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A collection of 37 fungi associated to Italian lemon plants with disease symptoms, was obtained. Ten genera including Aspergillus, Alternaria, Nigrospora, Lasiodiplodia, Dothideomycetes, Pleurostoma, Setosphaeria, Penicillium, Fusarium and Colletotrichum were identified by using ITS1–5.8S–ITS2, D1/D2 26S and COX1 loci. The last three genera were abundant on the damaged fruits, being Colletotrichum the more abundant (32.4 %). CaInt2 and CgInt primers support the identity of these isolates as C. gloeosporioides. Variability, inferred by rep-PCR and multilocus sequence analysis shows genetic differences among the C. gloeosporioides isolates. Infective profile evaluated in Colletotrichum isolates shows different leave infection percentages (26 to 60 %). SEM analysis showed mycelium, spores and appressoria on the leaves of selected Colletotrichum isolates. Specifically, the AL-05 and AL-13 isolates showed a high chitin deacetylase activity (CDA) peaking at 1.2 U/mg protein in AL-13. This is the first report on C. gloeosporioides infecting Italian lemon leaves in Mexico.

Competing Interests: Declaration of interests SA declares grants from the Indian Council for Medical Research (2022–25), the Government of Karnataka (2022–23), Rotary Bangalore Midtown (2022–23), Lowes Services India (2022–25), and Wellcome Trust (2023–26); payment for expert testimony received by Indian Council of Medical Research and Ashoka University; and a travel grant paid by University College London for being part of the Lancet Commission. SB declares grants from National Institute for Health and Care Research (NIHR), Economic and Social Research Council, Engineering and Physical Science Research Council, Canadian Institute for Health Research, the Alzheimer's Association, the Alzheimer's Society, Health Education England, Alzheimer's Association, Alzheimer's Society, and Health Education England. He has held the following positions: Non-Executive Director Somerset NHS Foundation Trust, Trustee of the Alzheimer's Society, Executive Dean of the University of Plymouth, and Pro-Vice Chancellor of the University of Nottingham. AB acts as a consultant for Lilly, TauRx Pharmaceuticals, and Eisai and carries out medico–legal work for solicitors. NCF declares consulting fees from F Hoffmann-La Roche, Eli Lilly, Ionis, Biogen, and Siemens; participation in data safety monitoring or advisory board for Biogen; and being a member of the Research Strategy Council for the Alzheimer's Society. LNG declares owning tailored activity programme licences. MK declares grants from Wellcome Trust (221854/Z/20/Z), the Medical Research Council (R024227), the National Institute on Aging (R01AG062553, R01AG056477), and the Academy of Finland (350426). KYL declared fellowship from Medical Research Council. EBL receives grants from the National Institutes of Health (NIH) and royalties from UpToDate. GL declares support for the manuscript from the Alzheimer's Society, the Alzheimer's Society UK, and UK Research and Innovation, who gave grants to pay for travel and accommodation. She is supported by the University College London Hospitals' NIHR Biomedical Research Centre, by North Thames NIHR Applied Research Collaboration, and as an NIHR Senior Investigator and has grants from NIHR Health Technology Assessment, NIHR Programme Grants for Applied Research, the Alzheimer's Association, the Norwegian Research Council, and Wellcome, outside of the submitted work. She works with the Alzheimer's Society as a member of the Research Strategy Council and is a trustee of Nightingale Hammerson care homes. KR declares grants from Canadian Institutes of Health Research, the Canadian Frailty Network, and Research Nova Scotia; royalties from Biotest, Qu Biologics, AstraZeneca UK, BioAge Labs, Congenica, Icosavax, KCR, Faraday Pharmaceuticals, Synairgen Research, Enanta Pharmaceuticals, Pfizer, Boehringer Ingelheim International, Fresenius Kabi Deutschland, Baycrest Geriatric Care, and Shanghai Ark Biopharmaceutical; payment or honoraria from University of British Columbia, Fraser Health Authority, McMaster University, Chinese Medical Association, Wake Forest University Medical School Centre, University of Omaha, and Atria Institute; participation on data safety or advisory board for EpiPharma; and leadership of the Canadian Consortium on Neurodegeneration in Dementia, Cap Breton University, and Nova Scotia Health. KS declares support from the Japan Society for the Promotion of Science fund (22H03352, 21KK0168, 16KK0059). LSS declares support from Della Martin Foundation, the NIH (P30 AG066530, R01 AG051346, R01 AG062687, R01 AG051346, R01 AG055444, P01 AG052350, R01 AG053267, R01 AG074983, R01 AG063826), Abbott, Biohaven, Biogen, Eisai, and Eli Lilly and consulting fees from AC Immune, Cortexyme, Alpha-cognition, BioVie, Athira, Eli Lilly/Avid, Corium, Lundbeck, Merck, Muna Therapeutics, Novo-Nordisk, Neurim, NeuroDiagnostics, Ono, Otsuka, Roche/Genentech, Cognition, Lighthouse, GW Research, ImmunoBrain, and Bristol Myers Squibb. AS declares grants from Wellcome Trust, the Alzheimer's Association, Brain Canada, and the NIHR. YY declares support from the National Natural Science Foundation of China (72374013) and the National Key R&D Program of China (2023YFB4603200, 2023YFC3606400). SW declares an NIHR doctoral training fellowship. GS has participated on advisory boards for the following pharmaceutical companies manufacturing drugs against Alzheimer's disease: Biogen, Roche, and Eisai. LG is an inventor of a training program for health and human service professionals in an evidence-based tailored activity intervention, the Tailored Activity Program; she and her respective universities are entitled to fees. All other authors declare no competing interests.

An in vitro system that evaluates pharmacokinetics in the small intestine is crucial for the development of oral drugs. We produced human induced pluripotent stem cell-derived small intestinal epithelial cells (hiSIECs) with high drug metabolizing enzyme and drug transporter activities. However, the gene expression of our hiSIECs partially differed from that of the human small intestine, with low drug metabolizing enzyme activities. Therefore, we used air-liquid interface (ALI) culture and 5-aza-2'-deoxycytidine (5AZA)-free medium to generate hiSIECs (novel hiSIECs). Novel hiSIECs showed enhanced gene expression of drug metabolizing enzymes, such as cytochrome P450 (CYP)3A4, CYP2C9, CYP2C19, and carboxylesterase 2 that are highly expressed in the small intestine. In addition, the expression of genes involved in nutrient absorption-one of the major functions of the small intestine-also increased. The novel hiSIECs expressed ZO-1 and E-cadherin. Moreover, the novel hiSIECs exhibited a barrier function that allowed low lucifer yellow permeation. The novel hiSIECs showed high activities of CYP3A4, CYP2C9, and CYP2C19, which are abundantly expressed in the small intestine. In conclusion, the novel hiSIECs have great potential as an in vitro system to evaluate pharmacokinetics in the small intestine., Competing Interests: Declarations of competing interest This study was funded by FUJIFILM Corporation., (© 2023 Published by Elsevier Ltd on behalf of The Japanese Society for the Study of Xenobiotics.)

Objectives: The sense of coherence refers to effectively using available resources to manage stress and promote overall health. Previous studies have linked it to various health outcomes; however, evidence regarding its association with the risk of incident dementia is limited. Hence, this study aimed to fill this research gap using data from a large-scale population survey., Design: Prospective cohort study., Setting and Participants: This study included 31,556 participants aged 65 years and older who were free from dementia and disabilities. The participants were enrolled in the 2010 baseline survey of the Japan Gerontological Evaluation Study, and were followed up until the end of 2019., Methods: The sense of coherence was assessed using a 6-item short-version questionnaire. Incident dementia cases were ascertained through the public long-term care insurance database in Japan., Results: During a median follow-up of 8.3 years, 4326 incident dementia cases were identified. After adjusting for conventional risk factors, the hazard ratios (95% CIs) for each quintile compared to the lowest quintile of the sense of coherence were 0.82 (0.75-0.90), 0.75 (0.68-0.83), 0.76 (0.68-0.84), and 0.78 (0.70-0.87), respectively. The multivariable hazard ratio (95% CI) per 1-SD increment was 0.91 (0.88-0.95). These inverse associations did not exhibit any gender differences (P for gender interaction = .11) and were further confirmed after excluding early incident cases. Similar associations were found for its components; the corresponding multivariable hazard ratios (95% CIs) per 1-SD increment were 0.94 (0.91-0.97) for comprehensibility, 0.92 (0.89-0.95) for manageability, and 0.93 (0.90-0.97) for meaningfulness., Conclusions and Implications: Moderate and above sense of coherence was associated with the lower risk of dementia among the older population, suggesting a beneficial role of stress management in maintaining the cognitive health of older adults., (Copyright © 2023. Published by Elsevier Inc.)

