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Abstract Background Cryptomeria japonica D. Don is one of the most commercially important conifers in Japan. However, the allergic disease caused by its pollen is a severe public health problem in Japan. Since large-scale analysis of expressed sequence tags (ESTs) in the male strobili of C. japonica should help us to clarify the overall expression of genes during the process of pollen development, we constructed a full-length enriched cDNA library that was derived from male strobili at various developmental stages. Results We obtained 36,011 expressed sequence tags (ESTs) from either one or both ends of 19,437 clones derived from the cDNA library of C. japonica male strobili at various developmental stages. The 19,437 cDNA clones corresponded to 10,463 transcripts. Approximately 80% of the transcripts resembled ESTs from Pinus and Picea, while approximately 75% had homologs in Arabidopsis. An analysis of homologies between ESTs from C. japonica male strobili and known pollen allergens in the Allergome Database revealed that products of 180 transcripts exhibited significant homology. Approximately 2% of the transcripts appeared to encode transcription factors. We identified twelve genes for MADS-box proteins among these transcription factors. The twelve MADS-box genes were classified as DEF/GLO/GGM13-, AG-, AGL6-, TM3- and TM8-like MIKCC genes and type I MADS-box genes. Conclusion Our full-length enriched cDNA library derived from C. japonica male strobili provides information on expression of genes during the development of male reproductive organs. We provided potential allergens in C. japonica. We also provided new information about transcription factors including MADS-box genes expressed in male strobili of C. japonica. Large-scale gene discovery using full-length cDNAs is a valuable tool for studies of gymnosperm species.

Abstract Background Populus is one of favorable model plants because of its small genome. Structural genomics of Populus has reached a breakpoint as nucleotides of the entire genome have been determined. Reaching the post genome era, functional genomics of Populus is getting more important for well-comprehended plant science. Development of bioresorce serving functional genomics is making rapid progress. Huge efforts have achieved deposits of expressed sequence tags (ESTs) in various plant species consequently accelerating functional analysis of genes. ESTs from full-length cDNA clones are especially powerful for accurate molecular annotation. We promoted collection and annotation of the ESTs from Populus full-length enriched cDNA clones as part of functional genomics of tree species. Results We have been collecting the full-length enriched cDNA of the female poplar (Populus nigra var. italica) for years. By sequencing P. nigra full-length (PnFL) cDNA libraries, we generated about 116,000 5'-end or 3'-end ESTs corresponding to 19,841 nonredundant PnFL clones. Population of PnFL cDNA clones represents 44% of the predicted genes in the Populus genome. Conclusion Our resource of P. nigra full-length enriched clones is expected to provide valuable tools to gain further insight into genome annotation and functional genomics in Populus.

This study attempted to classify a small amount of electroencephalogram (EEG) data on five states: four tasks involving right index-finger flexion (kinesthetic motor imagery, visual motor imagery, motor execution, and motor observation) and resting with eyes open. We employed a convolutional neural network (CNN) as a classifier and compared the classification accuracies of two types of CNNs: 1) a "single five-class CNN," which classified the aforementioned states with a single CNN and 2) "multiple two-class CNNs," wherein ten CNNs that classify pairs of states were combined. In addition, the classification accuracies were compared between two scenarios: one wherein the EEGs from all 19 scalp probe electrodes (19-channel EEG) were adopted as input data for the CNN, and the other wherein the EEGs of four regions closely related to the motor execution and observation of the index finger (4-channel EEG) were adopted. The classification accuracies of the single five-class CNN with 19- and 4-channel EEGs were 48.2 ± 5.9% and 46.6 ± 6.9%, respectively, and those of the multiple two-class CNNs with 19- and 4-channel EEGs were 52.8 ± 9.7% and 47.5 ± 9.4%, respectively. These results indicate the effectiveness of multiple two-class CNNs that utilize the EEGs of all scalp electrodes as input data for classifying motor imagery, execution, and observation, even in the case of the marginal dataset.

