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Abstract STAT3 is constitutively activated in many cancer types, including lung cancer, and can induce cancer cell proliferation and cancer stem cell (CSC) maintenance. STAT3 is activated by tyrosine kinases, such as JAK and SRC, but the mechanism by which STAT3 maintains its activated state in cancer cells remains unclear. Here, we show that PRMT5 directly methylates STAT3 and enhances its activated tyrosine phosphorylation in non-small cell lung cancer (NSCLC) cells. PRMT5 expression is also induced by STAT3, suggesting the presence of a positive feedback loop in cancer cells. Furthermore, methylation of STAT3 at arginine 609 by PRMT5 is important for its transcriptional activity and support of tumour growth and CSC maintenance. Indeed, NSCLC cells expressing the STAT3 mutant which R609 was replaced to alanine (R609K) show significantly impaired tumour growth in nude mice. Overall, our study reveals a mechanism by which STAT3 remains activated in NSCLC and provides a new target for cancer therapeutic approaches.

Abstract Background Laparoscopic adrenalectomy is widely performed for a number of hormone-producing tumors and postoperative management depends on the hormones produced. In the present study, we conducted a retrospective analysis to clarify the risk factors for postoperative complications, particularly postoperative fever after laparoscopic adrenalectomy. Methods We analyzed 406 patients who underwent laparoscopic adrenalectomy at our hospital between 2003 and 2019. Postoperative fever was defined as a fever of 38 °C or higher within 72 h after surgery. We investigated the risk factors for postoperative fever after laparoscopic adrenalectomy. Results There were 188 males (46%) and 218 females (54%) with a median age of 52 years. Among these patients, tumor pathologies included 188 primary aldosteronism (46%), 75 Cushing syndrome (18%), and 80 pheochromocytoma (20%). Postoperative fever developed in 124 of all patients (31%), 30% of those with primary aldosteronism, 53% of those with pheochromocytoma, and 8% of those with Cushing syndrome. A multivariate logistic regression analysis identified pheochromocytoma and non-Cushing syndrome as independent predictors of postoperative fever. Postoperative fever was observed in 42 out of 80 cases of pheochromocytoma (53%), which was significantly higher than in cases of non-pheochromocytoma (82/326, 25%, p

Summary:. The profunda artery perforator (PAP) flap, commonly used for small- to medium-sized breast reconstructions, offers easy harvest and inconspicuous donor-site scars. However, its shorter vascular pedicle compared with the deep inferior epigastric perforator flap limits its reach to lateral recipient vessels. This often requires strategic placement of perforators at the flap’s edge to extend reach, potentially causing congestion in the distal part of the flap. To address these challenges, using the posterior accessory saphenous vein (pASV) has proven effective. Using the pASV as a vein graft significantly extends the pedicle length of the PAP flap, enhancing anastomosis success with recipient vessels. Additionally, in cases of flap congestion, the proximal segment of the pASV can be used as an additional venous outflow pathway, while grafting the distal segment further extends its length. This dual approach improves overall flap viability and reduces venous congestion risks. This discussion highlights two cases demonstrating the innovative use of the pASV within the PAP flap. In case 1, the pASV extended the pedicle length, enhancing the flap’s placement flexibility and facilitating anastomosis with thoracodorsal vessels. In case 2, the pASV served as a secondary venous outflow pathway, with the distal segment grafted to extend the proximal portion. This adaptation provided additional venous drainage and effectively managed positioning constraints imposed by recipient vessel locations. These examples illustrate the significant benefits of utilizing the pASV in PAP flap breast reconstructions, offering a novel strategy to improve viability and expand its use in complex scenarios requiring extended vascular reach.

Abstract Tumor structure is heterogeneous and complex, and it is difficult to obtain complete characteristics by two‐dimensional analysis. The aim of this study was to visualize and characterize volumetric vascular information of clear cell renal cell carcinoma (ccRCC) tumors using whole tissue phenotyping and three‐dimensional light‐sheet microscopy. Here, we used the diagnosing immunolabeled paraffin‐embedded cleared organs pipeline for tissue clearing, immunolabeling, and three‐dimensional imaging. The spatial distributions of CD34, which targets blood vessels, and LYVE‐1, which targets lymphatic vessels, were examined by calculating three‐dimensional density, vessel length, vessel radius, and density curves, such as skewness, kurtosis, and variance of the expression. We then examined those associations with ccRCC outcomes and genetic alteration state. Formalin‐fixed paraffin‐embedded tumor samples from 46 ccRCC patients were included in the study. Receiver operating characteristic curve analyses revealed the associations between blood vessel and lymphatic vessel distributions and pathological factors such as a high nuclear grade, large tumor size, and the presence of venous invasion. Furthermore, three‐dimensional imaging parameters stratified ccRCC patients regarding survival outcomes. An analysis of genomic alterations based on volumetric vascular information parameters revealed that PI3K‐mTOR pathway mutations related to the blood vessel radius were significantly different. Collectively, we have shown that the spatial elucidation of volumetric vasculature information could be prognostic and may serve as a new biomarker for genomic alterations. High‐end tissue clearing techniques and volumetric immunohistochemistry enable three‐dimensional analysis of tumors, leading to a better understanding of the microvascular structure in the tumor space.

Abstract Background Due to an increase in life expectancy, the incidence of metastatic renal cell carcinoma (mRCC) in patients aged ≥75 years has been increasing. In this study we investigated the characteristics before treatment and the outcomes of systemic therapies for patients aged ≥75 years with mRCC and compared the results with those for patients aged

Abstract Keloids are benign fibroproliferative skin tumors caused by aberrant wound healing that can negatively impact patient quality of life. The lack of animal models has limited research on pathogenesis or developing effective treatments, and the etiology of keloids remains unknown. Here, we found that the characteristics of stem-like cells from keloid lesions and the surrounding dermis differ from those of normal skin. Furthermore, the HEDGEHOG (HH) signal and its downstream transcription factor GLI1 were upregulated in keloid patient–derived stem-like cells. Inhibition of the HH-GLI1 pathway reduced the expression of genes involved in keloids and fibrosis-inducing cytokines, including osteopontin. Moreover, the HH signal inhibitor vismodegib reduced keloid reconstituted tumor size and keloid-related gene expression in nude mice and the collagen bundle and expression of cytokines characteristic for keloids in ex vivo culture of keloid tissues. These results implicate the HH-GLI1 pathway in keloid pathogenesis and suggest therapeutic targets of keloids.

Abstract Tumor suppressor p53 plays a central role in response to DNA damage. DNA-damaging agents modulate nuclear actin dynamics, influencing cell behaviors; however, whether p53 affects the formation of nuclear actin filaments remains unclear. In this study, we found that p53 depletion promoted the formation of nuclear actin filaments in response to DNA-damaging agents, such as doxorubicin (DOXO) and etoposide (VP16). Even though the genetic probes used for the detection of nuclear actin filaments exerted a promotive effect on actin polymerization, the detected formation of nuclear actin filaments was highly dependent on both p53 depletion and DNA damage. Whilst active p53 is known to promote caspase-1 expression, the overexpression of caspase-1 reduced DNA damage-induced formation of nuclear actin filaments in p53-depleted cells. In contrast, co-treatment with DOXO and the pan-caspase inhibitor Q-VD-OPh or the caspase-1 inhibitor Z-YVAD-FMK induced the formation of nuclear actin filament formation even in cells bearing wild-type p53. These results suggest that the p53-caspase-1 axis suppresses DNA damage-induced formation of nuclear actin filaments. In addition, we found that the expression of nLifeact-GFP, the filamentous-actin-binding peptide Lifeact fused with the nuclear localization signal (NLS) and GFP, modulated the structure of nuclear actin filaments to be phalloidin-stainable in p53-depleted cells treated with the DNA-damaging agent, altering the chromatin structure and reducing the transcriptional activity. The level of phosphorylated H2AX (γH2AX), a marker of DNA damage, in these cells also reduced upon nLifeact-GFP expression, whilst details of the functional relationship between the formation of nLifeact-GFP-decorated nuclear actin filaments and DNA repair remained to be elucidated. Considering that the loss of p53 is associated with cancer progression, the results of this study raise a possibility that the artificial reinforcement of nuclear actin filaments by nLifeact-GFP may enhance the cytotoxic effect of DNA-damaging agents in aggressive cancer cells through a reduction in gene transcription.

