Itpr1 regulates the formation of anterior eye segment tissues derived from neural crest cells.

Mutations in ITPR1 cause ataxia and aniridia in individuals with Gillespie syndrome (GLSP). However, the pathogenic mechanisms underlying aniridia remain unclear. We identified a de novo GLSP mutation hotspot in the 3'-region of ITPR1 in five individuals with GLSP. Furthermore, RNA-sequencing and immunoblotting revealed an eye-specific transcript of Itpr1, encoding a 218amino acid isoform. This isoform is localized not only in the endoplasmic reticulum, but also in the nuclear and cytoplasmic membranes. Ocular-specific transcription was repressed by SOX9 and induced by MAF in the anterior eye segment (AES) tissues. Mice lacking seven base pairs of the last Itpr1 exon exhibited ataxia and aniridia, in which the iris lymphatic vessels, sphincter and dilator muscles, corneal endothelium and stroma were disrupted, but the neural crest cells persisted after completion of AES formation. Our analyses revealed that the 218-amino acid isoform regulated the directionality of actin fibers and the intensity of focal adhesion. The isoform might control the nuclear entry of transcriptional regulators, such as YAP. It is also possible that ITPR1 regulates both AES differentiation and muscle contraction in the iris.

Identification of Azalamellarin N as a Pyroptosis Inhibitor.

Pyroptosis is a form of regulated cell death that promotes inflammation; it attracts much attention because its dysregulation leads to various inflammatory diseases. To help explore the precise mechanisms by which pyroptosis is regulated, in this study, we searched for chemical compounds that inhibit pyroptosis. From our original compound library, we identified azalamellarin N (AZL-N), a hexacyclic pyrrole alkaloid, as an inhibitor of pyroptosis induced by R837 (also called imiquimod), which is an agonist of the intracellular multiprotein complex nucleotide-binding and oligomerization domain-like receptor (NLR) family pyrin domain containing 3 (NLRP3) inflammasome. However, whereas the effect of AZL-N on R837-induced pyroptosis was relatively weak, AZL-N strongly inhibited pyroptosis induced by extracellular ATP or nigericin, which are different types of NLRP3 inflammasome agonists. This was in contrast with the results that MCC950, a well-established NLRP3 inhibitor, consistently inhibited pyroptosis irrespective of the type of stimulus. We also found that AZL-N inhibited activation of caspase-1 and apoptosis-associated speck-like proteins containing a caspase activation and recruitment domain (ASC), which are components of the NLRP3 inflammasome. Analysis of the structure-activity relationship revealed that a lactam ring of AZL-N, which has been shown to contribute to the strong binding of AZL-N to its known target protein kinases, is required for its inhibitory effects on pyroptosis. These results suggest that AZL-N inhibits pyroptosis by targeting molecule(s), which may be protein kinase(s), that act upstream of NLRP3 inflammasome activation, rather than by directly targeting the components of the NLRP3 inflammasome. Further identification and analysis of target molecule(s) of AZL-N will shed light on the regulatory mechanisms of pyroptosis, particularly those depending on proinflammatory stimuli.

Factors associated with overweight/obesity of children aged 6-12 years in Indonesia.

