HTLV-1 cell-free DNA in plasma as a potential biomarker in HTLV-1 carriers and adult T-cell leukemia-lymphoma.

Viral cell-free DNA (cfDNA) in plasma has been widely evaluated for detecting cancer and monitoring disease in virus-associated tumors. We investigated whether the amount of cfDNA of human T-cell leukemia virus type 1 (HTLV-1) correlates with disease state in adult T-cell leukemia-lymphoma (ATL). HTLV-1 cfDNA in aggressive ATL was significantly higher than that in indolent ATL and asymptomatic carriers. Notably, patients with lymphoma type represented higher HTLV-1 cfDNA amount than chronic and smoldering subtypes, though they had no abnormal lymphocytes in the peripheral blood. HTLV-1 cfDNA can be a universal biomarker that reflects the expansion of ATL clones.

Targeting of BMI-1 expression by the novel small molecule PTC596 in mantle cell lymphoma.

Despite the development of the novel Bruton tyrosine kinase inhibitor ibrutinib, mantle cell lymphoma (MCL) remains an incurable B-cell non-Hodgkin lymphoma. BMI-1 is required for the self-renewal and maintenance of MCL-initiating stem cells. Upregulation of BMI-1 has been reported in MCL patients, especially in those with refractory/relapsed disease. We studied the effects of a novel small-molecule selective inhibitor of BMI1 expression, PTC596, in MCL cells. Eight MCL cell lines and patient-derived samples were exposed to PTC596. PTC596 induced mitochondrial apoptosis, as evidenced by loss of mitochondrial membrane potential, caspase-3 cleavage, BAX activation, and phosphatidylserine externalization. There was a positive correlation between baseline BMI-1 protein levels and PTC596-induced apoptosis. p53 status did not affect sensitivity to PTC596. PTC596 effectively decreased BMI-1-expressing and tumor-initiating side population MCL cells (IC50: 138 nM) compared with ibrutinib, which modestly decreased side population cells. Interestingly, PTC596, reported to target cancer stem cells, decreased MCL-1 expression levels and antagonized ibrutinib-induced increase in MCL-1 expression, leading to synergistic apoptosis induction in MCL cells. There are currently no drugs that specifically target cancer stem cell fractions, and a reduction in BMI-1 protein by PTC596 may offer a novel therapeutic strategy for MCL.

Ultrastructural and Cytological Studies on Mycosphaerella pinodes Infection of the Model Legume Medicago truncatula.

Ascochyta (Mycosphaerella) blight on cultivated peas is primarily caused by infection through asexual spores (pycnospores) of Mycosphaerella pinodes (Berk. et Blox.) Vestergren [recently renamed Peyronellaea pinodes (Berk. & A. Bloxam) Aveskamp, Gruyter & Verkley]. Using a model pathosystem involving Medicago truncatula and Mycosphaerella pinodes strain OMP-1, we examined the histology and ultrastructure of early infection events and fungal development including penetration by appressoria, vegetative growth of infection hyphae, and host responses. On the susceptible ecotype R108-1, pycnospores germinated and grew over the surface of the epidermis, then formed an appressoria and penetrated the cuticle. Beneath the cuticle, the infection peg expanded into a hyphae that grew within the outer wall of the epidermis. Subsequently, the hyphae penetrated down within mesophyll cells and proliferated vigorously, eventually, forming asexual fruiting bodies (pycnidia). In contrast, successful penetration and subsequent growth of infection hyphae were considerably restricted in the ecotype Caliph. Detected by its reaction with cerium chloride (CeCl3) to generate electron-dense cerium perhydroxides in transmission electron micrographs, hydrogen peroxide (H2O2) accumulated in epidermal and mesophyll cells of Caliph challenged with pycnospores of M. pinodes. This intracellular localization was confirmed by energy-dispersive X-ray spectroscopy. Our observations thus indicate that the oxidative burst reaction leading to the generation of reactive oxygen species is associated with a local host defense response in Caliph, since no clear H2O2 accumulation was detectable in susceptible R108-1. Indeed, aberrant hyphae such as intrahyphal hyphae and dead hyphae, probably due to a local defense elicited by the fungus, were abundant in Caliph but not in R108-1. Our results on the cellular interactions between the fungus and host cells provide additional insights to understand foliar infection by M. pinodes on cultivated peas.

