Association Between Tacrolimus Pharmacokinetics and Cytochrome P450 3A5 and Multidrug Resistance Protein 1 Exon 21 Polymorphisms.

BACKGROUND: Individual differences in the pharmacokinetics (PK) of tacrolimus (TAC), an immunosuppressive drug, are reportedly associated with single-nucleotide polymorphisms (SNPs) of cytochrome P450 (CYP) 3A5 and multidrug resistance protein 1 (MDR1). We determined the effect of SNPs in CYP3A5 and MDR1 exons 21 and 26 on TAC PK parameters. METHODS: Thirty-eight Japanese patients who underwent renal transplantation were genotyped for CYP3A5 and exons 21 and 26 of MDR1 with the use of polymerase chain reaction-restriction fragment length polymorphism analysis. TAC concentrations were determined 3 weeks after renal transplantation and PK parameters calculated. RESULTS: The area under the blood concentration-time curve (AUC) in CYP3A5 expressers was significantly higher than that in CYP3A5 nonexpressers (CYP3A5*3/*3). Patients with the MDR1 exon 21 A allele (G2677A) showed higher dose-adjusted AUC (AUC/D) and lower doses of TAC than those who did not possess that allele. Furthermore, patients with both CYP3A5*3/*3 and MDR1 G2677A showed significantly lower TAC doses and higher dose-adjusted trough levels (C/D) and AUC/D than those without those genotypes. There was no significant association between MDR1 exon 26 polymorphism and the PK of TAC. CONCLUSIONS: Patients with both CYP3A5*3/*3 and MDR1 G2677A had higher blood TAC concentrations than those without those genotypes. Japanese patients should be carefully monitored for consideration of lower TAC doses, because 24% of Japanese patients have double mutations.

A chromosome 16p13.11 microduplication causes hyperactivity through dysregulation of miR-484/protocadherin-19 signaling.

Chromosome 16p13.11 microduplication is a risk factor associated with various neurodevelopmental disorders such as attention-deficit/hyperactivity disorder, intellectual disabilities, developmental delay and autistic spectrum disorder. The underlying molecular mechanism of this genetic variation remained unknown, but its core genetic locus-conserved across mice and humans-contains seven genes. Here, we generated bacterial artificial chromosome-transgenic mice carrying a human 16p13.11 locus, and these mice showed the behavioral hyperactivity phenotype. We identified miR-484 as the responsible gene using a combination of expression and functional analyses. Mature miR-484 was expressed during active cortical neurogenesis, and overexpression of miR-484 decreased proliferation and increased neural progenitor differentiation in vivo. Luciferase screening identified the 3'-untranslated region of protocadherin-19 (Pcdh19) as a target of miR-484. The effect of miR-484 on neurogenesis was rescued by ectopic PCDH19 expression. These results demonstrate that miR-484 promotes neurogenesis by inhibiting PCDH19. Dysregulation of neurogenesis by imbalanced miR-484/PCDH19 expression contributes to the pathogenesis of 16p13.11 microduplication syndrome.

A proapoptotic effect of valproic acid on progenitors of embryonic stem cell-derived glutamatergic neurons.

Valproic acid (VPA) is a branched-chain saturated fatty acid with a long history of clinical use as an antiepileptic drug (AED). VPA is also known to inhibit histone deacetylases (HDACs) and to cause diverse effects on neural progenitor cells (NPCs) and neurons. Although the neuroprotective or neurodestructive effects of VPA have been investigated in heterogeneous cell populations, in this study, we used homogeneous populations of NPCs and glutamatergic cortical pyramidal neurons, which were differentiated from embryonic stem (ES) cells. At therapeutic concentrations, VPA had a proapoptotic effect on ES cell-derived NPCs of glutamatergic neurons, but not on their progeny. This effect of VPA most likely occurred through the inhibition of HDACs, because similar phenotypes were observed following treatment with other HDAC inhibitors (HDACis) such as trichostatin A and sodium butyrate. The proapoptotic phenotype was not observed when cells were exposed to a structural analog of VPA, valpromide (VPM), which has the same antiepileptic effect as VPA, but does not inhibit HDACs. Western blotting confirmed that treatment with HDACis, but not VPM, significantly increased the levels of histone H3 acetylation in NPCs. HDACi treatments did not affect the survival of neurons, although the acetylation levels were increased to a limited extent. These results, which are based on a homogeneous culture system, suggest that VPA inhibits HDAC activity and induces the apoptosis of NPCs that are fated to differentiate into glutamatergic neurons. The dose-dependent effects of VPA both on apoptosis and hyperacetylation of histone H3 in NPCs supported this notion. These cell type- and differentiation stage-specific effects of VPA imply that dysfunction of HDACs during pregnancy significantly increase the risk of congenital malformations associated with VPA administration.

