Convergent synthesis of phosphorodiamidate morpholino oligonucleotides (PMOs) by the H-phosphonate approach.

Phosphorodiamidate morpholino oligonucleotides (PMOs) are a promising type of antisense oligonucleotides, but their challenging synthesis makes them difficult to access. This research presents an efficient synthetic approach for PMOs using the H-phosphonate approach. The use of phosphonium-type condensing reagents significantly reduced coupling times compared with the current synthetic approach. Furthermore, phosphonium-type condensing reagents facilitated the fragment condensation of PMO, synthesizing up to 8-mer containing all four nucleobases with remarkable coupling efficacy. This is the first report on the convergent synthesis of PMOs. This approach would facilitate the large-scale synthesis of PMOs and accelerate their popularity and accessibility as a next-generation therapy.

Reconsideration of Leeway Space Calculated Using Models from Same Patients at Different Growth Stages.

In this study, original leeway space was determined using plaster models prepared from the same individuals over successive years and compared with that reported in an earlier study. The plaster models used for measurement were prepared from a normal dentition group comprising 64 patients and a crowded dentition group comprising 29. The mesiodistal crown width of the deciduous canines, deciduous first molars, and deciduous second molars, as well as that of the canines, first premolars, and second premolars was measured. Leeway space was then calculated and the results compared with that previously reported. The average maxillary and mandibular leeway spaces calculated in the same individuals were 1.46 and 2.66 mm, respectively, whereas those calculated from data collected from different individuals were 0.89 and 2.83 mm, respectively. No direct statistical comparison of leeway space between this and the earlier report was possible. The present results did indicate, however, that it was larger in the maxilla than previously reported. A larger leeway space is advantageous for the normal arrangement of the permanent teeth. While the mesiodistal crown width of some of the lateral segment teeth in the present study was significantly larger in boys than in girls, no significant difference was observed in leeway space between sexes. The mesiodistal width was larger in multiple permanent teeth such as canines in the crowded dentition group, and this difference was significant in boys. On the other hand, no difference was observed in mesiodistal width in the deciduous teeth between the two groups. These results showed a tendency toward larger teeth and less leeway space in the permanent dentition in the crowded dentition group, which may have been a factor in why such crowding occurred.

Surface Plasmon Resonance of Two-Dimensional Gold Colloidal Crystals Formed on Gold Plates.

The free electrons inside precious metals such as Au vibrate when the surface of the metal is irradiated with an electromagnetic wave of an appropriate frequency. This oscillation is referred to as surface plasmon resonance (SPR), and the resonance frequency varies with permittivity of the medium around the metal. SPR sensors are widely applied in the fields of bioscience and pharmaceutical sciences, including biosensing for drug discovery, biomarker screening, virus detection, and testing for food safety. Here, we fabricated a metal-insulator-metal (MIM) SPR sensor by constructing two-dimensional (2D) regular array of Au colloidal particles (2D colloidal crystals) on an insulator layer over a thin Au film coated on a glass substrate surface. The 2D crystals were fabricated by electrostatically adsorbing negatively charged three-dimensional crystals onto a positively charged thin insulator formed on Au film. The plasmon peaks/dips from the MIM structure were measured in aqueous solutions of ethylene glycol (EG) at various concentrations. Multiple plasmon peaks/dips were observed due to the localized SPR (LSPR) of the Au particles and the Fano resonance between the Au particles and thin film. The plasmon peaks/dips shifted to higher wavelengths on increasing EG concentrations due to an increase in the refractive index of the media. The observed peak/dip shift was approximately twice that of LSPR from an isolated Au particle. We expect the present MIM substrate will be useful as a highly sensitive sensor in the pharmaceutical field.

Homonymous quadrantanopia associated with hyperosmolar hyperglycemic syndrome.

We encountered a 64-year-old man with hyperosmolar hyperglycemic syndrome, having a sudden-onset homonymous right inferior quadrantanopia. This is the first documentation of such a phenomenon in hyperosmolar hyperglycemic syndrome. We believe this is a variant of hemianopia in patients with hyperglycemic hyperosmolar syndrome.

Solid-Phase Synthesis of Phosphate/Boranophosphate Chimeric DNAs Using the H-Phosphonate-H-Boranophosphonate Method.

