Expression of huntingtin-associated protein 1 in adult mouse dorsal root ganglia and its neurochemical characterization in reference to sensory neuron subpopulations.

Huntingtin-associated protein 1 (HAP1) is a polyglutamine (polyQ) length-dependent interactor with causal agents in several neurodegenerative diseases and has been regarded as a protective factor against neurodegeneration. In normal rodent brain and spinal cord, HAP1 is abundantly expressed in the areas that are spared from neurodegeneration while those areas with little HAP1 are frequent targets of neurodegeneration. We have recently showed that HAP1 is highly expressed in the spinal dorsal horn and may participate in modification/protection of certain sensory functions. Neurons in the dorsal root ganglia (DRG) transmits sensory stimuli from periphery to spinal cord/brain stem. Nevertheless, to date HAP1 expression in DRG remains unreported. In this study, the expression of HAP1 in cervical, thoracic, lumbar and sacral DRG in adult male mice and its relationships with different chemical markers for sensory neurons were examined using Western blot and immunohistochemistry. HAP1-immunoreactivity was detected in the cytoplasm of DRG neurons, and the percentage of HAP1-immunoreactive (ir) DRG neurons was ranged between 28-31 %. HAP1-immunoreactivity was comparatively more in the small cells (47-58 %) and medium cells (40-44 %) than that in the large cells (9-11 %). Double-immunostaining for HAP1 and markers for nociceptive or mechanoreceptive neurons showed that about 70-80 % of CGRP-, SP-, CB-, NOS-, TRPV1-, CR- and PV-ir neurons expressed HAP1. In contrast, HAP1 was completely lacking in TH-ir neurons. Our current study is the first to clarify that HAP1 is highly expressed in nociceptive/proprioceptive neurons but absent in light-touch-sensitive TH neurons, suggesting the potential importance of HAP1 in pain transduction and proprioception.

Geometric assessment of imaging methods for complete denture form: Comparisons among cone-beam computed tomography, desktop dental scanning, and handheld optical scanning.

PURPOSE: The present study aimed to examine three methods for imaging a complete denture form and to discuss the geometric assessment and availability of these methods. METHODS: Three methods were used: cone-beam computed tomography (CBCT), desktop dental scanning (DDS), and handheld optical scanning (HOS). Three sides of a cuboid resin block of a known size were measured on the captured images. Ten upper and lower complete dentures were digitized using each of the three methods. The denture forms in the standard triangulated language format were reconstructed using the computer aided design software with three resolutions: 0.1, 0.5 and 1.0. The two images were superimposed through a least-square algorithm, and the mean differences were examined among the three types of imaging methods and resolutions. RESULTS: The smallest mean differences and variations were obtained for CBCT and DDS, followed by HOS in the experiment using resin block. The lengths of longitudinal and horizontal sides were shortened in CBCT and DDS, whereas the height was shortened in HOS. The variations in each direction were very small by repetitive imaging within every imaging method. The difference between the CBCT and DDS was significantly smaller than that in the other two groups in experiment using actual dentures. The influence of resolution was comparatively low. CONCLUSIONS: The results suggested that CBCT and DDS had a similar geometric assessment, but that of the HOS was lower.

A single male auditory response test to quantify auditory behavioral responses in Drosophila melanogaster.

Many animals utilize auditory signals to communicate with conspecific individuals. During courtship, males of the fruit fly Drosophila melanogaster and related species produce a courtship song comprised of sine and pulse songs by vibrating their wings. The pulse song increases female receptivity and male courtship activity, indicating that it functions as a sexual signal. One song parameter, interpulse interval (IPI), varies among closely related species. In D. melanogaster, a song with a conspecific IPI induces a stronger behavioral response than heterospecific songs, indicating the ability of the flies to discriminate conspecific IPI. Traditionally, the fly's response to the song is measured under grouped conditions, in which the effect of sensory modalities other than audition cannot be excluded. Here, to quantify the individual ability to discriminate a conspecific song, we systematically analyzed the auditory response of single male flies to sound with various parameters. Moreover, we applied this method, termed SMART (Single Male Auditory Response Test), to two sister species for potential application in a comparative approach. By quantifying the locomotor activity of single D. melanogaster males during sound exposure, we detected increased locomotor activity in response to pulse songs, but not to white noise or pure tone. The conspecific song evoked stronger response than the heterospecific songs, and ablation of their antennal receivers severely suppressed the locomotor increase. A pulse song with a small IPI variation evoked a continuous response, while the response to songs with highly variable IPIs tends to be rapidly decayed. This provides the first evidence that fruit flies discriminate IPI variations, which possibly inform the age and social contexts of the singer. Sister species, D. sechellia, exhibited a locomotor response to pulse song, while D. simulans exhibited no behavioral response. This suggests that auditory and other stimuli that elicit this behavioral response are diversified among Drosophila species.

