Assessing late-life anxiety in Japanese older adults: psychometric evaluation of the Japanese version of the Geriatric Anxiety Scale.

BACKGROUND: This study developed a Japanese version of the Geriatric Anxiety Scale (GAS-J) and its short form (GAS-10-J) to evaluate anxiety in Japanese older adults and assess its psychometric properties using a cross-sectional design. METHODS: A total of 331 community-dwelling older adult participants (208 men, 116 women, seven unknowns; mean age = 73.47 ± 5.17 years, range = 60-88 years) recruited from two Silver Human Resources Centres in the Kanto region, Japan, answered a set of self-report questionnaires. Of these respondents, 120 participated in a follow-up survey to evaluate test-retest reliability. RESULTS: Confirmatory factor analysis suggested that, as with the original GAS, the GAS-J had a three-factor structure and the GAS-10-J had a unifactor structure with high standardised factor loadings. Test-retest correlations and internal consistency analyses indicated that these scales were reliable. Correlations between the GAS-J/GAS-10-J with the Geriatric Anxiety Inventory, Generalised Anxiety Disorder-7, Geriatric Depression Scale-15, World Health Organization-Five Well-Being Index, and Kihon Checklist were mostly consistent with our hypotheses, thereby supporting the construct validity of the GAS-J/GAS-10-J. CONCLUSIONS: The findings indicate that the GAS-J and GAS-10-J have robust psychometric properties for assessing late-life anxiety in Japanese older adults. Further GAS-J studies are required for clinical groups.

Contingent self-worth and depression in early adolescents: The role of psychological inflexibility as a mediator.

This study investigated whether lower psychological flexibility (psychological inflexibility) mediates the relationship between contingent self-worth and depressive symptoms among Japanese adolescents. A total of 210 Japanese junior high school students aged 12 to 15 years (106 boys and 104 girls) were recruited for this study. Participants completed the Japanese adaptations of the Self-Worth Contingency Questionnaire, the Avoidance and Fusion Questionnaire for Youth, and the Depression Self-Rating Scale for Children. Results indicated that psychological inflexibility mediated the association between contingent self-worth and depressive symptoms. Specifically, contingent self-worth affected lower psychological flexibility, which influenced higher depressive symptoms. The results highlight the importance of fostering autonomy and promoting psychological flexibility to reduce the risk of depression among adolescents.

Muscarinic M3 positive allosteric modulator ASP8302 enhances bladder contraction and improves voiding dysfunction in rats.

OBJECTIVES: Muscarinic M3 (M3 ) receptors mediate cholinergic smooth muscle contraction of the bladder. Current drugs targeting bladder M3 receptors for micturition disorders have a risk of cholinergic side effects due to excessive receptor activation and insufficient selectivity. We investigated the effect of ASP8302, a novel positive allosteric modulator (PAM) of M3 receptors, on bladder function in rats. METHODS: Modulation of carbachol-induced increases in intracellular Ca2+ was assessed in cells expressing rat muscarinic receptors. Potentiation of bladder contractions was evaluated using isolated rat bladder strips and by measuring intravesical pressure in anesthetized rats. Conscious cystometry was performed to investigate the effects on residual urine volume and voiding efficiency in rat voiding dysfunction models induced by the α1 -adrenoceptor agonist midodrine and muscarinic receptor antagonist atropine, and bladder outlet obstruction. To assess potential side effects, the number of stools and tracheal insufflation pressure were measured in conscious and anesthetized rats, respectively. RESULTS: ASP8302 demonstrated PAM effects on the rat M3 receptor in cell assays, and augmented cholinergic bladder contractions both in vivo and in vitro. ASP8302 improved voiding efficiency and reduced residual urine volume in two voiding dysfunction models as effectively as distigmine bromide, but unlike distigmine bromide did not affect the number of stools or tracheal insufflation pressure. CONCLUSIONS: Our results in rats indicate that ASP8302 improves voiding dysfunction by potentiating bladder contraction with fewer effects on cholinergic responses in other organs, and suggest a potential advantage over current cholinomimetic drugs for treating micturition disorders caused by insufficient bladder contraction.

Potentiation of Muscarinic M3 Receptor Activation through a New Allosteric Site with a Novel Positive Allosteric Modulator ASP8302.

