Atomic force microscopic analysis of the light-harvesting complex 2 from purple photosynthetic bacterium Thermochromatium tepidum.

Structural information on the circular arrangements of repeating pigment-polypeptide subunits in antenna proteins of purple photosynthetic bacteria is a clue to a better understanding of molecular mechanisms for the ring-structure formation and efficient light harvesting of such antennas. Here, we have analyzed the ring structure of light-harvesting complex 2 (LH2) from the thermophilic purple bacterium Thermochromatium tepidum (tepidum-LH2) by atomic force microscopy. The circular arrangement of the tepidum-LH2 subunits was successfully visualized in a lipid bilayer. The average top-to-top distance of the ring structure, which is correlated with the ring size, was 4.8 ± 0.3 nm. This value was close to the top-to-top distance of the octameric LH2 from Phaeospirillum molischianum (molischianum-LH2) by the previous analysis. Gaussian distribution of the angles of the segments consisting of neighboring subunits in the ring structures of tepidum-LH2 yielded a median of 44°, which corresponds to the angle for the octameric circular arrangement (45°). These results indicate that tepidum-LH2 has a ring structure consisting of eight repeating subunits. The coincidence of an octameric ring structure of tepidum-LH2 with that of molischianum-LH2 is consistent with the homology of amino acid sequences of the polypeptides between tepidum-LH2 and molischianum-LH2.

Electronic structure and thermal conductance of the MASnI3/Bi2Te3 interface: a first-principles study.

To develop high-performance thermoelectric devices that can be created using printing technology, the interface of a composite material composed of MASnI3 and Bi2Te3, which individually show excellent thermoelectric performance, was studied based on first-principles calculations. The structural stability, electronic state, and interfacial thermal conductance of the interface between Bi2Te3 and MASnI3 were evaluated. Among the interface structure models, we found stable interface structures and revealed their specific electronic states. Around the Fermi energy, the interface structures with TeII and Bi terminations exhibited interface levels attributed to the overlapping electron densities for Bi2Te3 and MASnI3 at the interface. Calculation of the interfacial thermal conductance using the diffuse mismatch model suggested that construction of the interface between Bi2Te3 and MASnI3 could reduce the thermal conductivity. The obtained value was similar to the experimental value for the inorganic/organic interface.

Local Cross-Coupling Activity of Azide-Hexa(ethylene glycol)-Terminated Self-Assembled Monolayers Investigated by Atomic Force Microscopy.

Azide-oligo(ethylene glycol)-terminated self-assembled monolayers (N3-OEG-SAMs) are promising interfacial structures for surface functionalization. Its many potential applications include chemical/bio-sensing and construction of surface models owing to its cross-coupling activity that originates from the azide group and oligo(ethylene glycol) (OEG) units for non-specific adsorption resistance. However, there are only a few studies and limited information, particularly on the molecular-scale structures and local cross-coupling activities of N3-OEG-SAMs, which are vital to understanding its surface properties and interfacial molecular design. In this study, molecular-scale surface structures and cross-coupling activity of azide-hexa(ethylene glycol)-terminated SAMs (N3-EG6-SAMs) were investigated using frequency modulation atomic force microscopy (FM-AFM) in liquid. The N3-EG6-SAMs were prepared on Au(111) substrates through the self-assembly of 11-azido-hexa(ethylene glycol)-undecane-1-thiol (N3-EG6-C11-HS) molecules obtained from a liquid phase. Subnanometer-resolution surface structures were visualized in an aqueous solution using a laboratory-built FM-AFM instrument. The results show a well-ordered molecular arrangement in the N3-EG6-SAM and its clean surfaces originating from the adsorption resistance property of the terminal EG6 units. Surface functionalization by the cross-coupling reaction of copper(I)-catalyzed azide-alkyne cycloaddition was observed, indicating a structural change in the form of fluctuating structures and island-shaped structures depending on the concentration of the alkyne molecules. The FM-AFM imaging enabled to provide information on the relationship between the surface structures and cross-coupling activity. These findings provide molecular-scale information on the functionalization of the N3-EG6-SAMs, which is helpful for the interfacial molecular design based on alkanethiol SAMs in many applications.

Contribution of Discharge Excited Atomic N, N2 *, and N2+ to a Plasma/Liquid Interfacial Reaction as Suggested by Quantitative Analysis.