Rapid Return for School Refusal: A School-Based Approach Applied With Japanese Adolescents.

Cognitive behavioral therapy (CBT) is often effective in the treatment of school refusal (SR). Its usefulness is limited, however, if youth displaying SR also refuse to attend treatment sessions. In these cases parents and school staff may consider using school-based interventions that do not rely on face-to-face assessment and treatment with the young person. The current study examined the effectiveness of a school-based intervention applied in Japan to achieve rapid return to school among adolescents displaying SR. Between 2009 and 2015, the parents of 62 adolescents displaying SR were invited to implement a school-based rapid return approach. Thirty-nine parents agreed to implement the approach and 23 decided to wait until their child spontaneously attended school. Of the 39 cases in which the approach was implemented, 28 adolescents (72%) resumed attendance at their original school, 2 (5%) transferred to another school, and 9 (23%) did not resume attendance. In contrast, all 23 non-intervention cases continued to refuse to attend school for 3 months or longer, and none of these adolescents returned to regular school attendance within 9 months. This study tentatively suggests that the rapid return approach may be an effective form of intervention for adolescents displaying SR and simultaneously refusing to attend individual therapy. Because this approach is ethically complex, involving forced school attendance in adolescence, it should only be employed under specific circumstances. These circumstances are discussed.

Dental students' levels of understanding normal panoramic anatomy.

BACKGROUND/PURPOSE: In dentistry, panoramic radiography is an important examination technique. The Faculty of Dentistry at Tokushima University educates students about panoramic radiographic anatomical landmarks. The purpose of this study was to investigate differences in the understanding of each panoramic anatomical landmark among students. MATERIALS AND METHODS: This study analyzed the results of 40 fifth- and 79 sixth-year faculty students who had taken a written examination to clarify their knowledge of anatomical landmarks in 28 panoramic radiographic regions. Anatomical landmarks were classified into 3 categories: , , and  to compare correct answer rates. RESULTS: The mean overall correct answer rate by the 119 students for the 28 regions was 53%. The rate did not vary between the 2 academic years. On comparison of the 3 categories, significant differences were only observed between  and , as the values for the latter were lower. Among the anatomical landmarks, the rates for the condylar head, hyoid bone, panoramic innominate line, styloid process, and nasopalatine line were higher, and those for ghost images of the contralateral nasopalatine line, the cervical vertebrae, mandibular foramen, mastoid air cell, and posterior pharyngeal wall were lower. The values for , such as the dorsum of tongue, middle and inferior nasal conche, and middle and inferior nasal meatuses, were also lower. CONCLUSION: These results indicate the necessity of improving educational approaches for regions with lower rates of correct answers.

Numerical Evaluation of Image Contrast for Thicker and Thinner Objects among Current Intraoral Digital Imaging Systems.

The purpose is to evaluate the performance of current intraoral digital detectors in detail using a precise phantom and new method. Two aluminum step wedges in 0.5 mm steps were exposed by two photostimulable phosphor plate (PSP) systems-one with automatic exposure compensation (AEC) and the other without AEC-and a CCD sensor. Images were obtained with 3 doses at 60 kV. The effect of metallic material also was evaluated. The contrast-to-noise ratio (CNR) for thinner steps and the low contrast value (LCV) for thicker steps were obtained. The CCD system was the best under all conditions (P < 0.001), although the Gray value was sensitive to the dose, and the Gray value-dose relation varied greatly. The PSP system with AEC was superior to that without AEC for the LCV (P < 0.001) but was inferior to it regarding the CNR (P < 0.001). CNR and LCV in the PSP system without AEC were not affected by the metallic plate. Intraoral digital imaging systems should be chosen according to their diagnostic purpose. PSP system with AEC may be the best for detecting molar proximal caries, whereas the PSP system without AEC may be better for evaluating small bone regeneration in periodontal disease. The CCD system provided the best performance.