The pathophysiological significance of PPM1D and therapeutic targeting of PPM1D-mediated signaling by GSK2830371 in mantle cell lymphoma.

PPM1D is a serine/threonine phosphatase that negatively regulates key DNA damage response proteins, such as p53, p38 MAPK, histone H2A.X, and ATM. We investigated the pathophysiological significance of PPM1D and its therapeutic targeting by the novel PPM1D inhibitor GSK2830371 in mantle cell lymphoma (MCL). Oncomine-based analyses indicated increased PPM1D mRNA levels in MCL cells compared with their normal counterpart cells. Higher PPM1D expression was associated with higher expression of the proliferation gene signature and poorer prognosis in patients. Eight MCL (three p53 wild-type and five mutant) cell lines were exposed to GSK2830371. GSK2830371 inhibited the cell growth, being prominent in p53 wild-type cells. GSK2830371 induced apoptosis in sensitive cells, as evidenced by induction of phosphatidylserine externalization and loss of mitochondrial membrane potential. p53 knockdown de-sensitized cell sensitivity. GSK2830371 increased the levels of total and Ser15-phosphorylated p53, and p53 targets p21 and PUMA. GSK2830371 and the MDM2 inhibitor Nutlin-3a acted synergistically in p53 wild-type cells. Interestingly, GSK2830371 sensitized MCL cells to bortezomib and doxorubicin in p53 wild-type and mutant cells; p38 signaling appeared to be involved in the GSK2830371/bortezomib lethality. PPM1D inhibition may represent a novel therapeutic strategy for MCL, which can be exploited in combination therapeutic strategies for MCL.

Preclinical activity of the novel B-cell-specific Moloney murine leukemia virus integration site 1 inhibitor PTC-209 in acute myeloid leukemia: Implications for leukemia therapy.

Curing patients with acute myeloid leukemia (AML) remains a therapeutic challenge. The polycomb complex protein B-cell-specific Moloney murine leukemia virus integration site 1 (BMI-1) is required for the self-renewal and maintenance of leukemia stem cells. We investigated the prognostic significance of BMI-1 in AML and the effects of a novel small molecule selective inhibitor of BMI-1, PTC-209. BMI-1 protein expression was determined in 511 newly diagnosed AML patients together with 207 other proteins using reverse-phase protein array technology. Patients with unfavorable cytogenetics according to Southwest Oncology Group criteria had higher levels of BMI-1 compared to those with favorable (P = 0.0006) or intermediate cytogenetics (P = 0.0061), and patients with higher levels of BMI-1 had worse overall survival (55.3 weeks vs. 42.8 weeks, P = 0.046). Treatment with PTC-209 reduced protein level of BMI-1 and its downstream target mono-ubiquitinated histone H2A and triggered several molecular events consistent with the induction of apoptosis, this is, loss of mitochondrial membrane potential, caspase-3 cleavage, BAX activation, and phosphatidylserine externalization. PTC-209 induced apoptosis in patient-derived CD34(+)CD38(low/-) AML cells and, less prominently, in CD34(-) differentiated AML cells. BMI-1 reduction by PTC-209 directly correlated with apoptosis induction in CD34(+) primary AML cells (r = 0.71, P = 0.022). However, basal BMI-1 expression was not a determinant of AML sensitivity. BMI-1 inhibition, which targets a primitive AML cell population, might offer a novel therapeutic strategy for AML.

Induction of CXCL2 and CCL2 by pressure force requires IL-1ß-MyD88 axis in osteoblasts.

