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Black photochromism, in which the photogenerated isomer has an absorption band over the whole visible region, is fascinating for applications such as information display, photochromic solar cells, or smart windows. However, black photochromism in a single organic photochromic component is quite rare. In this study, we tried to design and synthesize a photochromic diarylethene derivative that shows black photochromism in a single molecular backbone. By optimizing the molecular structure of a diarylethene derivative having a functionalized indole ring as the aryl group, we successfully prepared a black photochromic diarylethene derivative, which shows a reversible color change between pale yellow and black along with the photochromic reactions in solution as well as in solid state. Furthermore, with imitating the coloration system of some living organisms, optical patterning was successfully demonstrated based on the photoregulation of the contrast of structural color by changing the background color along with the photochromic reactions.

Purpose: Laparoendoscopic single-site donor nephrectomy (LESSDN) is a feasible and effective procedure because of its non-invasiveness and better cosmetic outcomes. However, there have been few multi-institutional studies conducted by multiple surgeons on LESSDN. We retrospectively compared the clinical data and outcomes between LESSDN and conventional laparoscopic donor nephrectomy (LDN) at multiple institutes in Japan. Materials and methods: From 2009 to 2015, the clinical data of 223 donors who underwent LESSDN and 151 donors who underwent LDN were collected from 10 institutes. All LESSDNs were performed transperitoneally, whereas LDNs were performed transperitoneally (P-LDN) in 75 patients and retroperitoneally (R-LDN) in 76 patients. Results: In the LESSDN group, the single-incision site was pararectal in 155 (69.5%) patients and umbilical in 65 (29.1%) patients. Multiple surgeons (one to eight per institute) performed the LESSDN. No significant differences were observed between the three groups regarding estimated blood loss and warm ischemic time. The operative time was significantly shorter in the LESSDN group than in the R-LDN group (p= 0.018). No significant differences were observed regarding the rates of blood transfusion, open conversion, visceral injuries, and postoperative complications. Furthermore, no significant differences were observed regarding the dose of analgesic and the rate of delayed graft function. One patient required open conversion due to injury to the renal artery. Selection of LESS procedure was not an independent risk factor for the median serum creatinine level of above 1.27 mg/dL in recipients at 1 year after kidney transplantation. Conclusion: The results showed the technical feasibility of LESSDN compared with the standard LDNs in a multi-institutional and multi-surgeon setting. A few observed non-negligible complications and the significantly higher levels of serum creatinine in patients who underwent LESSDN indicate that this procedure should be employed cautiously when performed by surgeons without ample experience in performing LESS procedures.

•We evaluated the relations of CYP3A phenoconversion with plasma levels of indoxyl sulfate and cytokines.•Higher plasma indoxyl sulfate levels were associated with CYP3A phenoconversion.•Dose adjustment may be needed in patients with CYP3A5*1and high indoxyl sulfate level.

Background: Phenoconversion is a phenomenon whereby some genotypic extensive metabolizers transiently exhibit drug metabolizing enzyme activity at similar level as that of poor metabolizers. Renal failure is known to decrease CYP3A activity in humans. Indoxyl sulfate, parathyroid hormone (PTH), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-a) have been reported to cause CYP3A downregulation in renal failure. We measured plasma concentrations of the above compounds in stable kidney transplant recipients, and evaluated their relations with phenoconversion of CYP3A evaluated by plasma concentration of 4ß-hydroxycholesterol, a biomarker of CYP3A activity. Phenoconversion was defined as a genotypic extensive/intermediate metabolizer exhibiting CYP3A activity below the cutoff value that discriminates extensive/intermediate from poor metabolizers. Methods: Sixty-three Japanese kidney transplant recipients who underwent transplantation more than 180 days prior to the study were included. Morning blood samples were collected, and CYP3A5 polymorphism as well as plasma concentrations of 4ß-hydroxycholesterol, indoxyl sulfate, intact-PTH, IL-6 and TNF-a were determined. Results: Significantly higher plasma 4ß-hydroxycholesterol concentration was observed in recipients with CYP3A5*1allele (n = 23) compared to those without the allele (n = 40), and the cut-off value was 40.0 ng/mL. Ten recipients with CYP3A5*1allele exhibited CYP3A activity below 40.0 ng/mL (phenoconversion). Only plasma indoxyl sulfate concentration was significantly higher in recipients with CYP3A phenoconversion compared to those without phenoconversion. Conclusions: These findings suggest that higher plasma indoxyl sulfate concentration may be involved in CYP3A phenoconversion. Dose adjustment of drugs metabolized by CYP3A may be needed in patients with CYP3A5*1allele and high blood indoxyl sulfate.

