Long-term intraocular pressure-lowering efficacy and safety of ripasudil-brimonidine fixed-dose combination for glaucoma and ocular hypertension: a multicentre, open-label, phase 3 study.

PURPOSE: To evaluate the long-term efficacy and safety of ripasudil-brimonidine fixed-dose combination (RBFC), a new intraocular pressure (IOP)-lowering medication for glaucoma and ocular hypertension (OHT). METHODS: This prospective, multicentre (23 sites in Japan), open-label study enrolled patients with primary open-angle glaucoma (POAG), OHT or exfoliative glaucoma and assigned them to one of four combination therapy cohorts, based on previous treatment(s) received: prostaglandin (PG) analogue (Cohort 1); PG analogue and beta-adrenoceptor blocker (ß-blocker) (Cohort 2); PG analogue, ß-blocker and carbonic anhydrase inhibitor (Cohort 3); or other/no treatment (Cohort 4). After a ≥ 4-week screening period, eligible patients received twice-daily RBFC for 52 weeks in addition to the treatments they were already receiving. Efficacy was assessed by change in IOP from baseline through week 52. Adverse events and adverse drug reactions (ADRs) were monitored throughout. RESULTS: In total, 179 patients from Cohort 1 (n = 48), Cohort 2 (n = 44), Cohort 3 (n = 41) and Cohort 4 (n = 46) entered the RBFC treatment period. For all cohorts, mean IOP was significantly reduced at 11:00 (2 h after instillation of RBFC) through week 52 with the changes from baseline at week 52 of - 2.7 to - 4.1 mmHg across cohorts; all p < 0.001. Common ADRs were conjunctival hyperaemia (58%), allergic conjunctivitis (18%) and blepharitis (17%), most of which were mild in severity. CONCLUSION: These data demonstrated the long-term efficacy and safety of RBFC, both alone and in combination with other anti-glaucoma agents. RBFC may offer a new treatment option for the long-term management of glaucoma and OHT. TRIAL REGISTRATION: Japan Registry of Clinical Trials Identifier: jRCT2080225063. DATE OF REGISTRATION: 17 February 2020.

Performance-Dependent Consolidation of Learned Vocal Changes in Adult Songbirds.

Motor skills learned through practice are consolidated at later time, which can include nighttime, but the time course of motor memory consolidation and its underlying mechanisms remain poorly understood. We investigated neural substrates underlying motor memory consolidation of learned changes in birdsong, a tractable model system for studying neural basis of motor skill learning. Previous studies in male zebra finches and Bengalese finches have demonstrated that adaptive changes in adult song structure learned through a reinforcement paradigm are initially driven by a cortical-basal ganglia circuit, and subsequently consolidated into downstream cortical motor circuitry. However, the time course of the consolidation process, including whether it occurs offline during nighttime or online during daytime, remains unclear and even controversial. Here, we provide in both species experimental evidence of virtually no consolidation of learned vocal changes during nighttime. We demonstrate instead that the consolidation occurs during daytime and the amount of consolidation is strongly correlated with the amount of learning, suggesting online, performance-dependent mechanisms of consolidation of learned vocal changes. Moreover, by using computer simulations based on our experimental results, we demonstrate that such online, performance-dependent consolidation can account for the contradicting conclusions concerning the time course of consolidation process reached by previous studies. These results thus reconcile a controversy in the study of vocal motor consolidation in songbirds, and illustrate the neural substrates through which newly learned motor skills initially implemented by cortical-basal ganglia circuits become encoded in the cortical motor circuitry.SIGNIFICANCE STATEMENT Motor skills learned through repetitive practice become stable and are consolidated into cortical motor circuits. We investigate neural substrates of this "motor memory consolidation" in adult songbirds, which produce songs that are complex motor skills learned and maintained through repetitive vocal practice. We demonstrate that learned changes in song acoustic structure are consolidated into the cortical motor circuits predominantly during daytime, but not during nighttime, depending on ongoing song performance. These consolidation mechanisms reconcile seemingly contradicting results of previous studies regarding the time course of vocal learning consolidation, and provide fundamental insights into the process through which learned performance of complex motor skills is consolidated and encoded in in motor circuits.

Baloxavir safety and clinical and virologic outcomes in influenza virus-infected pediatric patients by age group: age-based pooled analysis of two pediatric studies conducted in Japan.

