodystrophy 6 (HLD6)-associated mutations on the TUBB4A properties.

Hypomyelinating leukodystrophy (HLD) is genetic demyelinating or dysmyelinating disease and is associated with at least 13 responsible genes. The mutations seem likely cause the functional deficiency of their gene products. HLD4- and HLD5-associated HSPD1 and FAM126A mutations affect biochemical properties of the gene products (Miyamoto et al. (2015,2014) [[1], [2]]). Herein we provide the data regarding the effects of HLD6-associated tubulin beta 4A (TUBB4A) mutations on the properties.

Data supporting the role of Fyn in embryonic sciatic nerve fasciculation.

Fyn is the cytoplasmic tyrosine kinase that has critical roles in many aspects of biological functions. In the central [1] and peripheral nervous systems [2], [3], Fyn plays the key role in initiating myelination by myelin-forming glial cells (Schwann cells and oligodendrocytes). Herein we provide the data regarding the role of Fyn in fasciculation and branching of embryonic peripheral nerves.

Neuregulin-1 type III knockout mice exhibit delayed migration of Schwann cell precursors.

In an embryonic developmental stage of the peripheral nervous system (PNS), Schwann cell precursors migrate along neuronal axons to their final destinations. After birth, they eventually wrap around individual axons to form myelin sheaths, which insulate axons to increase the nerve conduction velocity. Some growth factors and adhesion molecules are known to control these developmental stages from in the fish to in the mammal. Neuregulin-1 (NRG1), which is composed of many alternative splicing variants, is such a growth factor. Among these variants, the type III isoform of NRG1, interacting with ErbB2 and ErbB3 receptors on Schwann cells, plays an essential role in myelination in the fish and the mammal. NRG1 type III is also known to promote migration of fish Schwann cell precursors; however, it still remains to be clarified whether mammalian type III isoform does it. We have therefore generated type III isoform-specific knockout mice in inbred strain. The mice result in delayed migration of the precursors from the dorsal to ventral root via a peripheral ganglion, comparing littermate controls. Similar results are observed in an in vitro migration assay using reaggregated Schwann cell precursors. Furthermore, the knockout mice exhibit reduced myelin thickness, consistent with the established role of NRG1 type III in myelination. These results indicate that in mice, NRG1 type III plays a key role not only in myelination but also in migration.

Data supporting the role of Fyn in initiating myelination in the peripheral nervous system.

Transgenic mice, which express active Fyn tyrosine kinase under the control of a glial fibrillary acidic protein promoter, have been produced. This promoter induces protein expression in the initiation stage of myelination in the peripheral nervous system (PNS) "Phosphorylation of cytohesin-1 by Fyn is required for initiation of myelination and the extent of myelination during development (Yamauchi et al., 2015 [1])". Herein we provide the data regarding myelination-related protein markers and myelin ultrastructure in transgenic mice.

Dock8 interacts with Nck1 in mediating Schwann cell precursor migration.

During embryonic development of the peripheral nervous system (PNS), Schwann cell precursors migrate along neuronal axons to their final destinations, where they will myelinate the axons after birth. While the intercellular signals controlling Schwann cell precursor migration are well studied, the intracellular signals controlling Schwann cell precursor migration remain elusive. Here, using a rat primary cell culture system, we show that Dock8, an atypical Dock180-related guanine-nucleotide exchange factor (GEF) for small GTPases of the Rho family, specifically interacts with Nck1, an adaptor protein composed only of Src homology (SH) domains, to promote Schwann cell precursor migration induced by platelet-derived growth factor (PDGF). Knockdown of Dock8 or Nck1 with its respective siRNA markedly decreases PDGF-induced cell migration, as well as Rho GTPase activation, in precursors. Dock8, through its unique N-terminal proline-rich motif, interacts with the SH3 domain of Nck1, but not with other adaptor proteins composed only of SH domains, e.g. Grb2 and CrkII, and not with the adaptor protein Elmo1. Reintroduction of the proline-rich motif mutant of Dock8 in Dock8 siRNA-transfected Schwann cell precursors fails to restore their migratory abilities, whereas that of wild-type Dock8 does restore these abilities. These results suggest that Nck1 interaction with Dock8 mediates PDGF-induced Schwann cell precursor migration, demonstrating not only that Nck1 and Dock8 are previously unanticipated intracellular signaling molecules involved in the regulation of Schwann cell precursor migration but also that Dock8 is among the genetically-conservative common interaction subset of Dock family proteins consisting only of SH domain adaptor proteins.