Discrimination between Conglomerates and Pseudoracemates Using Surface Coverage Plots in 2D Self-Assemblies at the Liquid-Graphite Interface.

The two-dimensional (2D) molecular ordering of two photochromic diarylethenes was investigated using scanning tunneling microscopy at the liquid-graphite interface. The racemic mixture of the closed-ring isomer of one of the diarylethenes showed spontaneous separation of its enantiomers upon 2D crystallization, whereas that of the other diarylethene formed a pseudoracemic crystal in which two enantiomers coexist in a 2D ordering domain. The mixing of enantiomers in 2D assemblies can be analyzed by the dependence of the surface coverage on the concentration of enantiomers at different enantiomeric ratios.

Availability of Japanese Government's supplemental texts on radiation reflecting the Fukushima Daiichi Nuclear Power Plant accident for elementary and secondary education from dental students' understanding.

Following the Fukushima Nuclear Power Plant accident, the Japanese government created two supplemental texts about radiation reflecting the accident for elementary, middle school, and high school students. These texts were made to explain radiation and consequently to obtain public consent for the continuation of the nuclear program. The present study aimed to evaluate the appropriateness of the content of the texts and to collect the basic data on the level of understanding necessary to improve radiation education. Lectures on radiology including nuclear energy and the Fukushima accident were given to 44 fourth-year dental students in 2013. The questionnaire was administered in 2014 when these students were in their sixth-year. The survey was also administered to 40 first-year students and 41 fourth-year students who hadn't any radiology lectures. Students rated their level of understanding of 50 phrases used in the texts on a four-point scale (understanding = 3, a little knowledge = 2, having heard = 1, no knowledge = 0). Questions on taking an advanced physics course in high school and means of learning about radiation in daily life were also asked. The level of understanding of phrases in the supplemental text for middle and high school students was significantly higher among sixth-year students (mean = 1.43) than among first-year (mean = 1.12) or fourth-year (mean = 0.93) students (p < 0.05). Overall, the level of understanding was low, with scores indicating that most students knew only a little. First-year students learning about radiation from television but four-year and six-year students learning about radiation from newspaper scored significantly higher (p < 0.05). It was concluded that radiation education should be improved by using visual material and preparing educators to teach the material for improving the public's understanding of radiation use-especially nuclear power generation because the phrases used in the supplementary texts are very difficult for students to understand.

Oral administration of monogalactosyl diacylglycerol from spinach inhibits colon tumor growth in mice.

Previously, we observed that purified monogalactosyl diacylglycerol (MGDG), a major glycoglycerolipid from spinach, selectively inhibits the activities of mammalian replicative DNA polymerases (α, δ and ε). However, the function of MGDG following ingestion is not well-known. In the present study, spinach MGDG suppressed the proliferation of Colon26 mouse colon cancer cells with an LD(50) of 24 µg/ml in vitro. γ-cyclodextrin (CD)-MGDG complex was prepared and administered orally following Colon26 mouse tumor adhesion for 26 days. It was observed that 20 mg/kg equivalent (eq.) of the CD-MGDG complex reduced tumor volume by ∼60% compared with that of the vehicle-treated controls. In immunohistochemical analysis, the CD-MGDG complex group showed a decreased number of proliferating cell nuclear antigen (PCNA)-positive cells and reduction of mitosis in the tumor cells compared with the control group. In addition, the CD-MGDG complex increased the number of terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL)-positive apoptotic cells and inhibited CD31-positive tumor blood vessel growth significantly. These results suggest that MGDG has the potential for cancer prevention and health promotion.

Plasmonic and Mie scattering control of far-field interference for regular ripple formation on various material substrates.

