Professional dental care provision systems for persons with disabilities by prefecture in Japan.

OBJECTIVE: The number of persons with disabilities has increased and aged. Although it is important to maintain good oral health to extend healthy life expectancy, it is difficult for such people. This study aimed to analyze regional disparities in dental care provision systems for disabled people and to propose measures for the establishment of an appropriate system. BASIC RESEARCH DESIGN: To examine regional disparities in dental care provision systems for persons with disabilities, the number of practicing dentists and dental clinics per 100,000 population, dentists certified by the Japanese Society for Disability and Oral Health, and institutions with certified dentists per 100,000 disabled persons for each prefecture were calculated. The Gini coefficient of each was also calculated to visualize and analyze regional disparities. RESULT: The Gini coefficients were 0.09 and 0.07 for practicing dentists and dental clinics and 0.32 and 0.28 for the certified dentists and institutions with the certified dentists, respectively. Dental institutions for the disabled abounded in the three metropolitan areas: Tokyo, Aichi, and Osaka, and their density tended to be lower in northern and southern Japan. In prefectures with few such institutions, there was no correlation between the number of institutions and prefectural residents' income, and some prefectures had similar incomes but had many institutions. CONCLUSION: The distribution of dental care to the disabled is highly uneven in Japan, therefore, a system needs to be established to address this issue.

Geographic distribution of dental specialists permitted to advertise dental practices in Japan.

OBJECTIVE: The purpose of this study was to determine the geographic distribution of dental specialists permitted to advertise dental practices in Japan. METHOD: We identified the populations of 349 secondary medical zones nationwide from the 2015 population census, as well as the number of dentists in five specialties, namely oral surgeons, pedodontists, periodontists, dental anesthesiologists, and dental radiologists, who had been permitted to advertise dental practices, from a 2016 survey of physicians, dentists, and pharmacists. We determined the placement rate, Lorenz curve, and Gini coefficient for dentists in each specialty in order to describe their geographic distributions. RESULTS: The placement rates of at least one of these types of dentist in each secondary medical zone were 73.9% for oral surgeons, 66.2% for pedodontists, 60.5% for periodontists, 31.8% for dental anesthesiologists, and 18.3% for dental radiologists. The Gini coefficients were 0.397, 0.400, 0.491, 0.650, and 0.761, respectively. CONCLUSION: The dentists in each specialty were few in number and were unequally distributed among the zones, but less so for oral surgeons and pedodontists. Dental anesthesiologists and radiologists were located primarily at university hospitals in urban areas and, therefore, were more unequally distributed.

3D niche microarrays for systems-level analyses of cell fate.

The behaviour of mammalian cells in a tissue is governed by the three-dimensional (3D) microenvironment and involves a dynamic interplay between biochemical and mechanical signals provided by the extracellular matrix (ECM), cell-cell interactions and soluble factors. The complexity of the microenvironment and the context-dependent cell responses that arise from these interactions have posed a major challenge to understanding the underlying regulatory mechanisms. Here we develop an experimental paradigm to dissect the role of various interacting factors by simultaneously synthesizing more than 1,000 unique microenvironments with robotic nanolitre liquid-dispensing technology and by probing their effects on cell fate. Using this novel 3D microarray platform, we assess the combined effects of matrix elasticity, proteolytic degradability and three distinct classes of signalling proteins on mouse embryonic stem cells, unveiling a comprehensive map of interactions involved in regulating self-renewal. This approach is broadly applicable to gain a systems-level understanding of multifactorial 3D cell-matrix interactions.

Trends in the geographic distribution of dental clinics in Japan.

AIM: In Japan, along with the increase in the number of dentists, the number of dental facilities has continuously increased as well. This study aimed to examine whether the increase in the number of dental clinics in Japan has led to an improvement in their geographic distribution. METHODS: We analysed the number of dental clinics and population in all municipalities in Japan as of 2000, 2005 and 2010. We obtained data on the population from the population census and data on the number of dental clinics from the Survey of Medical Institutions. The number of municipalities was 3,258 in 2000 but had dropped to 1,750 by 2010 due to municipal mergers so population and dental data for other years were recalculated based on 2010 municipal boundaries. Lorenz curves and Gini coefficients were used to assess the distribution of dental clinics per 100,000 persons. RESULTS: The mean number of dental clinics per 100,000 persons among all municipalities was 49.9 in 2000, 52.2 in 2005 and 53.4 in 2010. The Gini coefficient for the clinics in the whole country was 0.172 in 2000, 0.164 in 2005 and 0.153 in 2010. CONCLUSION: The results suggest that the regional inequalities in the availability of dentists have been reduced gradually as the number of dental clinics has increased.