Abstract In Japan, the number of elderly people in need of nursing care is increasing while the population of young people is decreasing, and the potential for labor shortages in the field of elder care is of great concern. This study aimed to estimate the behavior of the elderly by using sensors to monitor indoor air quality (IAQ), without placing undue burden on the elderly or their caregivers. Odor and carbon dioxide (CO2) concentrations were monitored in a private room of a nursing home in the Kanto Region of Japan, the behaviors of the resident and staff members were recorded, and the relationship between the two was analyzed. Both odor and CO2 concentrations were higher when the resident was present than when absent, indicating that the resident was one of the main sources of indoor odor and CO2. In addition, after the resident entered the room, the CO2 concentration increased and remained stable, whereas the odor concentration tended to vary after the resident entered the room, first increasing and later decreasing. This suggested that the increase or decrease in odor could be used to monitor the behavior of the resident and staff members. The relationship between the slopes of odor and CO2 in typical behavioral events suggest that if only odor increases and CO2 does not change, the likelihood of the event in which feces were observed during diaper changes is high. In addition, based on the behavior near the sensor, the rate of CO2 and odor emissions differed between the elderly resident and the younger staff members, suggesting that the ratio of odor slope to CO2 slope may be greater in the elderly than in younger people. Furthermore, the repeated number of increases and decreases in odor and CO2 suggested that multiple events could be distinguished. These results suggest that IAQ can be utilized to estimate the behavior of residents and staff in nursing care facilities for the elderly.

Background: The COVID-19 pandemic has revealed the fragility of the global food system, sending shockwaves across countries' societies and economy. This has presented formidable challenges to sustaining a healthy and resilient lifestyle. The objective of this study is to examine the food consumption patterns and assess diet diversity indicators, primarily focusing on the food consumption score (FCS), among households in 38 countries both before and during the first wave of the COVID-19 pandemic. Methods: A cross-sectional study with 37 207 participants (mean age: 36.70 ± 14.79, with 77 % women) was conducted in 38 countries through an online survey administered between April and June 2020. The study utilized a pre-tested food frequency questionnaire to explore food consumption patterns both before and during the COVID-19 periods. Additionally, the study computed Food Consumption Score (FCS) as a proxy indicator for assessing the dietary diversity of households. Findings: This quantification of global, regional and national dietary diversity across 38 countries showed an increment in the consumption of all food groups but a drop in the intake of vegetables and in the dietary diversity. The household's food consumption scores indicating dietary diversity varied across regions. It decreased in the Middle East and North Africa (MENA) countries, including Lebanon (p

Species extinction risk is a crucial indicator of the global impact of climate change on biodiversity. In life cycle impact assessment (LCIA), the relationship between the potentially extinct ratio and temperature is used to calculate characterization factors (CFs) that indicate additional biodiversity impact per unit greenhouse gas emissions, using the biodiversity loss indicator termed potentially disappeared fraction (PDF). However, current methods for calculating CFs that focus on terrestrial biodiversity do not show differences among taxa and regions. Here, we estimated taxon-level CFs based on a species-specific extinction risk approach for determining impact of CO2 emission on terrestrial biodiversity. The extinction risk of a species is defined as the reciprocal of the estimated waiting times for extinction caused by climate-derived habitat loss of the species. Based on the approach, CFs for five taxa—birds, reptiles, mammals, amphibians, vascular plants—were estimated as 0.017, 0.017, 0.025, 0.027, and 0.040 (PDF·km2·year·kton−1), respectively. In addition, CFs that use the number of potentially extinct species (species·kton−1) as indicators were also calculated based on the same species-specific extinction risk approach. The proposed method successfully calculates taxon-level CFs and has the potential to show their regional variations, which contributes to the precise impact assessment on climate-derived biodiversity in LCIA.

Abstract Background Previous clinical trials have demonstrated the potential efficacy of poly (ADP-ribose) polymerase (PARP) inhibitors (PARPis) in patients with cancer involving homologous recombination repair (HRR) gene-mutation. Moreover, HRR gene-mutated cancers are effectively treated with immune checkpoint inhibitors (ICIs) with the increase in tumor mutation burden. We have proposed to conduct a multicenter, single-arm phase II trial (IMAGENE trial) for evaluating the efficacy and safety of niraparib (PARPi) plus programmed cell death-1 inhibitor combination therapy in patients with HRR gene-mutated cancers who are refractory to ICIs therapy using a next generation sequencing-based circulating tumor DNA (ctDNA) and tumor tissue analysis. Methods Key eligibility criteria for this trial includes HRR gene-mutated tumor determined by any cancer gene tests; progression after previous ICI treatment; and Eastern Cooperative Oncology Group Performance Status ≤ 1. The primary endpoint is the confirmed objective response rate (ORR) in all patients. The secondary endpoints include the confirmed ORR in patients with HRR gene-mutation of ctDNA using the Caris Assure (CARIS, USA). The target sample size of the IMAGENE trial is 57 patients. Biomarker analyses will be performed in parallel using the Caris Assure, proteome analysis, and T cell repertoire analysis to reveal tumor immunosurveillance in peripheral blood. Expected outcome Our trial aims to confirm the clinical benefit of PARPi plus ICI combination therapy in ICI-resistant patients. Furthermore, through translational research, our trial will shed light on which patients would benefit from the targeted combination therapy for patients with HRR gene-mutated tumor even after the failure of ICIs. Trial registration The IMAGENE trial: jRCT, Clinical trial no.: jRCT2051210120, Registered date: November 9, 2021.

Abstract The livestock industry is a major source of atmospheric volatile organic compounds(VOCs), but details on these emissions are not well documented in Japan. In particular, it remains unclear how the rearing method affects the emissions of VOCs from livestock, which originate primarily from feces and urine. Here we aimed to estimate the amounts of VOCs emitted from the feces and urine of tethered Holstein dairy cattle in a cattle shed in Japan. Dimethyl sulfide and acetone accounted for about 60% of the total VOCs emitted from feces, followed by formaldehyde and acetaldehyde. Also, dimethyl sulfide and acetone were the dominant VOCs emitted from urine, accounting for 90% of the total VOCs. The VOCs from manure were considered to be emitted between the excretion and removal of the manure during the cleaning of the shed. As a result of analyzing images from three cameras installed in the shed, the average time between excretion and cleaning during the daytime (8:00 am–5:00 pm) was 80 min for feces and urine, whereas at night (5:00 pm–7:00 am), the average time between excretion and cleaning was 480 min. Based on the above findings, the emissions of VOCs in the interval between excretion and cleaning of the shed were estimated. As a result, the emissions of VOCs from feces and urine per head of cattle in the shed were estimated to be 1.75 and 1.52 g day−1, respectively. Furthermore, contribution of VOCs emitted from manure to odor activity value (OAV) and hydroxyl radical reactivity (OHR) were also estimated. Volatile fatty acids and sulfur compounds emitted from feces estimated to have high contribution to OAV, whereas aldehydes contributed mainly for OHR from manure.

