Catalytic Chemoselective O-Phosphorylation of Alcohols.

Phosphorylation of alcohols is a fundamentally important reaction in both life science and physical science. Product phosphate monoesters play key roles in living organisms, natural products, pharmaceuticals, and organic materials. Most of the chemical methods to date for synthesizing phosphate monoesters, however, require multistep sequences or are limited to specific types of substrates possibly due to harsh conditions. An alternative way to enable the simple production of phosphate monoesters from highly functionalized precursor alcohols is, thus, highly desired. We report herein a catalytic phosphorylation of alcohols with high functional group tolerance using tetrabutylammonium hydrogen sulfate (TBAHS) and phosphoenolpyruvic acid monopotassium salt (PEP-K) as the catalyst and phosphoryl donor, respectively. This method enables the direct introduction of a nonprotected phosphate group to the hydroxy group of a diverse menu of alcohol substrates, including functionalized small molecules, carbohydrates, and unprotected peptides. Nuclear magnetic resonance, mass spectrometric, and density functional theory analyses suggest that an unprecedented mixed anhydride species, generated from PEP-K and TBAHS, acts as an active phosphoryl donor in this reaction. This operationally simple and chemoselective catalytic phosphorylation allows for the efficient production of densely functionalized O-phosphorylated compounds, which are useful in diverse fields including biology and medicine.

Potential survival of the lichen Caloplaca flavovirescens under high helium-beam doses.

Testing the limits of survivability in space is the primary focus in astrobiological research. Although a number of previous studies have examined terrestrial life survival in an extraterrestrial environment, only a few have investigated how life systems respond to high doses of alpha cosmic ray, the main component of cosmic rays. We used respiration and photosynthetic rates as indicators of the vital signs of the lichen Caloplaca flavovirescens, which is a symbiotic life form including fungi and algae. Our experiment demonstrated that the photosynthetic rate decreased with increased helium-beam doses, whereas the respiration rate was relatively unaffected. Specifically, under a helium-beam dose greater than 10 Gy, the respiration rate remained nearly constant regardless of further increases in the radiation rate. Our results indicate that the different metabolic systems of terrestrial life forms might exhibit different survival characteristics when they are in space.

Clinical and Histological Study on Direct Pulp Capping With CO2 Laser Irradiation in Human Teeth.

The study aimed to histologically evaluate wound healing of exposed human pulp on direct pulp capping using super-pulsed CO2 laser preirradiation. In this single-blind clinical trial, 28 third molar teeth of 17 volunteers were randomly capped with either CO2 laser irradiation (n=14) or Dycal (calcium hydroxide cement; n=14) and restored using resin composite. The laser was operated in super-pulsed mode (pulse duration, 0.2 ms; interval, 5.8 ms; 0.003 J/pulse). The irradiation conditions were a power output of 0.5 W, an irradiation time of 15 seconds, repeat mode (10-ms irradiation and 10-ms intervals, for a total beam exposure time of 7.5 seconds), total applied energy of 3.75 J, and an activated air-cooling system. Each tooth was extracted at six or 12 months posttreatment and prepared for histological evaluation. We evaluated the parameters of pulp tissue disorganization, inflammatory cell infiltration, reparative dentin formation (RDF), and bacterial penetration. There were no significant differences between groups for all parameters at each postoperative period (Mann-Whitney U-test, p>0.05). CO2 laser irradiation completely controlled bleeding and exudate from the exposed pulp. The CO2 laser group had a tendency to delay RDF compared with the Dycal group, but 4 of 7 teeth from the CO2 laser group showed a complete dentin bridge at 12 months posttreatment.

Dermoscopic features of pseudoxanthoma elasticum.

Pseudoxanthoma elasticum (PXE) is a disease characterized by aberrant mineralization of soft tissue and fragmentation of elastic fibres. It is often difficult to distinguish PXE clinically from pseudoxanthoma elasticum-like papillary dermal elastolysis (PXE-like PDE). However, we have identified that the dermoscopic findings in PXE include coalescing and reticulated yellow-white clods on a light purple-red background, whereas the dermoscopic findings in PXE-like PDE lack such a coloured background. To our knowledge, this is the first detailed description of dermoscopic differences between PXE and PXE-like PDE.

Twenty-year trends in neonatal surgery based on a nationwide Japanese surveillance program.

