Glycation promotes pulp calcification in Type 2 diabetes rat model.

OBJECTIVES: Intrapulpal calcifications can occur in the dental pulp of patients with diabetes. We focused on the association between ectopic calcifications in the dental pulp and advanced glycation end products (AGEs) in Spontaneously Diabetic Torii (SDT)-fatty rats, an obese type 2 diabetic rat model, to determine the mechanism of calcification with pulp stone in the dental pulp. MATERIALS AND METHODS: Pathologic calcification in the dental pulp of SDT-fatty rats was observed using electron microscopy and immunohistochemical analysis. Moreover, mechanical analysis of periapical region of molar tooth against occlusal force was performed. RESULTS: In SDT-fatty rats, pathogenic pulpal calcifications occurred during blood glucose elevation after 6 weeks, and granular calcification was observed in the dental pulp after 11 weeks. Pentosidine, a major AGE, and the receptor for AGEs were strongly expressed in the dental pulp of SDT-fatty rats. S100A8, TNF-α, and IL-6 also showed positive response in the dental pulp of the SDT-fatty rat, which indicated pulpal inflammation. Blood flow disorder and hypoxic dental pulp cells were also observed. In silico simulation, strain from occlusal force concentrates on the root apex. CONCLUSIONS: Glycation makes blood vessels fragile, and occlusal forces damage the vessels mechanically. These are factors for intrapulpal calcification of diabetes.

Effects of advanced glycation end products on dental pulp calcification.

OBJECTIVE: The main aim of this study was to elucidate the effects of advanced glycation end products (AGEs) on the calcification of cultured rat dental pulp cells (RDPCs) and to investigate the crystallisation ability of glycated collagen. MATERIALS AND METHODS: AGEs were prepared via non-enzymatic glycation of a dish coated with type I collagen using dl-glyceraldehyde. To investigate the effects of AGEs on RDPCs, we performed WST-1 and lactate dehydrogenase assays; alkaline phosphatase, Alizarin Red S and immunohistochemical staining; and real-time quantitative reverse transcription PCR. In addition, we performed crystallisation experiments on glycated collagen. All microstructures were analysed using scanning electron microscopy/energy-dispersive X-ray spectroscopy and transmission electron microscopy/diffraction pattern analysis. RESULTS: AGEs did not affect the proliferation or differentiation of RDPCs, but enhanced the calcification rate and cytotoxicity. No major calcification-related genes or proteins were involved in these calcifications, and glycated collagen was found to exhibit a negative polarity and form calcium phosphate crystals. Cytotoxicity due to drastic changes in the concentration of pericellular ions led to dystrophic calcification, assumed to represent an aspect of diabetic pulp calcifications. CONCLUSION: Glycated collagen-containing AGEs provide a nurturing environment for crystallisation and have a significant effect on the early calcification of RDPCs.

Ultra-structural observation of human enamel and dentin by ultra-high-voltage electron tomography and the focus ion beam technique.

Human enamel and dentin are hard and brittle mineralized tissues. It is difficult to prepare thin specimens (under 200 nm) of these tissues for transmission electron microscope observation without demineralizing them. We present a novel method of creating three-dimensional ultra-structural images of human enamel and dentin, using the focus ion beam (FIB) method and ultra-high-voltage electron microscope tomography. Thin specimens (less than 2 µm) of enamel and dentin were prepared using the FIB method. This method is appropriate for nano-fabrication of thin specimens for brittle materials such as enamel and dentin. It allows penetration of an ultra-high-voltage electron beam of a 3000 kV maximal acceleration voltage into a specimen, enabling projections of the specimen to be taken. It facilitates tomography of the enamel rods, sheaths, dentinal tubules and collagen fibrils with a high resolution of 2 nm. The component percentages in ultra-structures of dentin can be expressed numerically by using three-dimensional reconstruction images of tomograms. The matrix of peritubular dentin differed from that of intertubular dentin by having relatively fewer collagen fibrils. The major advantage of this method is its ability to visualize ultra-structural tomograms of highly calcified specimens without demineralization.

Movement of the mandibular condyle and articular disc on placement of an occlusal splint.

