Spatiotemporal expression profiles of c-Mpl mRNA in the tooth germ: Comparative expression dynamics of vascularization-related genes.

BACKGROUND: Vascularization is an essential event for both embryonic organ development and tissue repair in adults. During mouse tooth development, endothelial cells migrate into dental papilla during the cap stage, and form blood vessels through angiogenesis. Megakaryocytes and/or platelets, as other hematopoietic cells, express angiogenic molecules and can promote angiogenesis in adult tissues. However, it remains unknown which cells are responsible for attracting and leading blood vessels through the dental papilla during tooth development. METHODS: Here we analyzed the spatiotemporal expression of c-Mpl mRNA in developing molar teeth of fetal mice. Expression patterns were then compared with those of several markers of hematopoietic cells as well as of angiogenic elements including CD41, erythropoietin receptor, CD34, angiopoietin-1 (Ang-1), Tie-2, and vascular endothelial growth factor receptor2 (VEGFR2) through in situ hybridization or immunohistochemistry. RESULTS: Cells expressing c-Mpl mRNA was found in several parts of the developing tooth germ, including the peridental mesenchyme, dental papilla, enamel organ, and dental lamina. This expression occurred in a spatiotemporally controlled fashion. CD41-expressing cells were not detected during tooth development. The spatiotemporal expression pattern of c-Mpl mRNA in the dental papilla was similar to that of Ang-1, which preceded invasion of endothelial cells. Eventually, at the early bell stage, the c-Mpl mRNA signal was detected in morphologically differentiating odontoblasts that accumulated in the periphery of the dental papilla along the inner enamel epithelium layer of the future cusp region. CONCLUSION: During tooth development, several kinds of cells express c-Mpl mRNA in a spatiotemporally controlled fashion, including differentiating odontoblasts. We hypothesize that c-Mpl-expressing cells appearing in the forming dental papilla at the cap stage are odontoblast progenitor cells that migrate to the site of odontoblast differentiation. There they attract vascular endothelial cells into the forming dental papilla and lead cells toward the inner enamel epithelium layer through production of angiogenic molecules (e.g., Ang-1) during migration to the site of differentiation. C-Mpl may regulate apoptosis and/or proliferation of expressing cells in order to execute normal development of the tooth.

Anatomical study of the zygomaticofacial foramen and zygomatic canals communicating with the zygomaticofacial foramen for zygomatic implant treatment: a cadaver study with micro-computed tomography analysis.

In the present study, anatomical assessment of zygomaticofacial foramina (ZFFs) and zygomatic canals communicating with ZFFs were performed using cadaver micro-computed tomography images. It was suggested that all ZFFs were located above the jugale (Ju)-zygomaxillare (Zm) line, which is the reference line connecting the Ju and Zm, and most were located in the zygomatic body area (ZBA). The anteroposterior position of the ZFF in the ZBA was within a middle to posterior region and was most often located slightly posteriorly in males and closer to the middle of the region in females. The mean distance from the Ju-Zm line to the ZFF in the ZBA was 12.36 mm (standard deviation [SD] 1.52 mm) in males and 11.48 mm (SD 1.61 mm) in females. In zygomatic canals communicating with ZFFs, most zygomatic canals were type I canals, communicating from the zygomaticoorbital foramen and harboring the zygomaticofacial nerve, and the others were type II canals, communicating from the zygomaticotemporal foramen and located near the posterior margin of the frontal process. These results provide useful anatomical information for preventing nerve injury during surgical procedures for zygomatic implant treatment.

Nisin lantibiotic prevents NAFLD liver steatosis and mitochondrial oxidative stress following periodontal disease by abrogating oral, gut and liver dysbiosis.