BACKGROUND: Anti-influenza treatment is important for children and is recommended in many countries. This study assessed safety, clinical, and virologic outcomes of baloxavir marboxil (baloxavir) treatment in children based on age and influenza virus type/subtype. METHODS: This was a post hoc pooled analysis of two open-label non-controlled studies of a single weight-based oral dose of baloxavir (day 1) in influenza virus-infected Japanese patients aged < 6 years (n = 56) and ≥ 6 to < 12 years (n = 81). Safety, time to illness alleviation (TTIA), time to resolution of fever (TTRF), recurrence of influenza illness symptoms and fever (after day 4), virus titer, and outcomes by polymerase acidic protein variants at position I38 (PA/I38X) were evaluated. RESULTS: Adverse events were reported in 39.0 and 39.5% of patients < 6 years and ≥ 6 to < 12 years, respectively. Median (95% confidence interval) TTIA was 43.2 (36.3-68.4) and 45.4 (38.9-61.0) hours, and TTRF was 32.2 (26.8-37.8) and 20.7 (19.2-23.8) hours, for patients < 6 years and ≥ 6 to < 12 years, respectively. Symptom and fever recurrence was more common in patients < 6 years with influenza B (54.5 and 50.0%, respectively) compared with older patients (0 and 25.0%, respectively). Virus titers declined (day 2) for both age groups. Transient virus titer increase and PA/I38X-variants were more common for patients < 6 years. CONCLUSIONS: The safety and effectiveness of single-dose baloxavir were observed in children across all age groups and influenza virus types. Higher rates of fever recurrence and transient virus titer increase were observed in children < 6 years. TRIAL REGISTRATION: Japan Pharmaceutical Information Center Clinical Trials Information JapicCTI-163,417 (registered 02 November 2016) and JapicCTI-173,811 (registered 15 December 2017).

Corticobasal ganglia projecting neurons are required for juvenile vocal learning but not for adult vocal plasticity in songbirds.

Birdsong, like human speech, consists of a sequence of temporally precise movements acquired through vocal learning. The learning of such sequential vocalizations depends on the neural function of the motor cortex and basal ganglia. However, it is unknown how the connections between cortical and basal ganglia components contribute to vocal motor skill learning, as mammalian motor cortices serve multiple types of motor action and most experimentally tractable animals do not exhibit vocal learning. Here, we leveraged the zebra finch, a songbird, as an animal model to explore the function of the connectivity between cortex-like (HVC) and basal ganglia (area X), connected by HVC(X) projection neurons with temporally precise firing during singing. By specifically ablating HVC(X) neurons, juvenile zebra finches failed to copy tutored syllable acoustics and developed temporally unstable songs with less sequence consistency. In contrast, HVC(X)-ablated adults did not alter their learned song structure, but generated acoustic fluctuations and responded to auditory feedback disruption by the introduction of song deterioration, as did normal adults. These results indicate that the corticobasal ganglia input is important for learning the acoustic and temporal aspects of song structure, but not for generating vocal fluctuations that contribute to the maintenance of an already learned vocal pattern.

Locomotor activity and histological changes observed in a mouse model of knee osteoarthritis.

[Purpose] This study aimed to elucidate the changes in locomotor activity in a mouse model of knee osteoarthritis (OA). [Materials and Methods] Fourteen 20-week-old mice were divided into control and OA groups. Knee OA was surgically induced under anesthesia by destabilizing the meniscus. The OA group was reared normally for 8 weeks following surgery, during which OA was induced. Locomotor activity was measured every hour for 8 weeks using an infrared locomotor activity measurement device. Histological changes were evaluated according to the classification-system of Glasson. [Results] Locomotor activity in the OA group significantly decreased up to 2 weeks after surgery. Histological findings in the control group revealed an irregular cartilage surface in a portion of the tibia with no other abnormalities. Contrastingly, those in the OA group had eburnation of the medial femoral condyle, as well as fibrillation and fissures in the medial tibial plateau. Histological scores in the OA group were significantly higher than the control group. [Conclusion] Locomotor activity evaluations, in addition to histological scores and findings, are imperative for studies aiming to clarify the disease state and effect of interventions using mice models.

