Functional Roles of the Edited Isoform of GluA2 in GluA2-Containing AMPA Receptor Channels.

GluA2R is the edited isoform at the Q/R site of the AMPA receptor GluA2 subunit. In vivo, GluA2R assembles with other AMPA receptor subunits into a high-conductance, tetrameric complex. Here we study the functional role of GluA2R in the context of GluA1/2R and GluA2Q/2R heteromeric receptors, using whole-cell recording and a laser-pulse photolysis technique. We find the incorporation of GluA2R with GluA1 or GluA2Q slows the channel-opening and channel-closing rates; for GluA2Q/2R, it further lowers EC50, compared to the value of the GluA2Q homomeric channels. When a conserved leucine-to-tyrosine (L → Y) mutation is "placed" in each of these subunits, the L → Y mutation exhibits a greater effect in GluA2R than in GluA1. Together, our results show GluA2R is the dominating isoform that shapes the overall functional properties of the GluA2R-containing channels. Our results further suggest that glutamate most likely binds to GluA2R with an EC50 of ∼0.5 mM, rather than a non-GluA2R subunit in a heteromeric AMPA receptor, and binding of two glutamate molecules to the two GluA2R subunits is sufficient to initiate a conformational change, leading to the opening of the channel.

High-performance liquid chromatography purification of chemically modified RNA aptamers.

2'-Fluoro modified RNAs are useful as potential therapeutics and as special substrates for studying RNA function. 2'-Fluoro modified RNAs generally need to be purified after they are prepared either enzymatically or by solid-phase synthesis. Here we introduce a protocol by which 2'-fluoro modified RNAs with 57 and 58 nucleotides can be resolved and purified using ion-pair, reverse-phase high-performance liquid chromatography (HPLC). Because the size of our RNA samples is in the range of many known RNA aptamers of therapeutic values, our protocol should be generally useful.

Effect of mindfulness-based intervention on endurance performance under pressure and performance-relevant mental attributes, an interdisciplinary perspective: Protocol for a Mindfulness-Based Peak Performance (MBPP) trial.

Performance under pressure is one of the primary features of competitive sports. Considering that increased competition levels are typically accompanied by elevated stress and anxiety, athletes' ability to cope with stress has gained even more importance in recent years. Accordingly, the current trial, entitled Mindfulness-based Peak Performance (MBPP), will take an interdisciplinary approach (e.g., sport psychology, sports training, and cognitive neuroscience), to more definitively examine whether a MBPP affects athletic performance under pressure and relevant mental attributes. This study is an 8-week, three-arm, randomized controlled trial (RCT). A total of 90 athletes, aged between 18 and 30 years will be recruited. Eligible participants will be randomly assigned into (1) an MBPP group, (2) a self-talk (ST) group, and (3) a wait-list control (WC) group. The MBPP and ST interventions consist of a 60-min session weekly for 8 weeks. Primary outcomes are endurance performance and performance-relevant mental attributes including behavior (i.e., stress response, emotion regulation, and engagement) and neurocognitive processes (e.g., attention, executive function, brain resting state), which will be assessed at baseline and post-intervention. Dispositional mindfulness and athletic psychological skills will be secondary outcomes, also assessed at baseline and post-intervention. The MBPP and ST are expected to improve performance under pressure, but MBPP is expected to show greater improvement than ST. Additionally, we expect the MBPP will improve the relevant mental attributes. The results from this trial might provide rigorous evidence and insight into MBI application in the sports context. ClinicalTrials.govregistration:NCT05612295.

The three-dimensional plasma structures and flows of the Earth's upper atmosphere due to the Moon's gravitational force.

The semidiurnal (12.42 h) and semimonthly (14.76 days) lunar tides have been well-known by fishermen for several centuries. The gravitational force of the relative positions between the Sun, the Moon, and the Earth results in two symmetrical tidal bulges (double bulges) appearing at equatorial latitudes directly under and opposite the Moon. We utilize ionospheric GNSS (Global Navigation Satellite System) radio occultation soundings to show the global three-dimensional structures and dynamics of the double bulges of ionospheric lunar tides for the first time. The double-bulge amplitude of ionospheric F2-peak height hmF2, lagging the sublunar or antipodal point by about 2-3 h, is about 3-5 km at the equator and 1.5-2.0 km at ± 35° magnetic latitude. The electron density further depicts global three-dimensional plasma flows in the ionosphere.

Cardiorespiratory fitness is associated with sustained neurocognitive function during a prolonged inhibitory control task in young adults: An ERP study.