OBJECTIVE: The aim of this study was to evaluate, with the use of magnetic resonance images, mandibular condyle and articular disc movement on occlusal splint application. STUDY DESIGN: Thirty-five subjects were classified into "Pain" and "No-pain" groups based on visual analog scale scoring of temporomandibular joint (TMJ) pain and were categorized as bilateral normal disc (ND), unilateral anterior disc displacement (ADD), or bilateral ADD position. t test and analysis of variance with multiple comparisons were used for statistical analyses. RESULTS: Pain and No-pain groups showed significant anteroinferior and rotated movement of the condyle on splint application, but the disc did not significantly move in the Pain group. Of all groups, anteroposterior movement of the condyle and disc with the splint was significantly less in the unilateral ADD group. CONCLUSIONS: TMJ pain was associated with decreased movement of the articular disc. Disc and condyle movement was different in the unilateral ADD group in the presence of a splint compared with the ND and bilateral ADD groups.

Dental haptic simulator to train hand skill of student - calibration method to realize visual haptic environment.

In prior work, we have developed dental training simulator to train hand skill of student (HHDTS). In the present study, we performed calibration between haptic device coordinate system and half-mirror coordinate system for our system to realize real clinical situation. As a result, the user can overlay the CG (turbine) onto the stylus of haptic device and intract to the CG (tooth) directly.

Virtual reality simulation training for dental surgery.

In dentistry the exquisite hand skill is required for daily treatments, However, dental students have little chance to treat patients in the clinical training So, present study constructed a process simulation to train the hand skill for tooth extraction, especially impacted wisdom tooth that was an example of the difficult case by using virtual reality haptic device.

A fermentation product of phytosterol including campestenone reduces body fat storage and body weight gain in mice.

Anti-obesity effects of a fermentation product of phytosterols including campestenone in ICR mice were investigated. Five-week-old male ICR mice were fed by the pair-feeding method for 8 wk. Experimental feed was prepared by adding TO-001, a phytostenone mixture produced by fermentation of phytosterols using Nocardioides simplex, at 0.25, 0.5, 1.0, or 2.0% or no additive to a high fat diet (fat 20%). Mice fed a stock feed (fat 5.6%) ad libitum were used as the standard growth group. In animals fed the high fat diet, control (no added TO-001) mice showed a weight gain that was about 10% higher than for the standard growth group. TO-001 reduced body weight dose-dependently. Final body weights of 0.5% and 1.0% TO-001-fed mice were lowered by about 9% and those of 2.0% TO-00 I-fed mice by about 12% compared with the control mice. Visceral and subcutaneous fat weight in mice fed TO-001 was significantly lower than that in mice fed the control diet. The concentrations of serum triglyceride (TG) and total cholesterol (TC) were significantly lower in the 1.0% and/or 2.0% TO-001-fed mice. Furthermore, levels of liver TG and TC were decreased in the TO-001-fed group. Increase of total lipid excretion in the feces was dose dependent. No obvious abnormalities due to consumption of TO-001 were detected by a blood biochemical examination, clinical observations or necropsy. The results suggested that TO-001, a fermentation product of phytosterols, may be a promising component of dietetic functional foods.

Skull metastasis of thyroid papillary carcinoma.

Skull metastasis of thyroid carcinoma is rare. The majority of skull metastases of thyroid carcinoma are of the follicular subtype, rather than thyroid papillary carcinoma. In this report, a 55-year-old woman with skull metastasis from thyroid papillary carcinoma is presented. The metastatic lesion of the skull was hypervascular and osteolytic, and the bleeding was profuse during resection. There have been only four reports of skull metastasis from thyroid papillary carcinoma. The mean period from the initial diagnosis of thyroid carcinoma until the detection of skull metastasis is 23.3 years, whereas in this patient, it was about 2 years. Therefore, in the clinical course of thyroid papillary carcinoma, skull metastasis should be considered, and the patient should be meticulously followed up.

Epidermoid tumor arising in the anterior interhemispheric fissure.

There have only been 18 cases reported in the literature of an epidermoid tumor arising in the interhemispheric fissure. We report a 45-year-old woman with an epidermoid tumor arising in the interhemispheric fissure. She presented with a focal convulsive seizure. Radiological imaging identified a tumor in the interhemispheric fissure, which was subtotally removed due to adherence to both anterior cerebral arteries and the cerebral cortex. Epidermoid tumor was confirmed by histopathology. Epidermoid tumors rarely arise in the interhemispheric fissure, but are often adherent to surrounding structures and thus, as in the presented case, complete resection may be unwise.

Analyses of the mechanism of intracellular transport and secretion of pituitary hormone, with an insight of the subcellular localization of pituitary hormone and its mRNA.