Muscarinic M3 (M3) receptors mediate a wide range of acetylcholine (ACh)-induced functions, including visceral smooth-muscle contraction and glandular secretion. Positive allosteric modulators (PAMs) can avoid various side effects of muscarinic agonists with their spatiotemporal receptor activation control and potentially better subtype selectivity. However, the mechanism of allosteric modulation of M3 receptors is not fully understood, presumably because of the lack of a potent and selective PAM. In this study, we investigated the pharmacological profile of ASP8302, a novel PAM of M3 receptors, and explored the principal site of amino-acid sequences in the human M3 receptor required for the potentiation of receptor activation. In cells expressing human M3 and M5 receptors, ASP8302 shifted the concentration-response curve (CRC) for carbachol to the lower concentrations with no significant effects on other subtypes. In a binding study with M3 receptor-expressing membrane, ASP8302 also shifted the CRC for ACh without affecting the binding of orthosteric agonists. Similar shifts in the CRC of contractions by multiple stimulants were also confirmed in isolated human bladder strips. Mutagenesis analysis indicated no interaction between ASP8302 and previously reported allosteric sites; however, it identified threonine 230 as the amino acid essential for the PAM effect of ASP8302. These results demonstrate that ASP8302 enhances the activation of human M3 receptors by interacting with a single amino acid distinct from the reported allosteric sites. Our findings suggest not only a novel allosteric site of M3 receptors but also the potential application of ASP8302 to diseases caused by insufficient M3 receptor activation. SIGNIFICANCE STATEMENT: The significance of this study is that the novel M3 receptor positive allosteric modulator ASP8302 enhances the activation of human M3 receptor by interacting with a residue distinct from the reported allosteric sites. The finding of Thr230 as a novel amino acid involved in the allosteric modulation of M3 receptors provides significant insight into further research of the mechanism of allosteric modulation of M3 and other muscarinic receptors.

Design, Synthesis, and Structure-Activity Relationships Study of N-Pyrimidyl/Pyridyl-2-thiazolamine Analogues as Novel Positive Allosteric Modulators of M3 Muscarinic Acetylcholine Receptor.

The M3 muscarinic acetylcholine receptor (mAChR) plays an essential pharmacological role in mediating a broad range of actions of acetylcholine (ACh) released throughout the periphery and central nerve system (CNS). Nevertheless, its agonistic functions remain unclear due to the lack of available subtype-selective agonists or positive allosteric modulators (PAMs). In the course of our extended structure-activity relationships (SARs) study on 2-acylaminothiazole derivative 1, a previously reported PAM of the M3 mAChR, we successfully identified N-pyrimidyl/pyridyl-2-thiazolamine analogues as new scaffolds. The SARs study was rationalized using conformational analyses based on intramolecular interactions. A comprehensive study of a series of analogues described in this paper suggests that a unique sulfur-nitrogen nonbonding interaction in the N-pyrimidyl/pyridyl-2-thiazolamine moiety enable conformations that are essential for activity. Further, a SARs study around the N-pyrimidyl/pyridyl-2-thiazolamine core culminated in the discovery of compound 3g, which showed potent in vitro PAM activity for the M3 mAChR with excellent subtype selectivity. Compound 3g also showed a distinct pharmacological effect on isolated smooth muscle tissue from rat bladder and favorable pharmacokinetics profiles, suggesting its potential as a chemical tool for probing the M3 mAChR in further research.

Psychometric properties of the Japanese version of the Geriatric Anxiety Inventory for community-dwelling older adults.