Background: Cohort evidence linking long-term survival with exposure to multiple air pollutants (e.g., fine particulate matter [PM 2.5 ] and ozone) was extensively sparse in low- and middle-income countries, especially among older adults. This study aimed to investigate potential associations of long-term exposures to PM 2.5 and ozone with all-cause mortality in Chinese older adults., Methods: A dynamic nationwide prospective cohort comprising 20,352 adults aged ≥65 years were enrolled from the Chinese Longitudinal Healthy Longevity Study and followed up through 2005-2018. Participants' annual exposures to warm-season ozone and year-round PM 2.5 were assigned using satellite-derived spatiotemporal estimates. A directed acyclic graph (DAG) was developed to identify confounding variables. Associations of annual mean exposures to PM 2.5 and ozone with mortality were evaluated using single- and two-pollutant Cox proportional hazards models, adjusting for time-dependent individual risk factors and ambient temperature., Results: During 100 thousand person-years of follow-up (median: 3.6 years), a total of 14,313 death events occurred. The participants were averagely aged 87.1 years at baseline and exposed to a wide range of annual average concentrations of warm-season maximum 8-hour ozone (mean, 54.4 ppb; range, 23.3-81.6 ppb) and year-round PM 2.5 (mean, 65.5 μg/m 3 ; range, 10.1-162.9 μg/m 3 ). Approximately linear concentration-response relationship was identified for ozone, whereas significant increases in PM 2.5 -associated mortality risks were observed only when concentrations were above 60 μg/m 3 . Rises of 10 ppb in ozone and 10 µg/m 3 in PM 2.5 above 60 µg/m 3 were associated with increases in all-cause mortality of 13.2% (95% confidence interval [CI]: 10.2-16.2%) and 6.2% (95% CI: 4.6-7.7%) in DAG-based single-pollutant model, and of 9.7% (95% CI: 6.6-13.0%) and 5.3% (95% CI: 3.7-6.9%) in DAG-based two-pollutant model, respectively. We detected significant effect modification by temperature in associations of mortality with ozone (P <0.001 for interaction), suggesting greater ozone-related risks among participants in warmer locations., Conclusions: This study provided longitudinal evidence that long-term exposure to ambient PM 2.5 and ozone significantly and independently contributed to elevated risks of all-cause mortality among older adults in China., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Ocean acidification can severely affect bivalve molluscs, especially their shell calcification. Assessing the fate of this vulnerable group in a rapidly acidifying ocean is therefore a pressing challenge. Volcanic CO 2 seeps are natural analogues of future ocean conditions that offer unique insights into the scope of marine bivalves to cope with acidification. Here, we used a 2-month reciprocal transplantation of the coastal mussel Septifer bilocularis collected from reference and elevated pCO 2 habitats to explore how they calcify and grow at CO 2 seeps on the Pacific coast of Japan. We found significant decreases in condition index (an indication of tissue energy reserves) and shell growth of mussels living under elevated pCO 2 conditions. These negative responses in their physiological performance under acidified conditions were closely associated with changes in their food sources (shown by changes to the soft tissue δ 13 C and δ 15 N ratios) and changes in their calcifying fluid carbonate chemistry (based on shell carbonate isotopic and elemental signatures). The reduced shell growth rate during the transplantation experiment was further supported by shell δ 13 C records along their incremental growth layers, as well as their smaller shell size despite being of comparable ontogenetic ages (5-7 years old, based on shell δ 18 O records). Taken together, these findings demonstrate how ocean acidification at CO 2 seeps affects mussel growth and reveal that lowered shell growth helps them survive stressful conditions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Purpose: Cerebellar hypoplasia and atrophy (CBHA) in children is an extremely heterogeneous group of disorders, but few comprehensive genetic studies have been reported. Comprehensive genetic analysis of CBHA patients may help differentiating atrophy and hypoplasia and potentially improve their prognostic aspects., Methods: Patients with CBHA in 176 families were genetically examined using exome sequencing. Patients with disease-causing variants were clinically evaluated., Results: Disease-causing variants were identified in 96 of the 176 families (54.5%). After excluding 6 families, 48 patients from 42 families were categorized as having syndromic associations with CBHA, whereas the remaining 51 patients from 48 families had isolated CBHA. In 51 patients, 26 aberrant genes were identified, of which, 20 (76.9%) caused disease in 1 family each. The most prevalent genes were CACNA1A, ITPR1, and KIF1A. Of the 26 aberrant genes, 21 and 1 were functionally annotated to atrophy and hypoplasia, respectively. CBHA+S was more clinically severe than CBHA-S. Notably, ARG1 and FOLR1 variants were identified in 2 families, leading to medical treatments., Conclusion: A wide genetic and clinical diversity of CBHA was revealed through exome sequencing in this cohort, which highlights the importance of comprehensive genetic analyses. Furthermore, molecular-based treatment was available for 2 families., Competing Interests: Conflict of Interest The authors declare no conflict of interest., (Copyright © 2022 American College of Medical Genetics and Genomics. Published by Elsevier Inc. All rights reserved.)

Background: Light after dusk disrupts the circadian rhythms and shifts the timing of sleep later; but it is unknown whether outdoor artificial light at night (ALAN) affects sleep quality. This study aimed to explore the association between residential outdoor ALAN and sleep duration in a nationally representative sample of Chinese older adults., Methods: We examined the cross-sectional associations of outdoor ALAN with self-reported sleep duration in 13,474 older adults participating in the 2017-2018 wave of the Chinese Longitudinal Healthy Longevity Survey (CLHLS). Outdoor ALAN exposure was estimated at the residence level using satellite images. We applied generalized linear mixed models to investigate the association between ALAN exposure and sleep duration. We performed stratified analyses by age, sex, education, and household income levels. Moreover, we used multi-level logistic regression models to investigate the effects of ALAN on the short sleep duration (≤6 h) and the long sleep duration (>8 h), respectively, in reference to sleep for >6-8 h per day., Results: We found a significant association between outdoor ALAN intensity and sleep duration. The highest quartile of ALAN was associated with 17.04 (95% CI: 9.42-24.78) fewer minutes of sleep as compared to the lowest quartile. The reductions in sleep duration per quartile change in ALAN were greater in the young old (≥65-85 years) and in those with higher levels of education, and those with higher household income, respectively. We did not detect a sex difference. In addition, those in the highest quartile of ALAN were more likely to report a 25% (95% CI: 10%-42%) increase in short sleep (<6 h), and a 21% (95% CI: 9%-31%) decrease in long sleep (>8 h)., Conclusions: Increasing outdoor nighttime light intensity surrounding residences was associated with shorter sleep duration in older residents in China. This finding implies the importance of urban outdoor artificial light management as a potential means to lower the public health burden of sleep disorders., (Copyright © 2022 Elsevier Inc. All rights reserved.)

In humans, atypical endometrial hyperplasia (AEH) is considered as a precancerous lesion of endometrial adenocarcinoma (EA), from which it must be distinguished. Precancerous lesions have not been reported in cats with EA. We now document the histopathological features of endometrial lesions in six cats, which histopathologically resembled human AEH and had a good prognosis following ovariohysterectomy. Grossly, one cat presented with papillomatous nodules and three cats had pyometra. Histopathologically, proliferation of endometrial epithelial cells without atypia was observed in all cases. In some regions of the endometrium, cells had increased atypia and were arranged in stratified layers, which formed irregular ducts and papillary structures. No invasive behaviour or vascular invasion was observed. On the basis of these findings, the cats were diagnosed with non-invasive or early-stage adenocarcinoma. Immunohistochemistry for oestrogen receptor and progesterone receptor revealed an inverse correlation with the severity of the endometrial lesions and degree of malignancy of tumour cells. Ki67 labelling revealed that mitotic activity increased as the lesion developed. All cats survived, with a median survival time of 385 days (range: 229-744 days). The distribution of the histopathological endometrial changes and the non-invasive behaviour in these feline cases resemble cases of AEH in humans., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Background and Purpose: An effect of dietary carotenes on risk of cardiovascular disease (CVD) is uncertain. We aimed to investigate whether the association between dietary carotenes intake and risk of CVD mortality will persist after controlling for the intakes of potential cardioprotective dietary factors that correlate with dietary alpha- and/or beta-carotenes., Methods and Results: We followed up a total of 58,646 Japanese between 1988 and 1990 and 2009. We used a food frequency questionnaire (FFQ) to determine the dietary intakes of carotenes, and estimated the hazard ratios (HRs) and 95% confidence intervals (CIs) of CVD mortality in relation to carotene intake by the proportional hazard regression developed by David Cox. During 965,970 person-years of follow-up (median 19.3 years), we identified 3388 total CVD deaths. After adjusting for demographic and lifestyle factors, dietary intakes of alpha-carotene were significantly associated with the reduced risk of mortality from coronary heart disease (CHD); adjusted HR (95% CI) in the highest versus lowest quintiles of intake was 0.75 (0.58-0.96; P-trend = 0.02) and dietary intakes of beta-carotene were significantly associated with the reduced risk of mortality from CVD, CHD, and other CVD; adjusted HRs (95% CIs) were 0.88 (0.79-0.98; P-trend = 0.04), 0.78 (0.61-0.99; P-trend = 0.01), and 0.81 (0.67-0.98; P-trend = 0.04), respectively. However, after further adjusting for the dietary intakes of potassium, calcium, vitamins C, E, or K, these associations disappeared., Conclusions: -Dietary alpha- and beta-carotene intakes were not associated with risk of CVD mortality after controlling for intakes of other potential cardioprotective nutrients., Competing Interests: Declaration of competing interest The authors declare that they have no conflicts of interest., (Copyright © 2021. Published by Elsevier B.V.)