We previously synthesized phosphoramidite monomers bearing Boc-protected 2,6-diaminopurine (D) and 2-methyl-4-methoxybenzyl-protected 2-thiouracil (sU) as building blocks for the preparation of pseudo-complementary serinol nucleic acids (SNAs). Since SNA is stable under acidic conditions, an acid-deprotection step could be inserted into the work-up. However, as the 4,4′-dimethoxytrityl group was concurrently removed at this step, purification of SNA by reversed-phase HPLC was difficult. Here, we report the syntheses of SNA and acyclic l-threoninol nucleic acid (l-aTNA) phosphoramidite monomers with bis(phenoxyacetyl)-protected D and 4-acetoxybenzyl-protected sU, both of which can be deprotected under mild basic conditions. Using these monomers, we prepared pseudo-complementary SNA and l-aTNA in high yield using conventional oligonucleotide synthesis protocols. These monomers can be used for large-scale syntheses of SNAs and l-aTNAs.

Neural damage is a devastating outcome of physical trauma. The glia are one of the main effectors of neuronal repair in the nervous system, but the dynamic interactions between peripheral neurons and Schwann cells during injury and regeneration remain incompletely characterized. Here, we combine laser microsurgery, genetic analysis, high-resolution intravital imaging and lattice light-sheet microscopy to study the interaction between Schwann cells and sensory neurons in a zebrafish model of neurotrauma. We found that chronic denervation by neuronal ablation leads to Schwann-cell death, whereas acute denervation by axonal severing does not affect the overall complexity and architecture of the glia. Neuronal-circuit regeneration begins when Schwann cells extend bridging processes to close the injury gap. Regenerating axons grow faster and directionally after the physiological clearing of distal debris by the Schwann cells. This might facilitate circuit repair by ensuring that axons are guided through unoccupied spaces within bands of Büngner towards their original peripheral target. Accordingly, in the absence of Schwann cells, regenerating axons are misrouted, impairing the re-innervation of sensory organs. Our results indicate that regenerating axons use haptotaxis as a directional cue during the reconstitution of a neural circuit. These findings have implications for therapies aimed at neurorepair, which will benefit from preserving the architecture of the peripheral glia during periods of denervation.

Several studies have shown that renal failure decreases CYP3A activity and that uremic toxins may play a role via transcriptional or translational modifications of cytochrome P450 (P450) enzymes and direct inhibition of P450-mediated metabolism. In this study, we evaluated the relationship between CYP3A activity (using plasma concentration of 4β-hydroxycholesterol as a biomarker) and clinical characteristics including plasma concentrations of indoxyl sulfate (3-INDS) and indole-3-acetic acid (3-IAA) in stable kidney transplant recipients. Forty-five Japanese kidney transplant recipients who underwent transplantation more than 90 days prior to the study were included. Morning blood samples were collected and plasma concentrations of 4β-hydroxycholesterol, 3-INDS, and 3-IAA were measured. Plasma concentrations of 4β-hydroxycholesterol were 57.1 ± 11.2, 42.1 ± 11.8, and 34.5 ± 7.3 ng/ml in recipients with CYP3A5*1/*1(n= 5), *1/*3(n= 15), and *3/*3(n= 25) genotypes, respectively, with significant differences between three genotypes. A significant correlation was observed between plasma concentrations of 4β-hydroxycholesterol and 3-INDS but not 3-IAA. Multiple regression analysis identified the number of CYP3A5*3alleles in genotype, plasma concentration of 3-INDS, and body weight as independent variables associated with plasma concentration of 4β-hydroxycholesterol. In conclusion, these results suggest that CYP3A5 polymorphism and plasma concentration of 3-INDS may account for the interindividual variability of CYP3A activity, and that plasma concentration of 3-INDS may partially explain the gap in CYP3A activity that cannot be explained by genetic contribution in patients with renal failure.

