Examination of the relationship between masseter muscle activity during sleep and wakefulness measured by using a wearable electromyographic device.

Purpose This study aimed to measure masseter muscle activity throughout the day in outpatients suspected of having awake bruxism (AB) and/or sleep bruxism (SB) and examine the relationship between AB and SB by comparing muscle activity during daytime wakefulness and nighttime sleep.Methods Fifty outpatients with suspected SB and/or AB participated in this study. A single-channel wearable electromyogram (EMG) device was used for EMG recording. The selected EMG bursts were divided into bursts during sleep (S-bursts) and bursts during awake state (A-bursts). The number of bursts per hour, average burst duration, and ratio of burst peak value to maximum voluntary contraction were calculated for both the S- and A-bursts. These values of the S- and A-bursts were then compared, and the correlations between them were analyzed. Additionally, the ratios of phasic and tonic bursts in the S- and A-bursts were compared.Results The number of bursts per hour was significantly higher for A-bursts than for S-bursts. No significant correlation was found between the numbers of S- and A-bursts. The ratio of phasic bursts was large and that of tonic bursts was small in both the S- and A-bursts. A comparison of the S- and A-bursts showed that the S-bursts had a significantly lower ratio of phasic bursts and higher ratio of tonic bursts than the A-bursts.Conclusions The number of masseteric EMG bursts during wakefulness did not show any association with that during sleep. It became clear that sustained muscle activity was not dominant in AB.

Frequency distribution of the number and amplitude of electromyographic waveforms of the masseter muscle during sleep in patients with a clinical diagnosis of sleep bruxism.

OBJECTIVE: This study aimed to clarify frequency distribution of number and peak amplitude of electromyographic (EMG) waveforms of sleep bruxism (SB) in outpatients with clinical diagnosis of SB (probable bruxer: P-bruxer). METHODS: Subjects were 40 P-bruxers. Masseteric EMG during sleep was measured at home using a wearable EMG system. EMG waveforms with amplitude of more than two times the baseline and with duration of 0.25 s were extracted as SB bursts. Clusters of bursts, i.e. SB episodes, were also scored. RESULTS: There were large variations among the subjects in numbers of SB bursts and episodes and in burst peak amplitude. As for burst peak amplitude within a subject, a wide right-tailed frequency distribution was shown with the highest frequency at the class of 5-10% maximum voluntary contraction. CONCLUSION: The number and amplitude of SB waveforms for P-bruxers were distributed over a wide range, indicating the existence of large individual differences.

Comparison of the occurrence of sleep bruxism under accustomed conditions at home and under polysomnography conditions in a sleep laboratory.

PURPOSE: We aimed to clarify the relationship between the number of sleep bruxism (SB) bursts at home and in a laboratory equipped with polysomnography with audio-video recording (PSG-AV). We applied an identical single-channel wearable electromyography (EMG) device for both types of SB burst scorings. METHODS: The subjects were 20 healthy student volunteers (12 men and 8 women; mean age, 21.9 years) who were clinically diagnosed with bruxism based on the criteria set forth by the International Classification of Sleep Disorders (ICSD-2). We used a wearable EMG device attached to the masseteric area (the FLA-500-SD [FLA]), for scoring SB bursts at home and in the laboratory. PSG-AV was set within the laboratory environment as well. The mean interval for both sleep studies was 28.8 days. EMG bursts with amplitudes greater than twice the baseline amplitude and with durations of longer than 0.25 s were selected. EMG bursts with amplitudes ≥5% MVC (maximum voluntary contraction), ≥10% MVC, and ≥20% MVC were selected as well. A cluster of bursts was defined as an episode. RESULTS: In all the conditions for selecting EMG bursts specified above, the number of SB bursts and episodes recorded under laboratory conditions was statistically significantly smaller than that recorded at home. There were no statistically significant differences between the data obtained on the first and second recording days. CONCLUSION: The results of this study suggest that the unfamiliar environment of a sleep laboratory equipped with PSG-AV affects the emergence of SB as compared with home conditions.

Identification of Transcription Factors and the Regulatory Genes Involved in Triacylglycerol Accumulation in the Unicellular Red Alga Cyanidioschyzon merolae.

Microalgal triacylglycerols (TAGs) are a good feedstock for liquid biofuel production. Improving the expression and/or function of transcription factors (TFs) involved in TAG accumulation may increase TAG content; however, information on microalgae is still lacking. In this study, 14 TFs in the unicellular red alga Cyanidioschyzon merolae were identified as candidate TFs regulating TAG accumulation using available transcriptome and phosphoproteome data under conditions driving TAG accumulation. To investigate the roles of these TFs, we constructed TF-overexpression strains and analyzed lipid droplet (LD) formation and TAG contents in the cells grown under standard conditions. Based on the results, we identified four TFs involved in LD and TAG accumulation. RNA-Seq analyses were performed to identify genes regulated by the four TFs using each overexpression strain. Among the TAG biosynthesis-related genes, only the gene encoding the endoplasmic reticulum-localized lysophosphatidic acid acyltransferase 1 (LPAT1) was notably increased among the overexpression strains. In the LPAT1 overexpression strain, TAG accumulation was significantly increased compared with the control strain under normal growth conditions. These results indicate that the four TFs positively regulate TAG accumulation by changing their target gene expression in C. merolae.