Mechanical stresses including pressure force induce chemokine expressions in osteoblasts resulting in inflammatory reactions and bone remodeling. However, it has not been well elucidated how mechanical stresses induce inflammatory chemokine expressions in osteoblasts. IL-1ß has been identified as an important pathogenic factor in bone loss diseases, such as inflammatory arthritis and periodontitis. Myeloid differentiation factor 88 (MyD88) is an essential downstream adaptor molecule of IL-1 receptor signaling. This study was to examine the gene expression profiles of inflammatory chemokines and the role of MyD88 in osteoblasts stimulated by pressure force. Pressure force (10g/cm(2)) induced significant mRNA increases of CXCL2, CCL2, and CCL5, as well as prompt phosphorylation of MAP kinases (ERK, p38 and JNK), in wild-type primary osteoblasts. The CXCL2 and CCL2 mRNA increases and MAP kinase phosphorylation were severely impaired in MyD88(-/-) osteoblasts. Constitutive low-level expression of IL-1ß mRNA was similarly observed in both wild-type and MyD88(-/-) osteoblasts, which was not altered by pressure force stimulation. Notably, neutralization of IL-1ß with a specific antibody significantly impaired pressure force-induced mRNA increases of CXCL2 and CCL2, as well as MAP kinase phosphorylation, in wild-type osteoblasts. Furthermore, pre-treatment with recombinant IL-1ß significantly enhanced MAP kinase phosphorylation and mRNA increases of CXCL2 and CCL2 by pressure force in wild-type but not MyD88(-/-) osteoblasts. These results have suggested that the activation of MyD88 pathway by constitutive low-level IL-1ß expression is essential for pressure force-induced CXCL2 and CCL2 expression in osteoblasts. Thus MyD88 signal in osteoblasts may be required for bone resorption by pressure force through chemokine induction.

Magnetic field effects on copper metal deposition from copper sulfate aqueous solution.

Effects of a magnetic field (≤0.5 T) on electroless copper metal deposition from the reaction of a copper sulfate aqueous solution and a zinc thin plate were examined in this study. In a zero field, a smooth copper thin film grew steadily on the plate. In a 0.38 T field, a smooth copper thin film deposited on a zinc plate within about 1 min. Then, it peeled off repeatedly from the plate. The yield of consumed copper ions increased about 2.1 times compared with that in a zero field. Mechanism of this magnetic field effect was discussed in terms of Lorentz force- and magnetic force-induced convection and local volta cell formation.

Changes in grafted autogenous bone during edgewise treatment in patients with unilateral cleft lip/palate or alveolus.

Objective : To examine the changes in autogenous bone from 6 to 12 months after alveolar bone grafting (ABG) (T1) through completion of edgewise treatment (T2). Design : Retrospective longitudinal study. Setting : Multidisciplinary long-term follow-up at Kagoshima University Hospital. Patients : Forty-three patients with unilateral cleft lip and palate or alveolus. Main Outcome Measures : At T1 and T2, the bone bridge and quantity of grafted bone were evaluated using the Chelsea scale and the ABG scale. The cleft-adjacent tooth angles before ABG and at T2, as well as the number of orthodontic space closures, were examined. Patients were classified as having either adequate (type A or C; adequate group) or poor bone bridges (type B, D, E, or F; poor group) by the assessment at T1. Results : At T1, the ABG scores for the cleft-adjacent central incisor side of patients in the adequate group were higher than those of patients in the poor group (P < .001). At T2, the adequate group had higher ABG scores for the cleft-adjacent central incisor side (P = .022) and the canine sides (P = .034). No significant differences in tooth angles or the number of orthodontic space closures were noted between the groups. Conclusions : These results suggest that the quantity of grafted bone in the cleft-adjacent central incisor at 6 to 12 months post-ABG may be an indicator of the quantity of grafted bone that will be present after edgewise treatment.

Outcome following secondary autogenous bone grafting before and after canine eruption in patients with unilateral cleft lip and palate.