Introduction and evaluation of computer-assisted education in an undergraduate dental hygiene course.

OBJECTIVE: This paper introduced newly developed computer-assisted learning materials and reports of a survey of junior college dental hygiene students who have used them. METHODS: We authored new educational material to promote students' basic dental hygiene practice skills using a simulation software generator. A set of five developed materials were tested by 43 female second-year dental hygiene students during the second semester at a college in Chiba, Japan. The evaluation was conducted in the form of a questionnaire including open-ended questions. Students' opinions were analysed using characteristic diagrams, a troubleshooting tool that can be used to visually illustrate the causes and effects of a problem. RESULT: The overall results of the evaluation were positive. The students were given five sets of simulation learning materials (SLMs). Eighty-three percent of the students felt that they could carry out independent study of clinical practice better after the virtual practice. Ninety-three percent of them felt that the exercises should be continued in the future, and eighty-eight percent of them felt that this virtual practice deepened their interest in other classes and training sessions. All of the students found the virtual practice beneficial for their learning. DISCUSSION: The present results suggest that the students became conscious of their lack of knowledge through SLMs. These findings indicate that SLMs for practicing basic clinical procedures is beneficial.

Lymphatic architecture of the human gingival interdental papilla.

Many studies have investigated the lymphatic architecture of head and neck using experimental animals, confirming the existence of lymphatic networks beneath the epithelium in gingival tissue. In this study, we investigated the use of these lymphatics as a drug delivery route by studying the architecture of lymphatic vessels in human interdental papilla. Serial cryosections were cut using the film-transfer method. To identify lymphatics, the sections were stained using enzyme histochemical and immunohistochemical techniques and three-dimensional images of lymphatics were reconstructed using 3D visualization software. Capillary lymphatic networks were observed in the lamina propria beneath the epithelium in human interdental papilla, and they joined with lymphatic networks beneath the epithelium in free gingiva. The networks consisted of a single layer of large irregular, hexagonal meshes and precollecting lymphatic vessels heading toward collecting lymphatic vessels that exited on the periosteum of the alveolar crest. These findings suggest that lymphatic flow from the interdental papilla drains into collecting lymphatic vessels running buccolingually on the alveolar crest of the interdental papilla. This may be an important anatomical feature during inflammation throughout the oral cavity in that the drainage function is maintained by part of lymphatic flow that is not impaired during the healing process.

AT7519, A novel small molecule multi-cyclin-dependent kinase inhibitor, induces apoptosis in multiple myeloma via GSK-3beta activation and RNA polymerase II inhibition.

Dysregulated cell cycling is a universal hallmark of cancer and is often mediated by abnormal activation of cyclin-dependent kinases (CDKs) and their cyclin partners. Overexpression of individual complexes are reported in multiple myeloma (MM), making them attractive therapeutic targets. In this study, we investigate the preclinical activity of a novel small-molecule multi-CDK inhibitor, AT7519, in MM. We show the anti-MM activity of AT7519 displaying potent cytotoxicity and apoptosis; associated with in vivo tumor growth inhibition and prolonged survival. At the molecular level, AT7519 inhibited RNA polymerase II (RNA pol II) phosphorylation, a CDK9, 7 substrate, associated with decreased RNA synthesis confirmed by [(3)H] Uridine incorporation. In addition, AT7519 inhibited glycogen synthase kinase 3beta (GSK-3beta) phosphorylation; conversely pretreatment with a selective GSK-3 inhibitor and shRNA GSK-3beta knockdown restored MM survival, suggesting the involvement of GSK-3beta in AT7519-induced apoptosis. GSK-3beta activation was independent of RNA pol II dephosphorylation confirmed by alpha-amanitin, a specific RNA pol II inihibitor, showing potent inhibition of RNA pol II phosphorylation without corresponding effects on GSK-3beta phosphorylation. These results offer new insights into the crucial, yet controversial role of GSK-3beta in MM and show significant anti-MM activity of AT7519, providing the rationale for its clinical evaluation in MM.