Oral microbiome dysbiosis mediates chronic periodontal disease, gut microbial dysbiosis, and mucosal barrier disfunction that leads to steatohepatitis via the enterohepatic circulation. Improving this dysbiosis towards health may improve liver disease. Treatment with antibiotics and probiotics have been used to modulate the microbial, immunological, and clinical landscape of periodontal disease with some success. The aim of the present investigation was to evaluate the potential for nisin, an antimicrobial peptide produced by Lactococcus lactis, to counteract the periodontitis-associated gut dysbiosis and to modulate the glycolipid-metabolism and inflammation in the liver. Periodontal pathogens, namely Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia and Fusobacterium nucleatum, were administrated topically onto the oral cavity to establish polymicrobial periodontal disease in mice. In the context of disease, nisin treatment significantly shifted the microbiome towards a new composition, commensurate with health while preventing the harmful inflammation in the small intestine concomitant with decreased villi structural integrity, and heightened hepatic exposure to bacteria and lipid and malondialdehyde accumulation in the liver. Validation with RNA Seq analyses, confirmed the significant infection-related alteration of several genes involved in mitochondrial dysregulation, oxidative phosphorylation, and metal/iron binding and their restitution following nisin treatment. In support of these in vivo findings indicating that periodontopathogens induce gastrointestinal and liver distant organ lesions, human autopsy specimens demonstrated a correlation between tooth loss and severity of liver disease. Nisin's ability to shift the gut and liver microbiome towards a new state commensurate with health while mitigating enteritis, represents a novel approach to treating NAFLD-steatohepatitis-associated periodontal disease.

Spatiotemporal distribution analyses of Wnt5a ligand and its receptors Ror2, Frizzled2, and Frizzled5 during tongue muscle development in prenatal mice.

BACKGROUND: The mammalian tongue is a highly specialized muscular organ. The Wnt5a ligand regulates muscle development by mediating the activation of several noncanonical Wnt signaling pathways in a receptor context-dependent fashion. However, there is poor information on the expression and behavior of Wnt5a proteins during muscle development of the embryonic tongue. METHODS: The spatiotemporal distribution profiles of the Wnt5a ligand and its receptors, receptor tyrosine kinase-like orphan receptor 2 (Ror2), Frizzled2 (Fzd2), and Frizzled5 (Fzd5), in the developing tongue muscles of prenatal mice from embryonic day 12.5-18.5 were analyzed using immunofluorescence (IF) double staining of a target protein and desmin, a marker protein of myogenic cells. Immunolabeling images were subjected to digital detection analysis using the WinROOF 2018 version 4.19.0 image processing software when needed. RESULTS: IF signals of the Wnt5a ligand protein and its receptors Ror2 and Fzd2 were detected in developing myoblasts and myotubes of the embryonic tongue, but they were undetectable in mature myofibers equipped with sarcomere structures. Fzd2 expression was specific for desmin-positive developing muscle cells, whereas those of Ror2 and the Wnt5a ligand were widespread and nonselective for desmin-positive cells and that of Fzd5 was predominant in desmin-negative cells of the epithelium and subepithelial mesenchyme. CONCLUSION: Developing muscle cells but not mature myofibers of the mouse embryonic tongue express the Wnt5a ligand and its receptors Ror2 and Fzd2, which may mediate Wnt5a signaling in the development processes of tongue muscle fibers.

Bimodal expression of Wnt5a in the tooth germ: A comparative study using in situ hybridization and immunohistochemistry.

BACKGROUND: During tooth development, Wnt5a, a member of the noncanonical Wnt ligand, is expressed prominently in the dental mesenchyme. However, the spatiotemporal profiles of Wnt5a protein production and distribution in tooth germs are largely unknown, which impairs elucidation of the Wnt5a-mediated regulatory mechanism of tooth development. METHODS: We performed analyzes of the spatiotemporal expression of Wnt5a in embryonic tooth germs (E11.5-E18.5) by using in situ hybridization and immunohistochemistry in parallel. The developmental stages of the embryonic tooth germs were determined by HE staining. In order to compare the spatiotemporal distribution patterns of Wnt5a mRNA-expressing cells and those of Wnt5a protein-expressing cells, serial frontal sections of paraffinized mouse embryo heads were used for the analyzes. When needed, the immunohistochemistry images were subjected to digital detection analysis of Wnt5a immunostaining signal using the WinROOF 2018 Ver. 4.19.0 image processing software program. RESULTS: Throughout the developmental process, cells expressing Wnt5a mRNA were found in various tissues including the dental follicle, dental papilla, inner and outer enamel epithelium, stratum intermediate, and stellate reticulum. However, odontoblasts differentiating and polarizing at E18.5 were the only cells representing an accumulation of Wnt5a protein in the apical region of the odontoblast process. The Wnt5a protein was undetectable in undifferentiated mesenchymal cells as well as any other cells positive for Wnt5a mRNA. CONCLUSION: Differentiating odontoblasts execute Wnt5a expression, the mode of which is distinct from that executed by the other cells constituting tooth germ. Change of the mode of Wnt5a expression begins to take place in the mesenchymal cells by E18.5, starting the elongation of the cytoplasmic process.