OBJECTIVE: To determine whether the long axis and eruption of the cleft-adjacent canine affect postoperative outcomes in secondary autogenous bone grafting (SABG). DESIGN: Retrospective longitudinal study. SETTING: Multidisciplinary long-term follow-up at Kagoshima University Hospital. SUBJECTS AND METHODS: Twenty-five patients with complete unilateral cleft lip and palate (11 male, 14 female) were compared between unerupted and erupted groups for canine developmental stage, canine angle, and vertical height at bone grafting at 1 year and more than 4 years after SABG. The interalveolar septal heights at 1 and more than 4 years were evaluated by orthopantomograms. RESULTS: All patients in both groups accomplished dental rehabilitation with orthodontic treatment alone without prosthetic appliances. Although the rate of an acceptable bone bridge tended to be lower in the unerupted group (62.5%) than in the erupted group (88.8%), the difference was not significant (P = .158). The canine angle at bone grafting was significantly different between acceptable (69.2° ± 12.2°) and poor cases (77.3° ± 6.2°) at more than 4 years in the unerupted group (P = .049). The acceptable bone bridge rate might reflect mechanical stress added by natural eruption and orthodontic force. CONCLUSIONS: We suggest that SABG should be planned in accordance with the canine angle, crown and root development, the eruption position of the cleft-adjacent canine, and the timing of added mechanical stress in the alveolar cleft, considering the bone formation in the alveolar cleft.

Association of problem behavior with sleep problems and gastroesophageal reflux symptoms.

BACKGROUND: There are few large-scale epidemiologic studies examining the associations between sleep problems, gastroesophageal reflux disease (GERD) symptoms, lifestyle and food habits and problem behaviors (PB) in adolescents. The aim of this study was to evaluate the associations among these factors in Japanese adolescents. METHODS: A cross-sectional survey of 1840 junior high school students was carried out using questionnaires. The subjects were classified into PB or normal behavior (NB) groups using the Pediatric Symptom Checklist (PSC). The scores of the sleep-related factors, sleep bruxism, lifestyle and food habits, and GERD symptoms were compared. Logistic regression analysis was used to determine the factors related to PB. RESULTS: Mean subject age was 13.3 ± 1.8 years. The PB group had significantly longer sleep latency and higher GERD symptom score (P < 0.001). Furthermore, the PB group was significantly more likely to experience absence of the mother at dinner time, skip breakfast, and have <30 min of conversation among family at dinner time. The PB group had significantly higher frequencies of sleep bruxism, difficulty falling asleep within 30 min, nightmares, feeling of low sleep quality, daytime somnolence, and daytime lack of motivation. Feelings of low sleep quality had the strongest association with PB, with an adjusted odds ratio of 12.88 (95% confidence interval: 8.99-18.46). CONCLUSIONS: PB in adolescents are associated with sleep problems, including sleep bruxism, as well as lifestyle and food habits and GERD symptoms.

Patient with oligodontia treated with a miniscrew for unilateral mesial movement of the maxillary molars and alignment of an impacted third molar.

This report describes the treatment of a 20-year-old woman with a dental midline deviation and 7 congenitally missing premolars. She had retained a maxillary right deciduous canine and 4 deciduous second molars, and she had an impacted maxillary right third molar. The maxillary right deciduous second molar was extracted, and the space was nearly closed by mesial movement of the maxillary right molars using an edgewise appliance and a miniscrew for absolute anchorage. The miniscrew was removed, and the extraction space of the maxillary right deciduous canine was closed, correcting the dental midline deviation. After the mesial movement of the maxillary right molars, the impacted right third molar was aligned. To prevent root resorption, the retained left deciduous second molars were not aligned by the edgewise appliance. The occlusal contact area and the maximum occlusal force increased over the 2 years of retention. The miniscrew was useful for absolute anchorage for unilateral mesial movement of the maxillary molars and for the creation of eruption space and alignment of the impacted third molar in a patient with oligodontia.