Background and Aims: The role of hobbies as a protective factor against cardiovascular diseases (CVD) has gained increasing attention; however, no large-scale studies were performed to confirm this. We aimed to examine the association between having hobbies and the risk of total CVD, coronary heart disease (CHD), and stroke in a large Japanese cohort., Methods: A total of 56,381 adults aged 45-74 years were divided into the non-hobby, having a hobby, and having many hobbies groups. We performed Cox proportional hazard models to estimate the hazard ratio (HRs) for incident CVD, CHD, and stroke after adjusting for potential confounding factors., Results: We identified 3685 incident CVDs (940 CHDs and 2839 strokes) during a median follow-up of 16.3 years. After multivariable adjustment, compared to the non-hobby group, participants having a hobby and many hobbies had a 10% (HR = 0.90 [0.83, 0.97]) and 20% (HR = 0.80 [0.69, 0.93]) lower risk of CVD incidence, respectively. The risk of CHD was lower in those with many hobbies, but the association between having many hobbies and CHD risk was not statistically significant. We also found a similar inverse association for stroke. Compared to the non-hobby group, participants having a hobby and many hobbies had a 13% (HR = 0.87 [0.80, 0.96]) and 20% (HR = 0.80 [0.68, 0.94]) lower risk of stroke., Conclusions: Having hobbies was associated with a lower risk of CVD. Engagement in hobbies may emerge as an important target for healthy lifestyle promotion for the primary prevention of CVD., (Copyright © 2021. Published by Elsevier B.V.)

We measured radiocesium in mussel tissue collected from the Pacific coast of Tohoku from 2011 to 2015 to investigate the temporal and spatial dynamics of radiocesium in the coastal area. Radioactive 137 Cs was detected in all the samples collected in 2011, but it was not found in samples from localities north of Sendai after 2012. In contrast, 137 Cs was detected in many sites in the Fukushima area even from 2012 to 2015. The fluctuation of 137 Cs concentration in mussel tissue seems to reflect the 137 Cs concentration in suspended particles in the seawater, suggesting that there was an influx of soil deposition and resuspension of seabed sediment. These results suggest that the 137 Cs concentration in mussel tissue sensitively indicates the 137 Cs concentration in the environment, and that the "mussel watch" approach is an effective way to understand the dynamics of radiocesium concentrations in coastal areas., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Immune checkpoint inhibitors (ICIs) are widely used for various malignancies. However, their safety and efficacy in patients with a kidney transplant have not been defined. To delineate this, we conducted a multicenter retrospective study of 69 patients with a kidney transplant receiving ICIs between January 2010 and May 2020. For safety, we assessed the incidence, timing, and risk factors of acute graft rejection. For efficacy, objective response rate and overall survival were assessed in cutaneous squamous cell carcinoma and melanoma, the most common cancers in our cohort, and compared with stage-matched 23 patients with squamous cell carcinoma and 14 with melanoma with a kidney transplant not receiving ICIs. Following ICI treatment, 29 out of 69 (42%) patients developed acute rejection, 19 of whom lost their allograft, compared with an acute rejection rate of 5.4% in the non-ICI cohort. Median time from ICI initiation to rejection was 24 days. Factors associated with a lower risk of rejection were mTOR inhibitor use (odds ratio 0.26; 95% confidence interval, 0.09-0.72) and triple-agent immunosuppression (0.67, 0.48-0.92). The objective response ratio was 36.4% and 40% in the squamous cell carcinoma and melanoma subgroups, respectively. In the squamous cell carcinoma subgroup, overall survival was significantly longer in patients treated with ICIs (median overall survival 19.8 months vs. 10.6 months), whereas in the melanoma subgroup, overall survival did not differ between groups. Thus, ICIs were associated with a high risk of rejection in patients with kidney transplants but may lead to improved cancer outcomes. Prospective studies are needed to determine optimal immunosuppression strategies to improve patient outcomes., (Copyright © 2020 International Society of Nephrology. Published by Elsevier Inc. All rights reserved.)

Objectives: The aim of this study was to investigate the association between different intensities and frequencies of non-occupational physical activity (PA) and the risk of dementia among Japanese older adults., Study Design: This was a prospective cohort study., Methods: A total of 2194 participants aged ≥65 years from the Japan Gerontological Evaluation Study were followed up between 2010 and 2016. The standardised dementia scale of the long-term care insurance system was used to identify incident dementia, whereas non-occupational PA (<2 or ≥2 times/week on each intensity: light, moderate and vigorous) was assessed using a questionnaire. Cox regression was used to compute the hazard ratios (HRs) and 95% confidence intervals (CIs) for incident dementia., Results: After adjustment for sociodemographic and medical characteristics, the following frequencies and intensities of non-occupational PA, compared with no non-occupational PA at all, were associated with a reduced risk of dementia: light PA ≥2 times/week (HR = 0.61, 95% CI: 0.38-0.97), moderate PA <2 times/week (HR = 0.46, 95% CI: 0.28-0.76), moderate PA ≥2 times/week (HR = 0.57, 95% CI: 0.36-0.91), vigorous PA <2 times/week (HR = 0.40, 95% CI: 0.21-0.74) and vigorous PA ≥2 times/week (HR = 0.29, 95% CI: 0.15-0.57). In the sex-specific analysis, moderate PA <2 times/week and vigorous PA ≥2 times/week were associated with a reduced risk of dementia in men, whereas light and moderate PA ≥2 times/week and all frequencies of vigorous PA were associated with a reduced risk of dementia in women., Conclusions: Practicing non-occupational PA was associated with a reduced risk of dementia among Japanese older adults., (Copyright © 2021 The Royal Society for Public Health. Published by Elsevier Ltd. All rights reserved.)