This study examined exercise intervention effects on older adults' brain structures and function. Brain data were analyzed from 47 healthy adults between 61 and 82 years of age who, in a previous study, showed cognitive improvement following a 3-month intervention. The participants were assigned to a motor exercise intervention group (n = 24), performing exercise training programs for a 12-week period, or a waiting control group (n = 23), abstaining from any exercise program. Structural analysis of the frontal cortex and hippocampus revealed increased gray matter volume and/or thickness in several prefrontal areas in the intervention group and reduced hippocampal gray matter volume in the control group. Importantly, the volume increase in the middle frontal sulcus in the intervention group was associated with a general cognitive improvement after the intervention. Functional analysis showed that the prefrontal functional connectivity during a working memory task differently changed in response to the intervention or waiting in the two groups. The functional connectivity decreased in the intervention group, whereas the corresponding connectivity increased in the control group, which was associated with maintaining cognitive performance. The current longitudinal findings indicate that short-term exercise intervention can induce prefrontal plasticity associated with cognitive performance in older adults., (© The Author(s) 2021. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

In this study, the feasibility of conducting a concurrent estimation of drowsiness, stress, and tiredness by heart rate variability (HRV) in a driving simulator environment was examined. Subjects were required to attend a 120-min driving session four times: two morning and two afternoon sessions. Blood pressure and salivary amylase were also recorded to assess acute stress. A set of estimators was prepared, and stepwise regression was conducted on two different models at p = 0.05. In this work, it was shown that the use of a stepwise method and additional estimator capable of extracting significant and relevant information for multiple emotions with average performance in the form of the correlation coefficient(root mean square error) can increase up to 0.68 ± 0.12 (0.66 ± 0.28), 0.72 ± 0.13 (0.43 ± 0.21), and 0.71 ± 0.13 (0.48 ± 0.21), corresponding to drowsiness, stress, and tiredness, respectively. The results suggest that a single time series of HRV can extract more than one emotion, thus enabling a concurrent model to be developed. It was also observed that physiological behavior while driving works in a more complex way. The current evidence indicates the feasibility of conducting concurrent emotion assessment during driving.

Studies that evaluate human emotions from biological signals have been actively conducted, with many using images or sounds to induce emotions passively. However, few studies utilized the action of working to elicit emotions (especially positive ones) actively. Hence, in this study, emotions were examined during working (a puzzle was used in this study) from the psychological viewpoint of the Profile of Mood States 2nd Edition and the physiological viewpoint of electroencephalograms (EEGs). As a result, different time-dependent changes of power change rate in the theta band in the frontal region were observed between the presence and absence of the emotion "fatigue-inertia." Those in the alpha band in the frontal region were observed between the existence and nonexistence of the emotion "vigor-activity." Therefore, it is suggested that we can evaluate the emotion of a subject while working by a spatiotemporal pattern of band power obtained by EEG.

Coherence between an electroencephalogram (EEG) and an electromyogram (EMG) of the soleus (SOL) muscle during an isometric contraction is observed in the beta-band (15 to 35 Hz) regardless of the contraction force. However, the dynamics on how a variation in coherence occurs over time in the head of the gastrocnemius (GLH) muscle, which is also known to have the same role as the soleus muscle, have yet to be considered. In this study, we focused on GLH and measured an EEG and EMG taken of the GLH muscle when executing an isometric contraction through the dorsiflexion of the right ankle joint for a 1-min period. Moreover, we investigated changes in the EEG-EMG coherence based on the contraction force and elapsed time. As a result, in most subjects, the peak coherence during a weak contraction force was continuously observed in the β-band, whereas the peak coherence during a strong contraction force was observed in the γ-band (35 to 60 Hz) for only the first 12 s. In addition, no significant coherence was observed. Therefore, it was suggested that muscle fatigue induced by a strong contraction force affects the peak coherence. Meanwhile, the inconsistencies observed between the properties of the peak coherence and SOL might be due to the differences in muscle composition.