Regulation of CCN1 via the 3'-untranslated region.

The 3'-untranslated region (UTR) is known to be a critical regulator of post-transcriptional events that determine the gene expression at the RNA level. The gene CCN1 is one of the classical members of the matricellular CCN family and is involved in a number of biological processes during mammalian development. In the present study, the 600-bp 3'-UTR of CCN1 was functionally characterized. Reporter gene analysis revealed that the entire 3'-UTR profoundly repressed gene expression in cis in different types of the cells, to which both the proximal and distal-halves of the 3'-UTR segments contributed almost equally. Deletion analysis of the 3'-UTR indicated a distinct functional element in the proximal half, whereas a putative target for microRNA-181s was predicted in silico in the distal half. Of note, the repressive RNA element in the proximal half was shown to be capable of forming a stable secondary structure. However, unexpectedly, a reporter construct with a tandem repeat of the predicted miR-181 targets failed to respond to miR-181a. In addition, the other major structured element predicted in the distal half was similarly characterized. To our surprise, the second element rather enhanced the reporter gene expression in cis. These results indicate the involvement of multiple regulatory elements in the CCN1 3'-UTR and suggest the complexity of the miRNA action as well as the 3'-UTR-mediated gene regulation.

Relationships between the root-crown ratio and the loss of occlusal contact and high mandibular plane angle in patients with open bite.

OBJECTIVE: To determine the root-crown (R/C) ratio and dental root length of teeth in patients with open bite and seek any relationships with occlusal contact (OC) and the mandibular plane (Mp) angle. MATERIALS AND METHODS: Thirty-one patients with open bite with negative overbite of at least four anterior teeth and 31 control patients with clinically normal overjet and overbite were enrolled. R/C ratios, dental root length, OC, and Mp angle were measured using panoramic radiographs, dental casts, and cephalograms, respectively. Mean differences between the groups, and variations between the R/C ratio or root length and Mp angle in patients with open bite were statistically analyzed. RESULTS: R/C and OC ratios from the incisors to premolars were significantly lower for patients with open bite than for controls, and some teeth had short dental roots. Relationships between low R/C ratio or root length and high Mp angle were significant in patients with open bite. CONCLUSION: Patients with open bite, especially those with a high Mp angle, have an unfavorable R/C ratio and short dental roots in some teeth, which may be related to the loss of OC.

Effects of the naturally-occurring disaccharides, palatinose and sucrose, on incretin secretion in healthy non-obese subjects.

AIMS/INTRODUCTION: Incretins might play some pathophysiological role in glucose metabolism in diabetes and obesity; it is not clear whether or not the amount and the pattern of incretin secretion vary with different types of sugars. To evaluate the effect of two types of disaccharides on glucose metabolism and the kinetics of incretin secretion, plasma levels were measured after palatinose or sucrose ingestion in non-obese healthy participants. MATERIALS AND METHODS: The study was carried out on healthy participants who were given a solution containing 50 g of palatinose or sucrose for ingestion. Blood samples were obtained before loading and after ingestion. Insulin, glucagon and incretins hormones were measured by the enzyme-linked immunosorbent assay method. RESULTS: When the data were compared between palatinose and sucrose ingestion, both plasma glucose values at 15, 30 and 60 min, and plasma insulin values at 15 and 30 min after palatinose loading were significantly lower than those after sucrose loading. Plasma levels of total glucose-dependent insulinotropic polypeptide at 15-90 min after palatinose loading were significantly lower than those after sucrose loading. Plasma levels of total and active glucagon-like peptide-1 at 90 min and the area under the curve (60-120 min) of the total glucagon-like peptide-1 were significantly higher with palatinose-loading than with sucrose loading. CONCLUSION: Compared with sucrose, palatinose appears to have a more favorable effect on glucose metabolism and protection of pancreatic islets as a result of less hyperglycemic and hyperinsulinemic potency.