Abstract Cancer stem cells (CSCs) have tumour initiation, self-renewal, and long-term tumour repopulation properties, and it is postulated that differentiated somatic cells can be reprogrammed to CSCs by oncogenic signals. We previously showed that oncogenic HRAS V12 conferred tumour initiation capacity in tumour suppressor p53-deficient (p53 −/− ) primary mouse embryonic fibroblasts (MEFs) through transcription factor NF-κB-mediated enhancement of glucose uptake; however, the underlying mechanisms of RAS oncogene-induced CSC reprogramming have not been elucidated. Here, we found that the expression of the reprogramming factor SOX2 was induced by HRAS V12 in p53 −/− MEFs. Moreover, gene knockout studies revealed that SOX2 is an essential factor for the generation of CSCs by HRAS V12 in mouse and human fibroblasts. We demonstrated that HRASV12-induced cyclin-dependent kinase 1 (CDK1) activity and subsequent enhancement of protein O-GlcNAcylation were required for SOX2 induction and CSC generation in these fibroblasts and cancer cell lines containing RAS mutations. Moreover, the CDK inhibitor dinaciclib and O-GlcNAcylation inhibitor OSMI1 reduced the number of CSCs derived from these cells. Taken together, our results reveal a signalling pathway and mechanism for CSC generation by oncogenic RAS and suggest the possibility that this signalling pathway is a therapeutic target for CSCs.

There is concern among the general public that information technology (IT) innovations may make existing jobs redundant. This may be perceived to pose a greater problem to future generations because new technologies, not limited to IT innovations, will be sophisticated in the future. Our previous work revealed that messages reminding people of familial support as a nudge can moderate risk-averse attitudes toward risks that are perceived to threaten future generations, which could be effective for other kinds of risks. Therefore, we conducted a randomized controlled trial to examine the message effects for information provision on IT innovations. The study was conducted via an online questionnaire survey in January 2020, before the COVID-19 pandemic, and more than 3,200 samples were collected from respondents aged 20 years or older living in Japan. The treatment groups received basic information supplemented with additional text or additional text and an illustration that highlighted IT innovations as support from previous generations. The control group received only the basic textual information. The effects of the intervention were evaluated by comparing changes in average subjective assessment of IT in the treatment groups with those in the control group. The intervention effect was statistically significant, and the sense of familial support after receiving the intervention messages was significantly increased in the treatment group that viewed the illustration compared with the control group. Additionally, we discuss how each component of the HEXACO personality traits influences responses to the intervention messages. Through a series of surveys, we demonstrated the potential of our framework for a wide variety of applications involving information provision perceived to involve future generations.

Abstract Volatile organic compounds (VOCs), CO2, temperature, and humidity in a private room in a care facility for the elderly were measured and the behavior of a resident and staff were recorded in order to clarify the effects of the resident’s behavior, especially defecation, on indoor air quality. Average indoor concentrations of total VOCs (in μg m−3) in summer, autumn, and winter were 40.9, 16.7, and 18.8, respectively. Average indoor concentrations of CO2 in summer, autumn, and winter were 813, 761, and 1144 ppm, respectively, revealing a tendency for the concentrations of CO2 to be higher in winter, in contrast to the VOC concentration. The concentrations of VOCs and CO2 were 1.1 to 1.5 times higher when the resident was present in the room than when the resident was absent. This result suggests that one of the main sources of VOC and CO2 emissions in indoor air was the resident. Acetic acid, 1-butanol, propanoic acid, hexanoic acid, and phenol, which are contained in human sweat, exhaled air, and excrement, were the predominant VOCs in the air of the room regardless of the season, and these five components accounted for more than 90% of the total VOCs. The concentrations of these components were higher when the resident was present in the room, suggesting that the resident was the main source of these components. Based on the changes in the VOC and CO2 concentrations over time and the records of the resident and the staff, it was noted that VOC concentrations decreased, in some cases, before and after diaper changes. Our research suggests that certain aspects of the behavior of residents can be inferred by monitoring changes in indoor air quality.

Abstract Several clinical studies have been conducted into the practicality and safety of regenerative therapy using hESC/iPSC-retinal pigment epithelium (RPE) as a treatment for the diseases including age-related macular degeneration. These studies used either suspensions of RPE cells or an RPE cell sheet. The cells can be injected using a minimally invasive procedure but the delivery of an intended number of cells at an exact target location is difficult; cell sheets take a longer time to prepare, and the surgical procedure is invasive but can be placed at the target area. In the research reported here, we combined the advantages of the two approaches by producing a quickly formed hiPSC-RPE strip in as short as 2 days. The strip readily expanded into a monolayer sheet on the plate, and after transplantation in nude rats, it showed a potency to partly expand with the correct apical/basal polarity in vivo, although limited in expansion area in the presence of healthy host RPE. The strip could be injected into a target area in animal eyes using a 24G canula tip.

Abstract Real-time onsite monitoring of indoor airborne microbes in a residential property in central Tokyo was carried out in 2020 and 2021, following the onset of the COVID-19 pandemic. A microbial sensor utilizing fluorescence emitted by microorganisms was used to measure bioaerosol concentrations in the living room and children’s bedroom as well as on the balcony. Indoor PM2.5 was also monitored simultaneously at certain time points using a PM2.5 sensor. The behavior of the residents was also recorded during some monitoring periods. The average number concentration of microbes as fungi in the living room was 15,100, 58,800, and 10,600 counts m−3 in spring, summer, and winter, respectively, increasing in summer when the outside temperature was high. Microbial number concentrations were closely related to human behavior, increasing rapidly during periods of physical activity, but decreasing again within 20–30 min of the activity ending. There was no clear correlation between indoor microbial number concentrations and PM2.5 concentrations, suggesting that indoor microorganisms are concentrated in coarse particles, such as dust, which are quickly removed via gravitational settling. The concentration of indoor airborne microorganisms decreased significantly after ventilation, and although an occasional increase was observed immediately after ventilation, concentrations decreased again rapidly within 10–20 min. These results suggest that even a short period of ventilation can significantly reduce the indoor bioaerosol.

Targeting the inhibitory receptors (IRs) LAG-3, TIM-3 and TIGIT is a promising immune-oncology approach and the identification of biomarkers of response is crucial. Here, the authors apply automated single-cell count for these IRs in human renal cell carcinoma and investigate the immunogenomic landscape of the disease.

Abstract Tumour-promoting inflammation is a hallmark of cancer, and chronic inflammatory disease increases the risk of cancer. In this context, MYD88, a downstream signalling molecule of Toll-like receptors that initiates inflammatory signalling cascades, has a critical role in tumour development in mice and its gene mutation was found in human cancers. In inflammation-induced colon cancer, tumour suppressor p53 mutations have also been detected with high frequency as early events. However, the molecular mechanism of MYD88-induced cancer development is poorly understood. Here, we demonstrated that MYD88 induced the protein accumulation of the transcription factor HIF-1α through NF-κB in p53-deficient cells. HIF-1α accumulation was not caused by enhanced protein stability but by NF-κB-mediated transcriptional activation, the enhanced translation of HIF-1α and JNK activation. In contrast, MYD88-induced mRNA expressions of HIF-1α and HIF-1-target genes were attenuated in the presence of p53. Furthermore, constitutively active forms of MYD88 induced tumour-initiating cell (TIC) generation in p53-deficient cells, as determined by tumour xenografts in nude mice. TIC generating activity was diminished by the suppression of NF-κB or HIF-1α. These results indicate that MYD88 signals induce the generation of TICs through the NF-κB-HIF-1α activation cascade in p53-deficient cells and suggest this molecular mechanism underlies inflammation-induced cancer development.