Motor imagery (MI) is divided into two types: kinesthetic (KMI) and visual (VMI). To estimate the MI that an examinee performs, event-related desynchronization (ERD) or event-related synchronization (ERS) is used to characterize KMI or VMI via electroencephalogram (EEG). However, no definitive method using ERD/ERS via EEG has been established yet to estimate the type of MI performed. This is because the MI performed by the examinee is not always the same as that instructed by the examiner. One of the reasons for this mismatch is the difficulty of MI, especially KMI. However, almost no reported studies have considered MI difficulty to estimate MI type. Therefore, in this study, we examined the relationship between MI difficulty and the ERD/ERS pattern corresponding to the type of MI in the case of single flexion of the right index finger (SFRIF). The results showed that for a subject who felt MI was less difficult, the α-band ERD value (αERD) at the electrode of the occipital area (O1 or O2 site) of the KMI instruction was significantly smaller than that of the VMI instruction. On the contrary, for a subject who felt MI was very difficult, αERD at the O1 or O2 site on the KMI instruction was similar to that of the VMI instruction. In addition, for the subject who felt MI was easy, the αERDs at the O1 or O2 site on the KMI and VMI instructions were similar to those on the movement execution (ME) and movement observation (MO) instructions, respectively. Therefore, in the case of SFRIF, it was suggested that MI difficulty could be estimated by ERD/ERS patterns in the occipital area. This was supported by referring to the ME and MO ERD/ERS patterns in the occipital area.

In this study, we proposed a measurement system that extracts heartbeat interval data from multiple piezoelectric sensors placed on a chair that eliminates noise generated by body movement. We asked five healthy males (21-24 years old) to sit in an arbitrary position on a chair that had eight piezoelectric sensors attached, and heartbeat signals were measured. The experiment consisted of four measurements (5 min measurements after adequate rest), while performing specific body movements at a specific time, with deliberately mixed noise of body movements. To remove body movement noise, bandpass filter processing (4 Hz-20 Hz) was applied to the signal obtained from the piezoelectric sensors, and the heartbeat component was extracted using independent component analysis (number of separations was eight) on the processed waveform. For verification, the error rate was obtained before and after the removal of body movement noise, respectively. The error rate after removal of body motion was 2.91 ± 0.75%. In our previous study with two piezoelectric sensors without body movement, the error rate was 2.47 ± 2.66%. Therefore, our proposed measurement system may improve the accuracy of heartbeat interval detection, which can be observed continuously by removing the body motion.

Traffic accidents remain one of the most critical issues in many countries. One of the major causes of traffic accidents is drowsiness while driving. Since drowsiness is related to human physiological conditions, drowsiness is hard to prevent. Several studies have been conducted in assessing drowsiness, especially in a driving environment. One of the common methods used is the electroencephalogram (EEG). It is known that drowsiness occurs in the central nervous system; thus, estimating drowsiness using EEG is the promising way to assess drowsiness accurately. In this study, we tried to estimate drowsiness using frequency-domain and time-domain analysis of EEG. To validate the physiological conditions of the subjects, the Karolinska sleepiness scale (KSS), a subject-based assessment of drowsiness condition; and an examiner-based assessment known as facial expression evaluation (FEE) were applied. Three categories were considered; alert (KSS <; 6; FEE <; 1), weak drowsiness (KSS 6-7; FEE 1-2) and strong drowsiness (KSS > 7; FEE > 2). The six parameters (absolute and relative power of alpha, ratio of β/α and (θ+α)/β, and Hjorth activity and mobility parameters) had statistically significant differences between the three drowsiness conditions (P <; 0.001). By using both KSS and FEE, these parameters showed high accuracy in detecting drowsiness (up to 92.9%). Taken together, we suggest that EEG parameters can be used in detecting the three drowsiness conditions in a simulated driving environment.

The purpose of this work was to investigate the information processing of passive joint motion of the rat hindlimb in the spinal nervous system in vivo. Action potentials using intracellular recordings and joint kinematics using video analysis were measured. The results show that the action potentials of the spinal nervous system-evoked passive joint motion were significantly changed. Therefore, physical therapy is one of the useful methods for the treatment of joint position and angular position sensitivity and spinal nervous system disorders.

