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PurposeOur preclinical studies showed that lycopene enhanced the anti-prostate cancer efficacy of docetaxel in animal models. A phase I trial (NCT0149519) was conducted to identify an optimum dose of synthetic lycopene in combination with docetaxel (and androgen blockade [androgen deprivation therapy, ADT]), and to evaluate its effect on the safety and pharmacokinetics of docetaxel in men with metastatic prostate cancer.MethodsSubjects were treated with 21-day cycles of 75 mg/m2 docetaxel (and ADT), plus lycopene at 30, 90 or 150 mg/day. A Bayesian model averaging continual reassessment method was used to guide dose escalation. Pharmacokinetics of docetaxel and multiple correlative studies were carried out.ResultsTwenty-four participants were enrolled, 18 in a dose escalation cohort to define the maximum tolerated dose (MTD), and six in a pharmacokinetic cohort. Docetaxel/ADT plus 150 mg/day synthetic lycopene resulted in dose-limiting toxicity (pulmonary embolus) in one out of 12 participants with an estimated probability of .106 and thus was chosen as the MTD. Lycopene increased the AUCinf and Cmax of plasma docetaxel by 9.5% and 15.1%, respectively. Correlative studies showed dose-related changes in circulating endothelial cells and vascular endothelial growth factor A, and reduction in insulin-like growth factor 1R phosphorylation, associated with lycopene therapy.ConclusionsThe combination of docetaxel/ADT and synthetic lycopene has low toxicity and favourable pharmacokinetics. The effects of lycopene on biomarkers provide additional support for the toxicity-dependent MTD definition.HighlightsThe maximum tolerated dose was identified as 150 mg/day of lycopene in combination with docetaxel/ADT for the treatment of metastatic prostate cancer patients. Small increases in plasma exposure to docetaxel were observed with lycopene co-administration. Mechanistically significant effects were seen on angiogenesis and insulin-like growth factor 1 signalling by lycopene co-administration with docetaxel/ADT.

Background: SHR-5 has been used as an "adaptogen" for enhancing physical and mental performance and for fighting stress in the healthy population. The purpose of this study is to determine the chemopreventive efficacy of SHR-5 for superficial bladder cancer and to investigate the underlying mechanisms of action. Methods: UPII-mutant Ha-ras bladder-cancer-transgenic mice, that developed low-grade and noninvasive papillary transitional urothelial cell carcinoma, were fed with 1.25 and 6.25 mg/mL SHR-5 in drinking water for 6 months. The survival of the mice, obstructive uropathy, tumor burden and morphology, and proliferation were evaluated by pathological, molecular, metabolic, and statistical analyses. Results: Approximately 95% or more of the male UPII-mutant Ha-ras mice that drank SHR-5 daily survived over 6 months of age, while only 33.3% of those mice that drank normal water survived over 6 months of age (p < 0.0001); SHR-5 drinking exposure also reduced tumor-bearing bladder weight and urinary tract obstruction and inhibited mTOR signaling in neoplastic tissues. Global metabolic analysis revealed that SHR-5 resulted in increased phenolic metabolites and decreased CoA, a critical metabolic cofactor for lipid metabolism. Conclusions: Our findings highlight the potential of SHR-5 as an anti-aging agent for bladder cancer prevention through reshaping tumor metabolism via the inhibition of the mTOR signaling. Global metabolomics profiling provides a unique and efficient tool for studying the mechanisms of complex herb extracts' action.