BACKGROUND: This study developed a Japanese version of the Geriatric Anxiety Inventory (GAI-J) and its short form (GAI-J-SF) to evaluate anxiety in older adults in Japan and assess these measures' psychometric properties with a cross-sectional design. METHODS: Participants (N = 400; mean age: 75 years) were community-dwelling older adults who answered a set of self-report questionnaires. They were recruited from a community centre for older persons in the Kanto region of Japan. Of the respondents, 100 participated in a follow-up survey to evaluate test-retest reliability. Item response theory was adopted to evaluate item parameters. RESULTS: Confirmatory factor analysis with categorical data suggested that, as with the original Geriatric Anxiety Inventory, the GAI-J/GAI-J-SF had a unifactor structure. Test-retest correlation and internal consistency analyses indicated that these scales had high reliability. Item response theory results showed that both measures' item parameters were acceptable. Correlations with the Penn State Worry Questionnaire, State Trait Anxiety Inventory-State Only, Generalized Anxiety Disorder-7 and Patient Health Questionnaire-9 were mostly consistent with our hypotheses. This supports the high convergent validity of the GAI-J/GAI-J-SF. CONCLUSIONS: The findings indicate that the GAI-J and the GAI-J-SF have robust psychometric properties for assessing late-life anxiety in older Japanese adults. Future GAI-J studies in clinical groups are needed.

Discovery and structure-activity relationships study of positive allosteric modulators of the M3 muscarinic acetylcholine receptor.

The M3 muscarinic acetylcholine receptor (mAChR) is a member of the family of mAChRs, which are associated with a variety of physiological functions including the contraction of various smooth muscle tissues, stimulation of glandular secretion, and regulation of a range of cholinergic processes in the central nerve system. We report here the discovery and a comprehensive structure--activity relationships (SARs) study of novel positive allosteric modulators (PAMs) of the M3 mAChR through a high throughput screening (HTS) campaign. Compound 9 exhibited potent in vitro PAM activity towards the M3 mAChR and significant enhancement of muscle contraction in a concentration-dependent manner when applied to isolated smooth muscle strips of rat bladder. Compound 9 also showed excellent subtype selectivity over other subtypes of mAChRs including M1, M2, and M4 mAChRs, and moderate selectivity over the M5 mAChR, indicating that compound 9 is an M3-preferring M3/M5 dual PAM. Moreover, compound 9 displayed acceptable pharmacokinetics profiles after oral dosing to rats. These results suggest that compound 9 may be a promising chemical probe for the M3 mAChR for further investigation of its pharmacological function both in vitro and in vivo.

Factor Structure and Psychometric Properties of a New Scale to Assess Alexithymia-Like Features in Japanese Youth.

BACKGROUND: Studies of alexithymia have primarily targeted adult populations. Although some recent studies of alexithymia have focused on children and young adolescents, the literature is not sufficient for development of an assessment tool. The aim of this study was to develop, and evaluate the psychometric properties of, a new scale to measure alexithymia-like features in young adolescents. METHODS: A total of 1,444 Japanese junior high school students (701 males, 743 females; age range 12-15; mean [SD] age, 13.37 [0.98] years) participated in 2 surveys conducted at their own schools. RESULTS: First, exploratory factor analysis of the first survey data (n=981) demonstrated that this new scale had a unifactor structure, as determined by minimum average partial analysis and parallel analysis. Second, confirmatory factor analysis of the second survey data (n=463) confirmed the unifactor structure of this new scale and acceptable goodness of model fit. The new scale had modest internal consistency. CONCLUSIONS: The correlations of this new alexithymia scale with related variables were weak but significant, in accordance with our hypothesis. The scale had acceptable reliability and convergent validity and thus might be useful for measuring alexithymic tendency in young adolescents.

Effect of ASP6432, a Novel Type 1 Lysophosphatidic Acid Receptor Antagonist, on Urethral Function and Prostate Cell Proliferation.

Current pharmacotherapies for lower urinary tract symptoms associated with benign prostate hyperplasia (LUTS/BPH) are in need of improvement. Lysophosphatidic acid (LPA) is a phospholipid with various biologic functions. However, its exact role in the lower urinary tract and its target receptor subtype have not been fully elucidated. We investigated the role of LPA and the type 1 LPA receptor (LPA1) in urethral/prostatic contractile function and prostate cell proliferation by pharmacologically characterizing ASP6432 (potassium 1-(2-{[3,5-dimethoxy-4-methyl-N-(3-phenylpropyl)benzamido]methyl}-1,3-thiazole-4-carbonyl)-3-ethyl-2,2-dioxo-2λ6-diazathian-1-ide), a novel LPA1 antagonist. ASP6432 exhibited potent and selective antagonistic activity against LPA1 in cells expressing LPA receptor subtypes. In isolated rat tissue strips and anesthetized rats, ASP6432 concentration-/dose-dependently inhibited LPA-induced urethra and prostate contractions. In addition, in anesthetized rats, ASP6432 maximally decreased the urethral perfusion pressure (UPP) in the absence of exogenous LPA stimulation by 43% from baseline, whereas tamsulosin, an α1-adrenoceptor antagonist, reduced UPP by 22%. Further, in human prostate stromal cells, ASP6432 significantly and concentration-dependently suppressed LPA-induced bromodeoxyuridine incorporation. These results demonstrate a pivotal role for LPA and LPA1 in the regulation of urethral tonus and prostate cell proliferation. The potent urethral relaxation and inhibition of prostatic stromal cell growth indicate the potential of ASP6432 as a novel therapeutic agent for LUTS/BPH.