Ascorbic acid inhibits antitumor activity of bortezomib in vivo.

Earlier studies have shown that ascorbic acid (vitamin C) inhibits bortezomib-induced cytotoxicity against cancer cells in vitro. However, the clinical significance of vitamin C on bortezomib treatment is unclear. In this study, we examined whether daily oral intake of vitamin C inhibits antimultiple myeloma (MM) activities of bortezomib. Vitamin C, at orally achievable concentrations, inhibited in vitro MM cell cytotoxicity of bortezomib and blocked its inhibitory effect on 20S proteasome activity. Specifically, plasma collected from healthy volunteers taking 1 g/day vitamin C reduced bortezomib-induced MM cell death in vitro. This antagonistic effect of vitamin C against proteasome inhibitors is limited to the boronate class of inhibitors (bortezomib and MG262). In vivo activity of this combination treatment was then evaluated using our xenograft model of human MM in SCID (severe combined immune-deficient) mice. Bortezomib (0.1 mg/kg twice a week for 4 weeks) significantly inhibits in vivo MM cell growth, which was blocked by oral vitamin C (40 mg/kg/day). Therefore, our results for the first time show that vitamin C can significantly reduce the activity of bortezomib treatment in vivo; and importantly, suggest that patients receiving treatment with bortezomib should avoid taking vitamin C dietary supplements.

[Coronary artery bypass in octogenarians; how to reduce operative mortality and enhance long-term quality of life].

Octogenarians are at increased risk for perioperative morbidity and mortality after coronary artery bypass. This study is aimed to elucidate the suitable operative strategy and perioperative management. A retrospective analysis was conducted of 54 consecutive patients with 80-years of age or older, who underwent elective isolated coronary artery bypass between May 1999 and May 2008. Mean follow-up was 43.3 months and 96.3% complete. Operavive mortality was 3.7% and the incidence of stroke was 3.7%. The 7-year cardiac survival was 80.4% and the 7-year cardiac event free was 65.0%. The use of arterial graft to the right coronary artery was identified as independent predictor of late cardiac event. Neither total arterial revascularization nor bilateral internal thoracic artery grafting was a significant cardiac event factor. This retrospective study suggests a benefit of the less invasive strategy in terms of operative mortality and morbidity. Application of fast-track treatment in octogenarians appears to be an effective approach to reduce perioperative morbidity and enhance long-term quality of life.

Preclinical activity of P276-00, a novel small-molecule cyclin-dependent kinase inhibitor in the therapy of multiple myeloma.

Cyclin D dysregulation and overexpression is noted in the majority of multiple myeloma (MM) patients, suggesting its critical role in MM pathogenesis. Here, we sought to identify the effects of targeting cyclin D in MM. We first confirmed cyclin D mRNA overexpression in 42 of 64 (65%) patient plasma cells. Silencing cyclin D1 resulted in >50% apoptotic cell death suggesting its validity as a potential therapeutic target. We next evaluated P276-00, a clinical-grade small-molecule cyclin-dependent kinase inhibitor as a way to target the cyclins. P276-00 resulted in dose-dependent cytotoxicity in MM cells. Cell-cycle analysis confirmed either growth arrest or caspase-dependent apoptosis; this was preceded by inhibition of Rb-1 phosphorylation with associated downregulation of a range of cyclins suggesting a regulatory role of P276-00 in cell-cycle progression through broad activity. Proliferative stimuli such as interleukin-6, insulin-like growth factor-1 and bone-marrow stromal cell adherence induced cyclins; P276-00 overcame these growth, survival and drug resistance signals. Because the cyclins are substrates of proteasome degradation, combination studies with bortezomib resulted in synergism. Finally, in vivo efficacy of P276-00 was confirmed in an MM xenograft model. These studies form the basis of an ongoing phase I study in the treatment of relapsed/refractory MM.