Although beneficial associations between cardiorespiratory fitness and cognitive function have been established, whether cardiorespiratory fitness is related to behavioral and neuroelectric indices of performance during a prolonged inhibitory control task remains unknown. Young adults, categorized into High and Low Fitness groups, completed a 60-min Stroop task, while the N1 and P3 components of event-related potentials were measured. The results showed that the High Fitness group demonstrated shorter response times, regardless of the Stroop task congruency or time-on-task, than Low Fitness group. The High Fitness group also exhibited larger P3 amplitudes than the Low Fitness group, but no differences in N1 amplitudes were observed. These findings suggest that cardiorespiratory fitness during young adulthood has beneficial effects on task performance and attention allocation during an inhibitory control task, and these benefits can be sustained for 60 min.

Ph2N-susbtituted ethylene-bridged p-phenylene oligomers: synthesis and photophysical and redox properties.

For a series of p-phenylene-based oligomers terminated with two triphenylamines, their absorption, photoluminescence, and band gaps show a pattern of extensive π-conjugation with increasing array size. Oligomers with large central arrays have greater quantum yields than their small analogues. Cyclic voltammetric (CV) measurements indicated two-step oxidations of the two diphenylamino groups for compounds 1-5 and one-step oxidations for the two amines of large oligomers 6 and 7.

The Combined Effects of Obesity and Cardiorespiratory Fitness Are Associated with Response Inhibition: An ERP Study.

Obesity and cardiorespiratory fitness exhibit negative and positive impacts, respectively, on executive function. Nevertheless, the combined effects of these two factors on executive function remain unclear. This study investigated the combined effects of obesity and cardiorespiratory fitness on response inhibition of executive function from both behavioral and neuroelectric perspectives. Ninety-six young adults aged between 18 and 25 years were recruited and assigned into four groups: the high cardiorespiratory fitness with normal weight (NH), high cardiorespiratory fitness with obesity (OH), low cardiorespiratory fitness with normal weight (NL), and low cardiorespiratory fitness with obesity (OL) groups. The stop-signal task and its induced P3 component of event-related potentials was utilized to index response inhibition. The participants with higher cardiorespiratory fitness (i.e., the NH and OH groups) demonstrated better behavioral performance (i.e., shorter response times and higher accuracy levels), as well as shorter stop-signal response times and larger P3 amplitudes than their counterparts with low cardiorespiratory fitness (i.e., the NL and OL groups). The study provides first-hand evidence of the substantial effects of cardiorespiratory fitness on the response inhibition, including evidence that the detrimental effects of obesity might be overcome by high cardiorespiratory fitness.

Relationship between Mindfulness, Psychological Skills, and Mental Toughness in College Athletes.

Numerous studies have shown that dispositional mindfulness is positively associated with many mental abilities related to sports performance, including psychological skills and mental toughness. The purpose of this study was to explore the relationship between dispositional mindfulness, psychological skills, and mental toughness among different types of athletes. For this cross-sectional study, 101 college athletes were recruited. Their dispositional mindfulness, psychological skills, and mental toughness were measured by the Mindfulness Attention Awareness Scale (MAAS), Athletic Psychological Skills Inventory (APSI), and Traits of Mental Toughness Inventory for Sports Scale (TMTIS). Pearson's correlation was used to calculate how dispositional mindfulness is associated with psychological skills and mental toughness. The results revealed that dispositional mindfulness is positively associated with comprehensive APSI (r = 0.21-0.36, p < 0.05), TMTIS overall (r = 0.27, p < 0.01), positive effort (r = 0.26, p = 0.01), and pressure (r = 0.30, p < 0.01). These findings suggest a positive linkage between mindfulness and the two examined psychological characteristics related to sports performance. Other approaches to increase mindfulness may be considered in the future.

Quantitative characterization of the interactions among c-myc transcriptional regulators FUSE, FBP, and FIR.

