Identification of a compound heterozygous inactivating ABCC8 gene mutation responsible for young‐onset diabetes with exome sequencing

Activating mutations in the ABCC8 gene cause diabetes and inactivating mutations usually cause hyperinsulinemic hypoglycemia in infancy. Patients with hypoglycemia in infancy due to a heterozygous inactivating mutation have been reported to occasionally progress to diabetes later in life. We explored the gene responsible for diabetes in two brothers, who were suspected to have diabetes at 15 and 18 years‐of‐age, respectively, with whole exome sequencing, and identified a compound heterozygous ABCC8 gene mutation (p.Arg168Cys and p.Arg1421Cys). Although their father and mother were heterozygous carriers of the p.Arg168Cys and the p.Arg1421Cys mutation, respectively, neither parent had diabetes. These mutations have been reported to be responsible for hypoglycemia in infancy and function as an inactivating mutation. Our results suggest that the inactivating ABCC8 gene mutation is also important in the etiology of diabetes.     

Nicotine inhibits mineralization of human dental pulp cells

Nicotine is a major component of tobacco smoke, and signals via nicotinic acetylcholine receptors (nAChR). However, little is known about the effects of nicotine on human dental pulp cells (HDPCs). In this study, we assessed the effects of nicotine on mineralization in HDPCs. We confirmed messenger RNA expression of nAChR subunits and examined the effects of nicotine on expression of extracellular matrices (ECMs), alkaline phosphatase (ALP) activity, and mineralized nodule formation by HDPCs. Gene expression of nAChR subunits alpha1, alpha2, alpha 4, alpha 5, alpha 6, alpha 7, beta1, beta2, and beta 4 was detected in HDPCs. Interestingly, the messenger RNA expression of dentin matrix acidic phosphoprotein-1, bone sialoprotein, and ALP activity were significantly reduced in nicotine-treated HDPC. In addition, mineralized nodule formation, which was examined by alizarin red staining, was also inhibited in HDPCs by the same treatment. These results indicate that nicotine suppresses the cytodifferentiation and mineralization of HDPCs, possibly via nAChR.     

Changes in response properties of periodontal mechanoreceptors after experimental orthodontic tooth movement in rats

Using an in vitro preparation, we investigated chronological changes in response properties of periodontal mechanoreceptors (PMRs) in the rat right mandibular first molar (M1) after experimental orthodontic tooth movement. Orthodontic force was applied to M1 for 14 days by activating 24.5 mN superelastic titanium-nickel alloy closed coil springs anchored to the mandibular incisors. Experiments were performed on days 3, 7, 10, and 14 during application of orthodontic force and on days 7, 14, 21, and 28 after removal of orthodontic force. The rats without application of orthodontic force were used as control group. In each group, direct mechanical stimulation using von Frey hairs and electrical stimulation was applied to the distal root of M1. Results showed that compared with controls (1) the mechanical thresholds were significantly lower during application of orthodontic force; however, no significant difference was found after removal of force application and (2) conduction velocities were significantly lower from day 7 during application of orthodontic force to day 14 after removal of orthodontic force; however, no significant difference was found on days 21 and 28 after removal of orthodontic force. These results suggest that the PMRs, although having some of their response properties altered during orthodontic force application, were able to recover and adapt to the newly acquired intraoral condition after removal of the orthodontic force.     

Binding specificity of Fusobacterium nucleatum to human erythrocytes, polymorphonuclear leukocytes, fibroblasts, and HeLa cells

Eighteen strains of Fusobacterium nucleatum were studied with respect to their adherence to human erythrocytes, polymorphonuclear leukocytes, fibroblasts, and HeLa cells. Four types of adherence were found: 1) sugar-sensitive adherence 2) amino acid-sensitive adherence 3) unknown adherence 4) weak adherence. There was no obvious correlation between the patterns of hemagglutination and adherence to other cells. The results indicate not only a complex network of bacterial receptors on human cells, but also a variety of types of bacterial adherence as expressed by the selectivity of the bacterial binding to specific cell surfaces.     

The pattern of alveolar bone activity during development and eruption of the molar in the rat

The pattern of alveolar bone activity around the developing and erupting mandibular first molar of the rat was studied by autoradiography. The rats were hilled at 1 day and 1, 2, 3 and 4 weeks after birth. One hour prior to sacrifice, they were injected with ³H-proline intraperitoneally. At one day after birth, the alveolar bone surfaces surrounding the tooth germ were unlabeled. At one week, the surface of the alveolar bone mesial to the developing tooth crown revealed heavy labeling, whereas the remaining surfaces surrounding the crown were unlabeled. The interdental septum between the first and second molars appeared at this stage. lts mesial surface was unlabeled troughout the four stages. Formation of the interradicular septum was ohserved at two weehs, and the future crestal area displayed very intense labeling. At three weeks, the crestal, mesial and distal surfaces of the interradicular septum showed heavy labeling. However, the apical one-third of the mesial surface of the septum was unlabeled at four weeks. The surfaces of the fundic bone just beneath the primitive pulp chamber and beneath the open ends of the roots were unlabeled throughout the entire period. At three and four weeks, the interdental and interradicular crests exhibited very heavy labeling, more intense than that at the crest of the alveolar bone mesial to the first molar.     

Microvascular response in the periosteum following mucoperiosteal flap surgery in dogs: angiogenesis and bone resorption and formation

BACKGROUND: When surgical stress reaches the periosteum, bone resorption and formation that occur as a periosteal response are closely related to angiogenesis and hemodynamics. Thus, we investigated bone remodeling in the healing process after mucoperiosteal flap surgery, focusing our attention on the microcirculation. METHODS: Mucoperiosteal flap surgery was performed on 12 adult beagle dogs. The periosteal vascular plexus was observed on days 7, 14, 21, and 28 after surgery, using three different techniques: in histological specimens into which India ink was injected into blood vessels, under a light microscope; in ultrathin sections, using a transmission electron microscope; and in acryl plastic-injected vascular corrosion cast specimens, under a scanning electron microscope. RESULTS: On day 7 after surgery, the interstitum of the elevated mucoperiosteal vascular plexus was filled with sinusoidal new blood vessels. Bone resorption by osteoclasts was observed around these new blood vessels and many highly permeable fenestrations were present in the vascular endothelium. On day 14 after surgery, sinusoidal new blood vessels were more markedly developed and some regions exhibited glomeruluslike morphology consistent with bone resorption cavities. Activated osteoblasts were present around these new blood vessels and highly permeable vesicles, which were considered to be possible vesiclo-vacuolar organelles (VVOs) and caveolae, were noted in the vascular endothelium. On days 21 and 28 after surgery, the mucoperiosteal vascular plexus was dissected through regression of endothelial cells and fibroblasts and reconstructed into a rough mesh structure, and simultaneously the bone surface became smooth. CONCLUSION: The morphology of the mucoperiosteal vascular plexus changed with bone metabolism and these changes contributed to transport of substances involved in periodontal repair.