Molecular mechanisms of the inhibitory effect of lipopolysaccharide (LPS) on osteoblast differentiation.

Osteoblasts express Toll like receptor (TLR) 4 and produce osteoclast-activating cytokines in response to the stimulation by lipopolysaccharide (LPS). It has recently been reported that LPS exerts an inhibitory effect on osteoblast differentiation into osteocytes. However, the molecular mechanisms of this inhibitory effect remain ambiguous. The downstream signals of TLR4 are mediated by adaptor molecules including myeloid differentiation factor 88 (MyD88), leading to the activation of mitogen-activated protein kinases (MAPKs), such as extracellular signal-regulated kinases (ERKs), whose activation by LPS requires the upstream serine/threonine kinase, Cot/Tpl2. To determine the signal molecules responsible for the inhibitory effects of LPS on osteoblast differentiation, we examined the in vitro differentiation of the primary osteoblasts from myd88(-/-) and cot/tpl2(-/-) mice. The matrix mineralization by the wild-type and cot/tpl2(-/-) osteoblasts was significantly inhibited by LPS, whereas that of myd88(-/-) was not affected. During differentiation, LPS suppressed the mRNA expression of runt related transcription factor 2 (Runx2), osterix (Sp7), and activating transcription factor 4 (ATF4) in the wild-type, but not in the myd88(-/-) osteoblasts. The inhibitory effect of LPS on the mRNA expression of these transcription factors was absent in the early phase but partially impaired in the late phase of differentiation in the cot/tpl2(-/-) osteoblasts. Thus, the inhibitory effect of LPS on osteoblast differentiation is Myd88-dependent, whereas the degree of its requirement for Cot/Tpl2 varies depending on the differentiation phase.

Dynein axonemal intermediate chain 2 is required for formation of the left-right body axis and kidney in medaka.

Ciliary defects lead to various diseases, such as primary ciliary dyskinesia (PCD) and polycystic kidney disease (PKD). We isolated a medaka mutant mii, which exhibits defects in the left-right (LR) polarity of organs, and found that mii encodes dynein axonemal intermediate chain 2a (dnai2a). Ortholog mutations were recently reported to cause PCD in humans. mii mutant embryos exhibited loss of nodal flow in Kupffer's Vesicle (KV), which is equivalent to the mammalian node, and abnormal expression of the left-specific gene. KV cilia in the mii mutant were defective in their outer dynein arms (ODAs), indicating that Dnai2a is required for ODA formation in KV cilia. While the mii mutant retained motility of the renal cilia and failed to show PKD, the loss of dnai2a and another dnai2 ortholog dnai2b led to PKD. These findings demonstrate that Dnai2 proteins control LR polarity and kidney formation through regulation of ciliary motility.

Reduction of orthodontic tooth movement by experimentally induced periodontal inflammation in mice.

Orthodontic therapy is known to have an aggravating effect on the progression of destructive periodontitis if oral hygiene is not maintained. However, it is largely unknown how active periodontitis affects the velocity of orthodontic tooth movement. In this study, we examined the effect of periodontal inflammation on orthodontic tooth movement using a mouse model. Orthodontic force was applied on the maxillary first molar of mice, with or without ligature wire to induce experimental periodontitis. The distance moved by the first molar was significantly reduced by the ligature-induced experimental periodontitis. Tartrate-resistant acid phosphatase staining revealed that the number of osteoclasts present during orthodontic treatment was lower in the pressure zone of alveolar bone in the presence of periodontal inflammation. Consistently, the expression level of receptor activator of nuclear factor-kappaB ligand (RANKL) in the pressure zone was decreased in the ligature group. By contrast, experimental periodontitis increased the expression of cyclooxygenase-2 mRNA in the periodontal tissues, while in vitro treatment with prostaglandin E(2) decreased extracellular signal-regulated kinase phosphorylation and RANKL expression induced by mechanical stress in osteoblasts. Taken together, these results suggest that the orthodontic force-induced osteoclastogenesis in alveolar bone was inhibited by the accompanying periodontal inflammation, at least partly through prostaglandin E(2), resulting in reduced orthodontic tooth movement.