Clinical guidelines for treating caries in adults following a minimal intervention policy--evidence and consensus based report.

OBJECTIVES: In 2002, FDI (World Dental Federation) published a policy advocating that caries be treated by minimal intervention (MI). This MI policy has been accepted worldwide and is taught in universities. But acceptance in general dental practice has been slower, especially in Japan where healthcare payment and practice favour drilling and filling. To help disseminate this MI policy into general practice, the Japanese Society of Conservative Dentistry developed an evidence-based clinical guideline for restoring carious permanent teeth in adult patients. METHODS: The guideline was developed by a committee of nine university clinicians and a librarian. The committee selected the most frequent clinical questions in treating caries and used electronic databases to search and assess the best scientific evidence for each. Members then added their clinical experience and discussed to reach consensus on each question on treating caries with MI policy. Graded recommendations and guidance were made for each clinical question. The provisional guideline was strengthened after review and discussion with university researchers and general practitioners. RESULTS: The guideline addresses the 16 most frequent clinical questions in treating adult caries, including restorative methods and how to tackle root caries. Recommendations for treatment using MI policy were developed using the best scientific evidence and consensus of experienced clinicians. CLINICAL SIGNIFICANCE: The guideline offers a practical expert view of treating caries with the MI policy that incorporates the best scientific evidence, the latest techniques, the most preferable materials and the general consensus of expert clinicians.

Rap1 is involved in the signal transduction of myelin-associated glycoprotein.

Myelin-associated glycoprotein (MAG) is a well-characterized axon growth inhibitor in the adult vertebrate nervous system. Several signals that play roles in inhibiting axon growth have been identified. Here, we report that soluble MAG induces activation of Rap1 in postnatal cerebellar granule neurons (CGNs) and dorsal root ganglion (DRG) neurons. The p75 receptor associates with activated Rap1 and is internalized in response to MAG. After MAG is applied to the distal axons of the sciatic nerves, the activated Rap1, internalized p75 receptor, and MAG are retrogradely trafficked via axons to the cell bodies of the DRG neurons. Rap1 activity is required for survival of the DRG neurons as well as CGNs when treated with MAG. The transport of the signaling complex containing the p75 receptor and Rap1 may play a role in the effect of MAG.

Temporary restorative resins using non-phthalate ester plasticizers.

OBJECTIVE: This study examined temporary restorative resins that use non-phthalate ester plasticizers. METHODS: Three non-phthalate plasticizers were tested: tri-ethyl citrate (TEC), tri-butyl citrate (TBC), and di-butyl sebacate (DBS). The experimental resins were compared with resins that use a phthalate ester plasticizer (di-butyl phthalate, DBP) and commercial products (Dura Seal, Plast Seal, and Fit Seal). For the experimental resins, the ratio of plasticizer to PMMA/MMA resin was varied from 30 to 60% (wt). The mechanical properties of the experimental resins were evaluated using a modified diametral compressive strength and the resistance to explorer insertion. RESULTS: The strengths of the experimental resins with plasticizer concentrations ranging from 40 to 50% (TEC, TBC, and DBP) or from 30 to 40% (DBS), by weight, were equivalent to the strengths of commercial temporary restorative resins. A similar tendency was seen for the resistance to explorer insertion. CONCLUSIONS: The results suggest that new temporary restorative resins can be prepared by choosing the appropriate proportions of resin monomer and non-phthalate ester plasticizer.