The Avian Basal Ganglia Are a Source of Rapid Behavioral Variation That Enables Vocal Motor Exploration.

The basal ganglia (BG) participate in aspects of reinforcement learning that require evaluation and selection of motor programs associated with improved performance. However, whether the BG additionally contribute to behavioral variation ("motor exploration") that forms the substrate for such learning remains unclear. In songbirds, a tractable system for studying BG-dependent skill learning, a role for the BG in generating exploratory variability, has been challenged by the finding that lesions of Area X, the song-specific component of the BG, have no lasting effects on several forms of vocal variability that have been studied. Here we demonstrate that lesions of Area X in adult male zebra finches (Taeniopygia gutatta) permanently eliminate rapid within-syllable variation in fundamental frequency (FF), which can act as motor exploration to enable reinforcement-driven song learning. In addition, we found that this within-syllable variation is elevated in juveniles and in adults singing alone, conditions that have been linked to enhanced song plasticity and elevated neural variability in Area X. Consistent with a model that variability is relayed from Area X, via its cortical target, the lateral magnocellular nucleus of the anterior nidopallium (LMAN), to influence song motor circuitry, we found that lesions of LMAN also eliminate within-syllable variability. Moreover, we found that electrical perturbation of LMAN can drive fluctuations in FF that mimic naturally occurring within-syllable variability. Together, these results demonstrate that the BG are a central source of rapid behavioral variation that can serve as motor exploration for vocal learning.SIGNIFICANCE STATEMENT Many complex motor skills, such as speech, are not innately programmed but are learned gradually through trial and error. Learning involves generating exploratory variability in action ("motor exploration") and evaluating subsequent performance to acquire motor programs that lead to improved performance. Although it is well established that the basal ganglia (BG) process signals relating to action evaluation and selection, whether and how the BG promote exploratory motor variability remain unclear. We investigated this question in songbirds, which learn to produce complex vocalizations through trial and error. In contrast with previous studies that did not find effects of BG lesions on vocal motor variability, we demonstrate that the BG are an essential source of rapid behavioral variation linked to vocal learning.

Sensory feedback independent pre-song vocalizations correlate with time to song initiation.

The song of the adult male zebra finch is a well-studied example of a learned motor sequence. Song bouts begin with a variable number of introductory notes (INs) before actual song production. Previous studies have shown that INs progress from a variable initial state to a stereotyped final state before each song. This progression is thought to represent motor preparation, but the underlying mechanisms remain poorly understood. Here, we assessed the role of sensory feedback in the progression of INs to song. We found that the mean number of INs before song and the progression of INs to song were not affected by removal of two sensory feedback pathways (auditory or proprioceptive). In both feedback-intact and feedback-deprived birds, the presence of calls (other non-song vocalizations), just before the first IN, was correlated with fewer INs before song and an initial state closer to song. Finally, the initial IN state correlated with the time to song initiation. Overall, these results show that INs do not require real-time sensory feedback for progression to song. Rather, our results suggest that changes in IN features and their transition to song are controlled by internal neural processes, possibly involved in getting the brain ready to initiate a learned movement sequence.

Daily and short-term application of joint movement for the prevention of infrapatellar fat pad atrophy due to immobilization.

[Purpose] To mobilize the knee joint during cast fixation and to determine whether infrapatellar fat pad changes can be prevented. [Materials and Methods] We randomly allocated Wistar rats into 3 groups as follows: normal group, raised in normal conditions (n=5); contracture group, immobilized with cast fixation (n=5); and prevention group, treated with joint movement during immobilization (n=5). We immobilized the right hindlimb using cast fixation. Joint movement in the prevention group was accomplished by repeatedly pulling the right hindlimb caudally and then returning the leg to the bent position for 10 minutes every day for 2 weeks. We used a metronome to maintain a constant speed, with one set lasting 2 seconds (1-second traction and 1-second return). [Results] The contracture group had adipose cells of various sizes and fibrosis in the infrapatellar fat pad. These changes were also found in milder forms in the prevention group. We found significant differences in the cross section of adipose cells and in knee extension restriction between the groups. [Conclusion] Promoting joint movement may not only have a therapeutic effect on adipose cells but also a preventative effect.

Effect of low-intensity pulsed ultrasound therapy on a rat knee joint contracture model.