Boranophosphate (PB) DNAs are promising antisense oligonucleotide candidates because of their attractive features, such as high nuclease resistance and low toxicity. However, a full boranophosphate backbone modification to antisense DNAs causes reduced duplex formation with complementary RNAs and reduced antisense activity. In this study, an efficient solid-phase synthesis of phosphate/boranophosphate (PO/PB) chimeric DNA was achieved by the combination of the H-phosphonate and H-boranophosphonate methods. The physiological and biological properties of the synthesized PO/PB chimeric DNAs were also evaluated. The strategy employed herein can facilitate the design and synthesis of PO/PB chimeric DNAs containing site-specific boranophosphate modifications.

Development of Jaw and Deciduous Teeth in Japanese Children -Comparing Size of Crown and Alveolar Area between Today and 40 Years Ago.

The purpose of the present study was to investigate change in the size of the jaw and deciduous teeth in Japanese children over the past 40 years by performing measurements on plaster models of the dental arch obtained from children born between 2007 and 2009 (2000s group) and children born between 1968 and 1974 (70s) group. A total of 61 children were enrolled in the 2000s group, comprising 25 boys (average age, 4 years 5 months) and 36 girls (average age, 4 years 4 months), and 93 children in the 70s group, comprising 45 boys (4 years 6 months) and 48 girls (4 years 4 months). The mesiodistal width of the crown, dental arch width, dental arch length, dental arch height, and available arch length in these groups were measured using a 3D measurement system and the sums of the mesiodistal width of the crown and tooth size-arch length discrepancies calculated. The results of the two groups were statistically compared and the following conclusions reached: the size of deciduous teeth has not changed statistically over the last 40 years; the size of the dental arch has decreased in boys (p<0.01), while remaining approximately the same in girls; and the tooth size-arch length discrepancy has decreased, especially in boys (p<0.01). These results indicate that the interdental spaces of the deciduous dentition have decreased, increasing the risk of crowding in Japanese children.

Alveolar Growth in Japanese Infants : A Comparison between Now and 40 Years ago.

To investigate differences in alveolar growth during the primary dentition period among different age groups, dental arch width, length, and height were measured in 93 dental arch plaster models obtained from 4-year-olds born between 1968 and 1974, 88 from 5-year-olds born between 1968 and 1974 (70s group), 61 from 4-year-olds born between 2007 and 2009, and 56 from 5-year-olds born between 2006 and 2008 (2000s group). A comparison was also performed to determine whether there was any difference in change between 4 and 5 years of age. The results showed age- and sex-dependent differences in growth patterns in primary dental arch width, length, and height. The amount of change was greater in the 2000s group than in the 70s group, suggesting that yearly growth between 4 and 5 years of age has increased over the last 40 years.

Influence of Lifestyle Factors on Risk of Dental Caries among Children Living in Urban China.

The prevalence of dental caries has been decreasing among kindergarten children in Shanghai, China, over recent years, although it still remains at an unacceptably high level. The purpose of this study was to identify which factors were important in providing oral health guidance and achieving further improvement in the oral health status of kindergarten children in urban China. A survey was conducted on dental caries in 128 Japanese and 368 Chinese kindergarten children and a questionnaire given to their parents/guardians on each child's lifestyle and dietary habits from birth to the present. Correlations between responses to each questionnaire item and the status of dental caries were statistically analyzed. The dft index score (p=0.0016), prevalence of dental caries (p=0.0002), and percentages of children with decayed (untreated caries-affected) teeth (p<0.0001) were significantly higher in the Chinese than in the Japanese children. Many differences were observed in lifestyle factors between the two groups. The percentage of parents failing to control the child's snacking habits between meals was higher in China, and weaning was significantly delayed in China compared with in Japan. These lifestyle factors were considered to be associated closely with the high risk of dental caries in Chinese kindergarten children. These findings indicate that oral health guidance for kindergarten children in Shanghai, China, should focus on control of dietary habits, including control of inter-meal snacking, and breastfeeding practices. The results of this study may help improve the status of dental caries among Chinese children.

A case of Bardet-Biedl syndrome complicated with intracranial hypertension in a Japanese child.

Bardet-Biedl syndrome (BBS) is a rare heterogeneous autosomal recessive disorder characterized by rod-cone dystrophy, postaxial polydactyly, truncal obesity, hypogonadism, learning disability, and renal anomaly that are caused by ciliary dysfunction. 16 genes have been associated with the BBS phenotype. Although recent pathophysiological studies using animal models have shown that ciliary dysfunction may induce hydrocephalus, there have been no reports of BBS with intracranial hypertension. We here describe a 9-year-old Japanese girl who was diagnosed as having BBS and later received renal transplantation due to chronic renal failure. She also exhibited intracranial hypertension, including papilledema and increased intrathecal pressure (260-300 mmH2O), but her brain magnetic resonance imaging was normal. No genetic abnormalities were detected by DNA chip analysis or exome sequencing. Her papilledema improved following administration of acetazolamide. This is the first report of a case of BBS complicated with intracranial hypertension and its treatment.