Cynomolgus monkey liver aldehyde oxidase: extremely high oxidase activity and an attempt at purification.

Aldehyde oxidase (EC 1.2.3.1) in monkey (Macaca fascicularis) liver was characterized. Liver cytosol exhibited extremely high benzaldehyde and phthalazine oxidase activities based on aldehyde oxidase, compared with those of rabbits, rats, mice and guinea pigs. Monkey liver aldehyde oxidase showed broad substrate specificity distinct from that of the enzyme from other mammals. Purified aldehyde oxidase from monkey liver cytosol showed two major bands and two minor bands in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). These bands were also observed in Western blotting analysis using anti-rat aldehyde oxidase. The molecular mass of the enzyme was estimated to be 130-151 kDa by SDS-PAGE, and to be about 285 kDa by HPLC gel filtration. The results suggest that isoforms of aldehyde oxidase exist in monkey livers.

Assessment of decontamination methods as pretreatment of silanization of composite glass fillers.

In terms of mechanical properties and durability, the surface of glass fillers should be decontaminated in order to optimize the silanization process for the production of resin composites. The objective of this study was to evaluate the decontamination efficiency of 18 cleaning methods on glass fillers as pretreatment of silane coupling. X-ray photoelectron spectroscopy revealed that SiO(2) boiled with a 5% sodium peroxodisulfate aqueous solution for 15 min, followed by ultrasonic rinsing with acetone for 30 min was most effective among all the decontamination methods investigated. In addition, nano-indentation measurements on SiO(2) treated by the above-mentioned method revealed that the surface was not significantly weakened as compared to untreated SiO(2). The results of this study should lead to an improved filler-matrix coupling and thus contribute to the development of better wear and fatigue-resistant composites. Therefore, sodium peroxodisulfate is proposed as a presilanization filler decontamination step in the production process of resin composites.

Regional distribution of parkinsonism-preventing endogenous tetrahydroisoquinoline derivatives and an endogenous parkinsonism-preventing substance-synthesizing enzyme in monkey brain.

1,2,3,4-Tetrahydroisoquinoline (TIQ) and 1-benzyl-1,2,3,4-tetrahydroisoquinoline (1BnTIQ), which exist in the brain of several mammalian species, are parkinsonism-inducing substances, and 1-methyl-1,2,3,4-tetrahydroisoquinoline (1MeTIQ), which is enzymatically synthesized in rat brain, is a parkinsonism-preventing substance. In this study, we examined the regional distribution of contents of TIQ, 1MeTIQ, and 1BnTIQ, and activity of 1MeTIQ-synthesizing enzyme in monkey brain. The TIQ and 1BnTIQ contents in cerebrum and substantia nigra, and the 1MeTIQ contents in striatum and substantia nigra were higher than those in other brain regions, and 1MeTIQ-synthetic activity was high in cerebrum and thalamus. We speculate that 1MeTIQ-synthesizing enzyme may play an important role in idiopathic Parkinson's disease.

Occlusal force distribution on the dental arch during various levels of clenching.

The purpose of this study was to explore the occlusal force distribution on the dental arch in the intercuspal position and to evaluate the relationship between the clenching strength and the occlusal force distribution. These variables were recorded using the Dental Prescale System in 16 healthy young adults. The number of tooth contacts, occlusal force and occlusal contact area increased linearly as clenching strength increased. The distribution of the occlusal force was greatest at the molar region followed by the premolar and anterior teeth region. The proportion of occlusal force (occlusal force at each region/total occlusal force) on molar regions increased as clenching strength increased. On the contrary, the proportion of occlusal force on the premolar and anterior teeth regions decreased as clenching strength increased. These findings suggest that control of occlusal force is important in diagnosis of the nature of occlusal contacts.

Short-term reaction of exposed monkey pulp beneath adhesive resins.

