Akito Tsuboi

Rare variant discovery by deep whole-genome sequencing of 1,070 Japanese individuals

The Tohoku Medical Megabank Organization reports the whole-genome sequences of 1,070 healthy Japanese individuals and construction of a Japanese population reference panel (1KJPN). Here we identify through this high-coverage sequencing (32.4 × on average), 21.2 million, including 12 million novel, single-nucleotide variants (SNVs) at an estimated false discovery rate of <1.0%. This detailed analysis detected signatures for purifying selection on regulatory elements as well as coding regions. We also catalogue structural variants, including 3.4 million insertions and deletions, and 25,923 genic copy-number variants. The 1KJPN was effective for imputing genotypes of the Japanese population genome wide. These data demonstrate the value of high-coverage sequencing for constructing population-specific variant panels, which covers 99.0% SNVs of minor allele frequency ≥0.1%, and its value for identifying causal rare variants of complex human disease phenotypes in genetic association studies.

Neurons of the trigeminal main sensory nucleus participate in the generation of rhythmic motor patterns

The trigeminal principal sensory nucleus (NVsnpr) contains both trigemino‐thalamic neurons and interneurons projecting to the reticular formation and brainstem motor nuclei. Here we describe the inputs and patterns of firing of NVsnpr neurons during fictive mastication in anaesthetized and paralysed rabbits to determine the role that NVsnpr may play in patterning mastication. Of the 272 neurons recorded in NVsnpr, 107 changed their firing patterns during repetitive stimulation of the left or right sensorimotor cortex to induce fictive mastication. Thirty increased their firing tonically. Seventy‐seven became rhythmically active, but only 31 fired in phase with mastication. The others discharged in bursts at more than twice the frequency of trigeminal motoneurons. Most rhythmic masticatory neurons were concentrated in the dorsal part, and those which fired during the jaw closing phase of the cycle were confined to the anterior pole of the nucleus. Most of these cells had inputs from muscle spindle afferents, whereas most of those firing during jaw opening had inputs from periodontal receptors. Non‐masticatory rhythmical neurons had receptive fields on the lips and face. The majority of rhythmical masticatory units were modulated during fictive mastication evoked by both the left and right cortices and only four changed their phase of firing when switching from one cortex to the other. When coupled with the finding that NVsnpr neurons exhibit spontaneous bursting in vitro[Sandler et al. (1998) Neuroscience, 83, 891], the results described here suggest that neurons of dorsal NVsnpr may form the core of the central pattern generator for mastication.

The Tohoku Medical Megabank Project: Design and Mission

The Great East Japan Earthquake (GEJE) and resulting tsunami of March 11, 2011 gave rise to devastating damage on the Pacific coast of the Tohoku region. The Tohoku Medical Megabank Project (TMM), which is being conducted by Tohoku University Tohoku Medical Megabank Organization (ToMMo) and Iwate Medical University Iwate Tohoku Medical Megabank Organization (IMM), has been launched to realize creative reconstruction and to solve medical problems in the aftermath of this disaster. We started two prospective cohort studies in Miyagi and Iwate Prefectures: a population-based adult cohort study, the TMM Community-Based Cohort Study (TMM CommCohort Study), which will recruit 80 000 participants, and a birth and three-generation cohort study, the TMM Birth and Three-Generation Cohort Study (TMM BirThree Cohort Study), which will recruit 70 000 participants, including fetuses and their parents, siblings, grandparents, and extended family members. The TMM CommCohort Study will recruit participants from 2013 to 2016 and follow them for at least 5 years. The TMM BirThree Cohort Study will recruit participants from 2013 to 2017 and follow them for at least 4 years. For children, the ToMMo Child Health Study, which adopted a cross-sectional design, was also started in November 2012 in Miyagi Prefecture. An integrated biobank will be constructed based on the two prospective cohort studies, and ToMMo and IMM will investigate the chronic medical impacts of the GEJE. The integrated biobank of TMM consists of health and clinical information, biospecimens, and genome and omics data. The biobank aims to establish a firm basis for personalized healthcare and medicine, mainly for diseases aggravated by the GEJE in the two prefectures. Biospecimens and related information in the biobank will be distributed to the research community. TMM itself will also undertake genomic and omics research. The aims of the genomic studies are: 1) to construct an integrated biobank; 2) to return genomic research results to the participants of the cohort studies, which will lead to the implementation of personalized healthcare and medicine in the affected areas in the near future; and 3) to contribute the development of personalized healthcare and medicine worldwide. Through the activities of TMM, we will clarify how to approach prolonged healthcare problems in areas damaged by large-scale disasters and how useful genomic information is for disease prevention.