Several behavioural studies have shown that the interplay between voice and face information in audiovisual speech perception is not universal. Native English speakers (ESs) are influenced by visual mouth movement to a greater degree than native Japanese speakers (JSs) when listening to speech. However, the biological basis of these group differences is unknown. Here, we demonstrate the time-varying processes of group differences in terms of event-related brain potentials (ERP) and eye gaze for audiovisual and audio-only speech perception. On a behavioural level, while congruent mouth movement shortened the ESs' response time for speech perception, the opposite effect was observed in JSs. Eye-tracking data revealed a gaze bias to the mouth for the ESs but not the JSs, especially before the audio onset. Additionally, the ERP P2 amplitude indicated that ESs processed multisensory speech more efficiently than auditory-only speech; however, the JSs exhibited the opposite pattern. Taken together, the ESs' early visual attention to the mouth was likely to promote phonetic anticipation, which was not the case for the JSs. These results clearly indicate the impact of language and/or culture on multisensory speech processing, suggesting that linguistic/cultural experiences lead to the development of unique neural systems for audiovisual speech perception.

Rationale: Various physiological and pathological conditions are correlated with cardiac autonomic function. Heart rate variability is a marker of cardiac autonomic modulation and can be measured by several methods. However, the available methods are sensitive to breathing patterns., Objectives: To quantify cardiac autonomic modulation by observing the potential force dynamics of the R-R interval time series in healthy individuals., Methods: We propose two "potentials of unbalanced complex kinetic" (PUCK) parameters to quantify the characteristics of the potential force dynamics of R-R interval time series: potential strength (slope) and fluctuation size (slope standard deviations [SSD1, SSD2]). We applied this method to the series of R-R intervals obtained from 30 healthy subjects in an experimental condition that elicited cardiac autonomic (i.e., sympathetic and vagal) activation (in supine, sitting, and standing positions). Subjects were categorized into three groups by decade (i.e., 20 s, 30 s, and 40 s) to verify the cardiac autonomic differences by age. Two respiration patterns were introduced to check the influence of the pattern into the analytical results., Measurements and Main Results: Sympathetic modulation activation significantly increased the slope and reduced SSD1 and SSD2; these trends were confirmed in all groups. The slope is concordant with the result of the low frequency/high frequency (LF/HF) ratio in frequency components as an indicator of sympathetic modulation. No trend was observed in slope among age groups. However, SSD1 and SSD2 in the 40 s group were significantly decreased in the supine and sitting positions. The results with respect to respiration frequency showed lower sympathetic modulation as shown in the LF/HF ratio and slope, whereas higher vagal modulation as shown in the HF appeared with a longer breathing rate., Conclusions: PUCK can quantify the cardiac autonomic modulation in the experimental conditions of different postures. SSD1 and SSD2 are more sensitive to age than frequency components and are unaffected by breathing patterns. This method may be an alternative method for observing cardiac autonomic modulation in clinical practice., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The human visual system seems to have a highly perceptual sensitivity to symmetry. However, where and when the discrimination of symmetrical properties begins in the context of visual information processing is largely unclear. This study investigates event-related potential (ERP) patterns in humans when perceiving symmetry-varied complex object images. ERP responses were derived from electroencephalography (EEG) data recorded from eight healthy subjects using 128-channel scalp electrodes. Visual stimulation was provided using gray-scaled photographs of a car with six different viewpoints, hence disrupting the vertical symmetry, where one of the stimuli was intentionally made symmetric by mirroring the image about its center vertical axis. The results show that discrimination of image symmetry is revealed by potential deflection in early ERP components recorded at occipito-temporal sites and can be significantly observed around 220 ms after stimulus onset.

In this paper, we propose a measurement system to extract the heartbeat and respiration signals from piezoelectric sensors located in a chair seat. Eight healthy male subjects (22-26 years old) sat on a chair with piezoelectric sensors. The experiment consisted of five acquisitions (300 s recording with 30 s resting). In the 1st acquisition, the subject was instructed to maintain a relax state. During the next three acquisitions, the subject maintained respiration rates of 3, 4, and 5 s. The order was randomly given for each subject. Finally, the subject was instructed to stay in a resting state for 300 s. Band-pass filters were used to separate the heartbeat and respiration signals from the output signal of the piezoelectric sensors (heartbeat: 0.7-7 Hz, respiration: 0.1-0.4 Hz). Then, the standard heartbeat interval was calculated using the autocorrelation function to generate a template. Heartbeat and respiration signals were extracted using a two-dimensional cross-correlation pattern matching method. For validation, the error ratio between the piezoelectric sensor data and the reference data was determined. The error ratios of the heartbeat interval and respiration interval were 2.89 ± 4.41% and 5.57 ± 4.42%, respectively. Therefore, it is suggested that heartbeat and respiration signals extracted from piezoelectric sensors in a sitting position can be used as an alternative method for extracting biological signals.