Arf6 mediates Schwann cell differentiation and myelination.

During development of the peripheral nervous system (PNS), Schwann cells wrap neuronal axons, becoming the myelin sheaths that help axonal functions. While the intercellular signals controlling the myelination process between Schwann cells and peripheral neurons are well studied, the transduction of these signals in Schwann cells still remains elusive. Here, we show that Arf6, an Arf protein of the small GTPase family, is involved in promoting the myelination process. Knockdown of Arf6 with the small-interfering (si)RNA in primary Schwann cells markedly decreases dibutyl-cyclic AMP-induced myelin marker protein expression, indicating that Arf6 plays a role in differentiation-like phenotypic changes. To obtain in vivo evidence, we generated small-hairpin (sh)RNA transgenic mice targeting Arf6 for Schwann cells. Transgenic mice exhibited reduced myelin thickness compared to littermate controls, consistent with the defective myelin formation observed in the transgenic mouse-derived Schwann cell and neuronal culture system. Transgenic mice also exhibited decreased phosphorylation of myelination-related signaling molecules such as Akt kinase cascade proteins as well as downregulation of myelin marker proteins. These results suggest that signaling through Arf6 is required for Schwann cell myelination, adding Arf6 to the list of intracellular signaling molecules involved in the myelination process.

Hypomyelinating leukodystrophy-associated missense mutation in HSPD1 blunts mitochondrial dynamics.

Myelin-forming glial cells undergo dynamic morphological changes in order to produce mature myelin sheaths with multiple layers. In the central nervous system (CNS), oligodendrocytes differentiate to insulate neuronal axons with myelin sheaths. Myelin sheaths play a key role in homeostasis of the nervous system, but their related disorders lead not only to dismyelination and repeated demyelination but also to severe neuropathies. Hereditary hypomyelinating leukodystrophies (HLDs) are a group of such diseases affecting oligodendrocytes and are often caused by missense mutations of the respective responsible genes. Despite increasing identification of gene mutations through advanced nucleotide sequencing technology, studies on the relationships between gene mutations and their effects on cellular and subcellular aberrance have not followed at the same rapid pace. In this study, we report that an HLD4-associated (Asp-29-to-Gly) mutant of mitochondrial heat shock 60-kDa protein 1 (HSPD1) causes short-length morphologies and increases the numbers of mitochondria due to their aberrant fission and fusion cycles. In experiments using a fluorescent dye probe, this mutation decreases the mitochondrial membrane potential. Also, mitochondria accumulate in perinuclear regions. HLD4-associated HSPD1 mutant blunts mitochondrial dynamics, probably resulting in oligodendrocyte malfunction. This study constitutes a first finding concerning the relationship between disease-associated HSPD1 mutation and mitochondrial dynamics, which may be similar to the relationship between another disease-associated HSPD1 mutation (MitCHAP-60 disease) and aberrant mitochondrial dynamics.

Arf6 guanine-nucleotide exchange factor cytohesin-2 regulates myelination in nerves.

In postnatal development of the peripheral nervous system (PNS), Schwann cells differentiate to insulate neuronal axons with myelin sheaths, increasing the nerve conduction velocity. To produce the mature myelin sheath with its multiple layers, Schwann cells undergo dynamic morphological changes. While extracellular molecules such as growth factors and cell adhesion ligands are known to regulate the myelination process, the intracellular molecular mechanism underlying myelination remains unclear. In this study, we have produced Schwann cell-specific conditional knockout mice for cytohesin-2, a guanine-nucleotide exchange factor (GEF) specifically activating Arf6. Arf6, a member of the Ras-like protein family, participates in various cellular functions including cell morphological changes. Cytohesin-2 knockout mice exhibit decreased Arf6 activity and reduced myelin thickness in the sciatic nerves, with decreased expression levels of myelin protein zero (MPZ), the major myelin marker protein. These results are consistent with those of experiments in which Schwann cell-neuronal cultures were treated with pan-cytohesin inhibitor SecinH3. On the other hand, the numbers of Ki67-positive cells in knockout mice and controls are comparable, indicating that cytohesin-2 does not have a positive effect on cell numbers. Thus, signaling through cytohesin-2 is required for myelination by Schwann cells, and cytohesin-2 is added to the list of molecules known to underlie PNS myelination.