Monkey pulpal responses to four adhesive resin systems used as direct pulp capping agents were histopathologically evaluated at 3, 7, 14, 30, and 60 days after operation. No serious inflammatory reaction of the pulp, such as necrosis or abscess formation, was observed. Slight inflammatory cell infiltration was the main inflammatory reaction of the exposed pulp, and the exposed area became occluded with dentin bridging as the observation period increased. Healing of exposed dental pulp beneath adhesive resin capping slightly differed depending on the materials used.

Monkey pulpal responses to conventional and adhesive luting cements.

Monkey pulpal responses to metal inlays luted with a combination of an adhesive resin and luting composite and conventional dental cements were histopathologically evaluated. Initial pulpal responses caused by re-exposure of the cut dentin surfaces and luting procedure under hydraulic pressure subsided at 90 days after final cementation. There was no significant difference among pulpal reactions to conventional dental cements and a combination of an adhesive resin and luting composite. The adhesive resin coating of freshly cut dentinal walls/floors immediately after cavity preparation seems to provide protection for the dentin and pulp in indirect restorations requiring temporary sealing.

Possible involvement of calpain in the growth of estrogen receptor positive breast cancer cells.

Calpain (Ca2(+)-activated neutral protease, EC 3.4.22.17) has been reported to hydrolyze the estrogen receptor (ER). However, there has been no report available regarding the role of calpain in the growth of breast cancer cells. To investigate the role of calpain in the growth of various breast cancer cell lines, we employed a synthetic peptide, calpeptin, which is a cell permeable specific inhibitor of calpain. Calpeptin inhibited the cell growth of ER positive breast cancer cells, such as MCF-7, T-47D, and ZR-75-1 in a dose dependent manner in the presence of E2. However, the growth of ER negative breast cancer cells, MDA-MB-231, was not inhibited by calpeptin. It is suggested that calpain plays an important role in the growth of ER positive breast cancer cells.

Monkey pulpal response to adhesively luted indirect resin composite inlays.

Monkey pulpal responses to resin-bonded indirect resin composite inlays were histopathologically evaluated by placing them in either etched-enamel, total-etched, or adhesive-resin-lined cavities. Initial pulpal responses caused by re-exposure of the cut dentin surfaces and luting procedure subsided if a tight marginal seal was secured by final cementation of the inlay. The adhesive resin coating of freshly cut dentinal walls/floors seems to provide a new technique to protect the dentin and pulp in indirect restorations requiring temporary sealing.

Studies on newly designed tooth surface treatments for adhesive composite restoration.

Recently, problems have occurred frequently such as cervical and exposed dentin surface lesions or root caries and subsequent hypersensitivity because of further increase of the elderly population. This study was projected to device a proper and simple treatment and materials for the adhesive composite restoration for such lesions. The 10-20Ca (10% citric acid containing 20% calcium chloride) solution was found to be a comparatively mild tooth surface conditioning agent and to be effective with a single application to both dentin and enamel. In addition, the use of the SA primer (3% N-methacryloyl 5-aminosalicylic acid in 80% ethanol) and the LVR (visible light-cured, 33% microfilled low viscous Bis-GMA resin) dramatically improved the adhesion and adaptability of the composite restoration in the saucer cavity at the cervical area.

Effect of acid etching on the dental pulp in adhesive composite restorations.

Pulpal response to marginal enamel etching or both enamel and dentine etching with 37% phosphoric acid for adhesive composite restorations was studied in the teeth of the Japanese monkey (Macaca fuscata). Dentine etching irritated the pulp and caused moderate to severe initial changes along the odontoblastic layer at 3 days. When no bacteria were present along the cavity walls, the subsequent pulpal reactions at 30 and 90 days decreased with time and large amounts of irritation dentine were formed. These inflammatory reactions were less than those caused by enamel etching only, or zinc oxide/eugenol cement. Significant correlation (P less than 0.01) was found between the intensity of the inflammatory reactions and the degree of bacterial infection. The pulpal irritation caused by acid etching of dentine for composite restorations is transitory only when the fillings have a good marginal seal and wall adaptation.