Decrease of lysyl hydroxylase 2 activity causes abnormal collagen molecular phenotypes, defective mineralization and compromised mechanical properties of bone.

Lysyl hydroxylase 2 (LH2) is an enzyme that catalyzes the hydroxylation of lysine (Lys) residues in fibrillar collagen telopeptides, a critical post-translational modification for the stability of intermolecular cross-links. Though abnormal LH2 activities have been implicated in various diseases including Bruck syndrome, the molecular basis of the pathologies is still not well understood. Since LH2 null mice die at early embryonic stage, we generated LH2 heterozygous (LH2+/-) mice in which LH2 level is significantly diminished, and characterized collagen and bone phenotypes using femurs. Compared to the wild-type (WT), LH2+/- collagen showed a significant decrease in the ratio of hydroxylysine (Hyl)- to the Lys-aldehyde-derived collagen cross-links without affecting the total number of aldehydes involved in cross-links. Mass spectrometric analysis revealed that, in LH2+/- type I collagen, the extent of hydroxylation of all telopeptidyl Lys residues was significantly decreased. In the helical domain, Lys hydroxylation at the cross-linking sites was either unaffected or slightly lower, but other sites were significantly diminished compared to WT. In LH2+/- femurs, mineral densities of cortical and cancellous bones were significantly decreased and the mechanical properties of cortical bones evaluated by nanoindentation analysis were compromised. When cultured, LH2+/- osteoblasts poorly produced mineralized nodules compared to WT osteoblasts. These data provide insight into the functionality of LH2 in collagen molecular phenotype and its critical role in bone matrix mineralization and mechanical properties.

Immunohistochemical localization of vascular factors in tooth germ of amphibian (Cynops pyrrhogaster) / Localización inmunohistoquímica de factores vasculares en germen dentario de anfibios (Cynops pyrrhogaster)

SUMMARY: Vascular endothelial growth factor (VEGF) and its receptor, VEGFR-2, are known to regulate blood vessel endothelium growth. They play important role in human and rodents teeth development. In newt jaws, there are sequential developmental teeth germs following behind the mature teeth. We examined the immunohistochemical localization of VEGF and its receptor and showed the specific expression pattern of VEGF and VEGF receptor in Cynops pyrrhogaster sequential tooth development. The intensity of immunoreactivity for VEGF in the inner enamel epithelium was weaker than that in the outer enamel epithelium in the dentine matrix formation and mineralization stages. Finally, at the enameloid maturation and enamel-like matrix formation stage, immunoreactivity for VEGF in inner enamel epithelium was stronger than in the outer enamel epithelium. The intensity of immunoreactivity for VEGFR-2 was positive for the outer enamel epithelium throughout tooth development. The crown sides of the odontoblasts were stained especially strongly for VEGF and VEGFR-2 during the dentine matrix formation and mineralization stage of the enameloid maturation and enamel- like matrix formation stage. We postulate that the expression of VEGF in the inner enamel epithelium and odontoblast widely effects tooth development in newts, as well as in human and rodents.

RESUMEN: Se sabe que el factor de crecimiento endotelial vascular (VEGF) y su receptor, VEGFR-2, regulan el crecimiento del endotelio de los vasos sanguíneos. Desempeñan un papel importante en el desarrollo de los dientes humanos y de los roedores. En las mandíbulas de tritón, hay gérmenes dentales de desarrollo secuenciales que siguen a los dientes maduros. Examinamos la localización inmunohistoquímica de VEGF y su receptor y mostramos el patrón de expresión específico de VEGF y receptor de VEGF en el desarrollo secuencial de dientes de Cynops pyrrhogaster. La intensidad de la inmunorreactividad para VEGF en el epitelio interno del esmalte era más débil que en el epitelio externo del esmalte en las etapas de formación y mineralización de la matriz de dentina. Finalmente, en la etapa de maduración del esmalte y de formación de la matriz similar al esmalte, la inmunorreactividad para VEGF en el epitelio interno del esmalte fue más fuerte que en el epitelio externo del esmalte. La intensidad de la inmunorreactividad para VEGFR- 2 fue positiva para el epitelio externo del esmalte durante el desarrollo del diente. Los márgenes de la corona de los odontoblastos se tiñeron especialmente para VEGF y VEGFR-2 durante la etapa de formación de la matriz de dentina y mineralización de la etapa de maduración del esmalte y la etapa de formación de la matriz similar al esmalte. Postulamos que la expresión de VEGF en el epitelio interno del esmalte y odontoblastos afecta ampliamente el desarrollo de los dientes en tritones, así como en humanos y roedores.