Orthodontic treatment combined with mandibular distraction osteogenesis and changes in stomatognathic function.

We performed an orthodontic treatment combined with mandibular distraction osteogenesis in a 15-year-old patient who wanted a correction of a chin deficiency and a protruding upper lip. The patient had an Angle Class II division 1 malocclusion with mandibular retrusion, a low mandibular plane angle, and scissors bite. First, a quad-helix appliance was applied to the mandibular dentition to correct the scissors bite in the bilateral premolar region. Later, a preadjusted edgewise appliance was applied to the maxillary and mandibular teeth. After 3 days, a mandibular distraction osteogenesis was performed. During and after the distraction, the open bite between the upper and lower dental arches was corrected using up and down elastics. The total treatment time with the edgewise appliance was 14 months. A skeletal Class I apical base relationship, good facial profile, and optimum intercuspation of the teeth were achieved with the treatment. The jaw-movement pattern on the frontal view did not change during gum chewing. However, the maximum gap without pain increased. The electromyographic (EMG) activity of the masseter and anterior temporalis muscles, and maximum occlusal force increased. The present case report suggests that an orthodontic treatment combined with mandibular distraction osteogenesis in a patient with mandibular retrusion in the late growth period might be effective for improving stomatognathic function.

Adenosine induces expression of glial cell line-derived neurotrophic factor (GDNF) in primary rat astrocytes.

Adenosine, which accumulates rapidly during ischemia due to the breakdown of ATP, has beneficial effects in many tissues. We examined whether adenosine induces the production of glial cell line-derived neurotrophic factor (GDNF) in cultured astrocytes. We evaluated GDNF mRNA expression and GDNF production in astrocytes cultured with adenosine and the adenosine selective receptor agonists 5-(N-ethylcarboxamido) adenosine (NECA), N(6)-cyclopentyladenosine (CPA) and 2-p-(2-carboxyethyl) phenethylamino-5'-N-ethylcarboxamindo-adenosine hydrochloride (CGS 21680). Moreover, we examined the possibility that the expression of GDNF is regulated differently in cultured astrocytes from the stroke-prone spontaneously hypertensive rat (SHRSP) than in those from Wistar Kyoto rats (WKY). In this study, we confirmed that adenosine and the selective A(2B) adenosine receptor agonist NECA induced the expression of GDNF in cultured astrocytes. The A(2B) receptor antagonist alloxazine was able to inhibit the increase in extracellular GDNF produced by adenosine. Furthermore, the amounts of GDNF produced were significantly reduced in astrocytes of the adenosine-treated SHRSP compared with those of WKY. These results indicate that adenosine induces the expression of GDNF, and adenosine A(2B) receptors participate in the regulation of GDNF levels in astrocytes. This expression was attenuated in astrocytes of SHRSP compared with those of WKY.

Design of highly efficient receptor sites by combination of cyclodextrin units and molecular cavity in TiO2 ultrathin layer.

A highly effective approach was developed for sensitive detection of organic substances in water. In order to achieve high sensitivity and selective detection for aromatic compounds, cyclodextrin (CD) hosts and imprinting effects were combined to fabricate binding sites within TiO(2) ultrathin layer. The electrode surface was modified with ultrathin TiO(2) gel film containing a 2:1 complex of beta-CD and bisphenol A (BPA), and then the BPA moiety was removed by washing. The resulting BPA-imprinted TiO(2)/beta-CD film showed specific and sensitive detection of BPA, as confirmed by quartz crystal microbalance (QCM) and cyclic surface-polarization impedance (cSPI) measurements. The high selectivity for BPA relative to structurally related guest molecules was estimated to be 1.6-3.4 at the guest concentration of 2x10(-7)M, and was considered to arise from the synergic effect of the binding site.