BACKGROUND: Globally, the prevalence of childhood obesity has increased considerably, including in Indonesia. Obesity results from multifactorial interactions at the personal, familial, and environmental levels. However, little is known about the factors associated with overweight/obesity among children in Indonesia. This study is intended to identify personal, familial, and environmental factors associated with overweight/obesity in children aged 6-12 years in Indonesia. METHODS: Study design was a secondary data analysis using the Indonesia Family Life Survey in 2014/2015, focusing on 6,090 children aged 6-12 years. The questions covered the child's body mass index and potential personal, familial, and environmental factors. Logistic regression analysis was performed to identify the personal, familial, and environmental factors. RESULTS: The mean age of participants was 8.9 years (SD = 2.0); 51.0% were boys; 9.4% were overweight; and 8.1% were obese. Overweight and obesity were associated with age [AOR 1.09 (95% CI 1.04-1.14)], having an overweight [AOR 1.93 (95% CI 1.58-2.36)] or obese [AOR 3.36 (95% CI 2.43-4.61)] father compared with a normal father, being of Chinese [AOR 9.51 (95% CI 1.43-79.43)] or Javanese [AOR 1.60 (95% CI 1.16-2.24)] ethnicity compared with Sundanese ethnicity, and residing in an urban area [AOR 1.36 (95% CI 1.10-1.70)]. A lower risk of child overweight/obesity was associated with the father's perception [AOR 0.56 (95% CI 0.38-0.80)] and mother's perception [AOR 0.66 (95% CI 0.43-0.98)] of the child's food consumption as being less than adequate compared with adequate. CONCLUSIONS: Risk factors in children for overweight/obesity were older age, having an overweight/obese father, membership of certain ethnic groups, and urban residence. The main protective factor was parents' perception that a child's food consumption was less than adequate. Health promotion programs focused on these factors could help control or prevent childhood obesity in Indonesia.

SH3P2 suppresses osteoclast differentiation through restricting membrane localization of myosin 1E.

Osteoclasts are multinucleated cells responsible for bone resorption. Src homology 3 (SH3) domain-containing protein-2 (SH3P2)/osteoclast-stimulating factor-1 regulates osteoclast differentiation, but its exact role remains elusive. Here, we show that SH3P2 suppresses osteoclast differentiation. SH3P2 knockout (KO) mice displayed decreased femoral trabecular bone mass and enhanced localization of osteoclasts on the tibial trabecular bone surface, suggesting that SH3P2 suppresses bone resorption by osteoclasts. Osteoclast differentiation based on cellular multinuclearity induced by macrophage colony-stimulating factor and receptor activator of nuclear factor-κB ligand (RANKL) was enhanced in bone marrow-derived macrophages lacking SH3P2. RANKL induced SH3P2 dephosphorylation, which increased the association of actin-dependent motor protein myosin 1E (Myo1E) with SH3P2 and thereby prevented Myo1E localization to the plasma membrane. Consistent with this, Myo1E in the membrane fraction increased in SH3P2-KO cells. Together with the attenuated osteoclast differentiation in Myo1E knocked down cells, SH3P2 may suppress osteoclast differentiation by preventing their cell-to-cell fusion depending on Myo1E membrane localization.

Association between the type of physical activity and metabolic syndrome in middle-aged and older adult residents of a semi-mountainous area in Japan.

BACKGROUND: Physical activity is reported to prevent metabolic syndrome. However, it is unclear whether exercise or daily physical activity is more beneficial for residents of semi-mountainous areas. This study aimed to identify whether daily physical activity is more beneficial than exercise for the prevention of metabolic syndrome among middle-aged and older residents in semi-mountainous areas. METHODS: We analyzed secondary data of 636 people who underwent a specific health checkup in a semi-mountainous area of Japan. Physical activity was classified into four types: inactivity (I-type; without exercise and without daily physical activity), only exercise (E-type; with exercise and without daily physical activity), only daily physical activity (D-type; without exercise and with daily physical activity), and full physical activity type (F-type; with exercise and with daily physical activity). We compared the means of risk factors for metabolic syndrome by these four types, followed by logistic regression analysis, to identify whether and to what extent the D-type was less likely to have metabolic syndrome than the E-type. RESULTS: The prevalence of metabolic syndrome was 28.5% (men 45.7%, women 15.8%). The proportions of men with exercise and daily physical activity were 38.7% and 52.8%, respectively. For women, the proportions were 33.0% and 47.1%, respectively. In women, the D-type had the significantly lowest BMI, smallest waist circumference, highest HDL-C, and lowest prevalence of metabolic syndrome of the four types; the same was not observed in men. Additionally, D-type activity was more strongly associated with a reduced risk of metabolic syndrome than E-type activity in women (adjusted odds ratio 0.24; 95% confidence interval 0.06-0.85, P = 0.028). CONCLUSIONS: Compared to middle-aged and older women residents with exercise in a semi-mountainous area of Japan, those with daily physical activity may effectively prevent metabolic syndrome.