AIM: To discuss the chronological changes observed in a national survey of neonatal surgery in Japan performed every 5 years by the Committee in the Japanese Society of Pediatric Surgeons. METHODS: We analyzed the data obtained for 20 years from 1993 to 2013 and herein report the chronological changes. RESULTS: The number of summarized cases was least in 1993, with 2806 cases, and subsequently increased to 3753 cases in 2013. The mortality rate among the patients with maternal transport linearly decreased (p = 0.0386). Although the proportion of extremely low birth weight infants linearly increased (p = 0.0014), with an annual rate of +0.39 %, the mortality rate linearly decreased (p = 0.0010), with an annual rate of -1.68 %. Moreover, the overall mortality rate linearly decreased (p = 0.0002), with an annual rate of -0.26 %. Most diseases were observed to exhibit a decline in the mortality rate with the same trend as overall mortality. The decline in the mortality rate was most robust with respect to congenital diaphragmatic hernia (CDH). The mortality rates, except for that of CDH, omphalocele, esophageal atresia, and intestinal perforation, declined to 5 % or lower by 2013. CONCLUSIONS: The present findings may be the result of remarkable progress in perinatal management.

Morphological characteristics of the temporomandibular joint in the pouch young of the Tasmanian devil.

We recently reported the absence of the articular disc, which is a constant structure in mammals, in the temporomandibular joint of the adult Tasmanian devil. However, whether the articular disc disappears with growth of the animal was unknown. The aim of this study was to determine whether a pouch young of the Tasmanian devil has the articular disc. The temporomandibular joint of a fresh carcass of the pouch young, whose crown-rump length was 43 mm, was examined microscopically and by microcomputed tomography. The absence of the articular disc in the pouch young temporomandibular joint was histologically confirmed. It is suggested that the articular disc of the Tasmanian devil is naturally absent.

Use of a replica graft tooth for evaluation before autotransplantation of a tooth. A CAD/CAM model produced using dental-cone-beam computed tomography.

A 33-year-old man was referred by a local dentist for autotransplantation of the right mandibular third molar to the site of the left mandibular first molar. Before the operation, dental-cone-beam computed tomography was performed to measure the dimensions of the graft tooth and the bone volume of the socket. The three-dimensional (3D) structure of the graft tooth was created from the 3D imaging data. A replica of the graft tooth was produced using a stereolithographic CAD/CAM system before autotransplantation. A socket of the appropriate size and structure was created and the graft tooth was extracted immediately before grafting. This procedure shortened the time needed to fit the graft tooth into the socket and reduced injury to the periodontal membrane. The postoperative outcomes of autotransplantation of the tooth were good.

Three-dimensional reconstruction of a fibro-osseous lesion using binary images transformed from histopathological images.

The aim of this report was to introduce a new method of three-dimensional (3D) reconstruction for fibro-osseous lesions (FOLs) using binary images transformed from histopathological images and to describe its usefulness. A sample of multiconfluent FOL was used (one of the five types of FOL according to a radiographic classification) which was diagnosed histopathologically as ossifying fibroma. Approximately 30 pathological images were assembled into a composite image of the slide using Tiling Boutique software version 3 for Windows (Sanyo Electric, Osaka, Japan). The tiling images were transformed into 8-bit scale images and then into binary images using ImageJ software ver.1.37 (National Institutes of Health, Bethesda, MD). These images were used for 3D reconstruction using ImageJ software. Images were loaded at the same matrix size and were reconstructed into layers of two-dimensional image stacks, adjusted so that contiguous images were aligned based on their centre points, and arranged with long axes horizontal. 3D findings aided the visual understanding of morphological features in the lesion. The 3D reconstruction can be displayed with arbitrary rotation. In this case, the 3D reconstruction, using Real Image software version 4.01 for Windows (KGT, Tokyo, Japan), was created from an arbitrary section. This allowed us to determine the pattern of calcification between groups of connected osteoids and to compare the internal structure of such lesions that are not visible on histopathological findings. Differentiation of features was even more pronounced with a two colour display indicating fibrous connective tissue and osteoid tissue. A 3D reconstruction of a multiconfluent ossifying fibroma was created using binary images transformed from histopathological images. The quality of the images depends above all on the functionality of the image-processing software. Comparison of each pattern of FOL might allow more simple assessment of the morphological features of FOLs.