Abstract Anti‐proinflammatory cytokine therapies against interleukin (IL)‐6, tumor necrosis factor (TNF)‐α, and IL‐1 are major advancements in treating inflammatory diseases, especially rheumatoid arthritis. Such therapies are mainly performed by injection of antibodies against cytokines or cytokine receptors. We initially found that the glycolytic inhibitor 2‐deoxy‐d‐glucose (2‐DG), a simple monosaccharide, attenuated cellular responses to IL‐6 by inhibiting N‐linked glycosylation of the IL‐6 receptor gp130. Aglycoforms of gp130 did not bind to IL‐6 or activate downstream intracellular signals that included Janus kinases. 2‐DG completely inhibited dextran sodium sulfate‐induced colitis, a mouse model for inflammatory bowel disease, and alleviated laminarin‐induced arthritis in the SKG mouse, an experimental model for human rheumatoid arthritis. These diseases have been shown to be partially dependent on IL‐6. We also found that 2‐DG inhibited signals for other proinflammatory cytokines such as TNF‐α, IL‐1β, and interferon ‐γ, and accordingly, prevented death by another inflammatory disease, lipopolysaccharide (LPS) shock. Furthermore, 2‐DG prevented LPS shock, a model for a cytokine storm, and LPS‐induced pulmonary inflammation, a model for acute respiratory distress syndrome of coronavirus disease 2019 (COVID‐19). These results suggest that targeted therapies that inhibit cytokine receptor glycosylation are effective for treatment of various inflammatory diseases.

To clarify the emissions of volatile organic compounds (VOCs) from hen rearing in Japan, we collected air samples from inside a hen shed for the four seasons in 2019 and analyzed 34 VOCs in the air samples by gas chromatography-mass spectrometry and high performance liquid chromatography. The temperature and humidity inside and outside of the shed were monitored simultaneously during each sampling campaign. The average concentrations of VOCs in the shed ranged from 150 to 427 μg m-3, the concentrations being higher in summer and lower in winter. Acetone, dimethyl sulfide, 2-butanone, 2-pentanone, and acetic acid were dominant throughout all the seasons and these five compounds accounted for 70-89% of the total VOCs. The reactivity of each VOC with hydroxyl radical was also calculated and dimethyl sulfide was found to be the most reactive VOC, accounting for 84-94% of the total hydroxyl radical reactivity. The emission rate (ER) for the total VOCs (μg h-1 kg-1) was 602 in winter, 7,900 in spring, 46,500 in summer and 37,600 in autumn, respectively. Acetone, dimethyl sulfide, 2-butanone, 3-pentanone and acetic acid had higher ERs throughout all the seasons, and these five components accounted for 70-90% of the ERs for the total VOCs. The ERs of the VOCs increased exponentially in accord with temperature increases inside the shed, indicating that the ERs of the VOCs depended on the ambient temperature. The annual VOC emission from one hen and from the hen shed was calculated to be 405 g y-1 and 121 kg y-1, respectively.

This study demonstrates the rapid fabrication and utility of 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer film for cell patterning. The film was obtained on a cell culture surface by microcasting MPC polymer ethanol solution into a degassed polydimethylsiloxane mold with a desired pattern. After removal of the mold, 293AD cells were cultured on the surface of the polymer film with the patterned microstructures. Patterned cell adhesion restricted by the film was successfully maintained during at least a 168-h cultivation. The microcast MPC polymer film can be prepared rapidly and used for efficient long-term cell confinement.

Background: Both pre-supplementary motor area (pre-SMA) and SMA-proper (SMA) must play important roles in visuomotor sequence learning. However, functional differences between the pre-SMA and SMA have not been well studied in humans. Objective: To elucidate the functional differences between the pre-SMA and SMA in sequence learning in humans. Methods: To induce LTP/LTD, we administered quadripulse transcranial magnetic stimulation (QPS) with an inter-stimulus interval of 5 or 50 ms (QPS-5/50) over the pre-SMA or SMA in healthy volunteers. The sham stimulation was also done as a control. We studied the effects of LTP/LTD in the pre-SMA/SMA on a new sequence learning and the performance of well-learned sequence by using sequence learning task called the “2 × 10 task”. Effects on the simple choice reaction time task were also studied for comparison. Results: QPS-5 over the pre-SMA increased the error rate without any changes in movement speed. When administered over the SMA, QPS-5 decreased, and QPS-50 increased the rate of reaction time reduction across trials without changes in the error rate. QPS over neither the pre-SMA nor SMA affected the performances of a well-learned sequence or a simple choice reaction time task. Conclusions: Our findings that QPS over the pre-SMA correlated with sequence learning performance and that over the SMA with execution speed are consistent with the previous results in animals and humans. Our results lend further support to the utility of QPS for modulating motor learning in humans.

Background The aims of this study were (1) to clarify the impact of tertiary lymphoid structure (TLS) status on the outcome and immunogenomic profile of human clear cell renal cell carcinoma (ccRCC) and (2) to determine phenotypic differences in TLSs between different types of genitourinary cancer, that is, urinary ccRCC and bladder cancer.Methods We performed a quantitative immunohistological analysis of ccRCC tissue microarrays and conducted integrated genome mutation analysis by next-generation sequencing and methylation array analysis. Since the tumor immune microenvironment of ccRCC often differs from that of other cancer types, we analyzed the phenotypic differences in TLSs between ccRCC and in-house bladder cancer specimens.Results Varying distribution patterns of TLSs were observed throughout ccRCC tumors, revealing that the presence of TLSs was related to poor prognosis. An analysis of genomic alterations based on TLS status in ccRCC revealed that alterations in the PI3K-mTOR pathway were highly prevalent in TLS-positive tumors. DNA methylation profiling also revealed distinct differences in methylation signatures among ccRCC samples with different TLS statuses. However, the TLS characteristics of ccRCC and bladder cancer markedly differed: TLSs had the exact opposite prognostic impact on bladder cancer as on ccRCC. The maturity and spatial distribution of TLSs were significantly different between the two cancer types; TLSs were more mature with follicle-like germinal center organization and likely to be observed inside the tumor in bladder cancer. Labeling for CD8, FOXP3, PD-1, and PD-L1 showed marked differences in the diversity of the immune microenvironment surrounding TLSs. The proportions of CD8-, FOXP3-, and PD-L1-positive cells were significantly higher in TLSs in bladder cancer than in TLSs in ccRCC; rather the proportion of PD-1-positive cells was significantly higher in TLSs in ccRCC than in TLSs in bladder cancer.Conclusion The immunobiology of ccRCC is unique, and various cancerous phenomena conflict with that seen in other cancer types; therefore, comparing the TLS characteristics between ccRCC and bladder cancer may help reveal differences in the prognostic impact, maturity and spatial distribution of TLSs and in the immune environment surrounding TLSs between the two cancers.

In computer-aided diagnosis systems for lung cancer, segmentation of lung nodules is important for analyzing image features of lung nodules on computed tomography (CT) images and distinguishing malignant nodules from benign ones. However, it is difficult to accurately and robustly segment lung nodules attached to the chest wall or with ground-glass opacities using conventional image processing methods. Therefore, this study aimed to develop a method for robust and accurate three-dimensional (3D) segmentation of lung nodule regions using deep learning. In this study, a nested 3D fully connected convolutional network with residual unit structures was proposed, and designed a new loss function. Compared with annotated images obtained under the guidance of a radiologist, the Dice similarity coefficient (DS) and intersection over union (IoU) were 0.845 ± 0.008 and 0.738 ± 0.011, respectively, for 332 lung nodules (lung adenocarcinoma) obtained from 332 patients. On the other hand, for 3D U-Net and 3D SegNet, the DS was 0.822 ± 0.009 and 0.786 ± 0.011, respectively, and the IoU was 0.711 ± 0.011 and 0.660 ± 0.012, respectively. These results indicate that the proposed method is significantly superior to well-known deep learning models. Moreover, we compared the results obtained from the proposed method with those obtained from conventional image processing methods, watersheds, and graph cuts. The DS and IoU results for the watershed method were 0.628 ± 0.027 and 0.494 ± 0.025, respectively, and those for the graph cut method were 0.566 ± 0.025 and 0.414 ± 0.021, respectively. These results indicate that the proposed method is significantly superior to conventional image processing methods. The proposed method may be useful for accurate and robust segmentation of lung nodules to assist radiologists in the diagnosis of lung nodules such as lung adenocarcinoma on CT images.