[Purpose] Histopathological investigation of the effects of low-intensity pulsed ultrasound (LIPUS) on joint components using a rat knee joint contracture model. [Subjects and Methods] Nineteen, 9-week-old Wistar male rats were divided into a control group (n=6) and an experimental group. Rats in the experimental group underwent cast immobilization of the right rear limb for 8 weeks. They were then randomly divided into a non-treatment group (n=6), which was raised under normal conditions for 4 weeks, and a treatment group (n=7), which underwent LIPUS for 4 weeks. LIPUS irradiation was performed at a frequency of 3 MHz, an intensity of 30 mW/cm2, and a pulse rate of 20% duty cycle. Irradiation was performed once daily for 10 min, 5 days per week. At the end of this period, tissue specimens in which the knee sagittal plane could be observed were prepared and observed using an optical microscope. [Results] The extension-limiting angle of the knee joint was significantly less in the treatment group compared with the non-treatment group. The posterior joint capsule was significantly thicker only in the non-treatment group, and the density was 53.5 ± 7.5% for the control group, 77.2 ± 5.7% for the non-treatment group, and 69.2 ± 2.9% for the treatment group, with significant differences existing across all groups. [Conclusion] LIPUS may widen the space between collagen fiber bundles of the joint capsule, thereby improving the range of motion.

Examination of Pyridazine as a Possible Scaffold for Nucleophilic Catalysis.

Pyridazines with amino groups positioned para to each aromatic ring nitrogen and fixed in six-membered rings were prepared. The representative symmetric amino N-Et derivative was found to slightly exceed DMAP in catalytic activity in the acetylation reaction of a tertiary alcohol in C6D6. Nucleophilicity eclipsing that of DMAP was established in competitive reactions using phenacyl bromide as the electrophile, and the unsymmetric N-Et derivative was revealed to have even higher nucleophilicity.

Task-related "cortical" bursting depends critically on basal ganglia input and is linked to vocal plasticity.

Basal ganglia-thalamocortical circuits are critical for motor control and motor learning. Classically, basal ganglia nuclei are thought to regulate motor behavior by increasing or decreasing cortical firing rates, and basal ganglia diseases are assumed to reflect abnormal overall activity levels. More recent studies suggest instead that motor disorders derive from abnormal firing patterns, and have led to the hypothesis that surgical treatments, such as pallidotomy, act primarily by eliminating pathological firing patterns. Surprisingly little is known, however, about how the basal ganglia normally influence task-related cortical activity to regulate motor behavior, and how lesions of the basal ganglia influence cortical firing properties. Here, we investigated these questions in a songbird circuit that has striking homologies to mammalian basal ganglia-thalamocortical circuits but is specialized for singing. The "cortical" outflow nucleus of this circuit is required for song plasticity and normally exhibits increased firing during singing and song-locked burst firing. We found that lesions of the striato-pallidal nucleus in this circuit prevented hearing-dependent song changes. These basal ganglia lesions also stripped the cortical outflow neurons of their patterned burst firing during singing, without changing their spontaneous or singing-related firing rates. Taken together, these results suggest that the basal ganglia are essential not for normal cortical firing rates but for driving task-specific cortical firing patterns, including bursts. Moreover, such patterned bursting appears critical for motor plasticity. Our findings thus provide support for therapies that aim to treat basal ganglia movement disorders by normalizing firing patterns.

Surveillance of antimicrobial susceptibility of Enterobacteriaceae pathogens isolated from intensive care units and surgical units in Russia.

A total of 473 strains of Enterobacteriaceae, including Escherichia coli, Klebsiella spp., Proteus spp., Citrobacter spp., Enterobacter spp., Serratia spp. and Providencia spp., were isolated from patients admitted to intensive care units and surgical units in Russia. About 90% of the isolates carried factors resistant to beta-lactams. The isolation rates of the extended-spectrum beta-lactamase (ESBL) producer defined in this study among E. coli, Klebsiella spp. and Proteus spp. were 45%, 48% and 17%, respectively. In the settings with high prevalence of the ESBL producer, flomoxef, which belongs to the oxacephem subgroup, and carbapenems retain their activity. The MIC50 of flomoxef, meropenem and imipenem against total isolates were 1 µg/mL, ≤ 0.063 µg/mL and 0.25 µg/mL, respectively. Fifty-five carbapenem-resistant strains were isolated in this study. The carbapenem resistant rates of E. coli, Klebsiella spp. and Proteus spp. were 3%, 16% and 29%, respectively

The temporal relationship of thresholds between muscle activity and ventilation during bicycle ramp exercise in community dwelling elderly males.