Microwave-mediated grafting of L-Arg onto naturally derived and stable multiradical poly(gallic acid) (PGAL) in aqueous media has been successfully achieved. This polymeric material has no adverse effect in human cells as there is no hemolytic activity upon MTT and Neutral Red assays. The analytical and computational characterization studies carried out in this study describe a helical molecular structure with random incorporation of L-Arginine pendant groups from PGAL's backbone. The antioxidant properties of the precursor polymer are preserved as proved by the elimination of stable DPPH and hydroxyl radical scavenging, as well as the FRAP and ORAC assays. Regarding the latter, the oxygen radical inhibition is enhanced compared to PGAL, which is attributed to the guanidyl moieties. PGAL-g-L-Arg displays antimicrobial activity against Gram (+) Listeria monocytogenes and Staphylococcus aureus strains with a MIC of 0.8 g/L and a bacteriostatic effect against Gram (-) Escherichia coli. Additionally, scanning electron and confocal fluorescence microscopies as well as crystal violet colorimetric assay demonstrate that the mechanism involved in the bacterial inhibition is related to the formation of porous channels on the membrane, which is discussed according to the helical secondary structure of the polymer and the amino acid guanidyl moieties interacting to bacterial membranes., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The Yesso scallop, Patinopecten yessoensis (Jay), is one of the most important bivalve species in the Japanese and Chinese mariculture industry. In recent years, however, high incidences of scallop shell deformity and mortality have occurred with increasing frequency, but timing of onset and underlying causes are often unclear. Here, we proposed a promising δ 18 O shell -based method for constraining the onset of shell deformity and mortality of P. yessoensis. Following six months of intermediate suspension culture in Funka Bay, Northern Japan, shells from healthy, deformed and dead scallops were randomly sampled. High-resolution seawater temperature time-series computed from healthy scallop shell δ 18 O profiles were precisely and temporally aligned to the instrumental temperature curve, thus allowing δ 18 O shell -derived temperature time-series from deformed and dead scallops to be contextualized and allowing timing of scallop deformity and death to be retrieved. Irrespective of scallop shell length, onsets of deformity were anchored in February, and since then deformed scallops grew slowly in comparison to healthy individuals. Without exception, however, dead scallops had already ceased their shell building and died before February, indicating different underlying causes of scallop deformity and mortality. Perhaps most promisingly, considering that shells do not have any isotopic turn-over and once formed, temperature information is locked in. Thus, this approach holds great promise for identifying time anchor points (onsets of deformity and death) in archived scallops collected over different time scales, especially during massive mortality events., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Introduction: Almonertinib (HS-10296) is a novel, third-generation EGFR tyrosine kinase inhibitor (EGFR TKI) that targets both EGFR-sensitizing and T790M resistance mutations. This first-in-human trial aimed to evaluate the safety, efficacy, and pharmacokinetics of almonertinib in patients with locally advanced or metastatic EGFR mutation-positive NSCLC that had progressed after pevious EGFR TKI therapy., Methods: This phase 1, open-label, multicenter clinical trial (NCT0298110) included dose-escalation (55, 110, 220, and 260 mg) and dose-expansion cohorts (55, 110, and 220 mg) with once daily oral administration of almonertinib. In each expansion cohort, tumor biopsies were obtained for the determination of EGFR T790M status. The safety, tolerability, antitumor activity, and pharmacokinetics of almonertinib were evaluated., Results: A total of 120 patients (26 patients in the dose-escalation cohort and 94 patients in the dose-expansion cohort) were enrolled. The maximum tolerated dose was not defined in the dose-escalation phase; the 260 mg regimen was not further evaluated in the dose-expansion phase owing to safety concerns and saturation of exposure. The most common treatment-related grade greater than or equal to 3 adverse events were increased blood creatine phosphokinase (10%) and increased alanine aminotransferase (3%). Among 94 patients with the EGFR T790M mutation in the dose-expansion cohort, the investigator-assessed objective response rate and disease control rate were 52% (95% confidence interval [CI]: 42-63) and 92% (95% CI: 84-96), respectively. Median progression-free survival was 11.0 months (95% CI: 9.5-not reached) months., Conclusions: Almonertinib is safe, tolerable and effective for patients with locally advanced or metastatic NSCLC harboring the EGFR T790M mutation who were pretreated with EGFR TKIs., (Copyright © 2020 International Association for the Study of Lung Cancer. Published by Elsevier Inc. All rights reserved.)

The gap junction beta-2 (GJB2) gene is the most common genetic cause of hereditary deafness worldwide. Especially, the 235delC mutation in GJB2 is most prevalent in East Asia. In this study, we generated two iPSC lines from PBMCs of siblings carrying homozygous 235delC mutation which exhibits an audiometric phenotype of profound hearing loss. These iPSC lines had normal karyotype, showed expression of pluripotency markers, and could differentiate into three germ layers. These disease specific iPSC lines may be useful for the construction of the disease models and for the elucidation of pathogenesis in GJB2-related deafness., (Copyright © 2020 The Author(s). Published by Elsevier B.V. All rights reserved.)

Background: There has been no report about outcome of pitavastatin versus atorvastatin therapy in high-risk patients with hypercholesterolemia., Methods: Hypercholesterolemic patients with one or more risk factors for atherosclerotic diseases (n = 664, age = 65, male = 54%, diabetes = 76%, primary prevention = 74%) were randomized to receive pitavastatin 2 mg/day (n = 332) or atorvastatin 10 mg/day (n = 332). Follow-up period was 240 weeks. The primary end point was a composite of cardiovascular death, sudden death of unknown origin, nonfatal myocardial infarction, nonfatal stroke, transient ischemic attack, or heart failure requiring hospitalization. The secondary end point was a composite of the primary end point plus clinically indicated coronary revascularization for stable angina., Results: The mean low-density lipoprotein cholesterol (LDL-C) level at baseline was 149 mg/dL. The mean LDL-C levels at 1 year were 95 mg/dL in the pitavastatin group and 94 mg/dL in the atorvastatin group. There were no differences in LDL-C levels between both groups, however, pitavastatin significantly reduced the risk of the primary end point, compared to atorvastatin (pitavastatin = 2.9% and atorvastatin = 8.1%, HR, 0.366; 95% CI 0.170-0.787; P = 0.01 by multivariate Cox regression) as well as the risk of the secondary end point (pitavastatin = 4.5% and atorvastatin = 12.9%, HR = 0.350; 95%CI = 0.189-0.645, P = 0.001). The results for the primary and secondary end points were consistent across several prespecified subgroups. There were no differences in incidence of adverse events between the statins., Conclusion: Pitavastatin therapy compared with atorvastatin more may prevent cardiovascular events in hypercholesterolemic patients with one or more risk factors for atherosclerotic diseases despite similar effects on LDL-C levels., Competing Interests: Declaration of competing interest Any potential conflicts of interest, including related consultancies, shareholding and funding grants: Dr. Moroi received a research grant from, Bayer Yakuhin, Ltd, Sanofi KK, MSD KK, Novartis Pharma, Ltd., Teijin Pharma Ltd., Mitsubishi Tanabe Pharma Corporation, Takeda Pharmaceutical Co., Ltd., and Nihon Mediphysics, Ltd. Dr. Shiba received grants and personal fees from Mitsubishi Tanabe Pharma, grants and personal fees from Daiichi Sankyo Company, Limited, grants and personal fees from Ono Pharmaceutical Co., Ltd., personal fees from Eli Lilly Japan K.K., personal fees from Boehringer Ingelheim, personal fees from Merck Sharp & Dohme (MSD), personal fees from Novartis Pharma, personal fees from Novo Nordisk Pharma Ltd., outside the submitted work. The remaining authors have nothing to disclose., (Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.)