Several previous studies have shown that renal failure decreases not only renal elimination but also metabolic clearance of drugs, particularly those metabolized by CYP3A. However, whether recovery of renal function results in recovery of hepatic CYP3A activity remains unknown. In this study, we evaluated the effect of renal function on CYP3A activity after kidney transplantation in patients with end-stage renal disease (ESRD) by measuring the change in CYP3A activity using plasma concentration of 4β-hydroxycholesterol as a biomarker. The study enrolled 13 patients with ESRD who underwent the first kidney allograft transplantation. Morning blood samples were collected before and 3, 7, 10, 14, 21, 30, 60, 90, 120, 150 and 180 days after kidney transplantation. Plasma concentration of 4β-hydroxycholesterol was measured using GC-MS. Compared with before kidney transplantation, creatinine clearance increased significantly from day 3 after kidney transplantation and stabilized thereafter. Plasma concentration of 4β-hydroxycholesterol was elevated significantly on days 90 and 180 after kidney transplantation. In conclusion, this study suggests the recovery of CYP3A activity with improvement in renal function after kidney transplantation in patients with ESRD.

Implantation serine protease ISP was first identified in the uteri of pregnant mice. It is thought that ISP may have an important role in the initiation of implantation. However, the expression status and detailed functions of ISP remain unclear. In this study, the expression of ISP was investigated in the rat uterus. The analysis of two rat genes registered in GenBank, accession nos. XM220240 and XM577076, exhibited high identities to the mouse ISP2 genes, respectively at an mRNA level. We labeled the former as rISP2aand the latter as rISP2b. Using RTPCR, we found that both genes were expressed in the uterus. Specifically, rISP2a mRNA was detected in the uterus throughout pregnancy, whereas rISP2b mRNA was only expressed in the uterus from day 5 of pregnancy until the end of gestation. Expression of both genes was observed specifically within the endometrial gland epithelium. Furthermore, rISP2awas also observed to be expressed in the fetus and placenta, whereas rISP2bexpression was observed in the fetus but not in the placenta. An expressional signal of the rISP2a gene was observed in the spongiotrophoblasts, giant cells and decidual endometrium in the placenta. In the embryo, the ventral specific region was positive in rISP2a and rISP2b gene expression. These findings indicate the possibility that the presently examined genes with high identity to mouse ISP2may play some role not only during the implantation phase, but also in the development of the placenta and embryo. J. Exp. Zool. Mol. Dev. Evol. 310B:642–649, 2008. © 2008 WileyLiss, Inc.

The Oxide Electrowinning method has been studied as one of the candidate dry reprocessing concepts of the future fuel cycle systems. On the MOX co-deposition process, the main process of that method, some fundamental experiments have been performed to confirm its feasibility. In the experiments, several parameters were set to study the suitable electrolysis condition to obtain desired granule of MOX. The concentrations of uranium, plutonium, fission products(FP) simulators, and corrosion products(CP) simulators were adopted as the parameters. The blowing gas composition (O2, Cl2, Ar) during the electrolysis was also set as the variable condition. Through these experiments, it was clarified that the partial pressure of chlorine gas during electrolysis was important to obtain MOX granule with high Pu concentration (about 30%) without generating bottom precipitation in melt. Finally, adequacy of the process control method for MOX co-electrolysis was confirmed through the test using spent fast reactor(FR) fuel.

A multifunctional growth and differentiation factor TGF‐β is expressed at various developmental stages, and its principle role may be involvement in organogenesis. The present study was performed to evaluate the temporal and spatial expression of TGF‐β2 mRNA in developing somites of chicken embryos during their early developmental periods. TGF‐βs were expressed in various tissues of the whole embryo obtained at stage 26 (5 days of incubation) as revealed by whole‐mount in situ hybridization. TGF‐β2 mRNA was predominantly expressed in somites as well as the head, branchial arch, wing buds, and leg buds. TGF‐β2 mRNA first appeared in the rostral somites on E4, and its expression sites expanded to the middle range of somites at stage 26. At stages 29–31 (6–7 days), expression in the rostral somites disappeared, and it appeared in the caudal somites. TGF‐β2 expression was also analyzed in sections of the embryo by in situ hybridization. The expression sites of TGF‐β2 were clearly observed in the myotomal somite tips as well as the neural tube. RT‐PCR analysis showed that TGF‐β2 expression was very low in the blastocyte stage embryo and thereafter increased linearly in the whole trunk until stage 26. These data indicate that TGF‐β2 may be a regulatory factor participating in the somitogenesis of chicken embryos. J. Exp. Zool. 303A:323–330, 2005. © 2005 Wiley‐Liss, Inc.