A Randomized Controlled Trial on Primigravid Women of Text Messaging Intervention Offering Pregnancy and Childbirth Support.

This study aimed to assess the efficacy of a text messaging intervention that offered pregnancy and childbirth support. Participants included 39 primigravid women who were less than 12 weeks pregnant. Text messages were sent twice weekly to the intervention group from week 13 of pregnancy until childbirth. Outcome measures were anxiety levels, lifestyle in the month before birth, pre-birth weight, pregnancy complications, delivery complications, birth weight, thoughts regarding the text messages, and the frequency of viewing of the text messages. For the item "I engage in body stretching," the average value in the intervention group was significantly higher than that in the control group. For the item "I have regular bowel movements," the average value in the intervention group was significantly lower. Most participants reported that the intervention was at least somewhat useful. This study indicates that text messaging intervention is practical and can be used to support numerous pregnant women simultaneously at a relatively low cost. Since this is a study pilot trial, large-scale studies are necessary to improve the method and allow for the generalization of the results.

Sensitivity and specificity of different antibody tests for detecting varicella-zoster virus.

INTRODUCTION: Antibody tests for detecting varicella-zoster virus include the fluorescent-antibody-to-membrane-antigen (FAMA) assay, immune adherence hemagglutination assay (IAHA), enzyme immunoassay (EIA), and the glycoprotein-based enzyme-linked immunosorbent assay (gpELISA). Although FAMA and gpELISA are highly sensitive, FAMA is not available commercially. Therefore, this study was performed to compare potential high-sensitivity tests with commercially available tests. METHODS: Four antibody tests, FAMA, gpELISA, EIA, and IAHA, were performed using sera collected from 32 children aged 7 months-10 years. Using FAMA as a reference, the sensitivity and specificity of gpELISA, EIA, and IAHA were assessed. Subsequently, using gpELISA as a reference, the positive agreement rate of EIA and IAHA was assessed. RESULTS: On a reference scale with FAMA set at 100%, the sensitivity and specificity of the antibody tests were as follows: gpELISA, 67% and 100%; EIA, 67% and 100%; and IAHA, 47% and 100%, respectively. The positive agreement rates of EIA and IAHA relative to gpELISA were 86% and 64%, respectively. CONCLUSIONS: gpELISA had a lower positive rate than did FAMA, and showed comparable sensitivity to that of EIA.

Homecare protective and risk factors for early childhood caries in Japan.

BACKGROUND: Early childhood caries (ECC) affects children across Japan and throughout the world. Thus, it is important to identify dietary and dental care habits that either promote oral health or cause ECC. The objective of this study was to identify protective and risk factors associated with ECC in Japan. METHODS: In a typical rural Japanese community, we selected children born between 2004 and 2008 who had received checkups at their community health center including oral examinations conducted by dentists. We obtained data from children's records and from a questionnaire filled out by parents. We enrolled only children who at their checkup for 18-month-olds had no caries, and we obtained data about them at their checkup for 3-year-olds. We classified children as either having caries (treated or untreated) or being caries-free. We conducted bivariate analyses using data on child/family demographic characteristics, child's dietary habits, and child/parental oral health habits. We also conducted logistic regression analysis to control for variables and identify predictors of the presence/absence of caries. RESULTS: Five hundred sixty six children (278 boys, 288 girls) were enrolled and followed. After 2 years, 173 children (30.6%) presented with caries. Logistic regression analysis predicting caries at follow-up identified the interaction term "bottlefed overnight and brushed irregularly" at 18 months of age as a highly significant predictor of developing caries-adjusted odds ratio (AOR) of 14.27, 95% confidence interval (CI) 1.02-199.71. Two variables measured at follow-up were also significant predictors: having low levels of dental plaque (AOR 2.41, 95% CI 1.34-4.35) and having a mother who had untreated caries (AOR 1.84, 95% CI 1.09-3.12). CONCLUSION: Public health efforts should encourage parents to eliminate bottle feeding overnight and promote brushing twice daily as children's teeth begin to erupt. Greater efforts should be made to teach parents and daytime caregivers how to brush effectively to remove all plaque. Health professionals should pay close attention to mothers' oral health status. Mothers with caries should receive prompt treatment and be assisted in developing better dietary and oral health habits that will benefit themselves and their children. Policies and programs should focus more on family oral health rather than just child oral health.