N-acetylglucosaminidase produced from Lecanicillium lecanii on submerged culture displayed hydrolytic and transglycosylation activities. The highest specific activity for the enzyme was 1.87 U/mg after 120 h of culture. The chromatographic purification for a single protein fraction showed a molecular weight of 50.4 kDa and hydrolytic N-acetylglucosaminidase activity of 17.59 U/mg at 37 °C and pH 6. This enzyme was able to transglycosylate and to synthesize oligosaccharides from 2 to 6 units with a degree of acetylation between 100 and 26% employing glucose, mannose, N-acetyl-D-glucosamine and N-acetyl-D-lactosamine as donor substrates. Optimal conditions of temperature and pH were determined for both types of enzymatic activities., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Pulau Redang and Pulau Tioman have experienced huge tourism growth over the last two decades, but minimal sewage treatment may threaten the resilience of their coral reefs. This study uses stable isotope techniques to identify suitable bioindicators of sewage nutrients (δ 15 N) at these islands by measuring macroalgae (Lobophora spp.), gastropods (Drupella spp.), scleractinian coral (Acropora spp.), and leather coral (Sinularia spp.). At tourist hubs using seepage septic tank systems, enrichment of Acropora δ 15 N (Redang, +0.7‰) and Sinularia δ 15 N (Tioman, +0.4‰) compared to pristine background levels indicate enhanced sewage nutrient discharge. Carbon isotopes and survey data suggest that sedimentation did not confound these δ 15 N trends. Potential damaging effects of sewage discharge on the coral reef communities at both islands are highlighted by strong correlations between Acropora δ 15 N and regional variation in coral reef community structure, and exclusive occurrence of degraded reefs at regions of high sewage influence., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Sea surface salinity (SSS) is a key parameter to understand and predict many physical, chemical and biological processes in dynamic coastal environments. Yet, in many regions, instrumental measurements are spatially sparse and insufficiently long, hindering our ability to document changes, causes, and consequences of SSS across different time scales. Therefore, there is an need to develop a robust proxy to extend SSS records back in time. Here, we test whether SSS can be reconstructed reliably and quantitatively from shell oxygen isotopic ratios (δ 18 O shell ) of the mussel Mytilus galloprovincialis (Lamarck, 1819) in Otsuchi Bay, Northern Japan. δ 18 O shell ratios vary spatially and temporally and exhibit strong linear correlations with both sea surface temperature (SST) and SSS measurements, indicating that the composite signal recorded by δ 18 O shell measurably responds to variations in both parameters. By combining contemporaneous variations of SST and δ 18 O shell , SSS records encoded into mussel shells are deconvolved that significantly correlate with in situ SSS values. To further validate the robustness of δ 18 O shell as a quantitative SSS proxy, high-resolution and temporally aligned time-series of δ 18 O shell -derived SSS are reconstructed that are highly synchronous with the instrumental records. In particular, two lowered SSS scenarios occur concomitantly with periods of the summer monsoon and typhoon events. δ 18 O shell -derived SSS time-series are also comparable to those from numerical modeling. In conclusion, our findings demonstrate that mussel δ 18 O shell signatures can be used as a useful tool to construct high-resolution records of SSS in the coastal regions., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Geographical traceability of marine bivalves is critical to guarantee their quality and safeguard the interest of both consumers and producers. The neodymium isotopic ratio ( 143 Nd/ 144 Nd) of the coastal water mainly reflects the geology of its neighboring watershed, displaying the distinct and systematic variability at high level of geographical detail and thereby shedding light on its potential as a geochemical tracer. For the first time, the present study investigated the utility and robustness of 143 Nd/ 144 Nd archived in mytilid mussel shells for geographical traceability purposes. The reproducibility of 143 Nd/ 144 Nd ratios maintained in mussels shells from the same cohort demonstrates that the Nd isotopic ratio meets the major requirement for an ideal geochemical tracer, i.e., the biologically induced variation should be rather minimal. The distribution and variability of mussel shell 143 Nd/ 144 Nd patterns were subsequently mapped along the Japanese and Chinese coastal waters. Neodymium isotopes of mussel shells record 143 Nd/ 144 Nd variations among local regions and between the two countries, which are rather compatible with the ages and lithology of the continental bedrocks. These findings highlight the great potential of 143 Nd/ 144 Nd for tracing the geographical origin of marine bivalves., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The ternary edible films based on commercial or bio-chemical chitosan (CCh or BCh) and starch (S) with different concentration of gallic acid (GA) were produced by casting. This work analyzed the effect of different type of Ch and GA concentrations (1.25 mg/g TB and 2.5 mg/g TB ) on the rheological properties of the film-forming solutions, as well as physicochemical (thickness, moisture content, swelling index, water vapor permeability (WVP), tensile strength, microstructure, opacity, and color) and antioxidant properties of the edible films themselves. The chemical cross-linking between the GA and Ch into the edible films of biopolymers could form of ester linkages between the carboxyl groups of GA and the hydroxyl groups of Ch. Results showed that the ternary edible films of BCh-S-GA 5.0 resulted with a positive effect on their rheological (η 0, λ, and p), physicochemical (WVP, swelling, tensile strength, microstructure) and antioxidant properties. The greatest incorporation of polyphenol into the BCh-S-GA 5.0 films greatly reduced WVP and swelling values in approximately 61.01% and 23%, respectively also the tensile strength value increased to 37.372 MPa. The ternary edible films developed in this study showed many desirable characteristics, which could potentially be used as bioactive packaging films for food applications., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Background: Since single lipid parameters are too weak to predict the risk of coronary artery disease, we examined whether the allocation of patients into four groups based on achievement of the target levels set by the Japan Atherosclerosis Guidelines at the time of percutaneous coronary intervention (PCI) would reveal different long-term (5 years) clinical outcomes in males and females., Methods: The results of a 5-year follow-up study are summarized as FU-Registry, Long-Term Clinical Outcome Results. The subjects consisted of 1158 patients who underwent elective PCI. The male and female patients were separately allocated into four groups: (1) high-density lipoprotein cholesterol (HDL-C≥40mg/dl as well as low-density lipoprotein-cholesterol (LDL-C)≥100mg/dl); (2) HDL-C≥40mg/dl as well as LDL-C<100mg/dl; (3) HDL-C<40mg/dl as well as LDL-C≥100mg/dl; (4) HDL-C<40mg/dl as well as LDL-C<100mg/dl, for a comparison of both patient as well as lesion characteristics and the endpoint of major adverse cardiac events (MACEs)., Results: Regarding lesion characteristics, significant differences (p<0.05) were detected in the usage rate of a drug-eluting stent (DES) as well as the bend, stent reference diameter, and stent minimum lumen diameter in females by ANOVA, and in severe calcification, the bend, and usage rate of DES (p<0.001) in males. In females, significant differences (p<0.05) were observed in MACEs and target lesion revascularization-PCI. In contrast, among males, the four groups had nearly equivalent outcomes. Uni- and multivariate analyses revealed that HDL-C as well as LDL-C in females were associated with MACEs [OR 3.29 (95% CI 1.05-8.57, p=0.04)], while no association was observed in male multivariate analysis., Conclusion: In female patients, HDL-C<40mg/dl and LDL-C≥100mg/dl were even more strongly related to MACEs, whereas the combination of LDL-C and HDL-C was not related to MACEs in male patients., (Copyright © 2017 Japanese College of Cardiology. Published by Elsevier Ltd. All rights reserved.)

A sclerochronological and radiocarbon-based study of life history traits of Stimpson's hard clam (Mercenaria stimpsoni), collected alive from Funakoshi Bay, northeast Japan, showed the lifespan of the species to be at least 92 years (determined from annual growth line counts). Three M. stimpsoni specimens exhibited the following synchronous growth pattern, suggestive of environmental control; annual increment width increasing after 1955 to a maximum value between 1970 and 1980, subsequently decreasing gradually until 2000, and thereafter remaining constant or increasing slightly. Variations on annual growth patterns, as well as standardized growth indices chronology, were relatively closely linked to the Atlantic Multidecadal Oscillation (AMO), but less so to Pacific Decadal Oscillation (PDO). Carbonate samples collected from ontogenetically younger shell portions, estimated from growth line counts to have been deposited before 1950, contained no nuclear bomb-test radiocarbon, thereby supporting the accuracy of annual growth line counts (versus overcounting from ventral margin). Together with the synchronous annual increment width patterns, this indicated that age and annual growth rate estimations for M. stimpsoni based on growth line counts were reliable and applicable to high-resolution sclerochronological analyses, which should contribute to a deeper understanding of multi-decadal northwest Pacific climate variability., (Copyright © 2017. Published by Elsevier Ltd.)

Background: The potassium voltage-gated channel subfamily Q member 2 (KCNQ2) gene has been reported to be associated with various types of epilepsy, including benign familial neonatal seizure (BFNS), early infantile epileptic encephalopathy (EIEE), and unclassified early onset encephalopathies. We herein report a patient with early myoclonic encephalopathy (EME) caused by a KCNQ2 mutation., Case Report: A male infant started to exhibit erratic myoclonus several days after birth and apnea attacks lasting for seconds with desaturation. One month after birth, his myoclonuses worsened in frequency. Electroencephalogram (EEG) showed a burst and suppression pattern, and myoclonuses occurred in the burst phase with diffuse polyspikes on EEG. At five months, inter-ictal EEG revealed hypsarrhythmia, but his attacks were still only myoclonuses. ACTH treatment was effective and the myoclonus frequency markedly decreased. At one year of age, whole-exome sequencing revealed a heterozygous mutation of the KCNQ2 gene (NM&#95;172107.2): c.601C>T; p.(Arg201Cys), which was confirmed as de novo by Sanger sequencing. This mutation lies within the extracellular portion of the S4 voltage sensor., Conclusion: Most patients with a KCNQ2 mutation present with seizures starting in the neonatal period with varying severity, ranging from BFNS to Ohtahara syndrome. Furthermore, KCNQ2 appears to be a causative gene for EME., (Copyright © 2017 The Japanese Society of Child Neurology. Published by Elsevier B.V. All rights reserved.)

Purpose: To clarify the relationship between macrocephaly and neurodevelopmental disorders, as well as identify the prevalence of PTEN mutations in autism spectrum disorders with macrocephaly in Japan., Subjects and Methods: Diagnostic and other medical information of children with macrocephaly younger than 4years (n=93) were collected for analysis. PTEN gene mutation analysis was conducted in another set of 16 macrocephalic individuals aged 3-22years., Results: Sixteen macrocephalic children were associated with neurodevelopmental disorders, including autism spectrum disorders (ASDs) (n=6), autistic traits (n=5), intellectual disability (n=5), attention deficit hyperactivity disorder (n=1), developmental coordination disorders (n=1), and language disorder (n=1). Male gender was significantly linked to these disorders, whereas a family history and degree of macrocephaly were not significantly linked to the diagnosis. A novel mutation in the PTEN gene was identified in a 16-year-old girl with autism, mental retardation, language delay, extreme macrocephaly (+4.7SD) with a prominent forehead, and digital minor anomalies., Conclusion: Children with macrocephaly, particularly males, are at a higher risk of neurodevelopmental disorders, rather than progressive etiologies, such as hydrocephalus and neurodegenerative disorders. The data provide a basis for routine health checks for young children in Japan, including the follow-up management and possible screening of PTEN mutations in children with ASDs and macrocephaly., (Copyright © 2017 The Japanese Society of Child Neurology. Published by Elsevier B.V. All rights reserved.)