Inhibition of RANKL-dependent cellular fusion in pre-osteoclasts by amiloride and a NHE10-specific monoclonal antibody.

The functions of Na(+) /H(+) exchangers (NHEs) during osteoclastic differentiation were investigated using the NHE inhibitor amiloride and a monoclonal antibody (MAb). Compared with sRANKL-stimulated control cells, amiloride decreased the number of large TRAP-positive osteoclast cells (OCs) with ≥10 nuclei and increased the number of small TRAP-positive OCs with ≤10 nuclei during sRANKL-dependent osteoclastic differentiation of RAW264.7 cells. NHE10 mRNA expression and OC differentiation markers were increased by sRANKL stimulation in dose- and time-dependent manners. NHEs 1-9 mRNA expression was not increased by sRANKL stimulation. Similar to amiloride, a rat anti-mouse NHE10 MAb (clone 6B11) decreased the number of large TRAP-positive OCs, but increased the number of small TRAP-positive OCs. These findings suggested that inhibition of NHEs by amiloride or an anti-NHE10 MAb prevented sRANKL-promoted cellular fusion. The anti-NHE10 MAb has the potential for use as an effective inhibitor of bone resorption for targeted bone disease therapy.

Data supporting Arf6 regulation of Schwann cell differentiation and myelination.

The data is related to the research article entitled "Arf6 mediates Schwann cell differentiation and myelination" [1]. To further investigate the role of Arf6 in promoting myelination by Schwann cells in vivo, we have characterized an another line (#2) of small-hairpin (sh)RNA transgenic mice targeting Arf6. The number of transgenes per one allele in this line was very low (2 transgenes), comparing with high copies in the previous line (#1, 20 transgenes) [1]. In 4 days of neonatal age, transgenic mice exhibited decreased myelin thickness; however, decreased levels were not as much as those in the line #1, likely depending on transgene copy number. In 60-day-old mice, the difference became smaller. On the other hand, transgene׳s effect was not related to cell proliferation and apoptosis. These data support the key role of Arf6 in Schwann cell myelination, especially in the initiation.

Bovine milk fermented with Lactobacillus rhamnosus L8020 decreases the oral carriage of mutans streptococci and the burden of periodontal pathogens.

AIM: The aim of this study was to find the oral isolate of lactobacilli, which has the potential to inhibit either periodontal, cariogenic, or fungal pathogens in vitro, and to examine the effects of bovine milk fermented with the isolate on the oral carriage of cariogenic and periodontal pathogens. METHODS: The inhibitory effects of the supernatant of Man-Rogosa-Sharpe broth, in which each of 42 oral isolates of lactobacilli grown, was examined. One isolate, Lactobacillus rhamnosus L8020, that showed the potential to inhibit either periodontal, cariogenic, or fungal pathogens in vitro, was used to examine the effects of fermented milk on the oral carriage of cariogenic and periodontal pathogens, which was examined by a placebo-controlled and cohort trial using 50 participants. RESULTS: Edible yogurt containing Lactobacillus rhamnosus L8020 significantly reduced the oral carriage of mutans streptococci (P < 0.01) and four periodontal pathogens examined: Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia, and Fusobacterium spp. (P < 0.01), but the phenomenon were not observed with the placebo yogurt (P > 0.05). CONCLUSION: These results suggest that yogurt with Lactobacillus rhamnosus L8020 could reduce the risk of dental caries and periodontal disease.

Characteristics of high-molecular-weight hyaluronic acid as a brain-derived neurotrophic factor scaffold in periodontal tissue regeneration.

