Oral environment and taste function of Japanese HIV-infected patients treated with antiretroviral therapy.

The aim of the study was to evaluate the oral environment and the taste function of Japanese HIV-infected patients treated with antiretroviral therapy. Their median age of 73 patients taking anti-HIV drugs was 46 years. The median period of taking anti-HIV drugs was 30 months. The oral condition was evaluated by measurement of oral moisture, amount of saliva secretion, the number of oral bacteria, presence of oral candida, a taste test, and the number of missing teeth. The levels of oral moisture and secreted saliva were significantly lower in the HIV-infected group than in the healthy volunteer (control) group. The HIV-infected group showed a more robust decrease in taste sensation than the control group. The number of missing teeth was significantly higher in the HIV-infected group than in the control group. Furthermore, all of the evaluated oral conditions were worse in the HIV-infected patients whose CD4+ T lymphocyte counts were less than 500/mm3 than in the control group. It became clear that the patients taking anti-HIV drugs, especially the CD4+ count < 500/mm3 group, had a deteriorated oral environment and dysgeusia, suggesting that the management of oral hygiene is necessary to maintain oral health, which leads to systemic health.

Comparison of sirolimus eluting stent with bioresorbable polymer to everolimus eluting stent with permanent polymer in bifurcation lesions: Results from CENTURY II trial.

OBJECTIVE: To demonstrate the safety and efficacy of a new sirolimus eluting stent with bioresorbable polymer, Ultimaster, (BP-SES) compared with everolimus-eluting, permanent polymer, Xience stent (PP-EES) in bifurcation lesions with respect to the freedom from Target Lesion Failure at 1-year. METHODS: Within 1,119 patients enrolled in the CENTURY II randomized controlled multicenter trial, 194 patients were treated for bifurcation lesions and randomized to either BP-SES (n = 95) or PP-EES (n = 99). The primary endpoint was freedom from target lesion failure (TLF) composite endpoint [cardiac death, MI not clearly attributable to a non-target vessel, and clinically driven target lesion revascularization (TLR)] at 1-year. RESULTS: Baseline patient demographic, angiographic, and stenting characteristics were similar in both study arms. A single stent technique with provisional or "cross over" stenting were the most widely used in both arms (93.2% BP-SES vs. 92.4% PP-EES). Freedom from TLF at 1-year was 94.7% for BP-SES and 91.9% for PP-EES (P for noninferiority 0.031). The rate of clinically driven target lesion revascularization (TLR) at 1-year was 3.2% for BP-SES and 3.0% for PP-EES (P = 0.95). There were no significant differences detected in any of the individual clinical endpoints or other secondary clinical endpoints between the study arms at 1-year follow up. CONCLUSIONS: The new bioresorbable polymer sirolimus-eluting stent showed safety and efficacy profiles similar to durable polymer everolimus-eluting in the treatment of patients with bifurcation lesions at 1-year follow up. © 2015 Wiley Periodicals, Inc.

Immunohistochemical expression of heat shock protein27 in the mouse dental pulp after immediate teeth separation.

AIM: After immediate teeth separation, expression of HSP27 in the mouse dental pulp was examined. Immunohistochemistry was performed to examine the incidence of HSP27 expression. MATERIALS AND METHODS: A total of 36 8-week-old ddY mice were used as experimental subjects and a wedge was inserted in between maxillary right molars. The wedge was removed 30 min or 3 h after insertion. Animals were immediately sacrificed after the removal of wedge or until 1 week later and serial sections from paraffin-embedded tissues were prepared. Immunohistochemistry was carried out to examine the expression of HSP27. The untreated side served as the control. RESULTS: In the control group, the endothelial cells and some pulp fibroblasts weakly expressed HSP27 suggesting that the expression is due to mechanical stress brought about by physiological masticatory force and pressure from the tongue. In both 30 min and 3 h experimental groups, HSP27 expression was highest at 24 h after wedge removal and the expression remained the same or started to decrease thereafter. The expression decreased at the same level as that of the control group 1 week after wedge removal. CONCLUSION: HSP27 may serve as an indicator of stimulus strong enough to show its expression.

Immunohistochemical expression of hard tissue related factors in the mouse dental pulp after immediate teeth separation.

