Regulation of thrombin activity by ligand-induced topological alteration in a thrombin-binding aptamer.

Thrombin-binding aptamer (TBA), which forms a G-quadruplex (G4) structure with anti-parallel topology, interacts with thrombin to inhibit its enzymatic activity. Here we show that the G4-topology-altering ligand L2H2-2M2EA-6LCO (6LCO) changes the anti-parallel topology of TBA G4 to the parallel topology, thereby abrogating the thrombin-inhibitory activity of TBA. This finding suggests that G4 ligands that alter topology may be promising drug candidates for diseases involving G4-binding proteins.

Mechanism of rate controllability of water-soluble bifunctional cyclooctadiynes through cation-anion interactions.

Click reactions are used for chemoselective functionalization in many research fields. Despite the utility of small, bioinert azide groups as a counterpart, applications of strain-promoted alkyne-azide cycloaddition (SPAAC) reactions for this purpose are still limited by slow reaction kinetics. Here, we report ion-pair-guided reaction rate enhancement by the use of water-soluble cyclooctadiynes (WS-CODYs) composed of bifunctional strained alkynes and polar side chains. Arrhenius plot analysis revealed that the rate enhancement by WS-CODYs is due to a kinetic salt effect between the polar substituents and the target azide. We demonstrate the utility of these compounds for rapid protein labelling and isoelectric point-dependent labelling.

Functions of neuronal Synaptobrevin in the post-Golgi transport of Rhodopsin in Drosophila photoreceptors.

Polarized transport is essential for constructing multiple plasma membrane domains in the cell. Drosophila photoreceptors are an excellent model system to study the mechanisms of polarized transport. Rab11 is the key factor regulating the post-Golgi transport of rhodopsin 1 (Rh1; also known as NinaE), a photoreceptive protein, to the rhabdomere, a photoreceptive plasma membrane. Here, we found that neuronal Synaptobrevin (nSyb) colocalizes with Rab11 on the trans-side of Golgi stacks and post-Golgi vesicles at the rhabdomere base, and nSyb deficiency impairs rhabdomeric transport and induces accumulation of Rh1 and vesicles in the cytoplasm; this is similar to the effects of Rab11 loss. These results indicate that nSyb acts as a post-Golgi SNARE toward rhabdomeres. Surprisingly, in Rab11-, Rip11- and nSyb-deficient photoreceptors, illumination enhances cytoplasmic accumulation of Rh1, which colocalizes with Rab11, Rabenosyn5, nSyb and Arrestin 1 (Arr1). Arr1 loss, but not Rab5 dominant negative (Rab5DN) protein expression, inhibits the light-enhanced cytoplasmic Rh1 accumulation. Rab5DN inhibits the generation of Rh1-containing multivesicular bodies rather than Rh1 internalization. Overall, these results indicate that exocytic Rh1 mingles with endocytosed Rh1 and is then transported together to rhabdomeres.

Single-molecule displacement assay reveals strong binding of polyvalent dendrimer ligands to telomeric G-quadruplex.

Binding between a ligand and a receptor is a fundamental step in many natural or synthetic processes. In biosensing, a tight binding with a small dissociation constant (Kd) between the probe and analyte can lead to superior specificity and sensitivity. Owing to their capability of evaluating competitors, displacement assays have been used to estimate Kd at the ensemble average level. At the more sensitive single-molecule level, displacement assays are yet to be established. Here, we developed a single-molecule displacement assay (smDA) in an optical tweezers instrument and used this innovation to evaluate the binding of the L2H2-6OTD ligands to human telomeric DNA G-quadruplexes. After measuring Kd of linear and dendrimer L2H2-6OTD ligands, we found that dendrimer ligands have enhanced binding affinity to the G-quadruplexes due to their polyvalent geometry. This increased binding affinity enhanced inhibition of telomerase elongation on a telomere template in a Telomerase Repeated Amplification Protocol (TRAP). Our experiments demonstrate that the smDA approach can efficiently evaluate binding processes in chemical and biological processes.

Differences in high trunk acceleration during single-leg landing after an overhead stroke between junior and adolescent badminton athletes.