WP1066 suppresses macrophage cell death induced by inflammasome agonists independently of its inhibitory effect on STAT3.

The compound WP1066 was originally synthesized by modifying the structure of AG490, which inhibits the activation of signal transducer and activator of transcription 3 (STAT3) by directly targeting Janus kinases (JAKs). WP1066 exhibits stronger anti-cancer activity than AG490 against malignant glioma and other cancer cells and is regarded as a promising therapeutic agent. By screening a small library of target-known compounds, we identified WP1066 as an inhibitor of macrophage cell death induced by agonists of the NLRP3 inflammasome, an intracellular protein complex required for the processing of the proinflammatory cytokine interleukin (IL)-1ß. WP1066 strongly inhibited cell death as well as extracellular release of IL-1ß induced by inflammasome agonists in mouse peritoneal exudate cells and human leukemia monocytic THP-1 cells that were differentiated into macrophagic cells by treatment with PMA. However, inflammasome agonists did not increase STAT3 phosphorylation, and another JAK inhibitor, ruxolitinib, did not inhibit cell death, although it strongly inhibited basal STAT3 phosphorylation. Thus, WP1066 appears to suppress macrophage cell death independently of its inhibitory effect on STAT3. In contrast, WP1066 itself induced the death of undifferentiated THP-1 cells, suggesting that WP1066 differentially modulates cell death in a context-dependent manner. Consistent with previous findings, WP1066 induced the death of human glioma A172 and T98G cells. However, neither ruxolitinib nor AG490, the former of which completely suppressed STAT3 phosphorylation, induced the death of these glioma cells. These results suggest that WP1066 targets cell death-modulating molecules other than those involved in JAK-STAT3 signaling.

TNF-α Induces Caspase-1 Activation Independently of Simultaneously Induced NLRP3 in 3T3-L1 Cells.

The intracellular cysteine protease caspase-1 is critically involved in obesity-induced inflammation in adipose tissue. A substantial body of evidence from immune cells, such as macrophages, has shown that caspase-1 activation depends largely on a protein complex, called the NLRP3 inflammasome, which consists of the NOD-like receptor (NLR) family protein NLRP3, the adaptor protein ASC, and caspase-1 itself. However, it is not fully understood how caspase-1 activation is regulated within adipocytes upon inflammatory stimuli. In this study, we show that TNF-α-induced activation of caspase-1 is accompanied by robust induction of NLRP3 in 3T3-L1 adipocytes but that caspase-1 activation may not depend on the NLRP3 inflammasome. Treatment of 3T3-L1 cells with TNF-α induced mRNA expression and activation of caspase-1. Although the basal expression of NLRP3 and ASC was undetectable in unstimulated cells, TNF-α strongly induced NLRP3 expression but did not induce ASC expression. Interestingly, inhibitors of the ERK MAP kinase pathway strongly suppressed NLRP3 expression but did not suppress the expression and activation of caspase-1 induced by TNF-α, suggesting that NLRP3 is dispensable for TNF-α-induced caspase-1 activation. Moreover, we did not detect the basal and TNF-α-induced expression of other NLR proteins (NLRP1a, NLRP1b, and NLRC4), which do not necessarily require ASC for caspase-1 activation. These results suggest that TNF-α induces caspase-1 activation in an inflammasome-independent manner in 3T3-L1 cells and that the ERK-dependent expression of NLRP3 may play a role independently of its canonical role as a component of inflammasomes. J. Cell. Physiol. 231: 2761-2767, 2016. © 2016 Wiley Periodicals, Inc.

Targeting the ERK signaling pathway as a potential treatment for insulin resistance and type 2 diabetes.