Cancer-associated retinopathy in a patient with seminoma.

PURPOSE: The purpose of this study was to report our findings on a patient with cancer-associated retinopathy who had seminoma. METHODS: A 43-year-old man, who had been diagnosed with Stage 1 seminoma, complained of blurred vision. Fundus examination revealed attenuated retinal arteries, mild optic disk pallor, and a thinning of the outer nuclear layer. Ring scotomas were found in the visual fields of both eyes. The amplitudes of the electroretinograms were moderately reduced. The patient's serum was analyzed for antibodies. RESULTS: Antibodies against recoverin, Hu, and Ma-2 antigens were not detected. However, Western blots of the serum detected 2 bands at 41 and 64 kDa. The 41-kDa band was an anti-retina-specific antibody. Immunoreactivity was identified in the photoreceptor layer of mice retinas. From these findings, we diagnosed the patient with cancer-associated retinopathy. The visual functions remained stable after the removal of the primary lesion at an early stage, treatment with steroids, and intravenous immunoglobulin. CONCLUSION: This is the first case of cancer-associated retinopathy because of seminoma. The progression of the disease was reduced by removing the seminoma at an early stage, treatment with steroids, and intravenous immunoglobulin.

Microperimetric determination of retinal sensitivity in areas of dissociated optic nerve fiber layer following internal limiting membrane peeling.

PURPOSE: To investigate the effects of a dissociated optic nerve fiber layer (DONFL) on retinal sensitivity. METHODS: The medical records of 17 eyes with an idiopathic macular hole that underwent vitrectomy and internal limiting membrane (ILM) peeling were examined. All patients underwent a complete ophthalmic examination, color fundus photography, and argon blue-filter photography to determine whether a DONFL was present. The retinal function was assessed by comparing the mean retinal sensitivities in the DONFL area to those in the surrounding normal retina by static microperimetry >3 months after the surgery. The retinal sensitivities of eight regions nearest the fovea and all at the same distance from it were determined in all patients. We divided the retinal sensitivities in the two areas, that is, inside and outside the area of the DONFL, and compared each mean value. RESULTS: Following the initial vitrectomy, the macular holes of all eyes were closed. The mean retinal sensitivities in the area of the DONFL did not differ significantly from those in the surrounding areas (P = 0.60). CONCLUSIONS: DONFL associated with ILM peeling does not alter retinal function in the area of the DONFL as it does with a nerve fiber layer defect.

{mu}-Crystallin, new candidate protein in endotoxin-induced uveitis.

PURPOSE. micro-Crystallin (CRYM) is a major taxon-specific lens protein. The purpose of this study was to investigate the function of CRYM in eyes of mice with endotoxin-induced uveitis (EIU). METHODS. EIU was induced by an injection of a lipopolysaccharide (LPS) into the footpad of male C57BL/6J, CRYM knockout (CRYM(-/-)), and wild-type (CRYM(+/+)) mice. The expression of CRYM in the iris-ciliary body (ICB) was investigated by Western blot analyses and real-time RT-PCR at 12 hours and 1, 3, and 5 days after the LPS injection. The number of cells that had infiltrated the anterior chamber (AC) of the CRYM(+/+) mice was compared to that in the CRYM(-/-) mice at 1, 3, 5, and 7 days. The expressions of the mRNA of interleukin (IL)-1alpha, IL-6, tumor necrosis factor (TNF)-alpha, and granulocyte macrophage-colony stimulating factor (GM-CSF) in the ICB of the two groups of mice were compared. RESULTS. The mRNA of CRYM was upregulated at 12 hours after LPS injection, and CRYM protein increased at 3 days. The number of inflammatory cells in the AC of the CRYM(-/-) mice was not significantly different on day 1 from that in the CRYM(+/+) mice, but was significantly lower (17.9 +/- 1.6 vs. 27.1 +/- 2.4 cells/section) on day 5. Expression of the mRNA of IL-1alpha and -6 in the CRYM(-/-) mice was significantly lower than that in the CRYM(+/+) mice on day 5. CONCLUSIONS. CRYM plays an important role in the development of the second peak of murine EIU.