Nudging is a method for eliciting a desired behavior. One approach to nudging involves information provision. When information presented for this purpose is designed from an evolutionary perspective, it may reveal a deeper level of rationality within human decision-making that might otherwise appear to be irrational. Based on insights from the evolution of altruism, we previously designed a message to remind people of the benefits they have received from the actions of relatives to realize industrialization. We then demonstrated that using this message in Japan was effective at moderating extreme risk-averse attitudes toward air pollution resulting from industrialization. However, the universality of the intervention effect, including whether it could be affected by exogenous factors, was not explored. Therefore, in the present study, we conducted a randomized controlled trial based on an online survey carried out in Japan, Canada, and the US. The intervention was shown to be effective in all the three countries, but the effect size varied according to segment. Although women showed more intervention effects than men in Japan and the US, no significant sex difference was observed in Canada. In terms of personality traits, higher agreeableness significantly contributed to the intervention effects. The influence of the COVID-19 pandemic, which necessitated many lifestyle changes, was found to weaken the intervention effect by increasing the message effect in the control group. We propose that this effect was caused by an increased perception of familial support in everyday life. These results suggest that the nudge message was universally effective, although the effect size might have been affected by cultural factors and social events.

Disposable plastics are drawing considerable attention as a source of environmental risk despite their benefits in daily life. Banning the use of disposable plastics could increase other types of risks, which may damage the public good in the long run. Considering the trade-off of the risks and benefits, one way to improve social welfare is to conduct proper recycling and to continue using plastics but limit them to essential use, avoiding an unnecessary ban. A potential barrier to such a policy might be risk-averse attitudes toward actions that are perceived to threaten future generations, which is a well-known phenomenon. We previously designed a framework for information provision using messages that remind individuals about familial support, which had significant effects in multiple countries on increasing positive attitudes toward air pollution caused by industrialization. We hypothesized that this information provision could also be effective for disposable plastic use. Thus, we conducted a randomized controlled trial via online surveys in Japan, Canada, and the US to identify the effects of our designed messages about recycling on increasing positive attitudes toward disposable plastics. The intervention effects were measured by the difference-in-difference method and panel analysis based on linear regression models using the respondents' attributes and personality traits. The effects were consistently correlated with a sense of familial support, with the effect sizes varying according to country (US > Japan > Canada). Attributes that positively contributed to the message being more effective were higher agreeableness, lower Machiavellianism, lower psychopathy, and being a woman. Although personal fear about COVID-19 moderated the message effects, concern about the threats to relatives and family boosted the effects. Although the effect sizes were influenced by external factors, the results suggested that our proposed framework for information provision has the potential to be applied to a wider variety of risk-related topics.

Greenhouse gas mitigation can involve land-use changes that alter the habitat available for wildlife. Here, Ohashi et al. perform an integrated assessment showing that climate mitigation can be beneficial for global biodiversity but may entail local biodiversity losses where land-based mitigation is implemented.

To clarify the nature and characteristics of volatile organic compounds (VOCs) emitted from dairy cattle within a cattle shed located in Chiba, Japan, air samples were collected and analyzed for the four seasons in 2017-2018. Thirty-four VOCs were determined by gas chromatography-mass spectrometry and high performance liquid chromatography. In addition, air temperature and relative humidity inside and outside of the shed were monitored during each sampling campaign to estimate the ventilation rate of the shed. The average concentrations of total VOCs (μg m-3) in the shed in each season were 50.5 (spring), 128.4 (summer), 168.8 (autumn), and 199.5 (winter). Ketones were always the most dominant components followed by alcohols and volatile fatty acids (VFAs). The sum of ketones, alcohols, and VFAs accounted for more than 80% of the total VOCs in all seasons. Acetone, 3-pentanone, 1-butanol, and acetic acid were the major components regardless of the season, accounting for more than 60% of the total VOCs. The average emission rates of total VOCs from the shed (μg h-1 kg-1) were calculated to be 623 (spring), 1520 (summer), 585 (autumn) and 469 (winter). The emission rates of almost all the VOCs except alcohols increased exponentially with increase of air temperature in the shed. The ranges of the emission rates for each class of chemical (μg h-1 kg-1) were 39-170 (VFAs), 247-913 (ketones), 65-134 (alcohols), 40-122 (phenols), 10-122 (aldehydes), 4.17-22.3 (sulfur compounds), and 0.0067-0.74 (indoles). Furthermore, the annual emissions of VOCs for a single dairy cattle and for the cattle shed were estimated to be 5.5 kg and 44 kg, respectively

Abstract Background The Rock Ptarmigan Lagopus muta japonica lives in the alpine zones of central Japan, which is the southern limit of the global distribution for this species. This species is highly dependent on alpine habitats, which are considered vulnerable to rapid climate change. This study aimed to assess the impact of climate change on potential L. muta japonica habitat based on predicted changes to alpine vegetation, to identify population vulnerability under future climatic conditions for conservation planning. We developed species distribution models, which considered the structure of the alpine ecosystem by incorporating spatial hierarchy on specific environmental factors to assess the potential habitats for L. muta japonica under current and future climates. We used 24 general circulation models (GCMs) for 2081–2100 as future climate conditions. Results The predicted potential habitat for L. muta japonica was similar to the actual distribution of the territories in the study area of Japan’s northern Alps (36.25–36.5°N, 137.5–137.7°E). Future potential habitat for L. muta japonica was projected to decrease to 0.4% of the current potential habitat in the median of occurrence probabilities under 24 GCMs, due to a decrease in alpine vegetation communities. Some potential habitats in the central and northwestern part of the study area were predicted to be sustained in the future, depending on the GCMs. Conclusions Our model results predicted that the potential habitats for L. muta japonica in Japan’s northern Alps, which provides core habitat for this subspecies, would be vulnerable by 2081–2100. Small sustainable habitats may serve as refugia, facilitating the survival of L. muta japonica populations under future climatic conditions. Impact assessment studies of the effect of climate change on L. muta japonica habitats at a nationwide scale are urgently required to establish effective conservation planning for this species, which includes identifying candidate areas for assisted migration as an adaptive strategy.

Bacillus Calmette–Guérin (BCG) is widely used in the clinic to effectively treat superficial urinary bladder cancer. However, a significant proportion of patients who fail to respond to BCG risk cystectomy or death. Though more than 3 million cancer treatments with BCG occur annually, surprisingly little is known about the initial signaling cascades activated by BCG. Here, we report that BCG induces a rapid intracellular Ca2+ (calcium ion) signal in bladder cancer cells that is essential for activating the transcription factor nuclear factor kappa‐light‐chain‐enhancer of activated B cells (NF‐κB) and for synthesizing and secreting proinflammatory cytokines, including interleukin 8 (IL‐8). A similar Ca2+ response was observed when cells were exposed to the supernatant of BCG. Studying cellular molecular mechanisms involved in the BCG signaling event, we found pivotal roles for phospholipase C and the Toll‐like receptor 4. Further assessment revealed that this signaling pathway induces synthesis of IL‐8, whereas exocytosis appeared to be controlled by global Ca2+ signaling. These results shed new light on the molecular mechanisms underlying BCG treatment of bladder cancer, which can help in improving therapeutic efficacy and reducing adverse side effects.