Human c-myc is critical for cell homeostasis and growth but is a potent oncogenic factor if improperly regulated. The c-myc far-upstream element (FUSE) melts into single-stranded DNA upon active transcription, and the noncoding strand FUSE recruits an activator [the FUSE-binding protein (FBP)] and a repressor [the FBP-interacting repressor (FIR)] to fine-tune c-myc transcription in a real-time manner. Despite detailed biological experiments describing this unique mode of transcriptional regulation, quantitative measurements of the physical constants regulating the protein-DNA interactions remain lacking. Here, we first demonstrate that the two FUSE strands adopt different conformations upon melting, with the noncoding strand DNA in an extended, linear form. FBP binds to the linear noncoding FUSE with a dissociation constant in the nanomolar range. FIR binds to FUSE more weakly, having its modest dissociation constants in the low micromolar range. FIR is monomeric under near-physiological conditions but upon binding of FUSE dimerizes into a 2:1 FIR(2)-FUSE complex mediated by the RRMs. In the tripartite interaction, our analysis suggests a stepwise addition of FIR onto an activating FBP-FUSE complex to form a quaternary FIR(2)-FBP-FUSE inhibitory complex. Our quantitative characterization enhances understanding of DNA strand preference and the mechanism of the stepwise complex formation in the FUSE-FBP-FIR regulatory system.

A novel ambipolar spirobifluorene derivative that behaves as an efficient blue-light emitter in organic light-emitting diodes.

A novel ambipolar spiro-configured D-A blue-light emitter bearing hole-transporting diphenylamino groups and electron-transporting phenylbenzimidazole groups was synthesized, characterized, and incorporated into an efficient single-layer organic light-emitting diode (OLED) device exhibiting blue-emission Commission International d'Eclairage (CIE) coordinates of 0.15 and 0.14, a turn-on potential of 4 V, a maximum brightness of 2800 cd/m2 at 830 mA/cm2 (19 V), and a maximum quantum efficiency of 0.53% (0.61 cd/A).

An unprecedented ambipolar charge transport material exhibiting balanced electron and hole mobilities.

2,7-Bis(4-tert-butylphenyl)-9,9'-spirobifluorene exhibits balanced electron and hole mobilities of up to 10(-3) cm(2) V(-1) s(-1), as measured using time-of-flight techniques.

R/G editing in GluA2Rflop modulates the functional difference between GluA1 flip and flop variants in GluA1/2R heteromeric channels.

In α-amino-3-hydroxy-5-methyl-4-isoxazole-propionate (AMPA) receptors, RNA editing and alternative splicing generate sequence variants, and those variants, as in GluA2-4 AMPA receptor subunits, generally show different properties. Yet, earlier studies have shown that the alternatively spliced, flip and flop variants of GluA1 AMPA receptor subunit exhibit no functional difference in homomeric channel form. Using a laser-pulse photolysis technique, combined with whole-cell recording, we measured the rate of channel opening, among other kinetic properties, for a series of AMPA channels with different arginine/glycine (R/G) editing and flip/flop status. We find that R/G editing in the GluA2 subunit modulates the channel properties in both homomeric (GluA2Q) and complex (GluA2Q/2R and GluA1/2R) channel forms. However, R/G editing is only effective in flop channels. Specifically, editing at the R/G site on the GluA2R flop isoform accelerates the rate of channel opening and desensitization for GluA1/2R channels more pronouncedly with the GluA1 being in the flop form than in the flip form; yet R/G editing has no effect on either channel-closing rate or EC50. Our results suggest R/G editing via GluA2R serve as a regulatory mechanism to modulate the function of GluA2R-containing, native receptors involved in fast excitatory synaptic transmission.

Glutamate-gated chloride channels inhibit juvenile hormone biosynthesis in the cockroach, Diploptera punctata.

Juvenile hormone (JH) synthesized and released from endocrine gland corpus allatum (CA) plays an important role in insect metamorphosis, vitellogenesis and reproduction. Glutamate is a major neurotransmitter in the nervous system and its activated receptors possess excitatory and inhibitory forms in muscle fibers of invertebrates. Previously, we have shown that the rise of intracellular calcium through excitatory glutamate receptors, N-methyl-d-aspartate (NMDA) and non-NMDA-type channels stimulates JH synthesis in the cockroach, Diploptera punctata. Here, we demonstrate the occurrence of inhibitory chloride permeable glutamate (GluCl) receptors on CA cell membranes. Application of the GluCl channel activators, ibotenic acid (Ibo) and ivermectin, but not gamma-aminobutyric acid caused a decline in JH synthesis in glands of either high or low activity during the gonadotrophic cycle. Also, while recording the membrane potential of the isolated whole CA glands intracellularly, Ibo induced a hyperpolarizated response. Both changes in the membrane potential and inhibition of JH synthesis could be abolished by the application of the chloride channel blocker picrotoxin. Finally, we found both excitatory and inhibitory glutamate receptors cause antagonistic effects on rates of JH synthesis. These results indicate a novel function of GluCl channels in the inhibition of JH synthesis that could be a potential pathway for developing a new generation of insecticides.