The combination of biological and electrochemical techniques enhances the bioremediation efficiency of treating oil-contaminated water. In this study a non-growing fungal whole cell biocatalyst (BC; Aspergillus brasiliensis attached to perlite) pretreated with an electric field (EF), was used to degrade a hydrocarbon blend (hexadecane-phenanthrene-pyrene; 100:1:1w/w) in an airlift bioreactor (ALB). During hydrocarbon degradation, all mass transfer resistances (internal and external) and sorption capacity were experimentally quantified. Internal mass transfer resistances were evaluated through BC effectiveness factor analysis as a function of the Thiele modulus (using first order reaction kinetics, assuming a spherical BC, five particle diameters). External (interfacial) mass transfer resistances were evaluated by k L a determination. EF pretreatment during BC production promoted surface changes in BC and production of an emulsifier protein in the ALB. The BC surface modifications enhanced the affinity for hydrocarbons, improving hydrocarbon uptake by direct contact. The resulting emulsion was associated with decreased internal and external mass transfer resistances. EF pretreatment effects can be summarized as: a combined uptake mechanism (direct contact dominant followed by emulsified form dominant) diminishing mass transfer limitations, resulting in a non-specific hydrocarbon degradation in blend. The pretreated BC is a good applicant for oil-contaminated water remediation., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Background: Upon skin injuries, dermal fibroblasts actively produce transforming growth factor-β (TGF-β), which leads to the formation of α-smooth muscle actin (αSMA)-positive granulation tissues. The hyperplasia or incomplete regression of these tissues subsequently causes scar formation in the skin, where sulfated glycosaminoglycans (GAGs), side chains of unique proteoglycans, are supposed to play important roles., Objective: The aim of this study is to clarify the effects of sulfated GAGs on dermal cell behaviors triggered by the TGF-β signaling, along with its possible regulators basic fibroblast growth factor (bFGF) and cell surface epimorphin. bFGF and epimorphin might regulate the TGF-β-induced αSMA expression, they could exert such effects only in specific cellular contexts, given that they lack conventional signal sequences for extracellular localization., Methods: Human scar-derived dermal fibroblasts (HSFs) were treated with TGF-β alone, TGF-β plus bFGF, and TGF-β plus cell surface expression of epimorphin. The effects of GAGs on the expression of αSMA and the cellular morphology were then investigated., Results: A highly sulfated chondroitin sulfate (CS-E) or its substitute (heparinoid) had marked inhibitory effects on TGF-β-mediated changes in HSF behaviors. We found that heparinoid can directly associate with TGF-β, bFGF and epimorphin. We also found that bFGF downregulated αSMA, which was attenuated by heparinoid, whereas epimorphin augmented αSMA expression, which was further amplified by heparinoid., Conclusions: TGF-β, bFGF and epimorphin in the extracellular microenvironment cooperatively affect HSF behaviors under the control of a highly sulfated chondroitin sulfate. These results provide important evidence towards understanding the regulation of TGF-β-induced HSF behaviors., (Copyright © 2017 Japanese Society for Investigative Dermatology. Published by Elsevier B.V. All rights reserved.)

Background: Percutaneous coronary intervention (PCI) with a drug-eluting stent (DES) is increasingly being used for the treatment of coronary artery diseases. However, the role and effectiveness of PCI with a bare metal stent (BMS) have not yet been established., Methods: Among the 2197 patients (2653 lesions) treated with PCI from 2003 to 2012 at three institutions in Fukuoka, 859 patients (1032 lesions) without acute coronary syndrome and in whom we were able to perform follow-up coronary angiography after stent placement and collect detailed data were selected for this study. Among the patients treated by BMS, the cut-off lesion reference for the development of in-stent restenosis (ISR) was calculated by comparing patients with and without ISR. In all patients with a lesion reference above/below the cut-off point of BMS-ISR, medium-term clinical and angiographic outcomes were compared between the BMS and DES groups., Results: In patients treated with a BMS, the lesion reference [odds ratio 0.68, 95% confidence interval (CI) 0.43-0.97, p=0.03] was strongly correlated with ISR, and the area under the receiver operating characteristic curve was 0.704, with moderate accuracy. Calculation of the cut-off lesion reference for BMS-ISR, using a relative cumulative frequency distribution and a sensitivity/specificity curve, showed that the cut-off lesion reference for BMS-ISR was 3.08mm, and the sensitivity/specificity was 60.4%. For the DES and BMS groups in cases with a lesion reference over 3.08mm, BMS placement did not correlate with ISR (odds ratio 0.98, 95% CI 0.85-1.12), and there was also no correlation between BMS and major adverse cardiac events., Conclusion: In patients with a lesion reference≧3.08mm, medium-term clinical and angiographic outcomes were equivalent between the use of a DES and BMS. Thus, BMS placement is well worth considering, especially in patients with a lesion reference≧3.08mm, in whom double-antiplatelet therapy needs to be stopped at an early stage., (Copyright © 2016 Japanese College of Cardiology. Published by Elsevier Ltd. All rights reserved.)

Detection of somatic mutations in non-small cell lung cancers (NSCLCs), especially adenocarcinomas, is important for directing patient care when targeted therapy is available. Here, we present our experience with genotyping NSCLC using the Ion Torrent Personal Genome Machine (PGM) and the AmpliSeq Cancer Hotspot Panel v2. We tested 453 NSCLC samples from 407 individual patients using the 50 gene AmpliSeq Cancer Hotspot Panel v2 from May 2013 to July 2015. Using 10 ng of DNA, up to 11 samples were simultaneously sequenced on the Ion Torrent PGM (316 and 318 chips). We identified variants with the Ion Torrent Variant Caller Plugin, and Golden Helix's SVS software was used for annotation and prediction of the significance of the variants. Three hundred ninety-eight samples were successfully sequenced (12.1% failure rate). In all, 633 variants in 41 genes were detected with a median of 2 (range of 0 to 7) variants per sample. Mutations detected in BRAF, EGFR, ERBB2, KRAS, NRAS, and PIK3CA were considered potentially actionable and were identified in 237 samples, most commonly in KRAS (37.9%), EGFR (11.1%), BRAF (4.8%), and PIK3CA (4.3%). In our patient population, all mutations in EGFR, KRAS, and BRAF were mutually exclusive. The Ion Torrent Ampliseq technology can be utilized on small biopsy and cytology specimens, requires very little input DNA, and can be applied in clinical laboratories for genotyping of NSCLC. This targeted next-generation sequencing approach allows for detection of common and also rare mutations that are clinically actionable in multiple patients simultaneously., (Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.)

Here we describe two patients with 5p- syndrome who suffered from epilepsy characterised by stimulus-induced epileptic spasms manifesting as head nodding. In patient 1, a series of spasms were exclusively triggered by eating, and were associated with diffuse high-voltage slow waves on ictal EEG, particularly presenting as a positive slow potential at the left mid-temporal area. Clusters of sharp waves with negative polarity emerged in the same area during the inter-spasm periods during eating. In patient 2, spasms were provoked by either eating or micturition. Ictal EEG of clustered spasms after micturition showed positive slow or triphasic waves, which correlated with each spasm, over the bifrontal and vertex areas. These findings suggest that the focal cortical areas act as trigger regions in reflex epilepsies, and that a spasm-generator responsible for the execution of reflex spasms exists either in other cortical areas or in the subcortical structures. Although epilepsy is an unusual complication of 5p- syndrome, this syndrome may have a propensity to develop reflex epilepsy, particularly epileptic spasms. However, identification of responsible genes and their roles in this phenotype requires further investigations., (Copyright © 2015 The Japanese Society of Child Neurology. Published by Elsevier B.V. All rights reserved.)