Indoor air quality (IAQ) in houses is often deteriorated by chemical substances emitted from heating, building materials, or other household goods. Since it is difficult for occupants to recognize air pollution, they rarely understand the actual conditions of the IAQ. An investigation into the actual condition of IAQ in houses was therefore conducted in this study. Carbon dioxide (CO2) concentrations in 24 occupied houses was measured, and the results from our analysis showed that the use of combustion heaters increased the concentration of CO2 and led to indoor air pollution. Results indicate that as outdoor temperature decreased, the frequency of ventilation decreased simultaneously, and CO2 concentration increased. Results of the questionnaire survey revealed that the actual IAQ in each house did not match the level of awareness its occupants had regarding ventilation. Along with this difficulty in perceiving air pollution, the lack of knowledge about ventilation systems and the effects of combustion heating may be additional barriers to IAQ awareness.

BackgroundAlzheimer's disease (AD) is assessed by carefully examining a patient's cognitive impairment. However, previous studies reported inadequate diagnostic accuracy for dementia in primary care settings. Many hospitals use the automated quantitative evaluation method known as the Voxel-based Specific Regional Analysis System for Alzheimer's Disease (VSRAD), wherein brain MRI data are used to evaluate brain morphological abnormalities associated with AD. Similarly, an automated quantitative evaluation application called the easy Z-score imaging system (eZIS), which uses brain SPECT data to detect regional cerebral blood flow decreases associated with AD, is widely used. These applications have several indicators, each of which is known to correlate with the degree of AD. However, it is not completely known whether these indicators work better when used in combination in real-world clinical practice.MethodsWe included 112 participants with mild cognitive impairment (MCI) and 128 participants with early AD in this study. All participants underwent MRI, SPECT, and the Mini-Mental State Examination (MMSE). Demographic and clinical characteristics were assessed by univariate analysis, and logistic regression analysis with a combination of MMSE, VSRAD and eZIS indicators was performed to verify whether the diagnostic accuracy in discriminating between MCI and early AD was improved.ResultsThe area under the receiver operating characteristic curve (AUC) for the MMSE score alone was 0.835. The AUC was significantly improved to 0.870 by combining the MMSE score with two quantitative indicators from the VSRAD and eZIS that assessed the extent of brain abnormalities.ConclusionCompared with the MMSE score alone, the combination of the MMSE score with the VSRAD and eZIS indicators significantly improves the accuracy of discrimination between patients with MCI and early AD. Implementing VSRAD and eZIS does not require professional clinical experience in the treatment of dementia. Therefore, the accuracy of dementia diagnosis by physicians may easily be improved in real-world primary care settings.

Although sensorineural hearing loss (SHL) is relatively common, its cause has not been identified in most cases. Previous studies have suggested that viral infection is a major cause of SHL, especially sudden SHL, but the system that protects against pathogens in the inner ear, which is isolated by the blood-labyrinthine barrier, remains poorly understood. We recently showed that, as audiosensory receptor cells, cochlear hair cells (HCs) are protected by surrounding accessory supporting cells (SCs) and greater epithelial ridge (GER or Kölliker's organ) cells (GERCs) against viral infections. Here, we found that virus-infected SCs and GERCs induce HC death via production of the tumour necrosis factor-related apoptosis-inducing ligand (TRAIL). Notably, the HCs expressed the TRAIL death receptors (DR) DR4 and DR5, and virus-induced HC death was suppressed by TRAIL-neutralizing antibodies. TRAIL-induced HC death was not caused by apoptosis, and was inhibited by necroptosis inhibitors. Moreover, corticosteroids, the only effective drug for SHL, inhibited the virus-induced transformation of SCs and GERCs into macrophage-like cells and HC death, while macrophage depletion also inhibited virus-induced HC death. These results reveal a novel mechanism underlying virus-induced HC death in the cochlear sensory epithelium and suggest a possible target for preventing virus-induced SHL.

Invasive alien tree species can exert severe impacts, especially in insular biodiversity hotspots, but have been inadequately studied. Knowledge of the life history and population trends of an invasive alien tree species is essential for appropriate ecosystem management. The invasive tree Bischofia javanica has overwhelmed native trees on Haha-jima Island in the Ogasawara Islands, Japan. We explored forest community dynamics 2 years after a typhoon damaged the Sekimon primary forests on Haha-jima Island, and predicted the rate of population increase of B. javanica using a logistic model from forest dynamics data for 19 years. During the 2 years after the typhoon, only B. javanica increased in population size, whereas populations of native tree species decreased. Stem diameter growth of B. javanica was more rapid than that of other tree species, including native pioneer trees. Among the understory stems below canopy trees of other species, B. javanica grew most rapidly and B. javanica canopy trees decreased growth of the dominant native Ardisia sieboldii. These competitive advantages were indicated to be the main mechanism by which B. javanica replaces native trees. The logistic model predicted that B. javanica would reach 30% of the total basal area between 2017 (in the eastern plot adjacent to a former B. javanica plantation) and 2057 (in the western plot distant from the plantation site), which is a maximum percentage allowing to eradicate under the present guideline of the National Forest. The results suggest immediate removal of B. javanica is required to preserve native biodiversity in these forests.

A well-known phenomenon is that humans perceive risks to threaten future generations as more dangerous in many cases. However, this tendency could be changed depending on certain conditions and could potentially be explained by the evolution of altruism. Our multi-agent simulation model, which was constructed to identify attributes contributing to subjective assessment of a risk source based on kin selection theory, showed that support from relatives can affect the agents' subjective risk assessment. We utilize this insight, which has never been explored in the context of nudge, to show that real-world messages reminding respondents that they are supported by their relatives can moderate the perception of a risk source as extremely dangerous. A randomized control trial based on an internet questionnaire survey was conducted to identify the intervention effect of such messages, using air pollution caused by industrialization as the risk source for the case study. Our analysis suggests that messages moderate extreme attitudes. Presentation of additional visual information can boost the sense of familial support and increase the effect of a message compared with a message comprising only textual information. The attributes and personality traits of the respondents who are responsive to the intervention message are also discussed.

Patterned cell culturing is one of the most useful techniques for understanding the interaction between geometric conditions surrounding cells and their behaviors. The authors previously proposed a simple method for cell patterning with an agarose gel microstructure fabricated by microcasting with a degassed polydimethylsiloxane (PDMS) mold. Although the vacuum pressure produced from the degassed PDMS can drive a highly viscous agarose solution, the influence of solution viscosity on the casting process is unknown. This study investigated the influences of micro-channel dimensions or solution viscosity on the flow of the solution in a micro-channel of a PDMS mold by both experiments and numerical simulation. It was found experimentally that the degassed PDMS mold was able to drive a solution with a viscosity under 575 mPa·s. A simulation model was developed which can well estimate the flow rate in various dimensions of micro-channels. Cross-linked albumin has low viscosity (1 mPa·s) in aqueous solution and can undergo a one-way dehydration process from solution to solid that produces cellular repellency after dehydration. A microstructure of cross-linked albumin was fabricated on a cell culture dish by the microcasting method. After cells were seeded and cultivated on the cell culture dish with the microstructure for 7 days, the cellular pattern of mouse skeletal myoblast cell line C2C12 was observed. The microcasting with cross-linked albumin solution enables preparation of patterned cell culture systems more quickly in comparison with the previous agarose gel casting, which requires a gelation process before the dehydration process.