The reciprocal relations between experiential avoidance, school stressor, and psychological stress response among Japanese adolescents.

The present study aimed to investigate the reciprocal relations between experiential avoidance, stressor, and psychological stress response (which consist of anger, depression, anxiety, helplessness, and physical complaints). In this study, 688 Japanese junior high school students (353 boys, 334 girls, 1 unidentified; mean age 13.28 years) completed three waves of questionnaires on experiential avoidance, stressor, and psychological stress response, with one-week intervals between measurement waves. Results from cross-lagged panel analyses showed that experiential avoidance predicted subsequent stressor and psychological stress response. Furthermore, the stressor and psychological stress response influenced by prior experiential avoidance affected subsequent occurrence of experiential avoidance. The findings suggest that reciprocal relations exist among the variables, and that the interaction between experiential avoidance and psychological stress was possible in adolescents.

Contingent self-worth moderates the relationship between school stressors and psychological stress responses.

This study examined the moderating role of contingent self-worth on the relationships between school stressors and psychological stress responses among Japanese adolescents. A total of 371 Japanese junior high school students (184 boys and 187 girls, Mage = 12.79 years, SD = 0.71) completed the Japanese version of the Self-Worth Contingency Questionnaire and a mental health checklist at two points separated by a two-month interval. Hierarchical multiple regression analyses were then used to determine whether contingent self-worth moderated the relationship between school stressors and psychological stress responses. The results indicated that, when psychological stress responses were controlled for at Time 1, contingent self-worth did not predict the psychological stress responses at Time 2. However, a two-way interaction between contingent self-worth and stressors was found to significantly influence psychological stress responses, thus indicating that stressors had a stronger impact on psychological stress responses among those with high contingent self-worth compared to those with low contingent self-worth.

Study on Physiological Roles of Stimulation of Prostaglandin E2 Receptor Subtype EP2 in Urethral Function in Rats.

OBJECTIVES: We investigated the relaxant effect of stimulation of prostaglandin E2 (PGE2 ) receptor subtype EP2 as well as the involvement of a cyclic AMP (cAMP)-dependent pathway related to stimulation of EP2 receptors in urethral function in rats by evaluating effects of PGE2 and selective EP2 receptor agonist CP-533,536. METHODS: Effects of PGE2 and CP-533,536 on cAMP accumulation were assessed in Chinese hamster ovary (CHO)-K1 cells expressing rat EP2 or EP4 receptors. Relaxant responses to PGE2 and CP-533,536 (0.01-10 µmol/L) in rat urethral tissue pre-contracted with 10 µmol/L phenylephrine were evaluated, and cAMP levels in isolated rat urethral tissue treated with these compounds were determined as well. The effects of PGE2 and CP-533,536 (0.003-0.3 mg/kg intravenously) on urethral perfusion pressure (UPP) in anesthetized rats were also evaluated. RESULTS: PGE2 concentration-dependently increased the accumulation of cAMP in cells expressing rat EP2 (EC50 value = 1.3 nmol/L) and EP4 receptors (EC50 value = 17 nmol/L). While CP-533,536 similarly increased the accumulation of cAMP in cells expressing rat EP2 receptors (EC50 value = 3.0 nmol/L), no such effects were noted in cells expressing rat EP4 receptors up to 10 µmol/L. Both PGE2 and CP-533,536 produced relaxation and increased cAMP levels in urethral tissues in a concentration-dependent manner. PGE2 and CP-533,536 both dose-dependently decreased UPP in anesthetized rats. CONCLUSIONS: Taken together, these results suggest that stimulation of EP2 receptors induces relaxation likely via activation of cAMP-dependent mechanisms in rat urethral tissue, leading to a reduction of UPP.