Temporal and dynamic changes in gingival blood flow during progression of ligature-induced periodontitis.

OBJECTIVES: To evaluate temporal changes in gingival blood flow (GBF) during progression of periodontitis in rats using a laser Doppler flowmeter (LDF) approach and to characterize morphological and biochemical features in the periodontium associated with GBF. MATERIALS AND METHODS: Forty-two Wistar rats were divided into a ligature-induced periodontitis group and a control group. To induce periodontitis, ligatures were tied around maxillary first molars bilaterally. GBF was measured in palatal gingiva at pretreatment and following ligature placement after 30 min, 1, 3, 7, 14, 21, and 28 days using LDF with a non-contact probe. Bone loss and gene expression in gingival tissues were assessed using micro-computed tomography (µCT) and quantitative polymerase chain reaction (PCR), respectively. Immunostaining for vascular endothelial growth factor (VEGF) in the maxilla was also histologically evaluated. RESULTS: GBF in the ligature group increased significantly compared with the control group 30 min after ligation. However, on days 3 and 7, GBF decreased in the ligature group. Also, after day 10, there was no difference in GBF between groups. The levels of alveolar bone loss, gene expression (interleukin-6, cluster of differentiation-31, VEGF-A, and lymphatic vessel endothelial hyaluronan receptor-1), and immunostained VEGF-positive vessels correlated well with changes in GBF. CONCLUSION PROGRESSION OF PERIODONTITIS: In rats was associated with a triphasic pattern of GBF, consisting of a short initial increase, followed by a rapid decrease, and then a gradual plateau phase.

Distribution of glutamate receptor, ionotropic, kainate 1 and neuropeptide calcitonin gene-related peptide mRNAs during formation of the embryonic and postnatal mouse molar in the maxilla.

The neuropeptide calcitonin gene-related peptide (CGRP) is a well-characterized neurotransmitter. Glutamate receptor, ionotropic, kainate 1 (Grik1) has also been demonstrated to generate high-affinity kainate receptors. However, little is known about the roles of CGRP and Grik1 during the developmental formation of teeth. In this study, we endeavoured to analyse the expression and localization of CGRP and Grik1 mRNAs using in situ hybridization on the mouse maxilla during development from the embryonic stage (E18.5) to after birth (P10, P15 and P20). We found that hybridization with an anti-sense probe for CGRP clearly localized in the maxilla at E18.5 in contrast to that of P15 and P20. Hybridization with an anti-sense probe for CGRP was not detected in the dental pulp of molars in the maxilla at P10, which is in contrast to Grik1 mRNA at the same developmental stage. Hybridization with an anti-sense probe for Grik1 mRNA was detected in the basal region of the dental pulp of molars at P10 and P15. Finally, these markers were not detected in molars in the mouse maxilla at P20. The ratio of positive cells for the hybridization signals of Grik1and CGRP in the dental pulp decreased from E18.5 (p<0.001). These features in CGRP and Grik1r mRNAs may indicate roles of function during tooth development between embryonic and postnatal stages with root formation and erupted movements.

Resveratrol Targets Urokinase-Type Plasminogen Activator Receptor Expression to Overcome Cetuximab-Resistance in Oral Squamous Cell Carcinoma.