HER3, a member of the receptor tyrosine kinase super family, is overexpressed in a number of cancers, and is associated with malignant phenotypes. Control of the protein stability of the membrane, as well as nuclear receptors, has been known to be an important process affecting tumor cells; however, their relationships have yet to be elucidated. In this study, we demonstrate that estradiol promotes rapid degradation of HER3 via the proteasome pathway in ER-positive breast cancer, MCF-7. ER prevented HER3 degradation, and knockdown of ER expression by si-RNA promoted rapid degradation of HER3. Breakdown of HER3 and ER were regulated by a ubiquitin ligase Nedd4-1 in the presence of estradiol stimulation. We speculate that estradiol quickly degrades ER, making HER3 accessible by Nedd4-1, and leads to the rapid degradation of HER3. In addition, knockdown of ubiquitin ligase Nedd4-1 enhances estradiol induced cell proliferation. These results indicate that HER3 and Nedd4-1 in ER-positive breast cancers might be an important therapeutic target. Keywords: HER3, Nedd4-1, Estradiol, ER, Degradation

The concentrations and chemical compositions of volatile organic compounds (VOCs), including volatile fatty acids, phenols, indoles, aldehydes, and ketones, which are the main organic compounds generated by swine, were investigated in July and October 2016 and January 2017. In addition, the emission rates and annual emissions of these components from the swine shed were estimated. The concentrations of VOCs in the swine shed averaged 511.3 μg m-3 in summer, 315.5 μg m-3 in fall and 218.6 μg m-3 in winter. Acetone, acetic acid, propionic acid, and butyric acid were the predominant components of the VOCs, accounting for 80-88% of the total VOCs. The hourly variations of VOC concentrations in the swine shed in fall and winter suggest that the VOC concentrations were related to the ventilation rate of the swine shed, the activity of the swine, and the temperature in the swine shed. Accordingly, the emission rates of VOCs from the swine shed were 1-2×103 μg (h kg-swine)-1.

Abstract Background Cisplatin is the most commonly used chemotherapeutic agent in the treatment of patients with metastatic and/or recurrent urothelial cancer. However, the effectiveness of these treatments is severely limited due to the development of cisplatin resistance. Cancer stem cells have been documented as one of the key hypotheses involved in chemoresistance. CD44v8–10 has been identified as one of the new cancer stem cells markers and was recently shown to enhance the antioxidant system by interaction with xCT, a subunit of the cystine transporter modulating intracellular glutathione synthesis. The aim of the present study was to investigate the clinical role of CD44v8–10 and the molecular mechanism underlying the acquisition of cisplatin resistance through CD44v8–10 in urothelial cancer. Methods We analyzed the clinical significance of the immunohistochemical CD44v9 expression, which detects the immunogen of human CD44v8–10, in 77 urothelial cancer patients treated with cisplatin-based systemic chemotherapy for recurrence and/or metastasis. We then evaluated the biological role of CD44v8–10 in the acquisition of cisplatin resistance using the urothelial cancer cell lines, T24 and T24PR, which were generated to acquire resistance to cisplatin. Results The 5-year cancer-specific survival rate was significantly lower in the CD44v9-positive group than in the CD44v9-negative group (P = 0.008). Multivariate analyses revealed that CD44v9 positivity was an independent risk factor of cancer-specific survival (P = 0.024, hazard ratio = 5.16) in urothelial cancer patients who had recurrence and/or metastasis and received cisplatin-based chemotherapy. The expression of CD44v8–10 and xCT was stronger in T24PR cells than in T24 cells. The amount of intracellular glutathione was significantly higher in T24PR cells than in T24 cells (p

Abstract Surface wettability is one of most important indexes for understanding the interfacial characteristics between a solid surface and a liquid, especially such physico-chemical properties as hydrophilicity and water repellency. Recently, non-contact wettability assessment by monitoring the behavior of liquid covering a sample surface by jet-flow application to the liquid has been proposed. This method is applicable to the in-liquid assessment of high wettability materials for which it is difficult to measure the contact angle, such as cultured cells in a liquid culture medium. However, when the flow is dirty, the contamination of the cell culture system by particles (e.g. microorganisms, virus, and aerosols) may occur. To solve this problem, we developed a contamination-free non-contact wettability assessment system that is composed of a clean-air source having a particle filter and an automated system for image processing instrumentation and motorized alignment device for sample positioning. We confirmed that the filtered air-jet contained no particles, although over ten thousand particles had been included in the air source. Furthermore, we detected no microorganisms on agar culture plates, which had been exposed to the air-jet before being incubated. With the image instrumentation, we automatically measured the liquid-squeezed size, which was the main index of surface wettability. The motorized alignment device allowed us to control the sample position without human direct operation. This developed system is compatible with wettability assessment in a high-level clean environment such as quality control of culture cells for regenerative medicine and in vitro assay of drug efficacy and safety.

Tumor-associated blood vessels and lymphatics are abnormal and dysfunctional. These are hallmarks of the tumor microenvironment, which has an immunosuppressive nature, such as through hypoxia. Treatment with anti-death receptor5 (DR5) monoclonal antibody MD5-1, which induces tumor cell death, is a potent anti-tumor immunotherapy. Generally, MD5-1 induces cell death mainly via antigen presenting cells (APCs) and generates tumor-specific effector T cells. To date, the effects of a simultaneous functional improvement of abnormal blood vessels and lymphatics on the immune microenvironment are largely unknown. A combination therapy using sunitinib, vascular endothelial growth factor (VEGF) and platelet-derived growth factor receptor inhibitor, and MD5-1 substantially inhibited tumor growth. Sunitinib improved pericyte coverage on endothelial cells and the expression levels of regulator of G-protein signaling 5, suggesting blood vessel normalization. Sunitinib also increased lymph flow from tumors to central lymph nodes, suggesting improved lymphatic function. In concordance with improved vasculature functions, sunitinib alleviated the tumor hypoxia, suggesting an improved tumor microenvironment. Indeed, the combination therapy induced strong activation of CD8+ T cells and dendritic cells in draining lymph nodes. The combination therapy reduced the ratio of immune-suppressive T regulatory cells in the tumors and draining lymph nodes. The combination therapy enhanced the numbers and activation of tumor-infiltrating CD8+ T cells. CD4 and/or CD8 depletion, or APC inhibiting experiments showed the contribution of CD8+ T cells and APCs to the combination therapy. These findings suggest that targeting blood vessels and lymphatics may have potential benefits for immunotherapy mediated by CD8+ T cells and APCs.

Abstract Ongoing climate change and land‐use change have the potential to substantially alter the distribution of large herbivores. This may result in drastic changes in ecosystems by changing plant–herbivore interactions. Here, we developed a model explaining sika deer persistence and colonization between 25 years in terms of neighborhood occupancy and habitat suitability. We used climatic, land‐use, and topographic variables to calculate the habitat suitability and evaluated the contributions of the variables to past range changes of sika deer. We used this model to predict the changes in the range of sika deer over the next 100 years under four scenario groups with the combination of land‐use change and climate change. Our results showed that both climate change and land‐use change had affected the range of sika deer in the past 25 years. Habitat suitability increased in northern or mountainous regions, which account for 71.6% of Japan, in line with a decrease in the snow cover period. Habitat suitability decreased in suburban areas, which account for 28.4% of Japan, corresponding to land‐use changes related to urbanization. In the next 100 years, the decrease in snow cover period and the increase in land abandonment were predicted to accelerate the range expansion of sika deer. Comparison of these two driving factors revealed that climate change will contribute more to range expansion, particularly from the 2070s onward. In scenarios that assumed the influence of both climate change and land‐use change, the total sika deer range increased by between +4.6% and +11.9% from the baseline scenario. Climate change and land‐use change will require additional efforts for future management of sika deer, particularly in the long term.