Brain-derived neurotrophic factor (BDNF), for which bovine collagen-derived atelocollagen is used as a scaffold, enhances periodontal tissue regeneration. However, a scaffold that does not contain unknown ingredients is preferable. Since the synthesized high-molecular-weight (HMW)-hyaluronic acid (HA) is safe and inexpensive, we evaluated the efficacy of HMW-HA as a BDNF scaffold. CD44, a major receptor of HA, was expressed in cultures of human periodontal ligament cells, and HMW-HA promoted the adhesion and proliferation of human periodontal ligament cells, although it did not influence the mRNA expression of bone (cementum)-related proteins. The in vitro release kinetics of BDNF from HMW-HA showed that BDNF release was sustained for 14 days. Subsequently, we examined the effect of BDNF/HMW-HA complex on periodontal tissue regeneration in dogs. A greater volume of newly formed alveolar bone and a longer newly formed cementum were observed in the BDNF/HMW-HA group than in the HMW-HA group, suggesting that HMW-HA assists the regenerative capacity of BDNF, although HMW-HA itself does not enhance periodontal tissue regeneration. Neither the poly (lactic-co-glycolic acid) group nor the BDNF/poly (lactic-co-glycolic acid) group enhanced periodontal tissue regeneration. In conclusion, HMW-HA is an adequate scaffold for the clinical application of BDNF.

Clonal expansion of double-positive intraepithelial lymphocytes by MHC class I-related chain A expressed in mouse small intestinal epithelium.

Expression of a distant homologue MHC class I molecule, MHC class I-related chain A (MICA), has been found to be stress inducible and limited to the intestinal epithelium. This nonclassical MHC molecule is associated with various carcinomas in humans. To understand the biological consequences of MICA expression in the gut, we generated transgenic (Tg) mice (T3(b)-MICA Tg) under the control of the T3(b) promoter. The T3(b)-MICA Tg mice expressed MICA selectively in the intestine and had an increased number of TCRalphabeta CD4CD8alphaalpha, double-positive (DP) intraepithelial lymphocytes (IELs) in the small bowel. These MICA-expanded DP IELs exhibited a bias to Vbeta8.2 and overlapped motifs of the complementarity-determining region 3 region among various Tg mice. Hence, the overexpression of MICA resulted in a clonal expansion of DP IELs. Studies in model of inflammatory bowel disease showed that transgenic MICA was able to attenuate the acute colitis induced by dextran sodium sulfate administration. Therefore, this unique in vivo model will enable investigation of possible influences of stress-inducible MICA on the gut immune surveillance.

Expression and intracellular localization of progesterone receptors in cultured human gingival fibroblasts.

OBJECTIVE: The aim of this immunocytochemical study was to characterize the expression and distribution of the progesterone receptor (PR) and estrogen receptor (ER) in gingival fibroblasts using culture cells derived from people at various ages. BACKGROUND: The reaction of female hormones is tissue or cell specific, and receptor availability in the cell is one of the major causes for the different reactions. Gingiva is a target tissue for female hormones; however, the characteristics of PR and ER in both the fibroblasts and the other component cells remain largely unknown. MATERIALS: Gingival tissue was obtained from six people at various ages and culture fibroblasts were established. At least three passages of each cell line were strained for PR and ER with monoclonal antibodies (Clone 1A6, Clone 1D5, respectively). RESULTS: PR positive cells were detected in all six cell lines through early passages to late ones, but ER were only observed in two of six samples with faint reactions. The staining intensity for PR was greater than for ER, but less than that shown in the MCF-7 breast cancer cells, positive control. In every positive control test, ER reactivity was equal to or higher than that of PR. During the interphase, significantly fewer positive fibroblasts occurred compared with negative fibroblasts, and positive nuclei were even fewer. Meanwhile, most of the mitotic cells were PR positive, showing intense localization around chromosomes and on microtubules. These findings suggest that gingival fibroblasts are fundamentally capable of expressing PR and transmit the signal to target genes. CONCLUSIONS: The present study may conclude that in either gender or at any age, gingival fibroblasts express PR rather low in level and do not necessarily localize PR in a nuclear dominant fashion, which is an essential feature for reproductive organ cells. The poor ER reactivity shown in the gingival fibroblasts was discussed in view of the receptor subtype.