Correlation between histopathological image and radiographic image pattern in fibro-osseous lesions in relation to bone complexity and distribution.

OBJECTIVES: This study examined correlations between radiographic patterns and the shape of osteoid tissue formations, as determined histopathologically. METHODS: 20 cases of fibro-osseous lesions were investigated, comprising 5 radiographic patterns: focal (n = 3), target (n = 6), lucent (n = 4), calcification (n = 3) and multiconfluent (n = 4). Histopathological images in the central area of a full-section specimen were transformed into binary images and then into 8-bit scale images. Bone complexity and density of bone distribution were calculated and compared between patterns. RESULTS: Bone complexity score was 7384.64 for lucent, 2029.85 for focal, 2713.40 for multiconfluent, 8388.63 for calcification and 1364.27 for target pattern. The results could be broadly separated into two types: small (target, focal and multiconfluent patterns), and large (lucent and calcification patterns). Density of bone distribution was relatively low in all areas for lucent and calcification patterns, and high for focal, multiconfluent and target patterns. No significant differences in bone complexity or density of bone distribution were seen between individual patterns. CONCLUSIONS: Correlations appear to exist between image patterns from radiography and the shape of osteoid tissue on histopathology, but reorganization of the five patterns may be warranted.

The views of doctors on registration trials in a Japanese rural area: a survey of medical institutions registered to the Tokushima Network for Clinical Trials.

Tokushima University Hospital has established the Tokushima Network for Clinical Trials (TNCT) to promote clinical trials in the area in collaboration with the Tokushima Medical Association. The present study investigated the views of doctors towards registration trials in the TNCT. A questionnaire was provided to 49 clinics/hospitals registered to the TNCT in 2006 and 38 (78%) responded. It revealed that 48% of doctors were aware of registration trials and 87% were favourable towards participating as investigators in them. They considered close contact with developmental drugs, advancement of therapy and the opportunity to learn about state-of-the-art treatment as benefits of participation. The main areas of difficulty included management of adverse reactions and patients' refusal to take part. Many doctors wanted more opportunity to learn about trial-related issues such as regulations. The survey indicates that the TNCT needs to develop the infrastructure and enlighten participants to promote registration trials in this rural regional area.

Breed differentiation among Japanese native chickens by specific skull features determined by direct measurements and computer vision techniques.

1. Inter-breed morphological comparisons were made among 11 breeds of Japanese native chickens (Gifujidori, Hinaidori, Shokoku, Totenko, Tomaru, Satsumadori, Shamo, Koshamo, Koeyoshi, Chabo and Nagoya), White Leghorn, broiler chickens (Chunky) and red junglefowl collected in the Philippines, based on results of direct measurements and analysis by computer vision techniques of the skull. 2. Analysis of direct measurements identified two groups of chicken: a small type that included the Chabo, Koshamo, red junglefowl, Gifujidori and Shokoku and a large type that included the remaining breeds studied. These groupings were made based on size determined both in the first (PC1) and second principal component (PC2). The greatest length of the cranium and condylobasal length greatly contributed to the morphological differences between these two groups. 3. Analysis by computer vision techniques, however, identified three groups: the Bantam group (which includes red junglefowl), Shokoku group and Shamo group. White Leghorn clustered within the Shokoku group while the broiler chicken belonged to the Shamo group. The region around the junction of the neural cranium and the visceral cranium contributed greatly to the morphological differences among breeds, both in the PC1 and PC2.

Serotonin induces vasoconstriction of smooth muscle cell-rich neointima through 5-hydroxytryptamine2A receptor in rabbit femoral arteries.