The magnitude of trunk acceleration reflects the ground reaction force. This study compared the frequency of high trunk accelerations during single-leg landing after an overhead stroke between junior and adolescent badminton players, and examined the difference in each directive magnitude according to age and landing leg. Thirty-eight female badminton players (17 junior and 21 adolescent athletes) played two singles games while wearing a tri-axial accelerometer on their upper back. The frequency and 95% confidence interval (CI) of single-leg landings that generated >4-G resultant acceleration, and each directive magnitude were calculated. A two-factorial analysis of variance (factor 1: group, factor 2: landing leg) was performed to determine the effects of age and different landing patterns. Frequency of single-leg landings following an overhead stroke in the adolescent athletes (mean, 1.71 cases/min; 95% CI, 1.59-1.83 cases/min) was higher than that in the junior athletes (mean, 1.13 cases/min; 95% CI, 1.01-1.25 cases/min). The adolescent athletes exhibited greater mediolateral acceleration in the movement towards racket-hand leg and anteroposterior acceleration in the movement towards the opposite leg than the junior athletes. This cross-sectional study suggests that the frequency and movement pattern associated with high-load landing in badminton games differ between junior and adolescent athletes.

Acceleration Profile of High-Intensity Movements in Basketball Games.

ABSTRACT: Koyama, T, Rikukawa, A, Nagano, Y, Sasaki, S, Ichikawa, H, and Hirose, N. Acceleration profile of high-intensity movements in basketball games. J Strength Cond Res 36(6): 1715-1719, 2022-This study aimed to elucidate movements that require greater acceleration during basketball games, their occurrence frequency, and compare acceleration components. Eighteen male collegiate basketball players (age: 19.5 ± 0.8 years) were enrolled. Triaxial accelerometer was used to measure acceleration and a synchronized video was recorded. Moments that generated resultant accelerations >4, 6, and 8 G, and their coincided movements were identified. Ratios and frequencies of the extracted movements were calculated and Jonckheere-Terpstra trend test was used to examine which movement rate increased when acceleration threshold increased. In addition, the top 7 movements that generated a resultant acceleration >6 G among the combination of basketball-specific movements were extracted. Their resultant, mediolateral, vertical, and anteroposterior accelerations were identified and compared using one-way analysis of variance. Cohen's d was used to calculate effect sizes. All p < 0.05 were considered statistically significant. The extracted frequencies were 33.6, 9.1, and 2.3 cases per minute for >4 G, >6 G, and >8 G, respectively. As the threshold increased, the rate of deceleration, landing, and physical contact increased. The mediolateral acceleration of physical contact was significantly greater than other movements, whereas the vertical acceleration of landing and deceleration was significantly greater than other movements. Thus, acceleration component analysis was performed to classify movement types. Greater acceleration appeared frequently in movement during defense. It is suggested that many defense movements involve a reaction to the ball and opponent. There are many passive movements during defense and speed changes rapidly. If many passive movements occur when defending, larger physical load is applied, and may lead to fatigue.

Neuromuscular responses of the hamstring and lumbopelvic muscles during unanticipated trunk perturbations.

Hamstring strain often occurs when an opponent unanticipatedly perturbs an athlete's movements. We examined the neuromuscular responses of hamstring and trunk muscles during unanticipated trunk perturbations in athletes with and without a history of hamstring strain injury. Male college athletes (11 with a history of a unilateral hamstring injury and 10 without prior injury) knelt while wearing a chest harness attached to a cable that was pulled backward. They were instructed to resist the force isometrically and maintain their position when the perturbations were applied. The pressure was released with or without a cue (CUE or NoCUE). We measured trunk acceleration, three-dimensional kinematic data, and surface electromyography (EMG) signals of the erector spinae, internal oblique, gluteus maximus, biceps femoris long head, and semitendinosus muscles. Maximum trunk acceleration and displacement were greater with NoCUE in both groups (p < 0.05). EMG amplitude did not differ after perturbation of any investigated muscle. The injured group demonstrated a delayed onset of the gluteus maximus and erector spinae muscles in NoCUE versus CUE stimuli (p < 0.05). Athletes with a history of hamstring strain injury exhibited a reduced neuromuscular coordination of the lumbopelvic muscles in response to unanticipated trunk movement.

High-Acceleration Movement, Muscle Damage, and Perceived Exertion in Basketball Games.