We present experimental and theoretical results on plasmonic control of far-field interference for regular ripple formation on semiconductor and metal. Experimental observation of interference ripple pattern on Si substrate originating from the gold nanosphere irradiated by femtosecond laser is presented. Gold nanosphere is found to be an origin for ripple formation. Arbitrary intensity ripple patterns are theoretically controllable by depositing desired plasmonic and Mie scattering far-field pattern generators. The plasmonic far-field generation is demonstrated not only by metallic nanostructures but also by the controlled surface structures such as ridge and trench structures on various material substrates.

Dehydroaltenusin is a specific inhibitor of mammalian DNA polymerase α.

INTRODUCTION: We carried out a screen for small molecule selective inhibitors of eukaryotic DNA polymerases (pols). Dehydroaltenusin, isolated from a fungus (Alternaria tenuis), was found to be a specific inhibitor of pol α. AREAS COVERED: We succeeded in chemically synthesizing dehydroaltenusin along with five analogs. Of these compounds, dehydroaltenusin was the strongest and most specific inhibitor of mammalian pol α, with an IC(50) value of 0.68 µM. The inhibitory mode of action of dehydroaltenusin against mammalian pol α activity was competitive with respect to the DNA template primer and non-competitive with respect to the 2'-deoxyribonucleoside 5'-triphosphate substrate. Dehydroaltenusin inhibited the cell proliferation of a human cervical cancer cell line, HeLa, by arresting the cells at the S-phase, and preventing the incorporation of thymidine into the cells. These observations indicate that dehydroaltenusin blocks in vivo DNA replication by inhibiting pol α. EXPERT OPINION: Dehydroaltenusin was effective in suppressing the growth of solid tumors and, therefore, is of interest as a candidate drug for anti-cancer treatment.

Cardiac arrest due to food asphyxiation in adults: resuscitation profiles and outcomes.

AIM: Food asphyxiation is uncommon but unignorable cause of sudden death in the elderly. Several autopsy studies, which identified those at particular risk, have been conducted on the subject. Resuscitation profiles and outcomes of food asphyxiation victims presenting with out-of-hospital cardiac arrest (OHCA) to the emergency department, however, have rarely been reported. METHODS: Data on adults (> or = 20 years) presenting with OHCA after witnessed food asphyxiation were retrieved from an institutional database. Clinical variables were evaluated to identify their demographic characteristics. Their outcomes, represented by return of spontaneous circulation (ROSC) and survival rate, were also investigated. RESULTS: Sixty-nine food asphyxiation victims presenting with OHCA were identified during the 4-year period. Food asphyxiation occurred most frequently in the age group of 71-80 years, followed by that of 81-90 years. The majority of victims had medical conditions that adversely affected mastication/swallowing, such as dementia. Bystander cardiopulmonary resuscitation (CPR) was performed only in 26%, although bystanders often attempted to clear the airway without performing CPR. Despite the high ROSC rate of 78%, only 7% survived to discharge. Asphyxiation-ROSC interval might play a crucial role in determining the outcomes: the interval was < or = 10 min in all survivors, while it was longer than 10 min in all non-survivors. CONCLUSION: Because of their advanced age and debilitating general condition, it may be difficult to substantially improve the outcomes of food asphyxiation victims. Effort should be directed to prevent food asphyxiation, and public education to perform standard CPR for food asphyxiation victims including the Heimlich manoeuvre is warranted.

Subarachnoid haemorrhage as a cause of out-of-hospital cardiac arrest: a prospective computed tomography study.