We examined change of Runx2 and ALP expression in mouse tooth pulp which exposed to teeth separation experiment by immunohistochemistry as a model for conservative dentistry treatment. 8-week-old 36 male ddY mice were used and wedge was inserted between upper 1st and 2nd molars. The wedge was removed 30 minutes as well as 3 hours after the insertion and the samples were prepared extending up to 1 week of time period for regular histopathological and immunohistochemical examinations for ALP and Runx2 expression. The opposite sides without wedge insertion were taken as controls. In the control group pulp, weak expressions of Runx2 and ALP in the vessel endothelial cells as well as the pulp cells were revealed, suggesting the appearance of these genes upon mechanical stress induced by mastication and tongue pressure etc. On the other hand in the experiment group, Runx2 expression increased both in 30-minute and 3-hour teeth separation group. The expression became maximum at 24 hours. Then it gradually decreased and became similar level with the control group at 1-week after the wedge insertion. Similarly ALP expression increased after the wedge insertion and was maximum at 24 hours and then gradually decreased to the levels similar with the control group. These results suggest that when immunohistochemical expression of Runx2 as well as ALP was used as an index, no severe damage occur upon clinical application of wedge insertion.

Association between lipoprotein subfraction profile and the response to hepatitis C treatment in Japanese patients with genotype 1b.

Pegylated interferon and ribavirin combination therapy is the standard treatment for patients with chronic hepatitis C (CHC). Some groups have reported a relation between lipid values and response while others have reported that microsomal triglyceride transfer protein, a key enzyme in the assembly and secretion of lipoproteins, was related to hepatitis C virus (HCV). The aim of this study was to investigate the association between the lipoprotein profiles, classified according to size, and hepatitis C treatment and the usefulness for predicting the outcome of treatment. Forty-four patients with CHC (27 men and 17 women) were included in the study. The serum cholesterol and triglyceride (TG) levels in the lipoprotein subclasses were determined using high-performance liquid chromatography with gel permeation columns, which classified lipoproteins into 20 subfractions based on particle size. According to a univariate analysis, those who achieved an sustained viral response (SVR) had a significantly higher serum total cholesterol level, higher cholesterol levels in the low-density lipoprotein subfraction (25.5 nm in diameter) and the very low-density lipoprotein (VLDL) subfraction (44.5 and 36.8 nm), and a higher serum TG level in the VLDL subfraction (44.5 nm), compared with the corresponding values in the non-SVR group. Higher serum cholesterol and TG concentrations in the lipoprotein subfractions were predictive of an SVR to therapy for HCV infection with genotype 1b prior to the start of interferon treatment.

Association between phospholipids and free cholesterol in high-density lipoprotein and the response to hepatitis C treatment in Japanese with genotype 1b.

Pegylated interferon and ribavirin combination therapy is the standard treatment for patients with chronic hepatitis C (CHC), but treatment failure can be difficult to predict. We and others have reported a relation between lipid values and sustained viral responses in patients with CHC. However, the relationship between lipid values and treatment failure has not been previously reported. The present study investigated the association between the profiles of phospholipids and free cholesterol (FC), the main constitutive ingredients of the surface of lipoprotein, classified according to particle size and hepatitis C treatment, and determined the usefulness of these parameters for predicting the outcome of treatment. Fifty-five patients with CHC (33 men and 22 women) were included in the study. The serum total cholesterol, triglyceride, phospholipids, and FC levels in the lipoprotein subclasses were determined using high-performance liquid chromatography with gel permeation columns, enabling the lipoproteins to be classified into 13 subclasses according to particle size. According to a univariate analysis, the treatment failure group had a significantly higher serum phospholipid level overall in the high-density lipoprotein (HDL) and medium HDL fractions as well as a higher serum FC level in the HDL fraction and all HDL subclass fractions compared with the corresponding values in the non-nonvirological response group. Higher serum phospholipid and FC concentrations in the HDL subclasses were predictive of a failure to respond in patients with genotype 1b.

IL-17 in synovial fluids from patients with rheumatoid arthritis is a potent stimulator of osteoclastogenesis.

IL-17 is a newly discovered T cell-derived cytokine whose role in osteoclast development has not been fully elucidated. Treatment of cocultures of mouse hemopoietic cells and primary osteoblasts with recombinant human IL-17 induced the formation of multinucleated cells, which satisfied major criteria of osteoclasts, including tartrate-resistant acid phosphatase activity, calcitonin receptors, and pit formation on dentine slices. Direct interaction between osteoclast progenitors and osteoblasts was required for IL-17-induced osteoclastogenesis, which was completely inhibited by adding indomethacin or NS398, a selective inhibitor of cyclooxgenase-2 (COX-2). Adding IL-17 increased prostaglandin E2 (PGE2) synthesis in cocultures of bone marrow cells and osteoblasts and in single cultures of osteoblasts, but not in single cultures of bone marrow cells. In addition, IL-17 dose-dependently induced expression of osteoclast differentiation factor (ODF) mRNA in osteoblasts. ODF is a membrane-associated protein that transduces an essential signal(s) to osteoclast progenitors for differentiation into osteoclasts. Osteoclastogenesis inhibitory factor (OCIF), a decoy receptor of ODF, completely inhibited IL-17-induced osteoclast differentiation in the cocultures. Levels of IL-17 in synovial fluids were significantly higher in rheumatoid arthritis (RA) patients than osteoarthritis (OA) patients. Anti-IL-17 antibody significantly inhibited osteoclast formation induced by culture media of RA synovial tissues. These findings suggest that IL-17 first acts on osteoblasts, which stimulates both COX-2-dependent PGE2 synthesis and ODF gene expression, which in turn induce differentiation of osteoclast progenitors into mature osteoclasts, and that IL-17 is a crucial cytokine for osteoclastic bone resorption in RA patients.