MOX deposition tests were continued in 2002 at RIAR, to increase the Pu concentration in the MOX deposit. The objective was achieved but a significant amount of PuO2precipitates was found. This phenomenon was investigated by evaluating a hypothetical quantity named “Quasi Equilibrium (Q.E.) concentration” of plutonyls in the melt and their solubility. It was found that the oxygen pressure was too high in the 2002 tests, which made the Q.E. concentration too high and the chlorine pressure was too low, which reduced the solubility of plutonyls in the melt. It may be necessary to increase the oxygen pressure to obtain MOX deposit with Pu concentration higher than 30%, but the pressure must be limited depending on the chlorine pressure to avoid the Pu precipitation.

Gene silencing using small interfering RNA (siRNA) is not established in avian species. The present study was performed to evaluate RNA interference (RNAi) in the chicken embryo by using a dual fluorescence reporter assay, a plasmid encoding green fluorescent protein (GFP) and a plasmid encoding red fluorescent protein (RFP). The siRNA targeting the GFP mRNA sequence (GFP‐siRNA) with both plasmids was introduced into cultured cells and whole embryos by lipofection and microelectroporation, respectively. GFP‐ and RFP‐expressed cells and embryos were observed under fluorescent microscopy and analyzed by flow cytometer, and their mRNAs were analyzed by reverse transcription‐PCR (RT‐PCR). The strong fluorescence was observed by introducing both plasmids into cells. The intensity of the green fluorescence generated by GFP was greatly suppressed by introducing GFP‐siRNA. RT‐PCR analysis showed that introducing GFP‐siRNA also decreased GFP mRNA levels. In contrast to GFP, the intensity of the red fluorescence generated by RFP and the RFP mRNA levels remained unchanged. In whole embryos, also, introducing GFP‐siRNA specifically suppressed GFP expression, and the suppression was maintained for at least 72 h. Consequently, it was concluded that the gene silencing using siRNA is applicable to analyzing the function of genes of interest during avian embryogenesis. J. Exp. Zool. 301A:820–827, 2004. © 2004 Wiley‐Liss, Inc.

An efficient solidphase synthetic method for 2,3,4,5Tetrahydro1,4benzodiazepin2,5diones, having amine derivatives on the benzene ring, was developed. This method has been successfully applied to the synthesis of several spatially separated druglike and informationrich smallmolecule libraries composed of 400 compounds using ACT496 automatic synthesizer and the IRORI radio frequencyencoded splitmix synthesis technology.

Asymmetric synthesis of β, β-disubstituted γ-butyrolactones was carried out in highly stereoselective and predictable way, by means of sequential dialkylation of (S) -γ-trityloxymethyl-γ-butyrolactone followed by lactone carbonyl transposition. Application to the synthesis of spirocyclic compound was also described.

Summary The normal development of elastin fibers in the thoracic aorta was studied in fetal, young, and adult monkeys. Tissue was examined by scanning electron microscopy (SEM) after NaOH treatment and by transmission electron microscopy (TEM). The NaOH treatment of fixed tissues effectively removed collagen fibers and enabled three-dimensional visualization of the elastin fibers. In intact fetal aortae, the internal elastic lamina (IEL) was situated immediately beneath the endothelium. This IEL consisted of superficial, longitudinally arranged bundles of elastin fibrils and an underlying solid sheet containing round fenestrations. In neonates, diffuse intimal thickening was observed. In the young and young-adult monkeys, the aortae exhibited intimal thickening with slender but split IEL. One of the most important findings of this study was that elastin fibers in the intimal thickening, as well as smooth muscle cells, ran in a longitudinal fashion. This was in contrast with the elastic laminae of the media which were mainly oriented circumferentially. Subendothelial elastin fibers in this intimal thickening combined with longitudinally arranged microfibrils which formed close associations with endothelial stress fibers. In some adult monkey aortae with well-developed intimal thickening, a complex meshwork of slender elastin fibers was also found beneath the endothelium. The development of the intimal elastin fibers is discussed in relation to hemodynamic forces.