AIM: Aneurysmal subarachnoid haemorrhage (SAH) is a relatively common cause of out-of-hospital cardiac arrest (OHCA). Early identification of SAH-induced OHCA with the use of brain computed tomography (CT) scan obtained immediately after resuscitation may help emergency physicians make therapeutic decision as quickly as they can. METHODS: During the 4-year observation period, brain CT scan was obtained prospectively in 142 witnessed non-traumatic OHCA survivors who remained haemodynamically stable after resuscitation. Demographics and clinical characteristics of SAH-induced OHCA survivors were compared with those with "negative" CT finding. RESULTS: Brain CT scan was feasible with an average door-to-CT time of 40.0 min. SAH was found in 16.2% of the 142 OHCA survivors. Compared with 116 survivors who were negative for SAH, SAH-induced OHCA survivors were significantly more likely to be female, to have experienced a sudden headache, and trended to have achieved return of spontaneous circulation (ROSC) prior to arrival in the emergency department less frequently. Ventricular fibrillation (VF) was significantly less likely to be seen in SAH-induced than SAH-negative OHCA (OR, 0.06; 95% CI, 0.01-0.46). Similarly, Cardiac Trop-T assay was significantly less likely to be positive in SAH-induced OHCA (OR, 0.08; 95% CI, 0.01-0.61). CONCLUSION: Aneurysmal SAH causes OHCA more frequently than had been believed. Immediate brain CT scan may particularly be useful in excluding SAH-induced OHCA from thrombolytic trial enrollment, for whom the use of thrombolytics is contraindicated. The low VF incidence suggests that VF by itself may not be a common cause of SAH-induced OHCA.

Structure and activity relationship of monogalactosyl diacylglycerols, which selectively inhibited in vitro mammalian replicative DNA polymerase activity and human cancer cell growth.

The glycoglycerolipid, monogalactosyl diacylglycerol (MGDG), containing two alpha-linolenic acids (C18:3), was isolated from bitter melon as a potent and selective inhibitor of mammalian DNA polymerase (pol) species such as pols alpha, gamma, delta, and epsilon with IC(50) values of 17.6-37.2muM. This MGDG also suppressed the growth of six human cancer cell lines, although normal human cell lines were not affected. This compound (i.e., MGDG-C18:3-C18:3) was a stronger inhibitor than both MGDG-C18:1-C18:0 and MGDG-C18:0-C18:0. In this study, we discussed the structure-function relationship in the selective inhibition of mammalian replicative pols and human cancer cell proliferation by MGDGs.

Anti-tumor effect of orally administered spinach glycolipid fraction on implanted cancer cells, colon-26, in mice.

We succeeded in purifying a major glycolipid fraction from a green vegetable, spinach. This fraction consists mainly of three glycolipids: monogalactosyl diacylglycerol (MGDG), digalactosyl diacylglycerol (DGDG), and sulfoquinovosyl diacylglycerol (SQDG). In a previous study, we found that the glycolipid fraction inhibited DNA polymerase activity, cancer cell growth and tumor growth with subcutaneous injection. We aimed to clarify oral administration of the glycolipid fraction, suppressing colon adenocarcinoma (colon-26) tumor growth in mice. A tumor graft study showed that oral administration of 20 mg/kg glycolipid fraction for 2 weeks induced a 56.1% decrease in the solid tumor volume (P < 0.05) without any side-effects, such as loss of body weight or major organ failure, in mice. The glycolipid fraction induced the suppression of colon-26 tumor growth with inhibition of angiogenesis and the expression of cell proliferation marker proteins such as Ki-67, proliferating cell nuclear antigen (PCNA), and Cyclin E in the tumor tissue. These results suggest that the orally administered glycolipid fraction from spinach could suppress colon tumor growth in mice by inhibiting the activities of neovascularization and cancer cellular proliferation in tumor tissue.

The inhibitory action of SQDG (sulfoquinovosyl diacylglycerol) from spinach on Cdt1-geminin interaction.

A human replication initiation protein, Cdt1, is a central player in the cell cycle regulation of DNA replication, and geminin down-regulates Cdt1 function by direct binding. It has been demonstrated that Cdt1 hyperfunction resulting from Cdt1-geminin imbalance, for example, by geminin silencing with small interfering RNA, induces DNA re-replication and eventual cell death in some cancer-derived cell lines. We established a high throughput screening system based on a modified enzyme linked immunosorbent assay to identify compounds that interfere with human Cdt1-geminin binding. Using this system, we screened inhibitors from natural materials containing food components, and found that a glycolipid, sulfoquinovosyl diacylglycerol (SQDG), from spinach can inhibit Cdt1-geminin interaction in vitro, with 50% inhibition observed at concentrations of 1.79mug/ml. Other major glycolipids, such as monogalactosyl diacylglycerol (MGDG) and digalactosyl diacylglycerol (DGDG) from spinach, had no influence. Surface plasmon resonance analysis demonstrated that SQDG bound selectively to Cdt1, but did not interact with geminin. Using three-dimensional computer modeling analysis, SQDG was considered to interact with the geminin interaction interface on Cdt1, and the sulfate group of SQDG was assumed to make hydrogen bonds with the residue of Arg346 of Cdt1. These data could help to further understanding of the structure and function of Cdt1. In addition, SQDG could be a clue to developing more appropriate inhibitors of Cdt1-geminin interactions.