This paper presents cardiac arrhythmia detection using the combination of a heart rate variability (HRV) analysis and a "potential of unbalanced complex kinetics" (PUCK) analysis. Detection performance was improved by adding features extracted from the PUCK analysis. Initially, R-R interval data were extracted from the original electrocardiogram (ECG) recordings and were cut into small segments and marked as either normal or arrhythmia. HRV analyses then were conducted using the segmented R-R interval data, including a time-domain analysis, frequency-domain analysis, and nonlinear analysis. In addition to the HRV analysis, PUCK analysis, which has been implemented successfully in a foreign exchange market series to characterize change, was employed. A decision-tree algorithm was applied to all of the obtained features for classification. The proposed method was tested using the MIT-BIH arrhythmia database and had an overall classification accuracy of 91.73%. After combining features obtained from the PUCK analysis, the overall accuracy increased to 92.91%. Therefore, we suggest that the use of a PUCK analysis in conjunction with HRV analysis might improve performance accuracy for the detection of cardiac arrhythmia.

Arabinoxylans may account for up to 25% of the mass of grass cell walls. The interactions of these polysaccharides with themselves and with cellulose and lignin is believed to affect the walls physical properties and increase the walls resistance to biochemical conversion to fermentable sugars. Arabinoxylans have a backbone composed of 1,4-linked β-D-xylosyl residues, some of which are substituted at O-2 or O-3 with single arabinofuranosyl (Araf) residues. The Araf residues are likely transferred from UDP-Araf to the xylan backbone by arabinofuranosyltransferases. UDP-Araf is itself formed from UDP-arabinopyranose (UDP-Arap) by UDP-arabinopyranose mutase (UAM). In this study, RNA interference (RNAi) was used to suppress UAM expression in rice plants and thereby reduce the amounts of UDP-Araf available for cell wall synthesis. Several of the transgenic plants had reduced proportions of Araf in their walls together with a decrease in the extent of substitution of the xylan backbone, and a reduction of between 25% and 80% in ferulic acid and p-coumaric acid contents of the cell walls. Those transgenic plants with >25% reduction in the amounts of Araf were dwarfed and infertile., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Anatomically distributed areas are dynamically linked to form functional networks for processing and integrating the different modalities of information in the human brain. A part of such networks is considered to be realized with synchronization of neuronal activities, which can generate correlated neural oscillation at the same and/or different frequency bands. To investigate the networks with the synchronization, analysis of connectivity between not only same frequency oscillation but also different frequency (i.e. cross-frequency) is needed. For source estimation with electroencephalogram (EEG) or magneto-encephalogram (MEG) signals, a spatial filtering technique is recently applied as an alternative method for equivalent current dipole (ECD) estimation technique. Non-adaptive type of spatial filtering technique, such as the Standardized low-resolution brain electromagnetic tomography (sLORETA), is reported to discriminate correlated sources. However, it may lead to inaccurate results due to its low spatial resolution. In the present study, we proposed a new systematic approach for localizing the sources of correlated cross-frequency oscillations. The method we propose can overcome the limitation of the non-adaptive spatial filtering technique by proactively using identified information in sensor level analysis (e.g. cross-correlation map and correlation topography), which allow us to focus on target sources. The performance of our proposed method is evaluated with simulated EEG signals, and is compared with traditional method.