BACKGROUND: Smooth muscle cell (SMC)-rich intima is a morphological feature of atherosclerotic lesions that is observed in eroded plaque and spastic arteries. Arteries with SMC-rich intima are susceptible to vasoconstriction or vasospasm against some vasoactive agents. OBJECTIVE: The present study evaluates the contribution of SMC-rich intima to thrombogenic vasoconstriction. METHODS: We established SMC-rich neointima by damaging rabbit femoral arteries using balloons and then measured the isometric tension of the femoral strips against 5-hydroxytryptamine (5-HT), adenosine diphosphate, adenosine triphosphate and thrombin. RESULTS: Among these agents, only 5-HT induced a hypercontractile response of the injured arteries with SMC-rich neointima, compared with non-injured arteries. Smooth muscle cells of both the neointima and media expressed 5-HT(2A) receptor, and sarpogrelate, a selective 5-HT(2A) receptor antagonist significantly inhibited the hypercontraction. Furthermore, 5-HT induced contraction of separated neointima and hypercontraction of separated media compared with non-injured media. Sarpogrelate and fasudil, a specific Rho-kinase inhibitor, significantly suppressed such contraction of both the neointima and media of injured arteries. CONCLUSIONS: These results suggest that 5-HT plays a crucial role in thrombogenic vasoconstriction, and that SMC-rich intima as well as media directly contributes to the hypercontractile response of atherosclerotic vessels through the 5-HT(2A) receptor and the Rho-kinase pathway.

Comparison of image validity between cone beam computed tomography for dental use and multidetector row helical computed tomography.

OBJECTIVES: To evaluate and compare the image validity of a cone beam CT machine for dental use (3DX) and the latest medical CT system, multidetector row helical CT (MDCT). METHODS: A dried right maxilla of an Asian adult was used as a phantom. It was cut from the zygomatic process towards the midline, parallel to the midline plane, into eight slices of 2 mm thickness. This phantom was imaged with the 3DX and MDCT machines. Images were evaluated by comparing them with one selected bone slice from the phantom. In this comparison, two types of MDCT images were used: one with the window level (WL) and window width (WW) suitable for observing teeth (MDCT tooth image), and the other appropriate for observing bone (MDCT bone image). Three dentists and one radiographer then used our reported subjective five-level scale to evaluate and compare images generated by the two systems in terms of validity. Cancellous bone as well as enamel, dentin, pulp cavity, periodontal ligament space, lamina dura and overall impression were evaluated. RESULTS: Statistically significant differences (P<0.05) were found for almost all observation items. Our subjective evaluation clarified that 3DX was superior to MDCT in terms of image validity. Few significant intra- or interevaluator errors were found. CONCLUSIONS: Our subjective evaluation of image validity clarified 3DX as being superior to MDCT. Taken together with the low skin dose we previously reported for 3DX, the results demonstrate 3DX to be beneficial for imaging diagnosis of hard tissues in the maxillofacial region.

Protective effect of endothelial nitric oxide synthase against induction of chemically-induced diabetes in mice.

Since activation of endothelial nitric oxide synthase has been shown to exert protective effects against the metabolic syndrome, while endothelial nitric oxide synthase knockout mice develop hyperinsulinemia and glucose intolerance, we hypothesised that endothelial nitric oxide might play a protective role against induction of diabetes. The role of endothelial nitric oxide in the development of chemically-induced diabetes has been determined using mice in which the bioavailability of endothelial nitric oxide was either increased, through upregulation of endothelial nitric oxide synthase, or absent, through deletion of endothelial nitric oxide synthase gene. Diabetes was induced intraperitoneally with either a single dose of alloxan, streptozotocin, or multiple low doses of streptozotocin and blood glucose monitored twice a week. The role of cyclic guanosine monophosphate was investigated in wildtype mice by treatment with the phosphodiesterase inhibitor, tadalafil, during diabetes induction. Results showed that the incidence of diabetes was markedly decreased in mice overexpressing endothelial nitric oxide synthase, compared to wildtype or endothelial nitric oxide synthase knockout mice, regardless of the method of diabetes induction. Under normal physiological conditions, or during diabetes induction with alloxan or multiple low doses of streptozotocin, blood glucose was significantly lower in mice overexpressing endothelial nitric oxide synthase compared to wildtype or knockout mice. Treatment with tadalafil had no effect on the incidence or severity of diabetes in wildtype mice. We conclude that upregulation of endothelial nitric oxide synthase exerts a protective action against diabetes induction through a direct effect of nitric oxide, independently of cyclic guanosine monophosphate.

Analysis of mtDNA sequences shows Japanese native chickens have multiple origins.