We report here a case of recurrent pheochromocytoma successfully managed by laparoscopic surgery in a patient with multiple endocrine neoplasia type 2A. A 25-year-old man presented with the contralateral adrenal mass after earlier right adrenalectomy. For the preoperative diagnosis of left adrenal pheochromocytoma, adrenal sparing surgery was considered. From the intraoperative laparoscopic view, we found that the tumor originated in periadrenal sympathetic ganglia, and laparoscopic excision of paraganglioma was successfully performed that preserved the integrity of normal adrenal gland. Extra-adrenal pheochromocytoma is rather rare in MEN 2A and it is important to identify precise intraoperative localization of pheochromocytoma with laparoscopic surgery.

In higher vertebrates, cephalic neural crest cells (NCCs) form craniofacial skeleton by differentiating into chondrocytes and osteoblasts. A subpopulation of cephalic NCCs, cardiac NCCs (CNCCs), migrates to the heart. However, CNCCs mostly do not yield skeletogenic derivatives, and the molecular mechanisms of this fate restriction remain elusive. We identify a disintegrin and metalloprotease 19 (Adam19) as a position-specific fate regulator of NCCs. Adam19-depleted mice abnormally form NCC-derived cartilage in their hearts through the upregulation of Sox9 levels in CNCCs. Moreover, NCC-lineage-specific Sox9-overexpressing mice recapitulate CNCC chondrogenesis. In vitroexperiments show that Adam19 mediates the cleavage of bone morphogenic protein (BMP) type I receptor Alk2 (Acvr1), whereas pharmacogenetic approaches reveal that Adam19 inhibits CNCC chondrogenesis by suppressing the BMP-Sox9 cascade, presumably through processing Alk2. These findings suggest a metalloprotease-dependent mechanism attenuating cellular responsiveness to BMP ligands, which is essential for both the positional restriction of NCC skeletogenesis and normal heart development.

Tumor-to-tumor metastasis is a rare phenomenon. From our review of the international literature, around 150 cases have been reported since it was first documented by Campbel in 1868. Renal clear cell carcinoma is well known to be the most common recipient of tumor-to-tumor metastasis in all tumors. However, renal chromophobe cell carcinoma has not been reported to be a recipient. Here, we report a first case of colorectal carcinoma metastatic to chromophobe renal cell carcinoma.

Chicken primordial germ cells (PGCs) dynamically migrate towards prospective gonadal area through germinal crescent region and bloodstream at early embryonic stages. So far, chicken PGC has been mainly identified by histochemical and immunohistochemical methods or its morphological characteristics. However, its origin, migration and differentiation are not fully understood, because of a lack of specific PGC molecular markers and difficulty in detection of chicken primordial germ cells. Recently, dead end, a novel vertebrate germ plasm component coding RNA binding protein was discovered in zebrafish and its expression was proved to be highly specific to germ cells. In the present study, we have isolated the chicken dead end homologue (CDH) to clone its full-length cDNA by Rapid amplification of cDNA ends (RACE)-PCR. Whole-mount in situ hybridization (WISH) analysis detecting CDH mRNA was also performed both in vivo and in vitro. The nucleotide sequence (1,465 bp) of full-length-cDNA revealed an open reading frame for 329 amino acids. The RNA binding motif (Prosite database profile PS50102) in N terminal half present in CDH at 54–133 was highly homologous to those in human (65%), mouse (66%), and Xenopus (66%) Dead end proteins, whereas its similarity to zebrafish Dead end protein was low (55%). There are also great variations in both the N-terminal and C-terminal amino acid sequences among these vertebrates. Our WISH analysis showed that CDH mRNA specifically expressed in primordial germ cells. CDH mRNA positive cells were also detected in embryonic stem cell (ES)-like colonies in vitro derived from blastderm at stage X. Consequently, detection of CDH expression is useful to identify chicken PGCs both in vivo and in vitro. This reseach was supported by Research Fellowships of the Japan Society for the Promotion of Science for Young Scientists (No. 09744) (to S.A.). (poster)

A modified NaOH maceration method for removing extracellular ground substance was applied to scanning electron microscopic (SEM) studies of the mammalian aorta and elastic cartilage. Fixed tissues were freeze-fractured in liquid nitrogen and then immersed in 2 N NaOH solution for 4–6 hr at room temperature. Since this method selectively digested proteoglycans, cellular and fibrous elements were observable clearly under the SEM. The elastic laminae of smooth muscle cells of the aortic tunica media were concealed by fine collagen fibrils, while chondrocytes in the elastic cartilage were encapsulated by a dense network of fine collagen fibrils and branching elastin fibers.