Extracellular signal-regulated kinase (ERK) has been implicated in the development of insulin resistance associated with obesity and type 2 diabetes mellitus. We have now examined the potential of pharmacological targeting of the ERK pathway with MEK (ERK kinase) inhibitors (PD184352 and PD0325901) for the treatment of obesity-associated insulin resistance. The effects of PD184352 and PD0325901 on the expression of adipocytokines and lipolysis activity were thus examined in 3T3-L1 adipocytes maintained in long-term culture as a model of adipocyte hypertrophy. Leptin receptor-deficient (db/db) mice and high-fat diet-fed KKAy mice, both of which are models of type 2 diabetes, were also treated orally with PD184352 to examine its effects on the diabetic condition. ERK activity was increased in hypertrophic 3T3-L1 adipocytes as well as in adipose tissue of db/db mice and high-fat diet-fed KKAy mice, and this enhanced ERK signaling was associated with dysregulation of adipocytokine expression and increased lipolysis activity. Specific blockade of the ERK pathway in hypertrophic 3T3-L1 adipocytes by MEK inhibitors ameliorated the dysregulation of adipocytokine expression and suppressed the enhanced lipolysis activity. Furthermore, repeated oral administration of PD184352 normalized hyperglycemia and hyperlipidemia and improved insulin sensitivity and glucose tolerance in the diabetic mice. These results suggest that sustained activation of the ERK pathway in adipocytes is associated with the pathogenesis of type 2 diabetes and that selective blockade of this pathway with MEK inhibitors warrants further study as a promising approach to the treatment of insulin resistance and type 2 diabetes.

Varicella zoster virus antibody detection: A comparison of four commonly used techniques.

BACKGROUND: Antibody tests for the varicella zoster virus (VZV) include neutralization, fluorescent antibody to membrane antigen (FAMA), immune adherence hemagglutination (IAHA), enzyme immunoassay (EIA), glycoprotein-based enzyme-linked immunosorbent assay (gpELISA), and complement fixation (CF) tests. Of these, FAMA is considered the most sensitive. However, in Japan, the EIA method is most frequently employed. OBJECTIVE: The VZV antibody detection rate of the FAMA, EIA, gpELISA, and IAHA methods was compared. METHODS: Four types of antibody tests were conducted with sera collected from 83 college students. The relationships between two antibody tests were examined using Pearson's correlation coefficients. RESULTS: All 83 subjects were observed to be VZV antibody-positive using the FAMA method. The Pearson correlation coefficients of gpELISA, EIA, and IAHA relative to FAMA were 0.808, 0.782, and 0.356, respectively. The positive agreement rate of IAHA relative to FAMA was 88.0% (73/83), whereas those of gpELISA and EIA were both 97.6% (81/83). Furthermore, EIA showed 100% positive agreement with gpELISA and a high correlation coefficient of 0.911, whereas these values for IAHA compared to gpELISA were much lower (90.1% and 0.530). The calculated Pearson correlation coefficient for comparison of the EIA and IAHA methods was 0.498, with a positive agreement rate of 90.1% (73/81). CONCLUSIONS: The EIA method should be employed in Japan based on the similarity of the positivity between EIA and gpELISA, as it is more available and practical than gpELISA.

Spatial Clusters of High Prevalences of Overweight and Obesity Among Children in Indonesia.