The two FLOWERING LOCUS T (FT)-like genes of apple (Malus x domestica Borkh.), MdFT1 and MdFT2, have been isolated and characterized. MdFT1 and MdFT2 were mapped, respectively, on distinct linkage groups (LGs) with partial homoeology, LG 12 and LG 4. The expression pattern of MdFT1 and MdFT2 differed in that MdFT1 was expressed mainly in apical buds of fruit-bearing shoots in the adult phase, with little expression in the juvenile tissues, whereas MdFT2 was expressed mainly in reproductive organs, including flower buds and young fruit. On the other hand, both genes had the potential to induce early flowering since transgenic Arabidopsis, which ectopically expressed MdFT1 or MdFT2, flowered earlier than wild-type plants. Furthermore, overexpression of MdFT1 conferred precocious flowering in apple, with altered expression of other endogenous genes, such as MdMADS12. These results suggest that MdFT1 could function to promote flowering by altering the expression of those genes and that, at least, other genes may play an important role as well in the regulation of flowering in apple. The long juvenile period of fruit trees prevents early cropping and efficient breeding. Our findings will be useful information to unveil the molecular mechanism of flowering and to develop methods to shorten the juvenile period in various fruit trees, including apple.

Background: Cryptomeria japonica D. Don is one of the most commercially important conifers in Japan. However, the allergic disease caused by its pollen is a severe public health problem in Japan. Since large-scale analysis of expressed sequence tags (ESTs) in the male strobili of C. japonica should help us to clarify the overall expression of genes during the process of pollen development, we constructed a full-length enriched cDNA library that was derived from male strobili at various developmental stages., Results: We obtained 36,011 expressed sequence tags (ESTs) from either one or both ends of 19,437 clones derived from the cDNA library of C. japonica male strobili at various developmental stages. The 19,437 cDNA clones corresponded to 10,463 transcripts. Approximately 80% of the transcripts resembled ESTs from Pinus and Picea, while approximately 75% had homologs in Arabidopsis. An analysis of homologies between ESTs from C. japonica male strobili and known pollen allergens in the Allergome Database revealed that products of 180 transcripts exhibited significant homology. Approximately 2% of the transcripts appeared to encode transcription factors. We identified twelve genes for MADS-box proteins among these transcription factors. The twelve MADS-box genes were classified as DEF/GLO/GGM13-, AG-, AGL6-, TM3- and TM8-like MIKCC genes and type I MADS-box genes., Conclusion: Our full-length enriched cDNA library derived from C. japonica male strobili provides information on expression of genes during the development of male reproductive organs. We provided potential allergens in C. japonica. We also provided new information about transcription factors including MADS-box genes expressed in male strobili of C. japonica. Large-scale gene discovery using full-length cDNAs is a valuable tool for studies of gymnosperm species.

Genes in the FLOWERING LOCUS T (FT) and TERMINAL FLOWER 1 (TFL1) family have been shown to be important in the control of the switch between vegetative and reproductive growth in several plant species. We isolated nine members of the FT/TFL1 family from Lombardy poplar (Populus nigra var. italica Koehne). Sequence analysis of the members of the FT/TFL1 family revealed considerable homology within their coding regions both among family members and to the members of the same family in Arabidopsis, tomato and grapevine. Moreover, members of this family in all four species examined display a common exon-intron organization. Phylogenetic analysis revealed that the genes fall into four different clades: two into the TFL1 clade; five into the FT clade; and one each into the MOTHER OF FT AND TFL1 and BROTHER OF FT AND TFL1 clades. One gene in the TFL1 clade, PnTFL1, is expressed in vegetative meristems, and transgenic Arabidopsis that ectopically expressed PnTFL1 had a late-flowering phenotype. The expression patterns of two genes in the FT clade, PnFT1 and PnFT2, suggested a role for them in the promotion of flowering, and transgenic Arabidopsis that ectopically expressed either PnFT1 or PnFT2 had an early-flowering phenotype.

Background: Populus is one of favorable model plants because of its small genome. Structural genomics of Populus has reached a breakpoint as nucleotides of the entire genome have been determined. Reaching the post genome era, functional genomics of Populus is getting more important for well-comprehended plant science. Development of bioresorce serving functional genomics is making rapid progress. Huge efforts have achieved deposits of expressed sequence tags (ESTs) in various plant species consequently accelerating functional analysis of genes. ESTs from full-length cDNA clones are especially powerful for accurate molecular annotation. We promoted collection and annotation of the ESTs from Populus full-length enriched cDNA clones as part of functional genomics of tree species., Results: We have been collecting the full-length enriched cDNA of the female poplar (Populus nigra var. italica) for years. By sequencing P. nigra full-length (PnFL) cDNA libraries, we generated about 116,000 5'-end or 3'-end ESTs corresponding to 19,841 nonredundant PnFL clones. Population of PnFL cDNA clones represents 44% of the predicted genes in the Populus genome., Conclusion: Our resource of P. nigra full-length enriched clones is expected to provide valuable tools to gain further insight into genome annotation and functional genomics in Populus.