A multilayer membrane amperometric glucose sensor fabricated using planar techniques for large-scale production.

This paper reports on a multilayer membrane amperometric glucose sensor fabricated using planar techniques. It is characterized by good reproducibility and suitable for large-scale production. The glucose sensor has 82 electrode sets formed on a single glass substrate, each with a platinum working electrode (WE), a platinum counter electrode (CE) and an Ag/AgCl reference electrode (RE). The electrode sets are coated with a membrane consisting of five layers: gamma-aminopropyltriethoxysilane (gamma-APTES), Nafion, glucose oxidase (GOX), gamma-APTES and perfluorocarbon polymer (PFCP), in that order. Tests have shown that the sensor has acceptably low dispersion (relative standard deviation, R.S.D.=42.9%, n=82), a wide measurement range (1.11-111 mM) and measurement stability over a 27-day period. Measurements of the glucose concentration in a control human urine sample demonstrated that the sensor has very low dispersion (R.S.D.=2.49%, n=10).

Percutaneous radio-frequency mandibular nerve rhizotomy guided by CT fluoroscopy.

We describe a new method for radio-frequency mandibular nerve rhizotomy under CT fluoroscopy. A patient with cancer had severe intractable and drug-resistant pain in his left mandibular region. Because he had an anatomic deformity due to cancer invasion and radiation therapy, we planned a mandibular nerve rhizotomy under CT fluoroscopic imaging. The needle was advanced to the mandibular nerve just caudal to the foramen ovale under real-time CT fluoroscopy, avoiding the cancer region. Pain scores of the patient were reduced after the nerve rhizotomy, without any complications.

Analysis of feeding behavior with direct linear transformation.

AIM: The sucking pattern of term infants is composed of a rhythmic alteration of expression and suction movements. The aim is to evaluate if direct linear transformation (DLT) method could be used for the assessment of infant feeding. SUBJECT AND METHODS: A total of 10 gnormalh infants and two infants with neurological disorders were studied using DLT procedures and expression/suction pressure recordings. Feeding pattern of seven gnormalh infants were evaluated simultaneously recording DLT and pressures. The other infants were tested non-simultaneously. We placed markers on the lateral angle of the eye, tip of the jaw, and throat. The faces of infants while sucking were recorded in profile. The jaw and throat movements were calculated using the DLT procedure. Regression analysis was implemented to investigate the relationship between suction and expression pressures and eye-jaw and eye-throat movement. All regression analyses investigated univariate relationships and adjusted for other covariates. RESULTS: Ten gnormalh infants demonstrated higher suction pressure than expression pressure, and their throat movements were larger than jaw movements. Two infants with neurological problems did not generate suction pressure and demonstrated larger movements in their jaw than throat. The simultaneous measurement (n=7) showed a significant correlation, not only between eye-jaw distance and the expression pressure, but also between eye-throat distance and suction pressure. The change in the eye-jaw distance was smaller than the changes in the eye-throat distance in gnormalh infants (p<0.001). CONCLUSIONS: The DLT method can be used to evaluate feeding performance without any special device.

Synthesis of procollagen by odontogenic cells of rabbit tooth germ.

Studies were performed to determine whether cultured odontogenic cells from rabbit tooth germ (RP cell) could synthesize dentine-like collagen. When cells were cultured with [14C]proline, 33% of the total incorporated proteins present were collagenous. Cultured RP cells were labeled with [14C]proline in the presence of beta-aminopropionitrile. The resulting fractions, on analysis by CM-cellulose chromatography, contained three radioactive protein peaks, alpha 1(I), [alpha 1(III)]3, alpha 2. From the radioactive measurements, RP cells synthesized a significant amount of type III collagen, comparable to type I collagen. DEAE-cellulose chromatography was used to separate collagen molecules from collagen precursors. The results showed that 60% of total collagen precursor was type III precursor and the remainder was type I precursor. CM-cellulose chromatography of CNBr peptides of collagen from culture medium and cell extract revealed the presence of type I and type III collagen. Thus, the RP cell, which is a diploid cell, is unique in the predominance of type III collagen in culture, differing thereby from the character of collagen in vivo.