Electric-discharge nitrogen comprises three main types of excited nitrogen species-atomic nitrogen (Natom ), excited nitrogen molecules (N2 *), and nitrogen ions (N2+ ) - which have different lifetimes and reactivities. In particular, the interfacial reaction locus between the discharged nitrogen and the water phase produces nitrogen compounds such as ammonia and nitrate ions (denoted as N-compounds generically); this is referred to as the plasma/liquid interfacial (P/L) reaction. The Natom amount was analyzed quantitatively to clarify the contribution of Natom to the P/L reaction. We focused on the quantitative relationship between Natom and the produced N-compounds, and found that both N2 * and N2+ , which are active species other than Natom , contributed to P/L reaction. The production of N-compounds from N2 * and N2+ was enhanced upon UV irradiation of the water phase, but the production of N-compounds from Natom did not increase by UV irradiation. These results revealed that the P/L reactions starting from Natom and those starting from N2* and N2+ follow different mechanisms.

Relationship between listening difficulty rating and objective measures in reverberant and noisy sound fields for young adults and elderly persons.

Listening difficulty ratings [Morimoto et al., J. Acoust. Soc. Am. 116, 1607-1613 (2004)] were obtained for 20 young adult listeners and 34 elderly listeners in reverberant and noisy sound fields simulated in an anechoic room. The listening difficulty ratings were compared with acoustical objective measures. The results and analyses showed the following: (i) The correlation between listening difficulty ratings and the revised speech transmission index (STI(r)), and that for the useful-detrimental ratio (U(50)) were high, regardless of the age of the listeners. (ii) STI(r) and U(50) need to be increased by 0.12 and 4.2 dB, respectively, to equalize the listening difficulty ratings for the elderly listeners with those for the young listeners. (iii) The estimation accuracies for STI(r) and U(50) can be improved by calculating them with the L(eq) of background noise linearly increased by 4 to 10 dB, which depends on the age of the listeners and the objective measures. However, the improvement was not statistically significant for the elderly listeners.

Acceptable range of speech level in noisy sound fields for young adults and elderly persons.

The acceptable range of speech level as a function of background noise level was investigated on the basis of word intelligibility scores and listening difficulty ratings. In the present study, the acceptable range is defined as the range that maximizes word intelligibility scores and simultaneously does not cause a significant increase in listening difficulty ratings from the minimum ratings. Listening tests with young adult and elderly listeners demonstrated the following. (1) The acceptable range of speech level for elderly listeners overlapped that for young listeners. (2) The lower limit of the acceptable speech level for both young and elderly listeners was 65 dB (A-weighted) for noise levels of 40 and 45 dB (A-weighted), a level with a speech-to-noise ratio of +15 dB for noise levels of 50 and 55 dB, and a level with a speech-to-noise ratio of +10 dB for noise levels from 60 to 70 dB. (3) The upper limit of the acceptable speech level for both young and elderly listeners was 80 dB for noise levels from 40 to 55 dB and 85 dB or above for noise levels from 55 to 70 dB.

Relationship between listening difficulty and acoustical objective measures in reverberant sound fields.

The previous work [Morimoto et al., J. Acoust. Soc. Am. 116, 1607-1613] showed that listening difficulty ratings can be used to evaluate speech transmission performance more exactly and sensitively than intelligibility. Meanwhile, speech transmission performance is usually evaluated using acoustical objective measures, which are directly associated with physical parameters of room acoustic design. However, the relationship between listening difficulty ratings and acoustical objective measures was not minutely investigated. In the present study, a total of 96 impulse responses were used to investigate the relationship between listening difficulty ratings and several objective measures in unidirectional sound fields. The result of the listening test showed that (1) the correlation between listening difficulty ratings and speech transmission index (STI) is the strongest of all tested objective measures, and (2) A-weighted D(50), C(50), and center time, which are obtained from the impulse responses passed through an A-weighted filter, also strongly correlate with listening difficulty ratings, and their correlations with listening difficulty ratings are not statistically different from the correlation between listening difficulty ratings and STI.

Effects of frequency characteristics of reverberation time on listener envelopment.

Spatial impression perceived in a listening space comprises at least two components: one is auditory (apparent) source width (ASW) and the other is listener envelopment (LEV). Both ASW and LEV are affected not only by temporal but also by spatial structures of reflections. It has been clarified that ASW for symphony music is significantly affected by low-frequency components of source signals and reflections, but not by their high-frequency components. The objective of this work is to investigate whether LEV is affected by the frequency characteristics of source signals and reverberation sounds, which are known to contribute to the creation of LEV. In this study, three experiments were performed to clarify the effects of reverberation time (RT) and its frequency characteristics on LEV. In contrast to the case of ASW, the experimental results show that RTs both at high and low frequencies affect LEV.

Acceptable range of speech level for both young and aged listeners in reverberant and quiet sound fields.