Cytokine and molecular analyses of intraocular lymphoma.

PURPOSE: The authors investigate the efficacy of using the cytokine levels and clonal heavy-chain immunoglobulin (IgH) gene rearrangements in the vitreous as adjunctive tools to diagnose intraocular lymphoma (IOL). METHODS: The IL-10 and IL-6 levels and IgH gene rearrangements were analyzed in vitreous samples from 8 cases of IOL and in 14 uveitis patients. RESULTS: The level of IL-10 with an IL-10/IL-6 ratio > 1 was significantly higher in all eyes with IOL. B-cell monoclonality was detected in only 5 of 8 eyes with IOL. CONCLUSIONS: The measurements of the levels of cytokines are valuable as a reliable biomarker.

[Two-year results following photodynamic therapy for age-related macular degeneration].

PURPOSE: To investigate the visual outcome 2 years after photodynamic therapy (PDT) in patients with exudative age-related macular degeneration (AMD) and polypoidal choroidal vasculopathy (PCV). METHODS: Thirty-six eyes undergoing PDT were retrospectively studied. Seventeen eyes of AMD (AMD group) and nineteen eyes of PCV (PCV group) were evaluated. RESULTS: In the AMD group, the mean pre-PDT visual acuity was 0.19. The mean post-PDT visual acuity was 0.16 two years after the PDT. Two years after PDT, the Log MAR visual acuity improved by 0.2 or more in 3 eyes (17.7%), but it decreased by 0.2 or more in 4 eyes (23.5%). In the PCV group, the mean pre-PDT visual acuity was 0.34. The mean post-PDT visual acuity was 0.20 two years after PDT. The Log MAR visual acuity improved by 0.2 or more in 5 eyes (26.3%), but it decreased by 0.2 or more in 7 eyes (36.8%). CONCLUSIONS: In this series of patients, more than half of the two groups were able to maintain their visual acuity for 2 years after PDT. Although the average visual acuity of the AMD group was worse than that of the PCV group, the AMD group was able to maintain their visual acuity for 2 years after PDT. The average visual acuity of the PCV group decreased 2 years after PDT.

The ligand activity of AGE-proteins to scavenger receptors is dependent on their rate of modification by AGEs.

The cellular interaction of proteins modified with advanced glycation end-products (AGEs) is believed to induce several different biological responses, which are involved in the development of diabetic vascular complications. We report here that the ratio of protein glycation is implicated in its ligand activity to scavenger receptors. Although highly-modified AGE-bovine serum albumin (high-AGE-BSA) was significantly recognized by human monocyte-derived macrophages and Chinese hamster ovary cells which overexpress such scavenger receptors as CD36, SR-BI (scavenger receptor class B type-I), and LOX-1 (Lectin-like Ox-LDL receptor-1), the mildly-modified-AGE-BSA (mild-AGE-BSA) did not show any ligand activity to these cells. Furthermore, when (111)In-labeled high- or mild-AGE-BSA were injected into the tail vein of mice, the high-AGE-BSA was rapidly cleared from the circulation whereas the clearance rate of the mild-AGE-BSA was very slow, similar to the native BSA. These results demonstrate the first evidence that the ligand activity of the AGE-proteins to the scavenger receptors and its pharmacokinetic properties depend on their rate of modification by AGEs, and we should carefully prepare the AGE-proteins in vitro to clarify the physiological significance of the interaction between the AGE-receptors and AGE-proteins.

The relation between tooth shape ratio and incisor arrangement in Japanese children.

To investigate whether the relationship between tooth shape ratio and incisor arrangement proposed by Peck et al. in 1972 was applicable to Japanese children, we compared two groups, one with normal permanent dentition and another with crowded permanent dentition, using chronologically constructed plaster dentition models. Tooth shape ratio was obtained by dividing the incisor mesio-distal width by the labio-lingual width and then multiplying the result by 100. The results revealed a significant difference in the coronal mesio-distal diameter of the incisors between the two groups, but the other items showed no significant differences, suggesting no clear relationship between tooth shape ratio and incisor arrangement. That was, our results suggest that there is no clear relationship between tooth shape ratio of the mandibular incisors and arrangement of the permanent incisors in Japanese children.

Total synthesis of epoxyquinols A, B, and C and epoxytwinol A and the reactivity of a 2H-pyran derivative as the diene component in the Diels-Alder reaction.