Nrf2 is a transcriptional activator of oxidative stress response genes. Here the authors show that Nrf2 binds to promoters of proinflammatory genes and interferes with their transcriptional upregulation in LPS-stimulated macrophages independently of its role in regulation of reactive oxygen species.

Alpinia galanga is a well-known medicinal plant in Southeast Asia and has been used for a long time as food and medicine. A large number of flavonoid phytochemical compounds have been identified in various parts of this medicinal herb. Flavonoids are commonly known as attractive compounds that can be applied to cosmetic or cosmeceutical product development because of their antioxidant, anti-aging and many other potential biological activities. This recent review aims to illustrate and update the taxonomic status as well as the species description that will be helpful for a rigorous identification and authenticate the raw material or living specimen from A. galanga. The flavonoid phytochemical compounds and the bioactivity of this medicinal plant are also provided. The future perspectives and research directions of A. galanga and its flavonoids are pointed out in this study as well.

The glioma-associated oncogene (GLI) family consists of GLI1, GLI2, and GLI3 in mammals. This family has important roles in development and homeostasis. To achieve these roles, the GLI family has widespread outputs. GLI activity is therefore strictly regulated at multiple levels, including via post-translational modifications for context-dependent GLI target gene expression. The protein arginine methyl transferase (PRMT) family is also associated with embryogenesis, homeostasis, and cancer mainly via epigenetic modifications. In the PRMT family, PRMT1, PRMT5, and PRMT7 reportedly regulate GLI1 and GLI2 activity. PRMT1 methylates GLI1 to upregulate its activity and target gene expression. Cytoplasmic PRMT5 methylates GLI1 and promotes GLI1 protein stabilization. Conversely, nucleic PRMT5 interacts with MENIN to suppress growth arrest-specific protein 1 expression, which assists Hedgehog ligand binding to Patched, indirectly resulting in downregulated GLI1 activity. PRMT7-mediated GLI2 methylation upregulates its activity through the dissociation of GLI2 and Suppressor of Fused. Together, PRMT1, PRMT5, and PRMT7 regulate GLI activity at multiple revels. Furthermore, the GLI and PRMT families have strong links with various cancers through cancer stem cell maintenance. Therefore, PRMT-mediated regulation of GLI activity would have important roles in cancer stem cell maintenance.

The extracellular matrix (ECM) surrounding cancer cells becomes stiffer during tumor progression, which influences cancer cell behaviors such as invasion and proliferation through modulation of gene expression as well as remodeling of the actin cytoskeleton. In this study, we show that MMP24 encoding matrix metalloproteinase (MMP)-24 is a novel target gene of Yes-associated protein (YAP), a transcription coactivator known as a mechanotransducer. We first examined the effect of substrate stiffness on MMP24 expression in MCF-7 human breast cancer cells and showed that the expression of MMP24 was significantly higher in cells grown on stiff substrates than that on soft substrates. The MMP24 expression was significantly reduced by knockdown of YAP. In contrast, the expression of constitutively active YAP increased MMP24 promoter activity. In addition, binding of YAP to the MMP24 promoter was confirmed by the chromatin immunoprecipitation (ChIP) assay. These results show that ECM stiffening promotes YAP activation, thereby inducing MMP24 expression. Based on the Human Protein Atlas database, breast cancer patients with lower MMP24 expression exhibit the worse survival rates overall. Thus, MMP24 may negatively regulate the aggressiveness of cancer cells under the stiff ECM environment during tumor progression.

Pine wilt disease (PWD) constitutes a serious threat to pine forests. Since development depends on temperature and drought, there is a concern that future climate change could lead to the spread of PWD infections. We evaluated the risk of PWD in 21 susceptible Pinus species on a global scale. The MB index, which represents the sum of the difference between the mean monthly temperature and 15 when the mean monthly temperatures exceeds 15°C, was used to determine current and future regions vulnerable to PWD (MB ≥ 22). For future climate conditions, we compared the difference in PWD risks among four different representative concentration pathways (RCPs 2.6, 4.5, 6.0, and 8.5) and two time periods (2050s and 2070s). We also evaluated the impact of climate change on habitat suitability for each Pinus species using species distribution models. The findings were then integrated and the potential risk of PWD spread under climate change was discussed. Within the natural Pinus distribution area, southern parts of North America, Europe, and Asia were categorized as vulnerable regions (MB ≥ 22; 16% of the total Pinus distribution area). Representative provinces in which PWD has been reported at least once overlapped with the vulnerable regions. All RCP scenarios showed expansion of vulnerable regions in northern parts of Europe, Asia, and North America under future climate conditions. By the 2070s, under RCP 8.5, an estimated increase in the area of vulnerable regions to approximately 50% of the total Pinus distribution area was revealed. In addition, the habitat conditions of a large portion of the Pinus distribution areas in Europe and Asia were deemed unsuitable by the 2070s under RCP 8.5. Approximately 40% of these regions overlapped with regions deemed vulnerable to PWD, suggesting that Pinus forests in these areas are at risk of serious damage due to habitat shifts and spread of PWD.

The geometrical confinement of small cell colonies gives differential cues to cells sitting at the periphery versus the core. To utilize this effect, for example to create spatially graded differentiation patterns of human mesenchymal stem cells (hMSCs) in vitro or to investigate underpinning mechanisms, the confinement needs to be robust for extended time periods. To create highly repeatable micro-fabricated structures for cellular patterning and high-throughput data mining, we employed here a simple casting method to fabricate more than 800 adhesive patches confined by agarose micro-walls. In addition, a machine learning based image processing software was developed (open code) to detect the differentiation patterns of the population of hMSCs automatically. Utilizing the agarose walls, the circular patterns of hMSCs were successfully maintained throughout 15 days of cell culture. After staining lipid droplets and alkaline phosphatase as the markers of adipogenic and osteogenic differentiation, respectively, the mega-pixels of RGB color images of hMSCs were processed by the software on a laptop PC within several minutes. The image analysis successfully showed that hMSCs sitting on the more central versus peripheral sections of the adhesive circles showed adipogenic versus osteogenic differentiation as reported previously, indicating the compatibility of patterned agarose walls to conventional microcontact printing. In addition, we found a considerable fraction of undifferentiated cells which are preferentially located at the peripheral part of the adhesive circles, even in differentiation-inducing culture media. In this study, we thus successfully demonstrated a simple framework for analyzing the patterned differentiation of hMSCs in confined microenvironments, which has a range of applications in biology, including stem cell biology.

Myanmar (Burma) constitutes a significant component of the Indo-Myanmar biodiversity hotspot, with elements of the Indian, the Indochina, and the Sino-Japanese floristic regions, yet thus far only a few reliable sources of the country's flora have been available. As a part of a contribution for the floristic inventory of Myanmar, since it is important in a floristic survey to obtain as much information as possible, in addition to previous two reports, here we present three more chromosome counts in the aquatic monocots of Myanmar: Limnocharis flava with 2n = 20, Sagittaria trifolia with 2n = 22 (Alismataceae), and Potamogeton distinctus × P. nodosus with 2n = 52 (Potamogetonaceae); the third one is new to science. A brief review of cytological researches in the floristic regions' 45 non-hybrid aquatic monocots plus well investigated two inter-specific hybrids that are recorded in Myanmar is given, indicating that the further works with a focus on species in Myanmar that has infra-specific chromosome variation in the floristic regions will address the precise evolutionary history of the aquatic flora of Myanmar.