To study the effects of external visual stimulation on motor cortex-muscle synchronization, coherence between electroencephalography (EEG) and electromyography (EMG) was measured in normal subjects under Before, Task (visual task: Ignore or Count, or arithmetic task) and After conditions. The control (Before and After) conditions required the subject to maintain first dorsal interosseous muscle contraction without visual stimulation. In the visual task, a random series of visual stimuli were displayed on a screen while the subjects maintained the muscle contraction. The subjects were asked to ignore the stimuli in the Ignore condition and to count certain stimuli in the Count condition. Also, in the arithmetic task, the subjects were asked to perform a simple subtraction. The EEG-EMG coherence found at C(3) site at 13-30 Hz (beta) was increased and sustained in magnitude during the Ignore and Count conditions, respectively. To examine the cause of the change of coherence, changes of EEG and EMG spectral power were computed for each frequency band. There was little change in the EMG spectral power in any frequency bands. While the spectral power of EEG unchanged in the beta band, it significantly increased and decreased in the range of 8-12 Hz and of 31-50 Hz, respectively, for both Ignore and Count conditions, not only at the C(3) site but at various sites as well. These results were in contrast to those obtained for the arithmetic task: the beta band EEG-EMG coherence was attenuated and the EEG spectral power at 4-7 Hz and at 31-50 Hz were significantly increased and decreased, respectively. As a conclusion, the present results are consistent with the idea that the enhanced 8-12 Hz/decreased 31-50 Hz oscillations affect strength of the beta band cortico-muscular synchronization by suppressing the visual processing.

The purpose of the present study was to develop a communication aid device utilizing event-related potentials (ERPs) for patients with severe motor disabilities. Preliminary experiments were performed on eight healthy volunteers to determine the optimal methods of presentation of visual stimuli and an off-line analysis of electroencephalography signals. Three clinical experiments were performed online on 17 healthy volunteers, 5 patients with amyotrophic lateral sclerosis, and 1 patient with olivopontocerebellar atrophy. First, one of four vertically aligned short expressions ("yes definitely," "yes," "no," and "don't know") was presented in a random sequence to determine the target. Second, a systematic table of 50 Japanese syllables, and third, 16 pictorial symbols in four columns were presented, with each column presented at random. The target was identified by the maximum sum of the 2-5 Hz power spectrum after fast Fourier transform of averaged ERPs. A high correct response rate was obtained in the clinical experiments of almost 100% in "yes or no" and pictorial symbol paradigms using seven averaged samples. These results indicate that such devices may be used to communicate with patients who have lost all ability to interact other than directly via the brain.

Recombinants were generated from the ectomycorrhizal basidiomycete, Suillus grevillei, through agroinfection using a binary vector carrying the hygromycin B resistance and the autofluorescent protein, DsRed2, markers. DsRed2 was driven by a cis-regulatory region of the glyceraldeyde-3-phosphate dehydrogenase gene (gpd) from the wood-rotting basidiomycete, Coriolus hirsutus, which contains promoters and 5' gpd sequences with first through fourth exons and expressed for the first time in Suillus spp. The transformation system and recombinants expressing an autofluorescent protein may be useful in genetic analysis of the symbiosis.

The effects of visual stimuli on cortico-spinal synchronization were investigated by measuring the coherence between electroencephalogram (EEG) and electromyogram (EMG) during isometric contraction of the first dorsal interosseous muscle of the right hand. Because a spinal motoneuron and the corresponding muscle fibers form a motor unit with one-to-one correspondence of their action potentials, the EMG indirectly measures the activity of the corresponding spinal neuronal group. The tasks were isometric contraction (Control condition); and isometric contraction with concurrent ignoring of visual stimuli (VS condition). By comparing the Control and VS conditions, the following results were obtained. The coherence increased significantly in magnitude, but was unchanged in frequency range (beta band) and scalp location; the EEG and EMG spectral power in the beta band were unchanged in amplitude; and the alpha and gamma bands of EEG spectral power were significantly increased and decreased, respectively. These findings suggest that the cortico-muscular coherence reflects the cognitive effort needed to maintain isometric muscle contraction. When visual stimuli need to be ignored, the cognitive effort and cortico-spinal coherence are enhanced.