PURPOSE: To evaluate the effect of the number of high-acceleration movements on muscle damage and the rating of perceived exertion (RPE) in basketball games. METHODS: Twenty-one male collegiate basketball players (mean age, 20.0 [1.0] y) were included. A triaxial accelerometer was used to measure acceleration in basketball-simulated scrimmages. To detect higher physical load during the actual game, the resultant acceleration was calculated, and 3 thresholds were set: >4G, >6G, and >8G resultant accelerations. The number of the extracted movements was calculated at each acceleration threshold. Plasma creatine kinase (CK) levels (marker of muscle damage) were estimated before and 24 hours after the match, and the session-RPE load was calculated within 30 minutes after the match. Pearson product-moment correlations with 95% confidence intervals were used to determine the relationships between the number of high-acceleration movements and plasma CK and session-RPE load. RESULTS: Significant correlations were observed between the number of high-acceleration movements >8G and CK level (r = .74; 95% confidence interval, 0.44-0.89; P < .0001). Furthermore, the correlation coefficient between acceleration and CK increased with increased acceleration threshold (>4G: r = .65; >6G: r = .69). Contrastingly, the correlation coefficient between acceleration and the session-RPE load decreased with increased acceleration threshold (>4G: r = .72; >6G: r = .52; >8G: r = .43). CONCLUSIONS: The session-RPE reflects the total amount of movement, while the high-acceleration movement reflects the momentary large impact load or intensity, and they evaluated different factors. Basketball coaching and conditioning professionals recommended combining acceleration and session-RPE when monitoring the load of athletes.

Acceleration profile of high-impact movements during young football games: a cross-sectional study involving healthy children.

Repetitive high-impact movements cause growth-related injuries in children. This study aimed to identify which movements during junior football games require >6 G and >8 G acceleration and the frequency at which they occur. Additionally, we compared the components of acceleration among movements with >8 G resultant acceleration. Eleven young male footballers (10.7 ± 0.4 years) played 8-a-side games while wearing a tri-axial accelerometer on their upper back. The number and frequency of the movements that generated >6 G and >8 G were calculated, and each directive acceleration of the top five items was compared using two-way ANOVA to examine the effect of movements. The frequency of movements that generated >6 G and >8 G acceleration during junior football games was 8.70 case/min and 2.62 case/min, respectively. The top five >8 G movements were braking and pre-braking in shuffle, slowdown, stop, and run/jog items. The vertical acceleration was significantly greater during braking in shuffle than during slowdown, stop, and run/jog and also greater during stop and pre-braking in shuffle than during run/jog movement. This pilot study suggests that decelerated movements mainly provoked high-impact situations and may be key actions for preventing overuse injury in young footballers.

Stabilization of telomeric G-quadruplex by ligand binding increases susceptibility to S1 nuclease.

The extent of thermodynamic stabilization of telomeric G-quadruplex (G4) by isomers of G4 ligand L2H2-6OTD, a telomestatin analog, is inversely correlated with susceptibility to S1 nuclease. L2H2-6OTD facilitated the S1 nuclease activities through the base flipping in G4, unlike the conventional role of G4 ligands which inhibit the protein binding to DNA/RNA upon ligand interactions.

High-Impact Details of Play and Movements in Female Basketball Game.

This study aimed to identify the high-impact details of play and movements with higher acceleration and their frequency during a female basketball match. Trunk acceleration was measured during a simulated basketball game with eight female players. The extracted instance was categorized, which generated at > 6 and 8 G resultant accelerations using a video recording and an accelerometer attached to the players' trunk, as details of play and movements. The frequency and ratio of the details of play and movements regarding all detected movements were calculated. A total of 1062 and 223 play actions were detected for the resultant acceleration thresholds of > 6 and 8 G, respectively. For these acceleration thresholds, in terms of details of play, positioning on the half-court was the most frequently observed (29.6 and 23.8%, respectively). In terms of movements, deceleration was the most frequently detected movement (21.5 and 23.3%, respectively), followed by landing (7.6 and 15.7%, respectively). Deceleration during positioning on the half-court and defense as well as landing mostly after a shot were detected as high-impact frequent basketball-specific movements. The results also showed that characteristics of movements or playing style and playing position may have an effect on acceleration patterns during a basketball game.

Selective alkylation of parallel G-quadruplex structure.

The selective alkylation of nucleic acids is important for a medicinal approach and biological study. We now report a novel selective alkylation of the parallel G-quadruplex structure using the conjugate of the macrocyclic hexaoxazole L2G2-6OTD-1M1PA and vinyl-quinazolinone-S(O)Me (6OTD-VQ-S(O)Me).