Drug resistance to anti-cancer agents is a major concern regarding the successful treatment of malignant tumors. Recent studies have suggested that acquired resistance to anti-epidermal growth factor receptor (EGFR) therapies such as cetuximab are in part caused by genetic alterations in patients with oral squamous cell carcinoma (OSCC). However, the molecular mechanisms employed by other complementary pathways that govern resistance remain unclear. In the current study, we performed gene expression profiling combined with extensive molecular validation to explore alternative mechanisms driving cetuximab-resistance in OSCC cells. Among the genes identified, we discovered that a urokinase-type plasminogen activator receptor (uPAR)/integrin ß1/Src/FAK signal circuit converges to regulate ERK1/2 phosphorylation and this pathway drives cetuximab-resistance in the absence of EGFR overexpression or acquired EGFR activating mutations. Notably, the polyphenolic phytoalexin resveratrol, inhibited uPAR expression and consequently the signaling molecules ERK1/2 downstream of EGFR thus revealing additive effects on promoting OSCC cetuximab-sensitivity in vitro and in vivo. The current findings indicate that uPAR expression plays a critical role in acquired cetuximab resistance of OSCC and that combination therapy with resveratrol may provide an attractive means for treating these patients.

Upregulation of Tumor Susceptibility Gene 101 (TSG101) by mechanical stress in podocytes.

Elevated mechanical stress in glomerular hypertension is thought to damage podocytes, the loss of which leads to development of glomerulosclerosis. Applying cDNA array analysis to mechanically stressed podocytes, we have recently identified TSG101 as a stretch-induced candidate gene among others. TSG101, which is part of the ESCRT-I complex, is involved in multivesicular body (MVB) formation. Here we demonstrate that TSG101 mRNA is strongly upregulated in conditionally immortalized mouse podocytes by cyclic mechanical stress. Differentiation of podocytes does not affect TSG101 mRNA levels. TSG101 immunofluorescence is distributed in a vesicular pattern in podocytes, the staining intensity being enhanced by mechanical stress. In DOCA/salt treated rats, a model of glomerular hypertension, glomerular TSG101 mRNA levels are elevated, and an increased number of MVBs is observed by electron microscopy in podocyte processes. Our data demonstrate that mechanical stress upregulates TSG101 in podocytes, suggesting that glomerular hypertension enhances sorting of cell surface proteins and their ligands into the degradative pathway in podocytes.

CBCT imaging of the alveolar bone structure in maxilla of elderly donor cadavers and PCA analysis.

There is an important bone matrix with remodelling between dentate and edentulous samples of the human maxilla for bone metabolism. Cone beam computed tomography (CBCT) is useful for structural analysis of bone. The objective of this study was to investigate morphological data of donor cadavers in detail using CBCT imaging and principal component analysis (PCA). We analysed 38 donor cadavers using a CBCT apparatus. The analytical results defined differences in skull measurement parameters and dentate and edentulous levels using PCA. We observed cortical bone, trabecular bone, and the distance from the bottom of the maxillary sinus to the oral mucosa at a right angle to the palatal plane of the first molar region between dentate and edentulous samples of the human maxilla using CBCT imaging. In the dentate sample of the maxilla, component 1 was defined by negative contributions from gender (-0.84) and age (-0.54) to positive contributions such as cortical bone structure (CBS, 0.68) and trabecular bone structure (TBS, 0.50). There was a difference in CBS between dentate and edentulous human maxilla samples. This study of CBCT data provides useful basal information for planning dental implant surgery using PCA.

Promoter motifs required for c-mpl gene expression induced by thrombopoietin in CMK cells.

Thrombopoietin (TPO) and its receptor, c-Mpl, are the central regulators of megakaryocyte development and platelet production and are also crucial to regulate megakaryocytopoiesis. TPO remarkably elevated c-mpl promoter activity, while the protein kinase C (PKC) inhibitors, GF109203, H7 and Calphostin C, clearly reduced the steady level of its promoter activity.  In the present study, motifs crucial for c-mpl promoter activity induced by TPO treatment have been analyzed using a human megakaryoblastic cell line, CMK. Destruction of the -107Sp1 and the -57Sp1 sites in the c-mpl promoter enhancer region resulted in decrease of the promoter activity by 53.1% and 64.4%, respectively, and destruction of -69Ets and -28Ets elements dramatically decreased the promoter activity by 96.4% and 87.8%, respectively, while mutation of -77GATA moderately reduced the activity by 31.4%. The result was in agreement with our previous report that showed the crucial motifs in the c-mpl promoter for the promoter activity induced by PMA-treatment. This indicates that TPO-induced activation of the c-mpl promoter activity is fully modulated by transcription through a PKC-dependent pathway and the two Sp1 and two Ets motifs are crucial for the activation of the c-mpl promoter activity rather than a GATA motif in the c-mpl promoter of CMK cells.