Combined in situ hybridization (ISH) and immunohistochemistry (IHC) under electron microscopy (EM-ISH&IHC) has sufficient ultrastructural resolution and provides two-dimensional images of subcellular localization of pituitary hormone and its mRNA in a pituitary cell. The advantages of semiconductor nanocrystals (Quantum dots, Qdots) and confocal laser scanning microscopy (CLSM) enable us to obtain three-dimensional images of subcellular localization of pituitary hormone and its mRNA. Both EM-ISH&IHC and ISH&IHC using Qdots and CLSM are useful for understanding the relation between protein and mRNA simultaneously in two or three dimensions. Another important issue is the intracellular transport and secretion of pituitary hormone. We have developed an experimental pituitary cell line, the GH3 cell, which has growth hormone (GH) linked to enhanced yellow fluorescein protein (EYFP). This stable GH3 cell secretes GH linked to EYFP upon stimulated by Ca2+ influx or Ca2+ release from storage. This GH3 cell is useful for real-time visualization of the intracellular transport and secretion of GH. These three methods enable us to visualize consecutively the process of transcription, translation, transport, and secretion of pituitary hormone.

Symptomatic pituitary metastasis of malignant thymoma.

Pituitary metastases from malignant thymoma are extremely rare. We report an 86 year-old-man with pituitary metastasis of malignant thymoma who complained of visual impairment and right ptosis. A pituitary lesion, which was partially removed by trans-sphenoidal surgery, was pathologically similar to the primary lesion and was diagnosed as pituitary metastasis of malignant thymoma. One autopsied case of pituitary metastasis of malignant thymoma has been found in the literature, and one case of this condition has been previously described in life. The present paper is the second report of symptomatic pituitary metastasis of malignant thymoma. Surgical removal of the pituitary lesion is a reasonable choice for the relief of progressive visual symptoms and for good quality of life in these patients.

Three-dimensional imaging of the intracellular localization of growth hormone and prolactin and their mRNA using nanocrystal (Quantum dot) and confocal laser scanning microscopy techniques.

Semiconductor nanocrystals (Quantum dots, Qdots) have recently been used in biological research, because they do not fade on exposure to light, and they enable us to obtain multicolor imaging because of a narrow emission peak that can be excited via a single wavelength of light. There have been no reports of simultaneous localization of mRNA and protein using Qdots. We successfully applied these advantages of Qdot and confocal laser scanning microscopy (CLSM) to three-dimensional images of the intracellular localization of growth hormone and prolactin and to their mRNA. In situ hybridization and immunohistochemistry using Qdots combined with CLSM can optimally illustrate the relationship between protein and mRNA simultaneously in three dimensions. Such an approach enables us to visualize functional images of proteins in relation with mRNA synthesis and localization.

A diagnostic pitfall: Angiosarcoma of the brain mimicking cavernous angioma.

Primary or secondary angiosarcoma in the central nervous system is rarely reported. We present a rare case of cerebral angiosarcoma, which comprised both poorly-differentiated solid areas and well-differentiated areas that led to the misdiagnosis of cavernous angioma. A 79-year old woman presented with an intracerebral hematoma in the left frontal lobe that was misdiagnosed as a hemorrhage from a cavernous angioma at initial operation. At a second surgery, the lesion was diagnosed as angiosarcoma involving the cerebellum, heart, femur, sacro-iliac bones and other locations. An autopsy suggested that the angiosarcoma of the heart was the primary lesion, which was occult at the time of the initial operation. Angiosarcoma may have areas with different degrees of differentiation and when a cavernous angioma is suspected histopathologically, the specimen should also be carefully explored for poorly-differentiated areas and the diagnosis of primary or secondary angiosarcoma considered.

Establishment of stable GH3 cell line expressing enhanced yellow fluorescein protein-growth hormone fusion protein.

To investigate, in real time, the transport and secretion of pituitary hormone, we have developed an experimental pituitary cell line, GH3 cell, which has secretory granules of growth hormone (GH) linked to enhanced yellow fluorescein protein (EYFP). This stable GH3 cell secretes secretory granules of GH linked to EYFP on stimulation by Ca2+ influx or Ca2 release from storage. This GH3 cell will be useful for the real-time visualization of the intracellular transport and secretion of GH.

Pituitary abscess in a patient with painless thyroiditis.

The preoperative diagnosis of pituitary abscess is difficult. The case of a 56-year-old man with a pituitary abscess and painless thyroiditis is presented. There has been no previous such report. The differential diagnosis includes pituitary abscess, lymphocytic adenohypophysitis or infundibuloneurohypophysitis, share clinical symptoms of panhypopituitarism and diabetes insipidus, and is of critical importance as the treatment of these conditions differ. The association with painless thyroiditis suggests a diagnosis of lymphocytic adenohypophysitis or infundibuloneurohypophysitis. However, the coincidence of pituitary abscess and painless thyroiditis was observed in our patient, and thus though rare, should be considered. Surgical exploration and histopathological examination are essential for the differential diagnosis of these diseases.