[Purpose] To compare the appearance time of the ventilatory threshold point and the electromyographic threshold in the activity of the vastus lateralis, rectus femoris, biceps femoris long head and gastrocnemius lateral head muscles during ramp cycling exercise in elderly males. [Subjects and Methods] Eleven community dwelling elderly males participated in this study. Subjects performed exercise testing with an expiratory gas analyzer and surface electromyography to evaluate the tested muscle activities during ramp exercise. [Results] The electromyographic threshold for rectus femoris was not valid because the slope after electromyographic threshold was not significant as compared to that before electromyographic threshold. The slope of the regression line for vastus lateralis was significantly decreased after electromyographic threshold while biceps femoris and gastrocnemius were increased. The electromyographic threshold appearance times for vastus lateralis and gastrocnemius were significantly earlier than ventilatory threshold point. There were no difference in electromyographic threshold appearance times among three muscles. [Conclusion] These results suggest that the increase in the slope of the regression line after electromyographic threshold for vastus lateralis was decreased, possibly indicating to postpone muscular fatigue resulting from the activation of biceps femoris and gastrocnemius as biarticular antagonists. This recruitment pattern might be an elderly-specific strategy.

Social performance reveals unexpected vocal competency in young songbirds.

Vocal ontogeny in songbirds provides a good model for understanding how complex motor behavior, including speech, is learned. For birdsong, as for other motor learning, it has generally been assumed that a subject's motor output at any point during learning represents what the subject has learned to produce by that time. Here, we show, however, that juvenile zebra finches partway through song learning, singing immature song, are capable of producing song with much more mature properties, depending on the behavioral context. In these birds, we were able to elicit courtship (female-directed) song, which young birds normally sing infrequently, and to compare it with the alone or "undirected" song (Undir) predominantly produced during learning as well as with the same bird's subsequent adult song. We found that the juvenile courtship song was much less variable than the immature Undir and as stereotyped as the adult song produced after a further month of practice. More strikingly, the juvenile courtship song was also acoustically much more similar than Undir to the adult song. This finding demonstrates that the Undir that juvenile birds usually produce underestimates the extent of learning and that song structure is learned faster than previously thought. Moreover, the rapid improvement in song quality in response to external social cues supports the idea that courtship singing is a state of motor "performance," in which the bird selects the best variants of the song learned during singing alone, and suggests that such performance states can reveal unappreciated progression of learning.

Experimental joint immobilization and remobilization in the rats.

[Purpose] The purpose of our study was to clarify temporal effects on restrictions to range of motion and the histopathological changes of joint components after joint immobilization in a rat knee-contracture model. [Subjects] Fifty-four male Wistar rats were randomly divided into two groups: a fixation group, and a control group. [Methods] In the fixation group, unilateral knee joints were immobilized at full flexion using a plaster cast for 4 weeks. At four weeks the animals were randomly divided into six subgroups, corresponding to the time of examination after cast removal: 0, 4, 8, 16, 24, and 32 weeks. For comparison, control group animals of corresponding age were also examined. [Results] Although movement restrictions of the knee joint had completely recovered 6 weeks after the cast removal, cartilage and synovial membrane structures did not completely recover. [Conclusion] These findings have not previously been reported, and as they form an addition to the fundamental scientific foundations of physical therapy, further research must examine these findings from a variety of perspectives.

Daily singing of adult songbirds functions to maintain song performance independently of auditory feedback and age.

Many songbirds learn to produce songs through vocal practice in early life and continue to sing daily throughout their lifetime. While it is well-known that adult songbirds sing as part of their mating rituals, the functions of singing behavior outside of reproductive contexts remain unclear. Here, we investigated this issue in adult male zebra finches by suppressing their daily singing for two weeks and examining the effects on song performance. We found that singing suppression decreased the pitch, amplitude, and duration of songs, and that those song features substantially recovered through subsequent free singing. These reversible song changes were not dependent on auditory feedback or the age of the birds, contrasting with the adult song plasticity that has been reported previously. These results demonstrate that adult song structure is not stable without daily singing, and suggest that adult songbirds maintain song performance by preventing song changes through physical act of daily singing throughout their life. Such daily singing likely functions as vocal training to maintain the song production system in optimal conditions for song performance in reproductive contexts, similar to how human singers and athletes practice daily to maintain their performance.