EMG study for perioral facial muscles function during mastication.

This study aimed to clarify the temporal and quantitative modulation in the orbicularis oris (OO) and buccinator (BUC) muscle activities during mastication. Ten healthy males (26.9 +/- 1.0 years) participated. Electromyograms (EMGs) of the facial muscles were recorded with fine wire electrodes when chewing the chewing gum (one to four sticks) and peanuts (one to five pieces). Surface EMGs of the masseter (MAS) and digastric muscles were recorded simultaneously. EMGs of the OO and BUC showed rhythmic single-peaked bursts corresponding to the jaw-opening phase of chewing cycles. The total cycle lengths were constant regardless of the food amount. Integrated EMGs of the OO changed significantly when the amount of both foods changed (anova: P < 0.05). Those of the BUC changed significantly with the amount of gum changed (P < 0.05), but did not change with the amount of peanuts changed. The burst duration of OO changed significantly when the amount of gum changed during ipsilateral chewing (P < 0.05). When the amount of peanuts changed during ipsilateral chewing, the onset of OO and the peak of BUC based on the onset of MAS activity changed significantly (P < 0.05). However, the onset, peak and offset of the OO and BUC based on the offset of MAS did not change regardless of the amounts chewed. The changes of the OO and BUC activities may derive from chewing-generated sensory inputs in accordance with the physical property of food in part, which would relate to the function of these muscles during mastication.

Possible Involvement of Histamine in Muscular Fatigue in Temporomandibular Disorders: Animal and Human Studies

As an approach to clarifying the molecular basis of pain and fatigue in muscles involved in temporomandibular disorders, we examined the activity of histidine decarboxylase (HDC), the enzyme which forms histamine, in the masseter muscles of mice. In the resting muscle, HDC activity was very low. Direct electrical stimulation of the muscle markedly elevated HDC activity. HDC activity rose within 3 hrs of the electrical stimulation, peaked at 6 to 8 hrs, and then gradually declined. Intraperitoneal injection of a small amount of interleukin-1 (IL-1) (from 1 to 10 μg/kg) produced a similar elevation of HDC activity in the masseter muscle. We also examined the effect of an antihistamine, chlorphenylamine (CP), on temporomandibular disorders in humans and compared it with that of an antiinflammatory analgesic, flurbiprofen (FB). Two groups received one or the other of the drugs daily for 7 days, and they were asked about their signs and symptoms before and after the treatment. A positive evaluation of their treatment was made by 74% of the CP group, but by only 48% of the FB group. Although the effects of CP on the limitation of mouth-opening and on joint noise were negligible, about 50% of the CP group answered positively concerning the drugs effect on spontaneous pain or pain induced by chewing or mouth-opening. The positive evaluation for CP (50%) in relieving associated symptoms (headache or shoulder stiffness) was significantly greater than for FB (13%). FB showed effectiveness similar to but sometimes weaker than that of CP on several symptoms. On the basis of these and previous results and the known actions of histamine, we propose that the histamine newly formed following the induction of HDC activity, which is itself mediated by IL-1, may be involved in inducing pain and, possibly, stiffness in muscles in temporomandibular disorders.

Inter-Individual Differences in the Oral Bacteriome Are Greater than Intra-Day Fluctuations in Individuals

Given the advent of massively parallel DNA sequencing, human microbiome is analyzed comprehensively by metagenomic approaches. However, the inter- and intra-individual variability and stability of the human microbiome remain poorly characterized, particularly at the intra-day level. This issue is of crucial importance for studies examining the effects of microbiome on human health. Here, we focused on bacteriome of oral plaques, for which repeated, time-controlled sampling is feasible. Eighty-one supragingival plaque subjects were collected from healthy individuals, examining multiple sites within the mouth at three time points (forenoon, evening, and night) over the course of 3 days. Bacterial composition was estimated by 16S rRNA sequencing and species-level profiling, resulting in identification of a total of 162 known bacterial species. We found that species compositions and their relative abundances were similar within individuals, and not between sampling time or tooth type. This suggests that species-level oral bacterial composition differs significantly between individuals, although the number of subjects is limited and the intra-individual variation also occurs. The majority of detected bacterial species (98.2%; 159/162), however, did not fluctuate over the course of the day, implying a largely stable oral microbiome on an intra-day time scale. In fact, the stability of this data set enabled us to estimate potential interactions between rare bacteria, with 40 co-occurrences supported by the existing literature. In summary, the present study provides a valuable basis for studies of the human microbiome, with significant implications in terms of biological and clinical outcomes.