Objective Childhood obesity has emerged as a pressing health concern in both high-income and lower-middle-income countries, including Indonesia. The prevalence of overweight and obesity among school children aged 5-12 years has been increasing in Indonesia, with 20% of Indonesian children identified as obese or overweight in 2018. Therefore, addressing this problem will be challenging. This study aims to identify district- and city-level clusters with high prevalences of overweight and obese children aged 5-12 years in Indonesia. Methodology This is an ecological study that utilizes secondary data from the 2018 Basic Health Research report conducted by the Indonesian Ministry of Health. We included 514 districts and cities to detect district- and city-level clusters. Spatial cluster analysis was performed using restricted flexible scan statistics to identify clusters with high prevalences of childhood overweight and obesity in Indonesian districts and cities. Results The findings reveal that childhood overweight and obesity are not randomly distributed. The study detected 20 clusters with high prevalences of childhood overweight and 36 clusters of obesity, with a particular concentration in Western Indonesia. A primary cluster of childhood overweight occurred in Sijunjung, Tanah Datar, Agam, Pasaman, South Solok, Dharmasraya, West Pasaman, Sawah Lunto City, Padang Panjang City, and Kampar. A primary cluster of obesity occurred in Mandailing Natal, South Tapanuli, Central Tapanuli, North Tapanuli, Labuhan Batu, North Padang Lawas, Padang Lawas, North Labuhan Batu, West Pasaman, and Rokan Hilir. Conclusions This study found 20 clusters with high prevalences of childhood overweight and 36 clusters of obesity in Indonesia. Implementing health promotion programs in the identified cluster regions will be crucial to effectively addressing the growing problem of childhood obesity in Indonesia.

Timing of Assessment of Humoral and Cell-Mediated Immunity after Influenza Vaccination.

Assessment of the immune response to influenza vaccines should include an assessment of both humoral and cell-mediated immunity. However, there is a lack of consensus regarding the timing of immunological assessment of humoral and cell-mediated immunity after vaccination. Therefore, we investigated the timing of immunological assessments after vaccination using markers of humoral and cell-mediated immunity. In the 2018/2019 influenza season, blood was collected from 29 healthy adults before and after vaccination with a quadrivalent inactivated influenza vaccine, and we performed serial measurements of humoral immunity (hemagglutination inhibition [HAI] and neutralizing antibody [NT]) and cell-mediated immunity (interferon-gamma [IFN-γ]). The HAI and NT titers before and after vaccination were strongly correlated, but no correlation was observed between the markers of cell-mediated and humoral immunity. The geometric mean titer and geometric mean concentration of humoral and cellular immune markers increased within 2 weeks after vaccination and had already declined by 8 weeks. This study suggests that the optimal time to assess the immune response is 2 weeks after vaccination. Appropriately timed immunological assessments can help ensure that vaccination is effective.

Up-regulation of pro-apoptotic protein Bim and down-regulation of anti-apoptotic protein Mcl-1 cooperatively mediate enhanced tumor cell death induced by the combination of ERK kinase (MEK) inhibitor and microtubule inhibitor.

Blockade of the ERK signaling pathway by ERK kinase (MEK) inhibitors selectively enhances the induction of apoptosis by microtubule inhibitors in tumor cells in which this pathway is constitutively activated. We examined the mechanism by which such drug combinations induce enhanced cell death by applying time-lapse microscopy to track the fate of individual cells. MEK inhibitors did not affect the first mitosis after drug exposure, but most cells remained arrested in interphase without entering a second mitosis. Low concentrations of microtubule inhibitors induced prolonged mitotic arrest followed by exit of cells from mitosis without division, with most cells remaining viable. However, the combination of a MEK inhibitor and a microtubule inhibitor induced massive cell death during prolonged mitosis. Impairment of spindle assembly checkpoint function by RNAi-mediated depletion of Mad2 or BubR1 markedly suppressed such prolonged mitotic arrest and cell death. The cell death was accompanied by up-regulation of the pro-apoptotic protein Bim (to which MEK inhibitors contributed) and by down-regulation of the anti-apoptotic protein Mcl-1 (to which microtubule and MEK inhibitors contributed synergistically). Whereas RNAi-mediated knockdown of Bim suppressed cell death, stabilization of Mcl-1 by RNAi-mediated depletion of Mule slowed its onset. Depletion of Mcl-1 sensitized tumor cells to MEK inhibitor-induced cell death, an effect that was antagonized by knockdown of Bim. The combination of MEK and microtubule inhibitors thus targets Bim and Mcl-1 in a cooperative manner to induce massive cell death in tumor cells with aberrant ERK pathway activation.

Functional characterization of the novel BRAF complex mutation, BRAF(V600delinsYM) , identified in papillary thyroid carcinoma.