Effect of Selective Prostaglandin E2 EP2 Receptor Agonist CP-533,536 on Voiding Efficiency in Rats with Midodrine-Induced Functional Urethral Obstruction.

OBJECTIVES: We investigated the effect of the selective prostaglandin E2 EP2 receptor agonist CP-533,536 on voiding efficiency in rats with midodrine-induced functional urethral obstruction. METHODS: The effect of CP-533,536 (0.03-0.3 mg/kg, intravenous [i.v.]) on urethral perfusion pressure (UPP) was investigated in anesthetized rats pre-treated with midodrine (1 mg/kg, i.v.), which forms an active metabolite that acts as an α1 -adrenoceptor agonist. The effect of CP-533,536 (0.03-0.3 mg/kg, i.v.) on cystometric parameters was also investigated in anesthetized rats. In addition, the effect of CP-533,536 (0.03-0.3 mg/kg, i.v.) on residual urine volume (RV) and voiding efficiency (VE) was investigated in conscious rats treated with midodrine (1 mg/kg, i.v.). RESULTS: CP-533,536 dose-dependently decreased UPP elevated by midodrine in anesthetized rats. In contrast, CP-533,536 did not affect maximum voiding pressure, intercontraction interval, or intravesical threshold pressure. In conscious rats, midodrine (1 mg/kg, i.v.) markedly increased RV and reduced VE. CP-533,536 dose-dependently ameliorated increases in RV and decreases in VE induced by midodrine. CONCLUSIONS: These results suggest that a selective EP2 receptor agonist could ameliorate the elevation of RV and improve the reduction of VE in rats with functional urethral obstruction caused by stimulation of α1 -adrenoceptors. The mechanism of action might be not potentiation of bladder contraction but rather preferential relief of urethral constriction.

[Relationship between self-evaluation of their emotions and subjective adaptation to school among junior high school students].

The effect of self-evaluation of emotions on subjective adaption to school was investigated among junior high school students (n = 217: 112 boys, 105 girls) who participated in a questionnaire survey. Hierarchical multiple regression analysis indicated that for boys "Infringement and maladjustment" differed based on their self-evaluation of anger and anxiety. For girls, on the other hand, the self-evaluation of anger alleviated psychological stress, worsened the "Relationship with the teacher" and the "Relationship with the class", whereas self-evaluation of anxiety played a role in increasing psychological stress and deteriorating the "Relationship with the class." Furthermore, negatively evaluating either anger or anxiety heightened the "Motivation for learning" in girls. These results suggest that the evaluation of emotions is different in boys and girls and for different emotions.

[Development of the Japanese version of the Emotion Awareness Questionnaire for junior high school students].

This study reports about the development of the Japanese version of the Emotion Awareness Questionnaire for junior high school students. Emotion Awareness Questionnaire (EAQ; Rieffe, Oosterveld, Miers, Meerum Terwogt, & Ly, 2008) for children and adolescents aims not only to monitor and differentiate emotions but also to measure attitudes about emotions. It consists of six factors: differentiating emotions, verbal sharing of emotions, not hiding emotion, bodily awareness, attending to others' emotion, analyses of emotions. To examine the reliability and validity of the Japanese version of the EAQ, junior high school students (7th to 9th grades) were requested to complete the questionnaires (n = 535 in time 1, n = 537 in time 2, n = 330 in time 3). The results showed that the Japanese version of the EAQ had almost the same six-factor structure as the original one. It also had moderate internal consistency and test-retest reliability (three weeks). The validity of the scale was examined in relation to emotional intelligence, social anxiety, depression, psychological stress responses, evaluation of emotions, self esteem and sense of authenticity. The results confirmed that the Japanese version of the EAQ had good validity.

[Vulnerability factors for school stress among junior high school students: over-adaptation and evaluation of emotion].