Bone-resorbing activity and prostaglandin E produced by human periodontal ligament cells in vitro.

Human periodontal ligament (PDL) cells were derived from healthy premolars extracted for orthodontic treatment and were utilized for in vitro experiments in passages 4-6. Human PDL cells were seeded in tissue culture tubes and incubated with interleukin-1 alpha (IL-1 alpha), IL-1 beta, tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), indomethacin, parathyroid hormone (PTH), or their combinations, for 1 h. The medium was then replaced with serum-free BGJb medium and incubated for 24 h without further additions. Prostaglandin E (PGE) concentrations in the conditioned media (CM) were measured by radioimmunoassay, and bone-resorbing activity was measured using 45Ca-labeled neonatal mouse calvariae. The results of this study indicated that (1) unstimulated cultured PDL cells produced PGE, and PDL CM stimulated bone resorption; (2) cytokine-treated (IL-1 alpha, IL-1 beta, and TNF-alpha) PDL cells had increased production of PGE and bone-resorbing activity compared to unstimulated PDL cells; (3) indomethacin completely inhibited PGE production from unstimulated PDL cells but only partially inhibited bone-resorbing activity, indicating that PDL cells produced nonprostaglandin bone-resorbing factor(s); (4) IFN-gamma did not change PGE or bone-resorbing activity production by cytokine-stimulated PDL cells; and (5) PTH treatment of PDL cells in addition to cytokines (IL-1 alpha, IL-1 beta, and TNF-alpha) had additive effects on the production of bone-resorbing activity and synergistic effects on PGE production compared to cytokine treatment alone.

Stimulation of DNA synthesis in trophoblasts and human umbilical vein endothelial cells by hepatocyte growth factor bound to extracellular matrix.

Hepatocyte growth factor (HGF) promotes the growth not only of hepatocytes but also of several other types of cells such as cytotrophoblasts and endothelial cells. Recent studies have revealed that HGF is trapped in the extracellular (ECM) matrix through heparan sulphate in vivo, thereby acting as a mitogen for hepatocytes in cooperation with heparan sulphate. In this study, we detected HGF protein in chorionic tissue and placental tissue extracts, and found that HGF and heparan sulphate were co-distributed in the endothelial basement membrane and trophoblast basement membrane on immunohistochemical examination. The rates of DNA synthesis in primary cultured cytotrophoblasts and human umbilical vein endothelial cells (HUVEC) cultured on HGF-bound Matrigeltrade mark were 6-8 times those of control cytotrophoblasts and HUVEC. When Matrigeltrade mark dishes were pretreated with heparinase and heparitinase prior to binding of HGF, stimulation of DNA synthesis was markedly decreased. A considerable decrease in stimulation of DNA synthesis was observed following washing of HGF-bound Matrigeltrade mark with 1 m acetic acid, 1 m NaCl and 0.1 per cent trypsin, but not following treatment with chondroitinase ABC. These observations suggest that HGF can be trapped in ECM in vivo, thereby acting as a mitogen for cytotrophoblasts and placental vein endothelial cells in cooperation with heparan sulphate.

Synthesis of procollagen by dental pulp cells from incisor in vitro.

Fibroblast-like cells were isolated from dental pulp of rabbit incisor, and were shown to synthesize and secrete procollagen. Analysis of the secreted procollagen and their CNBr peptides indicated that the prominent form was type I procollagen. Our results also suggested the presence of type III procollagen as a minor component synthesized by pulp cells in vitro.

Actin-binding specificity of marine macrolide toxins, mycalolide B and kabiramide D.

An actin-depolymerizing marine natural product, mycalolide B, and a related compound, kabiramide D, were labeled with biocytin, a biotin derivative, and used to specify target molecules in cultured rat 3Y1 fibroblasts. Mycalolide B exhibited the ability to bind to various intracellular proteins, probably through the Michael addition of a sulfhydryl group to C5 of mycalolide B. However, no intracellular proteins other than actin apparently reacted with biocytinylated kabiramide D, demonstrating that the binding of kabiramide D to actin was highly specific. Cells treated with biocytinylated kabiramide D followed by staining with fluorescein isothiocyanate-conjugated avidin showed that biocytinylated kabiramide D bound to stress fibers composed of F-actin, although the staining intensity was weaker than the fluorescent phalloidin staining. The assay for the binding of kabiramide D to actin, which had previously been treated with other actin-depolymerizing agents, showed that the actin-binding site for kabiramide D was the same as that for bistheonellide A, but not those for latrunculin A and cytochalasin D.