Induction of potent cell growth inhibition by schizophyllan/K-ras antisense complex in combination with gemcitabine.

Antisense oligonucleotides (AS-ODNs) specifically hybridize with target mRNAs, resulting in interference with the splicing mechanism or the regulation of protein translation. In our previous reports, we demonstrated that ß-glucan schizophyllan (SPG) can form a complex with AS-ODNs attached with oligo deoxyadenosine dA40 (AS-ODN-dA40/SPG), and that this complex can be recognized by ß-glucan receptor Dectin-1 on antigen presenting cells and lung cancer cells. In many types of cancer cell, activating K-ras mutations related to malignancy are frequently observed. In this study, we first designed 78 AS-ODNs for K-ras to optimize the sequence for highly efficient gene suppression. The selected AS-ODN (K-AS07) having dA40 made a complex with SPG. The resultant complex (K-AS07-dA40/SPG) showed an effect of silencing the ras gene in the cells (PC9: human adenocarcinoma differentiated from lung tissue) expressing Dectin-1, leading to the suppression of cell growth. Furthermore, the cytotoxic effect was enhanced when used in combination with the anticancer drug gemcitabine. Gemcitabine, a derivative of cytidine, was shown to interact with dA40 in a sequence-dependent manner. This interaction did not appear to be so strong, with the gemcitabine being released from the complex after internalization into the cells. SPG and the dA40 part of K-AS07-dA40 play roles in carriers for K-AS07 and gemcitabine, respectively, resulting in a strong cytotoxic effect. This combination effect is a novel feature of the AS-ODN-dA40/SPG complexes. These results could facilitate the clinical application of these complexes for cancer treatment.

Linear consecutive hexaoxazoles as G4 ligands inducing chair-type anti-parallel topology of a telomeric G-quadruplex.

G-quadruplex structures (G4s) in guanine-rich regions of DNA play critical roles in various biological phenomena, including replication, translation, and gene expression. There are three types of G4 topology, i.e., parallel, anti-parallel, and hybrid, and ligands that selectively interact with or stabilize a specific topology have been extensively explored to enable studies of topology-related functions. Here, we describe the synthesis of a new series of G4 ligands based on 6LCOs (6-linear consecutive oxazoles), i.e., L2H2-2M2EA-6LCO (2), L2A2-2M2EAc-6LCO (3), and L2G2-2M2EG-6LCO (4), which bear four aminoalkyl, acetamidealkyl, and guanidinylalkyl side chains, respectively. Among them, ligand 2 stabilized telomeric G4 and induced anti-parallel topology independently of the presence of cations. The anti-parallel topology induced by 2 was identified as chair-type by means of 19F NMR spectroscopy and fluorescence experiments with 2-aminopurine-labeled DNA.

Movements with greater trunk accelerations and their properties during badminton games.

This study aimed to elucidate the movements requiring greater trunk accelerations and its frequencies during badminton games, and compare the acceleration components among such movements. Trunk acceleration was measured using a triaxial accelerometer during badminton games. The moments that generated >4 G resultant acceleration were extracted, and movements consistent with the extracted moments were identified. We calculated the extracted movement ratio and frequency and compared the resultant, mediolateral, vertical and anteroposterior accelerations between the top five extracted movements. There were 1,342 movements that generated >4 G [mean, 7.72 (95% confidence interval, 7.31-8.14) cases/min]. The top five movements were lunging during underhand strokes with the dominant hand side leg, landing after overhand strokes on the dominant and non-dominant hand side leg, and cutting from a split step using the dominant and non-dominant hand side leg. Landing on the dominant hand side leg had a greater resultant acceleration than the other movements and had the greatest impact during the badminton game. Lunging during underhand strokes on the dominant hand side leg had greater mediolateral acceleration than the other movements. These results reflected the properties of badminton.

Exciton Polarization and Renormalization Effect for Optical Modulation in Monolayer Semiconductors.