Quantitative analysis of masticatory activity during unilateral mastication using muscle fMRI

OBJECTIVE Quantitative analysis of the activities of all masticatory muscles is required to elucidate the mechanism of stomatognathic dysfunction. Electromyography can be used to record the activity of masticatory muscles, but quantification of the overall activity of every masticatory muscle has not been accomplished because of methodological limitations. In this study, we used muscle functional magnetic resonance imaging for simultaneous quantification of the overall activities of the masseter, medial pterygoid and lateral pterygoid muscles during unilateral gum chewing. METHODS Seven healthy male volunteers participated in the study. We evaluated changes in the mean proton transverse relaxation time in the bilateral masseter, medial pterygoid and lateral pterygoid muscles before and after unilateral gum chewing, and to quantify the overall activity of these muscles simultaneously during unilateral gum chewing. RESULTS After 5 min of chewing, the activity of the ipsilateral masseter was highest among the six muscles, followed by the ipsilateral medial pterygoid, contralateral lateral pterygoid and contralateral masseter muscles. CONCLUSION These results affirm the importance of the ipsilateral masseter muscle and quantitatively demonstrate the important contribution of the ipsilateral medial pterygoid and contralateral lateral pterygoid muscles during unilateral mastication.

Association Between Tooth Loss, Receipt of Dental Care, and Functional Disability in an Elderly Japanese Population: The Tsurugaya Project

To examine whether number of remaining teeth and regular dental care affect incident functional disability in elderly Japanese adults.

Response properties of trigeminal ganglion mechanosensitive neurons innervating the temporomandibular joint of the rabbit

The primary mechanosensitive neurons innervating the temporomandibular joint (TMJ neurons) may play an important role in controlling mandibular movement and position. The purpose of the study was to investigate the neurophysiological properties of TMJ neurons during passive movement of the isolated condyle in 55 rabbits and the intact condyle in 29 rabbits. Discharges of TMJ neurons from the trigeminal ganglion were recorded with a microelectrode as the isolated condyle was moved manually and by a computer-regulated mechanostimulator and as the intact condyle was manually stimulated. A total of 237 TMJ neurons were recorded rostrocaudally from the mandibular nerve area lateral to the maxillary region in the dorsal half of the trigeminal ganglion. Of the recorded TMJ units, 97% were slowly adapting (SA) and 67% of the SA units had an accompanying ongoing discharge. The proportion of adaptation types and appearance of ongoing discharges for the isolated condyle did not differ significantly from those for the intact condyle. Most of the TMJ units (89%) responded multidirectionally to the rostral and ventral movements of the isolated condyle. The discharge frequencies of the TMJ units increased as the condylar displacement and velocity increased within a 5-mm anterior displacement of the isolated condyle. Displacement of the isolated condyle influenced the discharge frequency of the units to a greater extent than the velocity of the condyle movement. No responses of TMJ units were observed during the descending ramp. Based on these results, we conclude that sensory information is transmitted by TMJ neurons encoding joint position, displacement and velocity in a physiological range of mandibular displacement.

Response properties of periodontal mechanosensitive neurons in the trigeminal ganglion of rabbit and neuronal activities during grinding-like jaw movement induced by cortical stimulation

OBJECTIVE To examine the response properties of incisor- and molar-sensitive periodontal mechanosensitive (PM) neurons in the trigeminal ganglion of rabbit and the activities of the molar-sensitive PM neurons during the grinding-like jaw movement. DESIGN Discharges of PM units were recorded from the trigeminal ganglion with a microelectrode. The grinding-like jaw movement was induced by repetitive electrical stimulation of the cortical masticatory area. RESULTS Upper-incisor (UI) and upper-molar (UM) units were recorded from the rostromedial area of the trigeminal ganglion, and lower-incisor (LI) and lower-molar (LM) units were distributed in the caudolateral area. Most PM units were responsive to only one tooth, slowly adapting ones and responded to tooth stimulation of a force of less than 0.05 N. The optimal stimulus direction for most UI units was labio-lingual, axial or linguo-labial, and that for most LI units was linguo-labial or axial. The optimal stimulus direction of anterior UM and LM units was oriented predominantly mesio-distal or axial. The maximum frequency of spike discharges for UM units for which the optimal stimulus direction was axial or bucco-lingual was in the middle period of the grinding phase. However, UM units for which the optimal stimulus direction was mesio-distal or linguo-buccal were fired mostly in the early period. CONCLUSIONS Periodontal sensory information in the grinding phase of jaw movement is transmitted by PM neurons with various response properties encoding the magnitude and direction of a force at least, in a weaker range of force than a saturating response level.