HPLC purification of RNA aptamers up to 59 nucleotides with single-nucleotide resolution.

An RNA sample is usually heterogeneous. RNA heterogeneity refers to difference in length or size (i.e., number of nucleotides [nt]), sequence, or alternative but coexisting conformations. Separation and purification of RNA is generally required for investigating the structure and function of RNA, such as RNA catalysis and RNA structure determination by nuclear magnetic resonance or crystallography. Separation and purification of RNA is also required for using RNAs as functional probes and therapeutics as well as building blocks for RNA nanoparticles. Previously established protocols are limited in separating RNAs longer than 25 nt by single-nucleotide resolution. When the length of RNAs becomes longer, single-nucleotide separation of RNAs becomes more challenging. Here we describe protocols, by the use of ion-pair, reverse-phase high-performance liquid chromatography (HPLC), to extend our ability to separate regular RNAs up to 59 nt with single-nucleotide resolution. For chemically modified RNAs at 2' positions on the ribose, we can resolve RNAs of similar sizes even with a 26 Da difference. This is much less than 320 Da, an average single-nucleotide molecular weight difference.

Enhancing Protein Expression in HEK-293 Cells by Lowering Culture Temperature.

Animal cells and cell lines, such as HEK-293 cells, are commonly cultured at 37°C. These cells are often used to express recombinant proteins. Having a higher expression level or a higher protein yield is generally desirable. As we demonstrate in this study, dropping culture temperature to 33°C, but not lower, 24 hours after transient transfection in HEK-293S cells will give rise to ~1.5-fold higher expression of green fluorescent protein (GFP) and α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors. By following the time course of the GFP-expressing cells growing at 37°C and 33°C from 24 hours after transfection (including 19 hours recovery at 37°C in the normal growth medium), we found that a mild hypothermia (i.e., 33°C) reduces the growth rate of HEK-293S cells, while increasing cellular productivity of recombinant proteins. As a result, green cells remain undivided in a longer period of time. Not surprisingly, the property of a recombinant protein expressed in the cells grown at 33°C is unaffected, as shown by the use of AMPA receptors. We further demonstrate with the use of PC12 cells that this method may be especially useful when a recombinant protein is difficult to express using a chemical-based, transient transfection method.

Spirobifluorene-bridged donor/acceptor dye for organic dye-sensitized solar cells.

A new dye, SSD1, featuring two donor/acceptor chromophores aligned in a spiro configuration with two anchoring groups separated at a distance of 10.05 A (closely matching the distance between the adsorption sites of the anatase TiO(2) surface) undergoes efficient dye adherence on TiO(2) films. A dye-sensitized solar cell incorporating SSD1 exhibited a short-circuit current of 8.9 mA cm(-2), an open-circuit voltage of 0.63 V, a fill factor of 0.67, and a power conversion efficiency of 3.75%.

Areca (betel) nut extract activates mitogen-activated protein kinases and NF-kappaB in oral keratinocytes.

Areca (betel) was recently proved a carcinogenic substance by the International Agency for Research on Cancer. However, the signaling impact of areca in oral keratinocyte is still obscure. Mitogen-activated protein kinase superfamilies, including extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinases (JNK) and p38, together with transcription factor NF-kappaB, are important signaling elements. We examined the activation of these signaling pathways in OECM-1 and SAS oral keratinocytes, treated with ripe areca nut extract (ANE). In both cells, a rapid increase in JNK1 activity at 0.5 hr was noted following treatment of ANE. ERK was profoundly activated during 0.5-2 hr in OECM-1 cells. Contrasting p38 activity was noted in these 2 cells. In both cells, ANE also activated NF-kappaB pathway in a biphasic manner, particularly for SAS cells. NF-kappaB was activated by approximately 2- to 4-fold at 0.5-1 hr and a plateau or slight decrease of activity existed between 1 and 6 hr. Later, another higher episode of NF-kappaB activity was raised. This was accompanied with the rapid degradation in cytosolic IkappaBalpha as well as an increase of nuclear NF-kappaB in both cells. ANE treatment did not activate epidermal growth factor receptor signaling system, but blockage of NF-kappaB activation rendered the suppression of ANE-modulated COX-2 upregulation in OECM-1. This study identified that ANE affected interactive signaling systems in oral keratonocytes that could be the pathogenetic basis for areca.