This study investigated the role of over-adaptation and evaluation of emotion as stress vulnerability factors among Japanese junior high school students. Based upon the diathesis-stress model, 720 students (348 boys and 372 girls) completed questionnaires about stress responses, over-adaptation, and evaluation of emotion. Two weeks later, they completed the questionnaire about stress responses along a questionnaire about school stressors. Hierarchical multiple regression analysis indicated that boys with high over-adaptation tendencies reported a greater stress responses following the occurrence of negative school stressors than non-over-adaptive students. In addition, evaluation of emotion was found to interact with stressors to predict stress responses in boys. These results were not found in girls.

Rep68 protein of adeno-associated virus type 2 interacts with 14-3-3 proteins depending on phosphorylation at serine 535.

Rep78/68 proteins of adeno-associated virus type 2 (AAV-2) are involved in many aspects of the viral life cycle, including replication, gene expression, and site-specific integration. To understand the molecular mechanisms of the actions of Rep proteins, we searched for Rep68-interacting cellular proteins by utilizing a one-step affinity purification technique and identified two members of 14-3-3 proteins (14-3-3 epsilon and gamma). We found that phosphorylation of 535Ser at the carboxy terminus of Rep68 was critical for its association with 14-3-3. The association of 14-3-3 proteins to Rep68 resulted in reduction of the affinity of Rep68 for DNA. Furthermore, genome DNA replication of a recombinant mutant virus carrying a phosphorylation-deficient Rep68 (Ser535Ala) was more efficient than that of the wild-type virus. These results suggest that phosphorylation of Rep68 and subsequent association with 14-3-3 proteins regulates Rep-mediated functions during the AAV life cycle.

Simian virus 40 VP1 capsid protein forms polymorphic assemblies in vitro.

The simian virus 40 (SV40) capsid is composed of 72 pentamers of VP1, the major protein of SV40. These pentamers are arranged in a T=7d icosahedral surface lattice, which is maintained by three types of appropriately arranged, non-equivalent interactions between the pentamers. However, it remains unclear how these interactions are achieved. In this study, the in vitro assembly of recombinant VP1 was analysed. Electron microscopy observations revealed that these recombinant VP1 proteins assembled into structurally polymorphic particles depending on environmental conditions. VP1 pentamers assembled efficiently into virus-like particles (VLPs) when high concentrations of ammonium sulfate were present. However, in the presence of 1 M NaCl and 2 mM CaCl(2) at neutral pH, VP1 pentamers formed not only VLPs but also produced tiny T=1 icosahedral particles and tubular structures. The exclusion of CaCl(2) resulted in the exclusive formation of tiny particles. In contrast, in the presence of 150 mM NaCl at pH 5, the VP1 pentamers produced only extraordinarily long tubular structures. VP1 is thus quite unique in that it can assemble into such diverse structures. These observations provide clues that will help elucidate the mechanisms underlying SV40 capsid formation.

Importance of Vp1 calcium-binding residues in assembly, cell entry, and nuclear entry of simian virus 40.

For polyomaviruses, calcium ions are known to be essential for virion integrity and for the assembly of capsid structures. To define the role of calcium ions in the life cycle of the virus, we analyzed simian virus 40 (SV40) mutants in which structurally deduced calcium-binding amino acids of Vp1 were mutated singly and in combination. Our study provides evidence that calcium ions mediate not only virion assembly but also the initial infection processes of cell entry and nuclear entry. Mutations at Glu48, Glu157, Glu160, Glu216, and/or Glu330 are correlated with different extents of packaging defects. The low packaging ability of mutant E216R suggests the need to position the Glu216 side chain for proper virion formation. All other mutants selected for further analysis produced virus-like particles (VLPs) but were poorly infectious. The VLPs of mutant E330K could not attach to or enter the cell, and mutant E157A-E160A and E216K VLPs entered the cell but failed to enter the nucleus, apparently as a result of premature VLP dissociation. Our results show that five of the seven acidic side chains at the two calcium-binding sites-Glu48 and Glu330 (site 1), Glu157 and Glu160 (site 2), and Glu216 (both sites)-are important for SV40 infection. We propose that calcium coordination imparts not only stability but also structural flexibility to the virion, allowing the acquisition or loss of the ion at the two sites to control virion formation in the nucleus, as well as virion structural alterations at the cell surface and in the cytoplasm early during infection.