A morphological study of the foramina of the mandible in the Japanese macaque by cone-beam computed tomography.

The mandibular canal (MC) contains vessels and nerves in the mandible of the Japanese macaque (JM). The inferior alveolar nerves and vessels of the mandible insert from the mandibular foramen and then run through the MC, the mental foramen and spinal foramen to the outside of the mandible. However, the detailed morphological properties of multiple canals, such as the accessory canal (AC) of the mandible, are unknown in JMs. The purpose of this study was to describe the multiple canals of JMs and to determine the location and analyse the measurements of the JM mandible. In this study, we also showed the course of the lingual foramen in 17 JMs (male: n = 8; female: n = 9) using cone-beam computed tomography (CBCT). In our results, we classified multiple mental foramina and multiple lingual foramina found on the mandibular body at the premolar or molar region. However, there was no significance between the formation of mandibular properties and the lingual foramen. These multiple foramina contain nerves and blood vessels have a few branched canals; these branches run downward and connect with the inferior mandibular nerve and artery. These morphological features may provide useful information about surgical treatment of the alveolus in a human model.

A morphological study of the multi-posterior superior alveolar canals of maxilla in the Japanese macaque by cone-beam computed tomography.

The posterior superior alveolar canal (PSAC) composed of several canals which contains vessels and nerve in molar region of the maxilla of Japanese macaque. The PSAC of maxilla run to the maxillary sinus. However, the PSAC and accessory canal (AC) of the maxilla in the Japanese macaque (JM) is unknown in morphological features in the maxilla. The purpose of this study was to describe the PSAC of the primates and to determine whether this structure could be used as a model for the human clinical condition. In this study, we showed the course of PSAC structure of the 23 JMs (male: n = 15; female: n = 8) using a cone-beam computed tomography apparatus. In the results, we classified a type to have one AC toward, a type to have two ACs toward, and three ACs in a type to have in PSAC. The main canal have some bony branch canals (BBCs) composed of 3 types (no BBC, one BBC, two BBCs). These canals and they run downward and supply to MS, these roots of maxillary molar region of the craniofacial skeleton in contrast to numerous small accessory canals with no nerve and vessels observed in the posterior regions in maxilla. These morphology features may give useful information about MS in dental treatment human model.

Tenomodulin regulated the compartments of embryonic and early postnatal mouse masseter muscle.

The masseter muscle (MM) is a complex tendinous laminar structure during development; however, the stage of the laminar structure formation is unknown. Tenomodulin (TeM) is a useful marker of tendons and has an anti-angiogenic cysteine-rich C-terminal domain. Therefore, we analyzed mRNA of TeM and angiogenesis markers (CD31 and vascular endothelial growth factor (VEGF)) and performed in situ hybridization for the TeM genes in MM from on embryonic day 12.5 (E12.5) to postnatal day 5 (P5). The TeM expression is at first detectable in the middle region of the mesenchymal connective tissue in the MM at E 12.5. The expression domains of the TeM during development typically include the middle region of the MM, particularly surrounding the vascular regions. The level of TeM mRNA in the MM increased from E12.5 to E17.5 and decreased after birth. In contrast, the levels of CD31 and VEGF mRNAs were almost constant from E12.5 to E18.5 and then low from birth onward. Therefore, the development of the laminar tendinous structure in the middle region between superficial and deeper regions of the MM first occurs during the process of tendon formation at embryonic day 12.5. In our study of MM development, the laminar structure regulating TeM also prevents vascular invasion during the formation of compartment of the MM. The tendon may relate to the components of muscle mass of MM.

GATA-dependent regulation of TPO-induced c-mpl gene expression during megakaryopoiesis.