Background: It is not clear whether it is reasonable to use particular drugs for glycemic control in preference to other hypoglycemic agents in terms of the clinical outcome of percutaneous coronary intervention (PCI) in patients with diabetes mellitus (DM)., Methods and Results: Among 2148 patients (2568 lesions) in the FU-Registry, DM patients who underwent PCI (n=758; 922 lesions) were investigated to clarify the effects of various drugs for glycemic control on the clinical outcome [major adverse cardiac events (MACEs): death, myocardial infarction (MI), and target lesion revascularization (TLR)] over approximately 300 days of follow-up (UMIN000005679). The MACEs(+) group (n=165) had a higher usage of insulin (p<0.001) and a lower usage of biguanides (BG, p<0.05) and dipeptidyl peptidase-IV inhibitors (p<0.05) at PCI, compared to the MACEs(-) group (n=593). A multivariate logistic regression analysis showed that low-density lipoprotein cholesterol, insulin use, atherosclerosis obliterans, and lesion reference might be significantly associated with MACEs, while BG use was negatively correlated with MACEs (p=0.04). The cumulative frequency of MACEs in the insulin-treated group was significantly higher (p<0.05) than that in the non-insulin group, and the strongest association between insulin with MACEs was seen in the hemoglobin (Hb) A1c 6.5-7.5% group. There tended to be a negative correlation between the use of insulin and MACEs, with risk ratios of <1, for the HbA1c >8.5% groups., Conclusions: Among different hypoglycemic agents, treatment with insulin was associated with poor mid-term clinical outcomes in DM patients who underwent PCI, while BG use was negatively correlated with MACEs. It may be reasonable for patients with HbA1c >8.5% to avoid hyperglycemia and glucotoxicity, even through the use of insulin., (Copyright © 2014. Published by Elsevier Ltd.)

Introduction: The phosphotidylinositol-3 kinase (PI3K)/serine-threonine kinase (AKT)/mammalian target of rapamycin (mTOR) signaling pathway is frequently altered in head and neck squamous cell cancer (HNSCC). PX-866 is an oral, irreversible, pan-isoform inhibitor of PI3K. Preclinical models revealed synergy with docetaxel and a phase 1 trial demonstrated tolerability of this combination. This randomized phase 2 study evaluated PX-866 combined with docetaxel in patients with advanced, refractory HNSCC., Methods: Patients with locally advanced, recurrent or metastatic HNSCC who had received at least one and no more than two prior systemic treatment regimens were randomized (1:1) to a combination of docetaxel (75mg/m(2) IV every 21days) with or without PX-866 (8mg PO daily; Arms A and B, respectively). The primary endpoint was progression free survival (PFS). Secondary endpoints included objective response rate (RR), overall survival (OS), toxicity, and correlation of biomarker analyses with efficacy outcomes., Results: 85 patients were enrolled. There was a non-significant improvement in response rate in the combination arm (14% vs. 5%; P=0.13). Median PFS was 92days in Arm A and 82days in Arm B (P=0.42). There was no difference in OS between the two arms (263 vs. 195days; P=0.62). Grade 3 or higher adverse events were infrequent, but more common in the combination arm with respect to diarrhea (17% vs. 2%), nausea (7% vs. 0%), and febrile neutropenia (21% vs. 5%); grade 3 or higher anemia was more frequent in arm B (7% vs. 27%). PIK3CA mutations or PTEN loss were infrequently observed., Conclusion: The addition of PX-866 to docetaxel did not improve PFS, RR, or OS in patients with advanced, refractory HNSCC without molecular pre-selection., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Background: The phosphotidylinositol-3 kinase (PI3K)/serine-threonine kinase/mammalian target of rapamycin signaling pathway is frequently altered in head and neck squamous cell cancer (HNSCC). PX-866 is an oral, irreversible, pan-isoform inhibitor of PI3K. A phase I trial demonstrated tolerability of this combination. This randomized phase II study evaluated PX-866 combined with cetuximab in patients with advanced, refractory HNSCC., Methods: Patients with recurrent or metastatic HNSCC who had received at least one and no more than two prior systemic treatment regimens were randomized (1 : 1) to cetuximab with or without PX-866 (8 mg p.o. daily; arms A and B, respectively). The primary end point was progression-free survival (PFS). Secondary end points included objective response rate (ORR), overall survival (OS), toxicity, and correlation of key biomarkers with efficacy outcomes., Results: Eighty-three patients were enrolled. There was a similar response rate between arms (10% versus 7%). Of patients for whom tissue was assessable, 57% were human papillomavirus (HPV) positive. Median PFS was 80 days in both arms and there was no difference in OS between the two arms (211 versus 256 days). Overall toxicity was higher in arm A compared with arm B, especially in terms of nausea (53% versus 23%), vomiting (45% versus 15%), fatigue (43% versus 23%), diarrhea (40% versus 21%), and hypokalemia (25% versus 10%). Grade 3 or higher adverse events were infrequent, but more common in the combination arm although without a specific pattern. PIK3CA mutations were observed in 17% of the cases assessed, and PTEN loss was infrequently observed., Conclusion: The addition of PX-866 to cetuximab did not improve PFS, RR, or OS in patients with advanced, refractory HNSCC enrolled without molecular preselection. In this contemporary cohort, HPV-positive patients comprised the majority, and neither HPV-positive nor HPV-negative patients derived clinical benefit for the addition of cetuximab plus PX-866., (© The Author 2014. Published by Oxford University Press on behalf of the European Society for Medical Oncology. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Objective: To investigate the association of serum uric acid (SUA) with arterial stiffness assessed by cardio-ankle vascular index (CAVI)., Methods: We analyzed the cross-sectional data from 27,360 healthy Japanese subjects (12,910 males and 14,450 females) aged between 20 and 74 years without a past history of heart disease, stroke, hypertension, diabetes, nephritis or gout. We investigated whether SUA was independently associated with CAVI in a gender-specific manner., Results: BMI, CAVI, systolic/diastolic BP, GOT, GPT, γ-GTP, triglyceride (TG), creatinine and SUA were higher and HDL-C was lower in males than in females. Next, they were stratified by SUA into 3 groups: lower tertile (T1), middle tertile (T2) and upper tertile (T3) and by gender. CAVI increased progressive with increasing SUA tertile, after adjusting for age, BMI and systolic BP (sBP) identified in multiple regression analysis for CAVI. Multivariate analysis showed that the odds ratios (95% CI) relative to T1 for high CAVI (≥90(th) percentile) were 1.233 (0.928-1.638) in T2 and 1.352 (1.031-1.773) in T3 for males, and 1.133 (0.984-1.303) in T2 and 1.361 (1.098-1.687) in T3 for females, after adjusting for confounders. Furthermore, increase in adjusted CAVI was observed in a lower SUA range in females compared to that observed in males., Conclusion: We demonstrated an independent correlation between SUA and CAVI, and observed gender difference in the SUA range for increase in CAVI. These results may suggest the need to set different target SUA levels for men and women in anti-hyperuricemic treatment for atherosclerosis prevention., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)

Background: It is well known that percutaneous coronary intervention (PCI) in hemodialysis (HD) patients is associated with higher rates of in-stent restenosis and major adverse cardiovascular events (MACE) compared to that in non-HD patients, even if the target value in cholesterol management is achieved., Methods: To evaluate the factors that are associated with MACE in HD patients, we selected 142 HD patients (164 lesions) without acute coronary syndrome (ACS) from 2148 patients (2568 lesions) who underwent PCI in our database of the FU-Registry [UMIN000005679, Fukuoka University Hospital EC/IRB:10-1-08(09-105)], and compared 52 patients (53 lesions) with MACE [MACE(+)] to 90 patients (111 lesions) without MACE [MACE(-)]., Results: Total cholesterol (TC: 150±30mg/dL vs 166±39mg/dL, p<0.05) and high-density lipoprotein cholesterol (HDL-C: 40.1±14.7mg/dL vs 47.8±13.5mg/dL, p<0.01) levels were significantly lower in the MACE(+) group at follow-up. No significant differences were observed in other parameters, including triglyceride, low-density lipoprotein cholesterol (LDL-C; LDL-C/HDL-C ratio, and % changes in HDL-C, non-HDL-C, LDL-C), and hemoglobin A1c (US National Glycohemoglobin Standardization Program) between before and after PCI. TC, LDL-C, and non-HDL-C at the time of PCI and TC, and HDL-C at the 9-month follow-up were negatively correlated with MACE, while body mass index (BMI) [odds ratio (OR): 0.81; 95% confidence interval (CI): 0.68-0.95)], prior coronary artery bypass graft (CABG) (OR: 3.89; 95%CI: 1.29-12.6), and insulin use (OR: 3.17; 95%CI: 1.23-8.55) were strongly correlated with MACE in a multivariate analysis., Conclusion: BMI, CABG, and insulin use, but not LDL-C, are independent predictors of MACE in HD patients, suggesting that the application of lipid management for non-HD patients to HD patients at the time of PCI may not necessarily be beneficial for medium-term clinical outcomes., (Copyright © 2014. Published by Elsevier Ltd.)