The ideal quantum confinement structure of monolayer semiconductors offers prominent optical modulation capabilities that are mediated by enhanced many-body interactions. Herein, we establish an electrolyte-gating method for tuning the luminescence properties that are in transition metal dichalcogenide (TMDC) monolayers. We fabricate electric double-layer capacitors on TMDC/graphite heterostructures to investigate electric-field- and carrier-density-dependent photoluminescence. The exciton peak energy initially shows a slight quadratic red shift of ∼1 meV without carrier accumulations, which is caused by the quantum-confined Stark effect. In contrast, the exciton resonance exhibits a larger red shift up to 10 meV with the accumulated carrier density above 1013 cm-2. These results indicate that the optical transitions can be largely modulated by the carrier density control in S- and Se-based TMDCs, as triggered by the doping-induced band gap renormalization effect. To further inspire this modulation capability, we also apply our method to electrolyte-based TMDC light-emitting devices. Biasing solely in electrolyte-induced p-i-n junctions yields pronounced red shifts up to 40 meV for exciton and trion electroluminescence. Consequently, our approach reveals that the doping effects in the high-carrier-density regimes are potentially significant for efficient optical modulation in monolayer semiconductors.

Interband resonant high-harmonic generation by valley polarized electron-hole pairs.

High-harmonic generation in solids is a unique tool to investigate the electron dynamics in strong light fields. The systematic study in monolayer materials is required to deepen the insight into the fundamental mechanism of high-harmonic generation. Here we demonstrated nonperturbative high harmonics up to 18th order in monolayer transition metal dichalcogenides. We found the enhancement in the even-order high harmonics which is attributed to the resonance to the band nesting energy. The symmetry analysis shows that the valley polarization and anisotropic band structure lead to polarization of the high-harmonic radiation. The calculation based on the three-step model in solids revealed that the electron-hole polarization driven to the band nesting region should contribute to the high harmonic radiation, where the electrons and holes generated at neighboring lattice sites are taken into account. Our findings open the way for attosecond science with monolayer materials having widely tunable electronic structures.

Synthesis and Telomeric G-Quadruplex-Stabilizing Ability of Macrocyclic Hexaoxazoles Bearing Three Side Chains.

G-quadruplexes (G4s), which are structures formed in guanine-rich regions of DNA, are involved in a variety of significant biological functions, and therefore "sequence-dependent" selective G4-stabilizing agents are required as tools to investigate and modulate these functions. Here, we describe the synthesis of a new series of macrocyclic hexaoxazole-type G4 ligand (6OTD) bearing three side chains. One of these ligands, 5b, stabilizes telomeric G4 preferentially over the G4-forming DNA sequences of c-kit and K-ras, due to the interaction of its piperazinylalkyl side chain with the groove of telomeric G4.

Binding of a Telomestatin Derivative Changes the Mechanical Anisotropy of a Human Telomeric G-Quadruplex.

Mechanical anisotropy is an essential property for biomolecules to assume structural and functional roles in mechanobiology. However, there is insufficient information on the mechanical anisotropy of ligand-biomolecule complexes. Herein, we investigated the mechanical property of individual human telomeric G-quadruplexes bound to telomestatin, using optical tweezers. Stacking of the ligand to the G-tetrad planes changes the conformation of the G-quadruplex, which resembles a balloon squeezed in certain directions. Such a squeezed balloon effect strengthens the G-tetrad planes, but dislocates and weakens the loops in the G-quadruplex upon ligand binding. These dynamic interactions indicate that the binding between the ligand and G-quadruplex follows the induced-fit model. We anticipate that the altered mechanical anisotropy of the ligand-G-quadruplex complex can add additional level of regulations on the motor enzymes that process DNA or RNA molecules.

Random Formation of G-Quadruplexes in the Full-Length Human Telomere Overhangs Leads to a Kinetic Folding Pattern with Targetable Vacant G-Tracts.

G-Quadruplexes formed in the 3' telomere overhang (∼200 nucleotides) have been shown to regulate biological functions of human telomeres. The mechanism governing the population pattern of multiple telomeric G-quadruplexes is yet to be elucidated inside the telomeric overhang in a time window shorter than thermodynamic equilibrium. Using a single-molecule force ramping assay, we quantified G-quadruplex populations in telomere overhangs over a full physiological range of 99-291 nucleotides. We found that G-quadruplexes randomly form in these overhangs within seconds, which leads to a population governed by a kinetic, rather than a thermodynamic, folding pattern. The kinetic folding gives rise to vacant G-tracts between G-quadruplexes. By targeting these vacant G-tracts using complementary DNA fragments, we demonstrated that binding to the telomeric G-quadruplexes becomes more efficient and specific for telomestatin derivatives.