Lenalidomide inhibits osteoclastogenesis, survival factors and bone-remodeling markers in multiple myeloma.

Osteolytic bone disease in multiple myeloma (MM) is caused by enhanced osteoclast (OCL) activation and inhibition of osteoblast function. Lenalidomide and bortezomib have shown promising response rates in relapsed and newly diagnosed MM, and bortezomib has recently been reported to inhibit OCLs. We here investigated the effect of lenalidomide on OCL formation and osteoclastogenesis in comparison with bortezomib. Both drugs decreased alpha V beta 3-integrin, tartrate-resistant acid phosphatase-positive cells and bone resorption on dentin disks. In addition, both agents decreased receptor activator of nuclear factor-kappaB ligand (RANKL) secretion of bone marrow stromal cells (BMSCs) derived from MM patients. We identified PU.1 and pERK as major targets of lenalidomide, and nuclear factor of activated T cells of bortezomib, resulting in inhibition of osteoclastogenesis. Furthermore, downregulation of cathepsin K, essential for resorption of the bone collagen matrix, was observed. We demonstrated a significant decrease of growth and survival factors including macrophage inflammatory protein-alpha, B-cell activating factor and a proliferation-inducing ligand. Importantly, in serum from MM patients treated with lenalidomide, the essential bone-remodeling factor RANKL, as well as the RANKL/OPG ratio, were significantly reduced, whereas osteoprotegerin (OPG) was increased. We conclude that both agents specifically target key factors in osteoclastogenesis, and could directly affect the MM-OCL-BMSCs activation loop in osteolytic bone disease.

Novel etodolac analog SDX-308 (CEP-18082) induces cytotoxicity in multiple myeloma cells associated with inhibition of beta-catenin/TCF pathway.

We have reported previously that R-enantiomer of etodolac (R-etodolac), which is under investigation in phase 2 clinical trials in chronic lymphocytic leukemia, induces potent cytotoxicity at clinically relevant concentrations in multiple myeloma (MM) cells. In this study, we demonstrated that SDX-308 (CEP-18082), a novel analog of etodolac, has more potent cytotoxicity than R-etodolac against both MM cell lines and patient MM cells, including tumor cells resistant to conventional (dexamethasone, doxorubicine, melphalan) and novel (bortezomib) therapies. SDX-308-induced cytotoxicity is triggered by caspase-8/9/3 activation and poly (ADP-ribose) polymerase cleavage, followed by apoptosis. SDX-308 significantly inhibits beta-catenin/T-cell factor pathway by inhibiting nuclear translocation of beta-catenin, thereby downregulating transcription and expression of downstream target proteins including myc and survivin. Neither interleukin-6 nor insulin-like growth factor-1 protect against growth inhibition triggered by SDX-308. Importantly, growth of MM cells adherent to bone marrow (BM) stromal cells is also significantly inhibited by SDX-308. Our data therefore indicate that the novel etodolac analog SDX-308 can target MM cells in the BM milieu.

[Effects of dexmedetomidine hydrochloride on postoperative sedation in cardiovascular surgery].

BACKGROUND: Postoperative assessment of brain damage in cardiovascular surgery is often obscured by sedatives. Therefore, early postoperative detection of brain attack and its treatment are also hampered. A newly approved sedative, dexmedetomidine hydrochloride has weak analgesic effect and no respiratory depressive effect. These characteristics allow early assessment of brain damage after surgery. In this report, we compared 2 sedatives, propofol and dexmedetomidine hydrochloride, in cardiovascular settings. SUBJECT AND METHODS: Both sedatives were initiated right after admission to the intensive care unit (ICU), followed by titrimetric method targeting for the sedation agitation scale (SAS) from 1 to 4. Thirty-five cases were included in dexmedetomidine hydrochloride group (DEX group) and 16 cases were included in propofol group (Prop group). RESULTS: Preoperative and operative demographic data were the same between the 2 groups. Conversion rate to another sedatives, and incidence of vasopressor or hypotensor use were both in the same proportion in both groups. Intubated time was the same in both groups. Both systolic and diastolic blood pressures were kept lower in DEX group than Prop group until 8 hours after ICU admission. Other hemodynamic measurements, heart rate, pulmonary artery pressure and cardiac index showed no statistical difference. SAS and Ramsay score were better in DEX group early after ICU admission, and remained better until 10 hours later. CONCLUSION: Dexmedetomidine hydrochloride has no major hemodynamic nor other side effects after cardiovascular surgery. Dexmedetomidine hydrochloride could be used as an effective agent for postoperative sedation and analgesia in cardiovascular settings.