Serotonin (5-HT) is a known mitogen for vascular smooth muscle cells (VSMCs). The dedifferentiation and proliferation/apoptosis of VSMCs in the arterial intima represent one of the atherosclerotic changes. LR11, a member of low-density lipoprotein receptor family, may contribute to the proliferation of VSMCs in neointimal hyperplasia. We conducted an in vitro study to investigate whether 5-HT is involved in LR11 expression in human VSMCs and apoptosis of VSMCs induced by 7-ketocholesterol (7KCHO), an oxysterol that destabilizes plaque. 5-HT enhanced the proliferation of VSMCs, and this effect was abolished by sarpogrelate, a selective 5-HT2A receptor antagonist. Sarpogrelate also inhibited the 5-HT-enhanced LR11 mRNA expression in VSMCs. Furthermore, 5-HT suppressed the 7KCHO-induced apoptosis of VSMCs via caspase-3/7-dependent pathway. These findings provide new insights on the changes in the differentiation stage of VSMCs mediated by 5-HT., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Purpose: There is significant variation in recommendation for percutaneous endoscopic gastrostomy (PEG) tube placement in patients undergoing definitive chemoradiation therapy (CRT) for locally advanced squamous cell carcinoma of the head and neck (LAHNC), with some clinicians globally recommending prophylactic PEG and others waiting until toxicity has occurred. The present study was conceived to identify specific factors associated with PEG requirement, in a population of LAHNC patients who did not have up-front PEG placement., Methods and Materials: Using a quality assurance database, we identified patients with oropharyngeal (ORP) or laryngeal-hypopharyngeal (LHP) LAHNC who were treated with CRT for inclusion in a cohort study of factors impacting PEG placement. Eligibility included stage III/IV squamous cell carcinoma of ORP and LHP. Patients were excluded if they had a PEG placement prior to commencement of CRT. The primary endpoint compared across groups was PEG placement, and multivariate analysis of factors was performed., Results: We identified 107 patients with LAHNC who did not receive PEG tubes prior to treatment. After treatment initiation, 41% of patients with ORP tumors required PEG placement during treatment compared with 16% of LHP patients (P = .03). After adjusting for covariates, multivariate analysis revealed that the only predictor for PEG placement was ORP primary (odds ratio 4.77; 95% confidence interval 1.6-13.8, P = .009) using LHP as reference., Conclusions: Our findings suggest that the patients with ORP cancers are more likely to require PEG placement during treatment and should be considered for prophylactic PEG placement, while LHP sites were associated with lower likelihood of PEG requirement. The primary reason for this difference appears to be severity of pharyngitis; proactive nutritional monitoring and supplementation should be implemented early. Patients with pretreatment risk stratification for PEG placement in LAHNC may improve quality of care and avoid unnecessary treatment breaks., (Copyright © 2013 American Society for Radiation Oncology. Published by Elsevier Inc. All rights reserved.)

Objective: Obesity is associated with type 2 diabetes, dyslipidemia and hypertension, contributing to atherogenesis. Weight reduction is the fundamental therapy for obesity. Recently, a novel arterial stiffness parameter called cardio-ankle vascular index (CAVI) has been developed. We hypothesized that CAVI may be a candidate marker of increased vascular stiffness in obese patients. The aim of this study is to investigate the effect of weight reduction on CAVI., Subjects and Methods: Using CAVI as an indicator, we assessed the changes in arterial stiffness in 47 obese Japanese subjects (aged 46 ± 13 years) who underwent a 12-week weight reduction program consisting of a calorie restriction diet (20-25 kcal/day) and exercise therapy. Visceral fat area (VFA) was evaluated by CT., Results: At baseline, CAVI correlated positively with age (r = 0.70), blood pressure (r = 0.23), VFA (r = 0.26) and HbA1c (r = 0.39). After 12 weeks of weight reduction, mean BMI decreased from 33.3 ± 7.5 to 30.7 ± 6.4 kg/m(2) (p < 0.0001), and mean CAVI decreased from 8.3 to 7.9 (p < 0.01). The change in VFA correlated positively with change in CAVI in subjects with decrease in CAVI (r = 0.47). Furthermore, change in VFA was a significant independent predictor for change in CAVI. No significant correlation was observed between change in CAVI and clinical variables such as BMI, HbA1c and lipids., Conclusion: This study demonstrated that CAVI decreased after weight reduction, and was associated with a decrease in VFA. CAVI reduction maybe a marker of improved vascular stiffness after weight reduction in subjects with visceral adiposity., (© 2013 Asian Oceanian Association for the Study of Obesity . Published by Elsevier Ltd. All rights reserved.)

Aims: To clarify the usefulness of protein-sparing modified formula diet in obese type 2 diabetic patients, the effects of partial use of formula diet on weight reduction and changes in related metabolic variables, and the improving rates of risk factors per 1% body weight reduction, were compared with those of conventional subcaloric diet., Subjects and Methods: Obese patients [BMI >25 kg/m²] with diabetic mellitus were randomly assigned to a low-caloric diet with partial use of formula diet group (FD, n = 119) and a conventional low-caloric diet group (CD, n = 110). Subjects in FD took one pack of formula diet (MicroDiet®, 240 kcal/pack) in place of one of three daily low-caloric meals for 24 weeks. Total daily calorie prescribed was same., Result: Weight reduction was greater in FD than in CD (week 24: -3.5 vs -1.4 kg; all p < 0.001). Systolic blood pressure decreased significantly only in FD. HbA1c reduction was greater in FD than in CD. HDL-cholesterol increased significantly more in FD than in CD (week 24: +2.8 vs. +0.6 mg/dl, p < 0.001). Among several improving rates (%) of risk factors/1% body weight reduction, those of HbA1c at weeks 16 and 24, triglyceride at week 8 and HDL-cholesterol at week 24, were significantly higher in FD than CD. Doses of sulfonylurea and thiazolidinedione were significantly decreased in FD than in CD., Conclusion: Partial use of formula diet was much more effective in reducing body weight, and also in improving coronary risk factors than conventional diet in part due to reduced body weight through decreased energy diet intake and due to dietary composition of the formula diet., (© 2013 Asian Oceanian Association for the Study of Obesity . Published by Elsevier Ltd. All rights reserved.)

The common marmoset, Callithrix jacchus, is one of the smallest primates and is increasingly used for an experimental nonhuman primate model in many research fields. Analysis of T cell receptor (TCR) repertoires is a powerful tool to investigate T cell immunity in terms of antigen specificity and variability of TCR expression. However, monoclonal antibodies specific for many TCR Vα or Vβ chains have not been created. We have recently identified a large number of TCRα chain variable (TRAV) and TCRβ chain variable (TRBV) sequences from a cDNA library of common marmosets. The purpose of this study is to develop a new method for analysis of TCR repertoires in the common marmoset using this sequence information. This method is based on a microplate hybridization technique using 32 TRAV-specific and 32 TRBV-specific oligoprobes following an adaptor-ligation PCR. This enables the easy quantitation of the respective TRAV and TRBV expression levels. No cross-hybridization among specific-oligoprobes and very low variances in repeated measures of the same samples was found, demonstrating high specificity and reproducibility. Furthermore, this method was validated by an antihuman Vβ23 antibody which specifically bound to marmoset Vβ23. Using this method, we analyzed TCR repertoires from various tissue samples (PBMCs, spleen, lymph node and thymus) and isolated T cell subpopulations (CD4+CD8+, CD4+CD8− and CD4−CD8+) from the thymus of 10 common marmosets. Neither tissue-specific nor T cell subpopulation-specific differences was found in TRAV and TRBV repertoires. These results suggest that, unlike mice, TCR repertoires in the common marmoset are not affected by endogenous superantigens and are conserved among individuals, among tissues, and among T cell subpopulations. Thus, TCR repertoire analysis with high specificity and reproducibility is a very useful technique, with the potential to replace flow cytometric analysis using a panel of TRV-specific antibodies, many of which remain unavailable., (Copyright © 2012 Elsevier B.V. All rights reserved.)

We reported four patients (2 to 10 years) with Kawasaki disease complicated by clinically mild encephalitis/encephalopathy with a reversible splenial lesion (MERS). All were treated with γ-globulin (2 to 6 g/kg) after the diagnosis of Kawasaki disease, the fever being alleviated between day 6 and 25. One of two patients exhibiting a poor response to γ-globulin had a cardiac aneurysm as a sequela. Their neurological manifestations (delirious behavior and drowsiness), laboratorial hyponatremia, and radiological abnormalities completely disappeared. It is important for pediatricians to acknowledge that MERS can be observed in patients with Kawasaki disease, especially in older children, and that they might be at high risk for cardiac abnormalities., (Copyright © 2011 Elsevier B.V. All rights reserved.)