An activating mutation in the BRAF gene is the most common genetic alteration in papillary thyroid carcinomas (PTCs). The mutation in PTCs is almost a c.1799T>A transversion, resulting in a p.V600E amino acid substitution (BRAF(V600E) ). Here, we report a novel complex BRAF mutation identified in 4/492 Japanese PTC cases (0.81%). The mutation was comprised of one nucleotide substitution at position 1798, followed by an in-frame insertion of three nucleotides, c.1798delinsTACA in Exon 15, resulting in p.V600delinsYM. In silico three-dimensional protein structure prediction implied altered kinase activity of this mutant. In vitro kinase assay and western blotting revealed that this mutation conferred high kinase activity on the BRAF protein, leading to constitutive activation of the MAPK signaling pathway. The mutation also showed high transforming ability in focus formation assay using NIH3T3 cells. The degree of all the functional characteristics was comparable to that of BRAF(V600E) , and treatment with a BRAF inhibitor Sorafenib was also equally effective in this mutant. These findings suggest that the novel BRAF mutation, BRAF(V600delinsYM) , is a gain-of-function mutation and plays an important role in PTC development.

Blockade of the ERK pathway enhances the therapeutic efficacy of the histone deacetylase inhibitor MS-275 in human tumor xenograft models.

The ERK pathway is up-regulated in various human cancers and represents a prime target for mechanism-based approaches to cancer treatment. Specific blockade of the ERK pathway alone induces mostly cytostatic rather than pro-apoptotic effects, however, resulting in a limited therapeutic efficacy of the ERK kinase (MEK) inhibitors. We previously showed that MEK inhibitors markedly enhance the ability of histone deacetylase (HDAC) inhibitors to induce apoptosis in tumor cells with constitutive ERK pathway activation in vitro. To evaluate the therapeutic efficacy of such drug combinations, we administered the MEK inhibitor PD184352 or AZD6244 together with the HDAC inhibitor MS-275 in nude mice harboring HT-29 or H1650 xenografts. Co-administration of the MEK inhibitor markedly sensitized the human xenografts to MS-275 cytotoxicity. A dose of MS-275 that alone showed only moderate cytotoxicity thus suppressed the growth of tumor xenografts almost completely as well as induced a marked reduction in tumor cellularity when administered with PD184352 or AZD6244. The combination of the two types of inhibitor also induced marked oxidative stress, which appeared to result in DNA damage and massive cell death, specifically in the tumor xenografts. The enhanced therapeutic efficacy of the drug combination was achieved by a relatively transient blockade of the ERK pathway. Administration of both MEK and HDAC inhibitors represents a promising chemotherapeutic strategy with improved safety for cancer patients.

Atypical protein phosphatases: emerging players in cellular signaling.

It has generally been considered that protein phosphatases have more diverse catalytic domain structures and mechanisms than protein kinases; however, gene annotation efforts following the human genome project appeared to have completed the whole array of protein phosphatases. Ser/Thr phosphatases are divided into three subfamilies that have different structures from each other, whereas Tyr phosphatases and dual-specificity phosphatases targeting Tyr, Ser and Thr belong to a single large family based on their common structural features. Several years of research have revealed, however, the existence of unexpected proteins, designated here as "atypical protein phosphatases", that have structural and enzymatic features different from those of the known protein phosphatases and are involved in important biological processes. In this review, we focus on the identification and functional characterization of atypical protein phosphatases, represented by eyes absent (EYA), suppressor of T-cell receptor signaling (Sts) and phosphoglycerate mutase family member 5 (PGAM5) and discuss their biological significance in cellular signaling.

Factors influencing the recovery-oriented attitudes of nursing directors in psychiatric hospitals: A cross-sectional study in Japan.