[Aortic valve replacement in elderly patients with small aortic annulus; is the indexed effective orifice area important?].

We sought to determine whether the small indexed effective orifice area (EOAI) increased mortality and morbidity after aortic valve replacement (AVR) in patients over 75 years of age. From May 1999 to July 2005, 77 patients underwent isolated AVR for aortic stenosis. They were divided into 3 groups (S-EOAI : EOAI < or = 0.7 cm2/m2, M-EOAI : 0.7 cm2/m2

Invasive phenotype and apoptosis induction of Plesiomonas shigelloides P-1 strain to Caco-2 cells.

AIMS: The mechanism of the host cell invasion of Plesiomonas shigelloides and its capability to induce apoptosis were investigated. METHODS AND RESULTS: We performed a time course experiment on the bacterial adherence and invasion of the P. shigelloides P-1 strain into Caco-2 cells using an invasion assay and flow cytometry. The adherence of P. shigelloides to the Caco-2 cells was almost completed within 10 min after the infection. Thereafter, P. shigelloides starts internalization within the Caco-2 cells, which was completed within 60 min after the infection. Based on the invasion assay using nocodazole, cytochalasin D, and genistein, it became clear that the mechanism of the internalization depended on the signal transduction followed by the rearrangement of the cytoskeletal protein. Based on the DNA laddering and TUNEL methods, the cytotoxicity of the Caco-2 cells by the invasion of P. shigelloides occurred through the induction of apoptosis. CONCLUSIONS: This work demonstrated that the mechanism of invasion of P. shigelloides into Caco-2 cells and the invasion of P. shigelloides induces apoptotic cell death. SIGNIFICANCE AND IMPACT OF THE STUDY: This work revealed the virulence factor, which may be important for understanding of the pathogenesis of P. shigelloides.

The subjective dimension of functioning and disability: what is it and what is it for?

The International Classification of Functioning, Disability and Health (ICF, WHO 2001) made a great advancement over ICIDH of 1980 in the understanding of the human functioning and disability. However, in both of them there is an important 'missing' element. That is the subjective dimension of functioning and disability. One of the authors (S. Ueda) published on this topic in 1981 both in Japanese and English. It had originated from his clinical experience in rehabilitation medicine. The understanding of the inner world of the client has proved a great asset in clinical practice. This paper explains its importance and provides a definition. It also proposes a tentative framework of a classification of subjective dimension of functioning and disability as the starting point for more intensive and extensive discussion on this important problem, and for its future inclusion into ICF.

Purification and identification of monoubiquitin-phosphoglycerate mutase B complex from human colorectal cancer tissues.

Ubiquitin-conjugated proteins in human colorectal cancer tissues were analyzed by the immunoprecipitation with the antibody FK2 against conjugated ubiquitin followed with SDS-PAGE. In these immunoprecipitable proteins, a 38-kDa protein was abundant in the tumor regions but almost absent in the adjacent normal regions in 17/26 patients, thus we attempted to purify it. Using immunoaffinity chromatography with the antibody FK2 followed by gel filtration and SDS-PAGE, approximately 10 pmol of this protein was separated from 34 g of the pooled cancerous tissue and transferred onto a PVDF membrane. The 38-kDa protein was further digested with Achromobacter protease I, resulting in several peptide fragments. Amino acid sequences of these peptides showed complete sequence identity to those derived from either ubiquitin or phosphoglycerate mutase-B, suggesting that the 38-kDa protein is monoubiquitinated phosphoglycerate mutase-B, whose calculated mass is 37,369 Da. Western blot using an antibody against phosphoglycerate mutase-B revealed the presence of the 38-kDa protein in the anti-ubiquitin immunoprecipitates derived from the tumor regions, but not from normal counterparts. In addition, part of non-ubiquitinated phosphoglycerate mutase-B (29 kDa) was also found in the anti-ubiquitin immunoprecipitates, whose levels were higher in the tumor regions than in the adjacent normal regions. These results suggest that monoubiquitination of phosphoglycerate mutase-B as well as formation of a noncovalent complex containing ubiquitin and phosphoglycerate mutase-B increases in colorectal cancer and novel modification of phosphoglycerate mutase-B might have a pathophysiological role.