The speech level of verbal information in public spaces should be determined to make it acceptable to as many listeners as possible, while simultaneously maintaining maximum intelligibility and considering the variation in the hearing levels of listeners. In the present study, the universally acceptable range of speech level in reverberant and quiet sound fields for both young listeners with normal hearing and aged listeners with hearing loss due to aging was investigated. Word intelligibility scores and listening difficulty ratings as a function of speech level were obtained by listening tests. The results of the listening tests clarified that (1) the universally acceptable ranges of speech level are from 60 to 70 dBA, from 56 to 61 dBA, from 52 to 67 dBA and from 58 to 63 dBA for the test sound fields with the reverberation times of 0.0, 0.5, 1.0 and 2.0 s, respectively, and (2) there is a speech level that falls within all of the universally acceptable ranges of speech level obtained in the present study; that speech level is around 60 dBA.

Influence of fundamental frequency and source elevation on the vertical localization of complex tones and complex tone pairs.

This study investigates the vertical localization of single complex tones (monads) and simultaneous complex tone pairs (dyads), especially as it is affected by their fundamental frequency and source elevation. Two complex tone timbres are considered: one consisting of five low-order harmonics, and the other of all odd harmonics (a square wave). Sound sources were at -15, 0, 15, and 30 deg from the horizontal plane at ear height. For eight subjects, this source array was in the median plane, and for a further nine subjects, it was directly to the subject's left (lateral plane). The subjects localized the angle of the auditory image(s) of one or two complex tones around the vertical plane containing the sound sources. Mean responses for the five-harmonic complex tones show a systematic effect (referred to as Pratt's effect) of fundamental frequency on vertical localization--whereby high-frequency complex tones are localized to positions higher than low-frequency complex tones for equivalent source positions. For the square wave, the sound-source position dominates localization, although some effect of fundamental frequency is evident for median plane sources.

Effects of aging on word intelligibility and listening difficulty in various reverberant fields.

Listening difficulty ratings is a new measure to evaluate speech transmission performance in public spaces. The advantage is that it can evaluate sound fields with higher speech transmission performance more accurately and sensitively than intelligibility scores. The effect of aging, or hearing level on listening difficulty ratings has not been clarified yet. In this paper, the results of the listening tests demonstrated that hearing levels above 2 kHz significantly affect both word intelligibility scores and listening difficulty ratings for the aged. For the aged, both of them can be estimated from the results for the young plus the average of hearing levels at 2 and 4 kHz. The equations for estimating the word intelligibility scores and the listening difficulty ratings for the aged are obtained from the multiple linear regression analyses. The estimated scores and ratings generally coincide with the measured scores and ratings, and the averaged difference between them is 5.2% for the word intelligibility scores and 3.8% for the listening difficulty ratings, in the range of listening tests performed in this paper.

Optimum speech level to minimize listening difficulty in public spaces.

For ideal speech communication in public spaces, it is important to determine the optimum speech level for various background noise levels. However, speech intelligibility scores, which is conventionally used as the subjective listening test to measure the quality of speech communication, is near perfect in most everyday situations. For this reason, it is proposed to determine optimum speech levels for speech communication in public spaces by using listening difficulty ratings. Two kinds of listening test were carried out in this work. The results of the tests and our previous work [M. Morimoto, H. Sato, and M. Kobayashi, J. Acoust. Soc. Am. 116, 1607-1613 (2004)] are jointly discussed for suggesting the relation between the optimum speech level and background noise level. The results demonstrate that: (1) optimum speech level is constant when background noise level is lower than 40 dBA, (2) optimum speech level appears to be the level, which maintains around 15 dBA of SN ratio when the background noise level is more than 40 dBA, and (3) listening difficulty increases as speech level increases under the condition where SN ratio is good enough to keep intelligibility near perfect.

Caffeine biosynthesis and adenine metabolism in transgenic Coffea canephora plants with reduced expression of N-methyltransferase genes.

In anti-sense and RNA interference transgenic plants of Coffea canephora in which the expression of CaMXMT1 was suppressed, caffeine biosynthesis from [8-(14)C]adenine was investigated, together with the overall metabolism of [8-(14)C]adenine. Compared with wild type control plants, total purine alkaloid biosynthesis from adenine and conversion of theobromine to caffeine were both reduced in the transgenic plants. As found previously, [8-(14)C]adenine was metabolised to salvage products (nucleotides and RNA), to degradation products (ureides and CO(2)) and to purine alkaloids (theobromine and caffeine). In the transgenic plants, metabolism of [8-(14)C]adenine shifted from purine alkaloid synthesis to purine catabolism or salvage for nucleotides. HPLC analysis revealed a significantly reduced caffeine content in the transgenic plants. A small quantity (less than 20 nmol g(-1) fresh weight) of xanthosine had accumulated in at least one of the transgenic plants.

Caffeine production in tobacco plants by simultaneous expression of three coffee N-methyltrasferases and its potential as a pest repellant.