Thrombopoietin (TPO) and its receptor, c-Mpl, play the crucial role during megakaryocytopoiesis. Previously, we have shown that the promoter activity of c-mpl induced by TPO is modulated by transcription through a PKC-dependent pathway and that GATA(-77) is involved as a positive regulatory element in TPO-induced c-mpl gene expression in the megakaryoblastic CMK cells. In this research, to examine participating possibility of GATA promoter element in TPO- induced c-mpl gene expression through a PKC-independent pathway, the promoter activity of site-directed mutagenesis and the effect of potein kinase C modulator were measured by a transient transfection assay system. Together with our previous results on the TPO-induced c-mpl promoter, this study indicates destruction of -77GATA in c-mpl promoter decreased the activity by 47.3% under existence of GF109203. These results suggest that GATA promoter element plays significant role in TPO-induced c-mpl gene expression through a PKC-independent pathway.

Differences in the morphology of the maxillary sinus and roots of teeth between Macaca fuscata and Macaca fuscata yakui determined using cone beam computed tomography.

The Japanese macaque is an endemic species consisting of two subspecies: Macaca fuscata fuscata (MFF) and Macaca fuscata yakui (MFY). The MFY is indigenous to Yakushima Island and represents a subspecies of MFF that lives from Honshu to Shikoku and Kyushu, Japan. However, the differences in the skulls of the MFY and MFF are unknown, despite these subspecies having different skull sizes. The maxillary sinus (MS) indicates that the features of the frontal view reflect the transversal growth of the maxilla of the skull. In this study, we show the MS structures of the MFF (n = 9, 18 sides) and MFY (n = 10, 20 sides) using a cone-beam computed tomography instrument. Base on three-dimensional (3D) reconstructed images the MS and nasal cavity were found to present almost to no significant differences between MFF and MFY. However, we designated three classifications of the sinus floor based on the 3D MS images of these Japanese macaques: a round-like shape (type a, MFF = 66.7% (12/18), MFY = 45% (9/20)), a flat-like shape (type b, MFF = 22.2% (4/18), MFY = 35% (7/20)), and an irregular shape (type c, MFF = 11.1% (2/18), MFY = 20.0% (4/20)). The sinus floor shapes of the MFF were mostly type a, while those of the MFY were mostly type b. The prevalence of a root contacting the cortical bone is higher in the canine (26.7%, (8/30)) and second premolar (20%, (6/30) of the MFY at the nasal cavity, moreover, this value is higher in the third molar (42.9%, (9/21)) of the MS in the MFY. These results suggest that the features of the floor of the MS are related to the differences in maxillary root apices teeth between MMF and MMF.

Role of promoter element in c-mpl gene expression induced by TPO.

Thrombopoietin (TPO) and its receptor, c-Mpl, play the crucial role for the development of megakaryocyte and considered to regulate megakaryocytopoiesis. Previously we reported that TPO increased the c-mpl promoter activity determined by a transient expression system using a vector containing the luciferase gene as a reporter and the expression of the c-mpl gene is modulated by transcription through a protein kinase C (PKC)-dependent pathway in the megakaryoblastic cells. In this research, to elucidate the required elements in c-mpl promoter, the promoter activity of the deletion constructs and site-directed mutagenesis were measured by a transient transfection assay system. Destruction of -77GATA in c-mpl promoter decreased the activity by 22.8%. Our study elucidated that -77GATA involved in TPO-induced c-mpl gene expression in a human megakaryoblastic cell line, CMK.

Distributions of calcitonin gene-related peptide and substance P in the human maxillary sinus of Japanese cadavers.

BACKGROUND: Substance P (SP) and calcitonin gene-related peptide (CGRP) are released by the nociceptive sensory nerve and are involved in blood flow, pain and inflammation in the nasal mucosa. The purpose of this study was to assess the distribution of the SP and CGRP nerve fibres related to blood supply within human Schneiderian membrane of the maxillary sinus (MS). MATERIAL AND METHODS: In this study, the MS from Japanese cadavers was examined by whole-mount immunohistochemistry. Human male cadavers (ranging in age from 80 to 90 years) were used in this study. RESULTS: SP- and CGRP-positive fibres were found around large vessels of the medialis superior alveolar branches and also within the floor region of the MS. The floor region of the MS was composed of complex branches of these fibres. CONCLUSION: Our results give useful information for surgical sinus floor elevation in this region of the MS. These anatomical features may assist in the execution of a successful surgical procedure.