Clinical studies of bioabsorbable poly-L-lactide sternal coaptation pins.

BACKGROUND: Median sternotomy has become the most commonly used incision in cardiac surgery. Since sternal dehiscence, however, is a major complication, we used bioabsorbable poly-L-lactide (P-L-LA) sternal coaptation pins for sternal closure to prevent it. METHODS: From February 1998 to October 1999, 99 patients (64 men, 35 women; mean age, 63+/-1.2 years) underwent median sternotomy for cardiac surgery using sternal coaptation pins. Nineteen patients had diabetes mellitus and seven had renal failure. In closure, two sternal pins were inserted into the bone marrow of the sternum, one into the manubrium, the other into the body, and the sternum was sutured with five stainless steel wires. RESULTS: Five patients died in the hospital. The causes of death were cardiac failure in two patients, respiratory problem in two and perforation of the stomach in one. The average length of hospitalization was 2 4.5+/-2.5 days. Sternal dehiscence occurred in one patient and mediastinitis in four. There was no bleeding from the bone marrow and no complication related to the use of the sternal pins. CONCLUSIONS: P-L-LA sternal pins were easy to insert and may be effective in preventing dehiscence of the sternum.

Identification and characterization of the human serotonin-4 receptor gene promoter.

The human serotonin-4 (5-HT(4)) receptor gene expression is highly regulated in various tissues. We isolated the human 5-HT(4) receptor gene containing the 5'-flanking region and characterized its promoter. By 5'-RACE (5'-rapid amplification of the cDNA ends) and inverse PCR, multiple transcription initiation sites were identified. The most 5' one (assigned to +1) was 5135 bp upstream to the translation start site. The 500-bp 5'-flanking region contained potential binding sites for transcription factor Sp-1, AP-2, AP-4, and GATA. However, this region lacked TATA- and CAAT-boxes. Transient transfection analyses in human choriocarcinoma T3M-3 (5-HT(4) receptor-positive) and HepG2 (5-HT(4) receptor-negative) cells revealed that the region (-210 to -105) is necessary for the basic and cell-type specific 5-HT(4) receptor gene expression. In addition, untranslated exon 1 contained negative (+112 to +182) as well as positive (+1 to +111) modulators, indicating that exon 1 plays a regulatory role in the 5-HT(4) receptor gene expression.

Isolation of ubiquitin-E2 (ubiquitin-conjugating enzyme) complexes from erythroleukaemia cells using immunoaffinity techniques.

A variety of ubiquitin-associated (or conjugated) proteins, including substrates and enzymes for the ubiquitin system, are present in eukaryotic cells. In the present study we developed a simple method for their isolation, consisting of immunoaffinity chromatography using the monoclonal antibody FK2, which recognizes the conjugated ubiquitin molecule. Using this method followed by gel filtration, we isolated multi-ubiquitinated proteins with high molecular masses (>30 kDa) and also ubiquitinthioester-linked and mono-ubiquitinated forms of ubiquitin-conjugating (E2) enzymes, UbcH7 and UBE2N, together with mono-, di- and tri-ubiquitin molecules, from the cytoplasmic extract of heat-shock-treated K562 erythroleukaemia cells. We also demonstrated that the FK2 antibody was capable of precipitating a ubiquitin-UbcH7 thioester, but not free UbcH7, which enabled the measurement of the respective cellular levels separately. The immunoprecipitable ubiquitin-UbcH7 thioester was found only when the cells were treated with heat-shock. These results suggest the usefulness of the immunoaffinity techniques for identifying and analysing the cellular enzyme/protein-ubiquitin complexes.