WHAT IS KNOWN ON THE SUBJECT?: Recovery-oriented services have been shown to promote recovery in people with mental illness; their implementation is important for current psychiatric practice and is being considered by many professional institutions. The Japanese government released the policy 'A Vision for Reform of Mental Health and Medical Welfare' (Ministry of Health, Labour and Welfare, 2004; https://www.mhlw.go.jp/topics/2004/09/dl/tp0902-1a.pdf), aiming to update public consciousness, reorganize the mental healthcare welfare system and strengthen the foundation of mental healthcare to promote a basic policy focusing on the transition 'from hospital to community life'. WHAT THE PAPER ADDS TO EXISTING KNOWLEDGE?: This is the first study to clarify recovery-oriented attitudes among psychiatric nursing directors and related factors in Japan. It shows an association between recovery-oriented attitudes and hospital size. WHAT ARE THE IMPLICATIONS FOR PRACTICE?: A relationship exists between the recovery knowledge and recovery-oriented attitudes of psychiatric nursing directors. It may thus be valuable for directors to see and hear about the experiences of patients living in the community during their recovery process. As hospital size and organizational climate may hinder the successful implementation of recovery-oriented practices, these factors must be considered when introducing such practices. Further research is needed on the relationship between psychiatric nursing directors' recovery-oriented attitudes and practices. Moreover, to develop intervention studies that uncover success factors for enhancing recovery-oriented attitudes, organizational factors that enable recovery-oriented practices must be further investigated. ABSTRACT: INTRODUCTION: Recovery-oriented practices in mental health services have become a global priority. Despite their proven effectiveness, Japanese hospitals have not widely implemented such practices. AIM: This study clarified psychiatric nursing directors' recovery-oriented attitudes and factors regarding promoting recovery-oriented practices to identify methods for developing these practices in Japan. METHOD: This cross-sectional study used a questionnaire survey with 250 nursing directors from 1287 Japanese psychiatric hospitals. Multivariable regression analysis assessed how socio-demographic variables and Recovery Knowledge Inventory (RKI) scores affected Recovery Attitudes Questionnaire (RAQ-7) scores. RESULTS: The analysis revealed that more clinical and managerial experience was associated with more negative recovery-oriented attitudes. High RKI scores, knowledge of strength, hospital size and outpatient work experience were associated with more positive recovery-oriented attitudes. DISCUSSION: Implementing recovery practices requires consideration of knowledge and experience in recovery and hospital size alongside recovery training based on a hospital's organizational structure. Although the low response rate could influence the study's generalisability, it could also indicate low interest in recovery-oriented practices among professionals. IMPLICATIONS FOR PRACTICE: Acquiring more knowledge and experience regarding recovery promotes recovery-oriented attitudes among nursing directors. An appropriate organizational culture and consideration of hospital size are required when introducing such recovery practices.

Types and frequency of social participation and comprehensive frailty among community-dwelling older people in Japan.

Although research has established social participation as important for preventing frailty in older people, the association between the type and frequency of social participation and comprehensive frailty remains unclear. This study aimed to reveal the associations between types and frequency of social participation and comprehensive frailty among community-dwelling older people. This was a cross-sectional study conducted in four cities and towns (Inabe City, Nabari City, Odai Town, and Kiho Town) of Mie Prefecture, Japan, among adults who were: (i) aged ≥65 years and (ii) not certified as needing long-term care. We measured comprehensive frailty using the participants' total scores on the Kihon Checklist, developed by Ministry of Health, Labour and Welfare of Japan, which divides frailty status into three categories: robust (0-3 points), prefrail (4-7), and frail (8-25). Types and frequency of social participation were explanatory variables, and ordered logistic regression analysis adjusted for potential confounding factors identified the associations. The frailty status of the 296 participants (age 65-74 years: 44.3 %; female: 74.0 %) was as follows: frail, 21.3 %; prefrail, 40.2 %; and robust, 38.5 %. Lower level of frailty was associated with interaction using smartphones 2-3 times per month, participating in sports ≥4 times per week, participating in local improvement activities several times per year, and engaging in activities for children 2-4 times per month. Social participation among older adults at appropriate frequencies were associated with the lower level of comprehensive frailty. However, future longitudinal studies are needed using populations from more diverse countries or regions and from different cultures.