Caffeine (1,3,7-trimethylxanthine) is derived from xanthosine through three successive transfers of methyl groups and a single ribose removal in coffee plants. The methyl group transfer is catalyzed by N-zmethyltransferases, xanthosine methyltransferase (XMT), 7-methylxanthine methyltransferase (MXMT) and 3,7-dimethylxanthine methyltransferase (DXMT). We previously cloned three genes encoding each of these N-methyltransferases from coffee plants, and reconstituted the final sequence of the caffeine synthetic pathway in vitro. In the present study, we simultaneously expressed these coffee genes in tobacco plants (Nicotiana tabacum), using a multiple-gene transfer method, and confirmed successful caffeine production up to 5 microg g(-1) fresh weight in leaves of the resulting transgenic plants. Their effects on feeding behavior of tobacco cutworms (Spodoptera litura), which damage a wide range of crops, were then examined. Leaf disc choice test showed that caterpillars selectively fed on the wild-type control materials, or positively avoided the transgenic materials. The results suggest a novel approach to confer self-defense by producing caffeine in planta. A second generation of transgenic crops containing caffeine may save labor and agricultural costs and also mitigate the environmental load of pesticides in future.

Using listening difficulty ratings of conditions for speech communication in rooms.

The use of listening difficulty ratings of speech communication in rooms is explored because, in common situations, word recognition scores do not discriminate well among conditions that are near to acceptable. In particular, the benefits of early reflections of speech sounds on listening difficulty were investigated and compared to the known benefits to word intelligibility scores. Listening tests were used to assess word intelligibility and perceived listening difficulty of speech in simulated sound fields. The experiments were conducted in three types of sound fields with constant levels of ambient noise: only direct sound, direct sound with early reflections, and direct sound with early reflections and reverberation. The results demonstrate that (1) listening difficulty can better discriminate among these conditions than can word recognition scores; (2) added early reflections increase the effective signal-to-noise ratio equivalent to the added energy in the conditions without reverberation; (3) the benefit of early reflections on difficulty scores is greater than expected from the simple increase in early arriving speech energy with reverberation; (4) word intelligibility tests are most appropriate for conditions with signal-to-noise (S/N) ratios less than 0 dBA, and where S/N is between 0 and 15-dBA S/N, listening difficulty is a more appropriate evaluation tool.

Application of RNAi to confirm theobromine as the major intermediate for caffeine biosynthesis in coffee plants with potential for construction of decaffeinated varieties.

The caffeine biosynthetic pathway in coffee plants has been proposed to involve three distinct N -methyltransferases, xanthosine methyltransferase (XMT), 7- N -methylxanthine methyltransferase (MXMT; theobromine synthase), and 3,7-dimethylxanthine methyltransferase (DXMT; caffeine synthase). We previously isolated all corresponding cDNAs designated as CaXMT1 , CaMXMT1 , CaMXMT2 and CaDXMT1 , respectively, and showed that caffeine was indeed synthesized in vitro by the combination of their gene products. In order to regulate caffeine biosynthesis in planta , we suppressed expression of CaMXMT1 by the double stranded RNA interference (RNAi) method. For this purpose, we first established a protocol for efficient somatic embryogenesis of Coffea arabica and C. canephora , and then Agrobacterium -mediated transformation techniques. The RNAi transgenic lines of embryogenic tissues derived from C. arabica and transgenic plantlets of C. canephora demonstrated a clear reduction in transcripts for CaMXMT1 in comparison with the control plants. Transcripts for CaXMT1 and CaDXMT1 were also reduced in the most cases. Both embryonic tissues and plantlets exhibited a concomitant reduction of theobromine and caffeine contents to a range between 30% and 50% of that of the control. These results suggest that the CaMXMT1 -RNAi sequence affected expression of not only CaMXMT1 itself, but also CaXMT1 and CaDXMT1 , and that, since the reduction in theobromine content was proportional to that for caffeine, it is involved in the major synthetic pathway in coffee plants. The results also indicate that the method can be practically applied to produce decaffeinated coffee plants.

Listening difficulty as a subjective measure for evaluation of speech transmission performance in public spaces.

Listening difficulty ratings, using words with high word familiarity, are proposed as a new subjective measure for the evaluation of speech transmission in public spaces to provide realistic and objective results. Two listening tests were performed to examine their validity, compared with intelligibility scores. The tests included a reverberant signal and noise as detrimental sounds. The subject was asked to repeat each word and simultaneously to rate the listening difficulty into one of four categories: (1) not difficult, (2) a little difficult, (3) fairly difficult, and (4) extremely difficult. After the tests, the four categories were reclassified into, not difficult [response (1)] and some level of difficulty, (the other 3 responses). Listening difficulty is defined as the percentage of the total number of responses indicating some level of difficulty [i.e. not (1)]. The results of two listening tests demonstrated that listening difficulty ratings can evaluate speech transmission performance more accurately and sensitively than intelligibility scores for sound fields with higher speech transmission performance.