Full details of two versions of the total synthesis of epoxyquinols A, B, and C and epoxytwinol A (RKB-3564D) are described. In the first-generation synthesis, the HfCl(4)-mediated diastereoselective Diels-Alder reaction of furan with Corey's chiral auxiliary has been developed. In the second-generation synthesis, a chromatography-free preparation of an iodolactone, by using acryloyl chloride as the dienophile in the Diels-Alder reaction of furan, and the lipase-mediated kinetic resolution of a cyclohexenol derivative have been developed. This second-generation synthesis is suitable for large-scale preparation. A biomimetic cascade reaction involving oxidation, 6pi-electrocyclization, and then Diels-Alder dimerization is the key reaction in the formation of the complex heptacyclic structure of epoxyquinols A, B, and C. Epoxytwinol A is synthesized by the cascade reaction composed of oxidation, 6pi-electrocyclization, and formal [4 + 4] cycloaddition reactions. A 2H-pyran, generated by oxidation/6pi-electrocyclization, acts as a good diene, reacting with several dienophiles to afford polycyclic compounds in one step. An azapentacyclic compound is synthesized by a similar cascade reaction composed of the four successive steps: oxidation, imine formation, 6pi-azaelectrocyclization, and Diels-Alder dimerization.

Different reaction modes for the oxidative dimerization of epoxyquinols and epoxyquinones. Importance of intermolecular hydrogen-bonding.

An oxidative dimerization reaction, involving the three successive steps of oxidation, 6 pi-electrocyclization, and Diels-Alder reaction, has been experimentally and theoretically investigated for the three 2-alkenyl-3-hydroxymethyl-2-cyclohexen-1-one derivatives epoxyquinol 3, epoxyquinone 6, and cyclohexenone 10. Of the sixteen possible modes of the oxidation/6 pi-electrocylization/Diels-Alder reaction cascade for the epoxyquinone 6, and eight for the cyclohexenone 10, only the endo-anti(epoxide)-anti(Me)-hetero and endo-anti(Me)-hetero modes are, respectively, observed, while both endo-anti(epoxide)-anti(Me)-hetero and exo-anti(epoxide)-anti(Me)-homo reaction modes occur with the epoxyquinol 3. Intermolecular hydrogen-bonding is found to be the key cause of formation of both epoxyquinols A and B with 3, although epoxyquinone 6 and cyclohexenone 10 both gave selectively only the epoxyquinol A-type product. In the dimerization of epoxyquinol 3, two monomer 2H-pyrans 5 interact with each other to afford intermediate complex 28 or 29 stabilized by hydrogen-bonding, from which Diels-Alder reaction proceeds. Theoretical calculations have also revealed the differences in the reaction profiles of epoxyquinone 6 and cyclohexenone 10. Namely, the rate-determining step of the former is the Diels-Alder reaction, while that of the latter is the 6 pi-electrocyclization.

Ocular manifestations in Blau syndrome associated with a CARD15/Nod2 mutation.

PURPOSE: To report cases of Blau syndrome with a CARD15/Nod2 mutation. DESIGN: Observational and interventional case report. PARTICIPANTS: A 10-year-old Japanese boy (proband) was seen with secondary angle-closure glaucoma (iris bombe), uveitis, skin rashes, and camptodactyly. His sister had posterior synechia and camptodactyly. She had iritis in both eyes during the follow-up period. Both eyes of the father were phthisical because of granulomatous uveitis and secondary glaucoma. The father also had camptodactyly. METHODS: Surgery was performed to release the iris bombe. Ocular inflammation was treated by topical and systemic steroids. Biopsy specimens from the skin rash and from the iris (from iridectomy) were obtained from the proband. Genetic analyses were performed on the proband, his sister, and their mother for a CARD15/Nod2 mutation. MAIN OUTCOME MEASURES: Clinical features, pathologic findings of the skin and iris specimens, and genetic analysis of the CARD15/Nod2 gene. RESULTS: Phacoemulsification, intraocular lens implantation, and peripheral iridectomy released the iris bombe. The biopsy specimen from the skin rash showed noncaseating, granulomatous infiltration with epithelioid cells and lymphocytes. The iridectomy specimen showed nonspecific inflammation. Systemic and topical steroid therapy partly reduced the ocular inflammation. Genetic analyses showed that the proband and his sister had an R334W mutation in the CARD15/Nod2 gene, but their mother was of the wild type. CONCLUSIONS: Blau syndrome should be considered in the differential diagnosis of childhood uveitis. Genetic analysis of the CARD15/Nod2 gene is helpful in the diagnosis.