Real-World Effectiveness of Ensitrelvir in Reducing Severe Outcomes in Outpatients at High Risk for COVID-19.

INTRODUCTION: This study aimed to evaluate the effectiveness of ensitrelvir, an oral antiviral, in reducing hospitalization risk in outpatients at high-risk for severe COVID-19 during the Omicron era. METHODS: This was a retrospective study using a large Japanese health insurance claims database. It included high-risk outpatients for severe symptoms who received their first COVID-19 diagnosis between November 2022 and July 2023. The study included outpatients aged ≥ 18 years. The primary endpoint was all-cause hospitalization during the 4-week period from the date of outpatient diagnosis and medication, comparing the ensitrelvir group (n = 5177) and the no antiviral treatment group (n = 162,133). The risk ratio and risk difference were evaluated after adjusting patient background distribution by the inverse probability of treatment weight (IPTW) method. Secondary endpoints were incidence of respiratory and heart rate monitoring, oxygen therapy, ventilator use, intensive care admission, and all-cause death. RESULTS: The risk ratio for all-cause hospitalization between the ensitrelvir group (n = 167,385) and the no antiviral treatment group (n = 167,310) after IPTW adjustment was 0.629 [95% confidence interval (CI) 0.420, 0.943]. The risk difference was - 0.291 [95% CI - 0.494, - 0.088]. The incidence of both respiratory and heart rate monitoring and oxygen therapy was lower in the ensitrelvir group. Ventilator use, intensive care admission, and all-cause death were difficult to assess because of the limited events. CONCLUSIONS: The incidence of all-cause hospitalization was significantly lower in the ensitrelvir group than in the no antiviral treatment group, suggesting ensitrelvir is an effective treatment in patients at risk of severe COVID-19.

COVID-19 still poses a risk for patients with serious health conditions and weakened immune systems, who are more likely to develop severe illness. Several studies have indicated that some oral antiviral medications might be effective in preventing severe disease. This study aimed to evaluate if ensitrelvir, an oral antiviral medication, can help prevent hospitalization in outpatients who are at risk of developing severe symptoms from the Omicron variant of the SARS-CoV-2 virus. The hospitalization rates of patients who received ensitrelvir was compared with those who did not receive any antiviral treatment, using medical records from a large health insurance database in Japan focused on outpatients who were at risk of severe symptoms and were diagnosed with COVID-19 between November 2022 and July 2023. Respiratory and heart rate monitoring, oxygen therapy, ventilator use, intensive care admission, and all-cause death were also evaluated. The study found that patients who received ensitrelvir had a lower risk of being hospitalized compared to those who did not receive any antiviral treatment. The ensitrelvir group also had lower rates of respiratory and heart rate monitoring and oxygen therapy. However, it was challenging to assess the effects on ventilator use, intensive care admission, and all-cause death due to the small number of events in the population under evaluation. Based on these findings, ensitrelvir appears to be an effective treatment for reducing the risk of hospitalization in patients at risk of severe COVID-19.

Screening Cutoff Values for the Detection of Aldosterone-Producing Adenoma by LC-MS/MS and a Novel Noncompetitive CLEIA.