Many secretory and several vacuolar proteins in higher plants contain hydroxylated proline residues. In many cases, hydroxyprolines in proteins are glycosylated with either arabinogalactan or oligoarabinose. We have previously shown that a sporamin precursor is O-glycosylated at the hydroxylated proline 36 residue with an arabinogalactan-type glycan when this protein is expressed in tobacco BY-2 cells (Matsuoka et al., 1995). Taking advantage of the fact that this is the only site of proline hydroxylation and glycosylation in sporamin, we analyzed the amino acid requirement for proline hydroxylation and arabinogalactosylation. We expressed several deletion constructs and many amino acid substitution mutants in tobacco cells and analyzed glycosylation and proline hydroxylation of the expressed sporamins. Hydroxylation of a proline residue requires the five amino acid sequence [AVSTG]-Pro-[AVSTGA]-[GAVPSTC]-[APS or acidic] (where Pro is the modification site) and glycosylation of hydroxyproline (Hyp) requires the seven amino acid sequence [not basic]-[not T]-[neither P, T, nor amide]-Hyp-[neither amide nor P]-[not amide]-[APST], although charged amino acids at the -2 position and basic amide residues at the +1 position relative to the modification site seem to inhibit the elongation of the arabinogalactan side chain. Based on the combination of these two requirements, we concluded that the sequence motif for efficient arabinogalactosylation, including the elongation of the glycan side chain, is [not basic]-[not T]-[AVSG]-Pro-[AVST]-[GAVPSTC]-[APS].

Poplar, whose genome is the first to be sequenced among woody plants, is a favorable model for plant biologists to enable them to understand molecular processes of growth, development and responses to environmental stimuli in trees. The sequence will allow the development of a strategy for improving environmental stress tolerance in forest trees. In this study, we have generated a full-length enriched cDNA library from leaves of axenically grown poplar (Populus nigra var. italica) subjected to environmental stress treatments by dehydration, high salinity, chilling, heat, abscisic acid (ABA) and H2O2. We sequenced >30,000 expressed sequence tags (ESTs) from the cDNA library and consequently collected approximately 4,500 non-redundant clones. We further analyzed cDNAs encoding an ERF/AP2-domain transcription factor which is specific in plants and plays an important role under stress. Thirteen candidates containing the ERF/AP2 domain were found within our EST resource. Some of them showed stress-responsive gene expression. We report here the first collection of full-length enriched stress-related ESTs of poplar and discuss environmental stress responses of forest trees in the light of comparative genomics.

We had developed a communication system using a theory of P300 through a computer. 17 healthy subjects participated in all 3 experiments, while 4 ALS and 1 OPCA patients took part in the first 2 experiments only. In the 1st experiment, 4 short sentences of 'yes very much', 'yes' 'no' and 'can't answer' were shown on the display. The subjects were asked to select one of them as target. During the experiment 1, color of background was randomly changed for each sentence as the stimulation for the period of 300 msec and the interstimulus interval was 1,500 msec. In the 2nd experiment, 46 letters of Japanese character and several symbols were shown on the display (5 columns x 10 rows). In the 3rd experiment, we used 16 pictures together with X (4 columns x 5 rows). During recordings, the color of background was randomly changed for each row and then for each letter/picture of column containing target in experiment 2 as well as in experiment 3. The subjects were asked to start counting and when the target was stimulated, they were instructed to stop the counting process and repeat counting again until they encountered the stimulation of target again. This was repeated for the entire recordings. Computer defined a target by a biggest positive voltage between 300 and 600 msec and/or by the most powerful spectrum in 2-5 Hz after Fast Fourier transform in an averaged P300 (averaging number = 5-7). We then calculated the percentage of correct target by comparing the result and actual target. Results indicate that this communication aids can be used conveniently in patients with severe motor disabilities.