Actin-depolymerizing effect of dimeric macrolides, bistheonellide A and swinholide A.

We compared the effects of dimeric marine toxins, bistheonellide A, and swinholide A, on actin polymerization. Bistheonellide A and swinholide A possess two identical side chains with similar structures to those of other marine toxins, mycalolide B, and aplyronine A. By monitoring changes in fluorescent intensity of pyrenyl-actin, bistheonellide A was found to inhibit polymerization of G-actin and to depolymerize F-actin in a concentration-dependent manner. The relationship between the concentration of bistheonellide A and its inhibitory activity on actin polymerization suggested that one molecule of bistheonellide A binds two molecules of G-actin. We demonstrated by SDS-PAGE that the complex of G-actin with bistheonellide A, swinholide A, or mycalolide B could not interact with myosin. No evidence was found that bistheonellide A severs F-actin at the concentrations examined (molar ratio to actin; 0. 025-2.5), while swinholide A showed severing activity, although it was weaker than that of mycalolide B. We also demonstrated that the depolymerizing effect of bistheonellide A or mycalolide B is irreversible. Bistheonellide A increased, while swinholide A decreased, the rate of nucleotide exchange in G-actin, suggesting that binding of these toxins induces different conformational changes in the actin molecule. These results suggest that bistheonellide A intervenes between two actin molecules, forms a tertiary complex with each of its side chains bound to G-actin, and inhibits polymerization by sequestering G-actin from incorporation into F-actin. A difference in structure at the end of the side chain between dimeric macrolides and mycalolide B may account for the weak severing activity of the former.

Geographic clustering of myeloma in Tokushima.

Between 1978-1988, seven cases of multiple myeloma were found in T Town (population 9,000), which is located at the mouth of a large river within the boundaries of Tokushima City, Japan. This is a significantly high incidence, 7.06 per 100,000 as compared with an incidence of 1.20 in Tokushima City itself and 1.53 in Japan as a whole. The clinical and laboratory features of these patients were typical of myeloma. Although the causes of this small but apparent cluster of multiple myeloma remain obscure, the fact that five of the seven patients had been working for long periods as fishermen was notable.

Measurement of somatostatin release in rat brain by microdialysis.

We determined the most suitable conditions for measuring the somatostatin (SRIF) level by brain microdialysis and investigated its release from the hypothalamus. The relative recovery rate of SRIF was 8.4 +/- 0.5% (mean +/- SE) using a polycarbonate (PC) membrane with the push-pull method at a flow rate of 2 microliters/min. Using tubes with an internal diameter of 0.28 mm and lengths of 5, 25, 50 and 100 cm, the relative recovery rates using a PC membrane with the push method were 8.2 +/- 0.5%, 7.3 +/- 0.6%, 6.2 +/- 0.5% and 4.1 +/- 0.6%, respectively. When using tubes with an internal diameter of 0.1 mm and lengths of 5, 25, 50 and 100 cm, the relative recovery rates were 7.3 +/- 0.7%, 5.6 +/- 1.0%, 3.5 +/- 1.1% and 1.4 +/- 0.7%, respectively. The relative recovery rate was 5.2 +/- 0.5% with a polysulfone (PS-F, Fresenius) membrane, 4.5 +/- 0.4% with a PS-H (Hospal) membrane, 2.6 +/- 0.2% with an ethylenevinyl alcohol membrane (EVAL), 5.1 +/- 0.8% with a polyvinyl alcohol (PVA) membrane and 10.4 +/- 0.8% with a PS-K (Kaneka) membrane. With the push method, the extracellular SRIF level in rat pituitary was 42.8 +/- 1.8 pg/ml with a PC membrane, 23.1 +/- 2.9 pg/ml with an EVAL membrane at a flow rate of 2 microliters/min. With the push-pull method, it was 52.7 +/- 5.2 pg/ml using a PC membrane, 33.5 +/- 2.8 pg/ml using a PVA membrane and 54.4 +/- 3.2 pg/ml using a PS-K membrane.(ABSTRACT TRUNCATED AT 250 WORDS)

Dental bar tracheopexy in a neonate.

Double aortic arch often presents with tracheoesophageal compression. Management requires division of the vascular ring and, if tracheomalacia is present, some form of tracheopexy. We report here the use of a dental bar as an external splint for tracheopexy in a neonate. Follow-up to 18 months shows good tracheal growth and no erosion.