Establishment of a firefly luciferase reporter assay system in the unicellular red alga Cyanidioschyzon merolae.

The firefly luciferase (Luc) reporter assay is a powerful tool used to analyze promoter activities in living cells. In this report, we established a firefly Luc reporter assay system in the unicellular model red alga Cyanidioschyzon merolae. A nitrite reductase (NIR) promoter-Luc fusion gene was integrated into the URA5.3 genomic region to construct the C. merolae NIR-Luc strain. Luc activities in the NIR-Luc strain were increased, correlating with the accumulation of endogenous NIR transcripts in response to nitrogen depletion. Luc activity was also significantly increased by the overexpression of the MYB1 gene, which encodes a transcription factor responsible for NIR promoter activation. Thus, our results demonstrate the utility of the Luc reporter system in C. merolae.

Loss of p57KIP2 expression confers resistance to contact inhibition in human androgenetic trophoblast stem cells.

A complete hydatidiform mole (CHM) is androgenetic in origin and characterized by enhanced trophoblastic proliferation and the absence of fetal tissue. In 15 to 20% of cases, CHMs are followed by malignant gestational trophoblastic neoplasms including choriocarcinoma. Aberrant genomic imprinting may be responsible for trophoblast hypertrophy in CHMs, but the detailed mechanisms are still elusive, partly due to the lack of suitable animal or in vitro models. We recently developed a culture system of human trophoblast stem (TS) cells. In this study, we apply this system to CHMs for a better understanding of their molecular pathology. CHM-derived TS cells, designated as TSmole cells, are morphologically similar to biparental TS (TSbip) cells and express TS-specific markers such as GATA3, KRT7, and TFAP2C. Interestingly, TSmole cells have a growth advantage over TSbip cells only after they reach confluence. We found that p57KIP2, a maternally expressed gene encoding a cyclin-dependent kinase inhibitor, is strongly induced by increased cell density in TSbip cells, but not in TSmole cells. Knockout and overexpression studies suggest that loss of p57KIP2 expression would be the major cause of the reduced sensitivity to contact inhibition in CHMs. Our findings shed light on the molecular mechanism underlying the pathogenesis of CHMs and could have broad implications in tumorigenesis beyond CHMs because silencing of p57KIP2 is frequently observed in a variety of human tumors.

Accelerated triacylglycerol production without growth inhibition by overexpression of a glycerol-3-phosphate acyltransferase in the unicellular red alga Cyanidioschyzon merolae.

Microalgae accumulate triacylglycerols (TAGs), a promising feedstock for biodiesel production, under unfavorable environmental or stress conditions for their growth. Our previous analyses revealed that only transcripts of CmGPAT1 and CmGPAT2, both encoding glycerol-3-phosphate acyltransferase, were increased among fatty acid and TAG synthesis genes under TAG accumulation conditions in the red alga Cyanidioschyzon merolae. In this study, to investigate the role of these proteins in TAG accumulation in C. merolae, we constructed FLAG-fused CmGPAT1 and CmGPAT2 overexpression strains. We found that CmGPAT1 overexpression resulted in marked accumulation of TAG even under normal growth conditions, with the maximum TAG productivity increased 56.1-fold compared with the control strain, without a negative impact on algal growth. The relative fatty acid composition of 18:2 in the TAGs and the sn-1/sn-3 positions were significantly increased compared with the control strain, suggesting that CmGPAT1 had a substrate preference for 18:2. Immunoblot analysis after cell fractionation and immunostaining analysis demonstrated that CmGPAT1 localizes in the endoplasmic reticulum (ER). These results indicate that the reaction catalyzed by the ER-localized CmGPAT1 is a rate-limiting step for TAG synthesis in C. merolae, and would be a potential target for improvement of TAG productivity in microalgae.

Derivation of Human Trophoblast Stem Cells.

Trophoblast cells play an essential role in the interactions between the fetus and mother. Mouse trophoblast stem (TS) cells have been derived and used as the best in vitro model for molecular and functional analysis of mouse trophoblast lineages, but attempts to derive human TS cells have so far been unsuccessful. Here we show that activation of Wingless/Integrated (Wnt) and EGF and inhibition of TGF-ß, histone deacetylase (HDAC), and Rho-associated protein kinase (ROCK) enable long-term culture of human villous cytotrophoblast (CT) cells. The resulting cell lines have the capacity to give rise to the three major trophoblast lineages, which show transcriptomes similar to those of the corresponding primary trophoblast cells. Importantly, equivalent cell lines can be derived from human blastocysts. Our data strongly suggest that the CT- and blastocyst-derived cell lines are human TS cells, which will provide a powerful tool to study human trophoblast development and function.