Megacolon in an adult case of hypoganglionosis, a pseudo-Hirschsprung's disease: an autopsy study.

We report an autopsied 20-year-old man case of intestinal necrosis associated with megacolon from hypoganglionosis, a pseudo-Hirschsprung's disease. The patient had suffered from severe constipation since two years of age, and presented abdominal distention from age ten. Autopsy revealed marked dilatation and necrosis of the entire large intestine. Although ganglion cells in the intestinal plexus were found throughout the large intestine, their number was reduced to 12-20% of that in the normal control. In pseudo-Hirschsprung's disease, there are occasional cases where an acute abdomen first presents itself in adulthood after running its course as chronic constipation.

Anti-tumor effects of the glycolipids fraction from spinach which inhibited DNA polymerase activity.

We succeeded in purifying the fraction of monogalactosyl diacylglycerol (MGDG), digalactosyl diacylglycerol (DGDG), and sulfoquinovosyl diacylglycerol (SQDG) containing the major glycolipids from a green vegetable, spinach (Spinacia oleraceaL.). This glycolipids fraction inhibited the activities of replicative DNA polymerases (pols) such as alpha, delta, and epsilon, and mitochondrial pol gamma with IC50 values of 44.0-46.2 microg/ml, but had no influence on the activity of repair-related pol beta. The fraction also inhibited the proliferation of human cervix carcinoma (HeLa) cells with LD50 values of 57.2 microg/ml. In an in vivo anti-tumor assay on nude mice bearing solid tumors of HeLa cells, the fraction was shown to be a promising suppressor of solid tumors. Histopathological examination revealed that tumor necrosis with hemorrhage was significantly enhanced with the glycolipids fraction in vivo. The spinach glycolipids fraction might be a potent anti-tumor compound, and this fraction may be a healthy food substance with anti-tumor activity.

Inhibitory effect on replicative DNA polymerases, human cancer cell proliferation, and in vivo anti-tumor activity by glycolipids from spinach.

We succeeded in purifying a major glycolipids fraction (i.e., Fraction-II) in the class of monogalactosyl diacylglycerol (MGDG), digalactosyl diacylglycerol (DGDG) and sulfoquinovosyl diacylglycerol (SQDG) from spinach using hydrophobic column chromatography. Fraction-II inhibited the activities of replicative DNA polymerases (pols) such as alpha, delta and epsilon, and mitochondrial pol gamma with IC(50) values of 43-79 microg/ml, but had no influence on the activity of repair-related pols beta and lambda. MGDG, DGDG, SQDG were purified from Fraction-II of spinach using silica gel column chromatography, and SQDG was the strongest inhibitor of mammalian pols in the three glycolipids. Therefore, SQDG and its related compounds were chemically synthesized, and the sulfate group and fatty acid moiety of the compound were suggested to be important for pol inhibition. These glycolipids showed no effect even on the activities of plant pols, prokaryotic pols and other DNA metabolic enzymes such as T4 polynucleotide kinase, T7 RNA polymerase and deoxyribonuclease I. Fraction-II also inhibited the proliferation of human cervix carcinoma (HeLa) cells with LD(50) values of 57 microg/ml, and could halt the cell cycle at the G1-phase, and subsequently induced severe apoptosis. In an in vivo anti-tumor assay on nude mice bearing solid tumors of HeLa cells, Fraction-II was shown to be a promising suppressor of solid tumors. Histopathological examination revealed that tumor necrosis with hemorrhage was significantly enhanced with Fraction-II in vivo. The spinach Fraction-II containing SQDG might be a potent anti-tumor compound, and may be a healthy food substance with anti-tumor activity.