Context: Detecting patients with surgically curable aldosterone-producing adenoma (APA) among hypertensive individuals is clinically pivotal. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is the ideal method of measuring plasma aldosterone concentration (PAC) because of the inaccuracy of conventional chemiluminescent enzyme immunoassay (CLEIA). However, LC-MS/MS is expensive and requires expertise. We have developed a novel noncompetitive CLEIA (NC-CLEIA) for measuring PAC in 30 minutes. Objective: This work aimed to validate NC-CLEIA PAC measurements by comparing them with LC-MS/MS measurements and determining screening cutoffs for both measurements detecting APA. Methods: We retrospectively measured PAC using LC-MS/MS and NC-CLEIA in 133 patients with APA, 100 with bilateral hyperaldosteronism, and 111 with essential hypertension to explore the accuracy of NC-CLEIA PAC measurements by comparing with LC-MS/MS measurements and determined the cutoffs for detecting APA. Results: Passing-Bablok analysis revealed that the values by NC-CLEIA (the regression slope, intercept, and correlation coefficient were 0.962, -0.043, and 0.994, respectively) were significantly correlated and equivalent to those by LC-MS/MS. Bland-Altman plot analysis of NC-CLEIA and LC-MS/MS also demonstrated smaller systemic errors (a bias of -0.348 ng/dL with limits of agreement of -4.390 and 3.694 within a 95% CI) in NC-CLEIA than LC-MS/MS. The receiver operating characteristic analysis demonstrated that cutoff values for aldosterone/renin activity ratio obtained by LC-MS/MS and NC-CLEIA were 31.2 and 31.5 (ng/dL per ng/mL/hour), with a sensitivity of 91.0% and 90.2% and specificity of 75.4% and 76.8%, respectively, to differentiate APA from non-APA. Conclusion: This newly developed NC-CLEIA for measuring PAC could serve as a clinically reliable alternative to LC-MS/MS.

Impact of diradical character on two-photon absorption: bis(acridine) dimers synthesized from an allenic precursor.

The two-photon absorption (TPA) of a bis(acridine) dimer (8) having singlet diradical character in its ground state was found to be enhanced by more than 2 orders of magnitude as compared to its closed-shell counterpart (12), which has the same structural backbone and atom composition. The dimer, a tetracation species consisting of two connected acridinium cation moieties with high coplanarity, was obtained during our attempts to synthesize triplet carbenes by double oxidation of an allenic precursor (3b). High conjugation over the two aromatic rings connected by dimerization was revealed by X-ray analysis, and a small HOMO-LUMO gap was found in the visible-near-infrared one-photon absorption spectrum in solution and in the crystalline state, exhibiting that the ground state of 8 has singlet diradical nature. Ab initio molecular orbital calculations of the ground state also suggested that 8 has an intermediate diradical character (y) of 0.685. Interconversion between diradical tetracation dimer 8 and closed-shell dication dimer 12 was achieved by oxidation/reduction in good yields and was accompanied by formation of monoradical trication dimer 13 as an intermediate. TPA measurements at near-infrared wavelengths revealed that diradical dimer 8 has large TPA cross sections (3600 GM at 1200 nm), while closed-shell 12 has TPA cross sections of <21 GM. This result represents a straightforward comparison between the TPA activity of molecules with the same structural backbone and atom composition but with different degree of the diradical character, supporting the theoretical prediction that enhanced TPA intensity can be observed in the intermediate y region (0 < y < 1).

Highly active catalysts for the transfer dehydrogenation of alkanes: synthesis and application of novel 7-6-7 ring-based pincer iridium complexes.

A series of Ir-PCP pincer precatalysts [(7-6-7-(R) PCP)Ir(H)(Cl)] and [(7-6-7-(Ar) PCP)Ir(H)(Cl)(MeCN)] bearing a novel "7-6-7" fused-ring skeleton have been synthesized based upon the postulate that the catalytic species would have durability due to their rather rigid structure and high activity owing to the low but sufficient flexibility of their backbones, which are not completely fixed. Treatment of these precatalysts with NaOtBu gave rise to the active 14 electron (14e) species [(7-6-7-(iPr) PCP)Ir] and [(7-6-7-(Ph) PCP)Ir], which can trap hydrogen and were spectroscopically characterized as the tetrahydride complexes. Both [(7-6-7-(iPr) PCP)Ir] and [(7-6-7-(Ph) PCP)Ir] were found to be highly effective in the transfer dehydrogenation of cyclooctane with tert-butylethylene as the hydrogen acceptor, the initial reaction rate at high temperature (230 °C) being higher for [(7-6-7-(iPr) PCP)Ir] than [(7-6-7-(Ph) PCP)Ir], and the turnover number (TON) of the overall hydrogen transfer being higher for the latter. Nonetheless, the estimated TONs were as high as 4600 and 4820 for the two complexes at this temperature, respectively, which are unprecedented absolute values. In terms of durability, the [(7-6-7-(Ph) PCP)Ir] complex is the catalyst of choice for this reaction. Structural analysis and computational studies support the importance of the low flexibility of the ligand core.