In this study, we analysed the mitochondrial DNA D-loop region of Japanese native chickens to clarify their phylogenetic relationships, possible maternal origin and routes of introduction into Japan. Seven haplogroups (Types A-G) were identified. Types A-C were observed in Jidori, Shokoku and related breeds. However, Type C was absent in Shokoku, which was introduced from China, while most Indonesian native chickens were included in the Type C haplogroup. Types D-G were observed in Shamo and related breeds. Type E had a close genetic relationship with Chinese native chickens. Our results indicate that some breeds were not introduced into Japan as suggested in conventional literature, based on low nucleotide diversity of certain chicken breeds. Sequences originating from China and Korea could be clearly distinguished from those originating from Southeast Asia. In each group, domestic chickens were divided into the Jidori-Shokoku and Shamo groups. These results indicate that Chinese and Korean chickens were derived from Southeast Asia. Following the domestication of red junglefowl, a non-game type chicken was developed, and it spread to China. A game type chicken was developed in each area. Both non-game and game chickens formed the foundation of Japanese native chickens.

Relationships between nitric oxide-mediated endothelial function, eNOS coupling and blood pressure revealed by eNOS-GTP cyclohydrolase 1 double transgenic mice.

Endothelium-dependent relaxation in conduit vessels is mediated largely by nitric oxide (NO), produced by the enzyme endothelial nitric oxide synthase (eNOS) in the presence of the cofactor tetrahydrobiopterin (BH4) and mediated through a cGMP-dependent downstream signalling cascade. Endothelial NOS regulates blood pressure in vivo, and impaired endothelial NO bioactivity in vascular disease states may contribute to systemic hypertension. In the absence of sufficient levels of the cofactor BH4, NO becomes uncoupled from arginine oxidation and eNOS produces superoxide rather than NO. The enzymatic uncoupling of eNOS is an important feature of vascular disease states associated with increased oxidative stress. However, whether eNOS coupling, rather than overall eNOS activity, has specific effects on endothelium-dependent vasorelaxation in vitro, or on blood pressure regulation in vivo, remains unclear. In this study, we evaluate the relationships between blood pressure and endothelial function in models of eNOS uncoupling, using mice with endothelium-targeted transgenic eNOS overexpression (eNOS-Tg), in comparison with littermates in which eNOS coupling was rescued by additional endothelium-targeted overexpression of GTP cyclohydrolase 1 (eNOS/GCH-Tg) to increase endothelial BH4 levels. Despite the previously characterized differences in eNOS-dependent superoxide production between these animals, we find that blood pressure is equally reduced in both genotypes, compared with wild-type animals. Furthermore, both eNOS-Tg and eNOS/GCH-Tg mice exhibit similarly impaired endothelium-dependent vasorelaxation. We show that reduced vasorelaxation responses result from desensitization of cGMP-mediated signalling and are associated with increased NO production rather than changes in superoxide production.

Single air contrast arthrography for temporomandibular joint disorder using limited cone beam computed tomography for dental use.

MRI has the problem of artefacts caused by metal or body motion and is also incompatible for patients with claustrophobia. Arthrography is invasive and involves the risk of perforation or allergy to a contrast medium. This report discusses a patient with temporomandibular joint (TMJ) disorder who required soft tissue imaging of the TMJ. As she had claustrophobia and a reaction to iodine, air contrast arthrography and pumping manipulation therapy using limited cone beam computed tomography for dental use (3DX) was performed. We conclude that the 3DX examination method used in the study is practical as a diagnostic procedure and thus recommend this method to be used for patients with TMJ disorder in the presence of iodine contraindication.

Therapeutic efficacy of sustained drug release from chitosan gel on local inflammation.

The model anti-inflammatory drug prednisolone (PS) was retained in chitosan (CS) gel beads, which were prepared in a 10% aqueous amino acid solution (pH 9.0). Sustained release of PS from the CS gel beads was observed. Carrageenan solution was injected into air pouches (AP), which were prepared subcutaneously on the dorsal surface of mice, in order to induce local inflammation. CS gel beads retaining PS were then implanted into the AP to investigate the therapeutic efficacy of sustained PS release against local inflammation. In vivo PS release from CS gel beads was governed by both diffusion of the drug and degradation of the gel matrix. Sustained drug release by CS gel beads allowed the supply of the minimum effective dose and facilitated prolonged periods of local drug presence. Inflammation indexes were significantly reduced after implantation of CS gel beads when compared with injection of PS suspension. Thus, extension of the duration of drug activity by CS gel beads resulted in improved therapeutic efficacy. These observations indicate that CS gel beads are a promising biocompatible and biodegradable vehicle for treatment of local inflammation.