Secretion of high-molecular-weight adrenocorticotropic hormone from a pituitary adenoma in a patient without Cushing stigmata. Case report.

The authors report a case in which a patient harbored a corticotroph macroadenoma that secreted biologically inactive high-molecular-weight adrenocorticotropic hormone (ACTH) as well as authentic ACTH 1-39. The secretion of the high-molecular-weight ACTH was determined using gel chromatography. The authors believe that these two molecules competed with each other at the ACTH receptor and, thus, the bioactivity of ACTH 1-39 was masked and Cushing features were not manifested in the patient. This type of silent corticotroph adenoma may be categorized as a clinically nonfunctioning adenoma. Plasmas from patients with silent corticotroph adenomas, which are identified by positive immunohistochemical staining of ACTH, should be frozen, stored, and analyzed using gel chromatography to examine whether the tumors produce and secrete high-molecular-weight ACTH.

Specific gene suppression using antisense strategy for growth suppression of glioma.

Antisense strategy using synthetic oligodeoxynucleotides has been applied to the suppression of specific gene expression, the modulation of various gene expression, and its biological activity. Antisense strategy is applicable for the growth suppression of glioma cells. Several genes, including transforming growth factor-alpha, basic fibroblast growth factor, fibroblast growth factor receptor 1, vascular endothelial growth factor, telomerase, topoisomerase II alpha-subunit, protein kinase C-alpha, and microtubule-associated protein 1A, have been targeted by antisense strategy in glioma cells. These antisense strategies provide a potential novel antitumor therapy for gliomas.

Microsurgical excision of a retro-odontoid disc hernia via a far-lateral approach: successful treatment of a rare cause of myelopathy: case report.

OBJECTIVE AND IMPORTANCE: Among mass lesions causing myelopathy at the craniovertebral junction, retro-odontoid intervertebral disc hernias are very rare, with only four such cases reported in the literature. CLINICAL PRESENTATION: A 77-year-old woman with this rare condition complained of motor and sensory disturbances in her extremities. Magnetic resonance imaging scans demonstrated an extradural mass lesion at the craniovertebral junction, compressing the lower medulla oblongata and the upper cervical cord posteriorly. INTERVENTION: The lesion, which was partly mucinous cartilaginous and partly fatty and fibrous, was meticulously removed via a left far-lateral approach. The lesion was not neoplastic but was determined to be composed of fibrocartilaginous tissue, consistent with disc material. Postoperatively, the patient's sensory disturbances and motor weakness improved, and magnetic resonance imaging scans demonstrated marked shrinkage of the lesion. CONCLUSION: Sagittal, T1-weighted, magnetic resonance imaging scans demonstrated a low-intensity band between the odontoid process and the body of the axis, which suggested a persistent cartilaginous band. Although upward migration of a herniated disc from the lower cervical spine and degeneration of retro-odontoid ligaments might be possible causes, a persistent cartilaginous band extending between the odontoid process and the body of the axis was considered to be the more likely origin of the retro-odontoid intervertebral disc hernia. Because the far-lateral surgical approach does not require retraction of the cervical cord and provides safe access to the lesion at the craniovertebral junction, it is a suitable surgical method for this condition.

In vivo growth suppression of rat C6 glioma transplanted in rat brain using antisense oligonucleotide for microtubule-associated protein 1A messenger ribonucleic acid.

Microtubule-associated proteins (MAPs) are essential for various cellular processes such as mitosis. The aim of this study was to verify that the suppression of MAP 1A using antisense oligonucleotide can suppress the in vivo proliferation of C6 glioma cells transplanted in rat brain. After 7 days from transplantation, antisense, sense or scramble phosphorothioate oligodeoxynucleotide for MAP 1A mRNA was gradually delivered into the established tumours through amini-osmotic pump. The mean diameters of the tumours from rats treated with antisense, sense and scramble phosphorothioate oligodeoxynucleotide for MAP 1A mRNA were 2.33, 4.625 and 5.25 mm. There was statistically significant difference in diameters of tumours between rats treated with antisense oligodeoxynucleotide, and those treated with sense or scramble oligodeoxynucleotide. This study strongly suggests the important role of MAP 1A in cell proliferation and its suppressionmay serve as a novel antitumour therapy for gliomas.