Abstract Purpose Our preclinical studies showed that lycopene enhanced the anti‐prostate cancer efficacy of docetaxel in animal models. A phase I trial (NCT0149519) was conducted to identify an optimum dose of synthetic lycopene in combination with docetaxel (and androgen blockade [androgen deprivation therapy, ADT]), and to evaluate its effect on the safety and pharmacokinetics of docetaxel in men with metastatic prostate cancer. Methods Subjects were treated with 21‐day cycles of 75 mg/m2 docetaxel (and ADT), plus lycopene at 30, 90 or 150 mg/day. A Bayesian model averaging continual reassessment method was used to guide dose escalation. Pharmacokinetics of docetaxel and multiple correlative studies were carried out. Results Twenty‐four participants were enrolled, 18 in a dose escalation cohort to define the maximum tolerated dose (MTD), and six in a pharmacokinetic cohort. Docetaxel/ADT plus 150 mg/day synthetic lycopene resulted in dose‐limiting toxicity (pulmonary embolus) in one out of 12 participants with an estimated probability of .106 and thus was chosen as the MTD. Lycopene increased the AUCinf and Cmax of plasma docetaxel by 9.5% and 15.1%, respectively. Correlative studies showed dose‐related changes in circulating endothelial cells and vascular endothelial growth factor A, and reduction in insulin‐like growth factor 1R phosphorylation, associated with lycopene therapy. Conclusions The combination of docetaxel/ADT and synthetic lycopene has low toxicity and favourable pharmacokinetics. The effects of lycopene on biomarkers provide additional support for the toxicity‐dependent MTD definition. Highlights The maximum tolerated dose was identified as 150 mg/day of lycopene in combination with docetaxel/ADT for the treatment of metastatic prostate cancer patients. Small increases in plasma exposure to docetaxel were observed with lycopene co‐administration. Mechanistically significant effects were seen on angiogenesis and insulin‐like growth factor 1 signalling by lycopene co‐administration with docetaxel/ADT.

Abstract Background Kawasaki disease (KD) is usually treated with high-dose intravenous immunoglobulin (IVIg) as severe infectious and other diseases. Due to issues that are associated with immunoglobulin preparation, such as the risk of possible contamination by infectious agents and limited blood banking resources, recombinant immunoglobulins are required. We developed a novel recombinant antibody drug candidate, “VasSF,” based on the therapeutic effects it exerted on a mouse spontaneous crescentic glomerulonephritis model (SCG/Kj). Apolipoprotein A-2 (ApoA2) has been identified as one of VasSF’s target molecules. Methods Here, we tested the potential of anti-apolipoprotein A-2 antibodies (anti-ApoA2) as a new therapeutic drug against KD by examining its effect on a mouse model, in which KD was induced via Candida albicans water-soluble fraction (CAWS). CAWS (2 mg/mouse) was injected intraperitoneally into C57BL/6NCrSlc mice for five consecutive days. The incidence and histological severity of vasculitis in CAWS-induced coronary arteritis in mice administered anti-ApoA2 was examined. The following experimental groups were tested: solvent (only PBS (−) injection); anti-ApoA2 antibodies at dosages of 0.05 mg, 0.1 mg, and 0.5 mg/kg/day; human IgG at 0.1 mg/kg/day. Results The group treated with anti-ApoA2 0.5 mg/kg/day showed a lower incidence of panvasculitis induced by CAWS, less inflammation of the coronary arteries and aortic roots, and lower levels of serum IL-6, M-CSF, and MIP-1α and 32 cytokines/chemokines compared with those in the solvent group. Conclusions The anti-ApoA2 treatment suppressed the development of coronary arteritis in an animal KD model and anti-ApoA2 shows potential as an effective therapeutic candidate for the treatment of KD vasculitis. The use of specific antibodies that display higher vasculitis-suppressing effects, such as anti-ApoA2, may attenuate KD as well as other infectious diseases, with less severe adverse side effects than treatment with IVIg.

Abstract Ramp phenomena caused by abrupt changes in wind speed may confound the stable operation of correlated electrical power supply systems, yet accurate numerical predictions are challenging, as the wind is affected by complex interactions between large‐scale weather patterns and local geographical conditions. Further, optimal numerical weather prediction (NWP) methods and physics schemes vary as a function of weather patterns. The present study proposed a new real‐time wind power ramp forecast framework based on the flexible selection of optimal NWP models, which were derived via principal component analysis (PCA). The novelty of this analysis lies in that statistical methods were employed for NWP optimization, compared with their more conventional use during an NWP postprocessing. Here, a weather pattern was classified by PCA using outcomes from the global‐scale prediction models, and the optimum regional NWP system settings were acquired according to the weather patterns for further wind field dynamical downscaling. The performance of the developed prediction system was verified with wind power at wind turbine hub‐heights for three areas in eastern Japan, and the Critical Success Index (CSI) indicated an improvement of prediction accuracy over benchmark predictions by ≤0.184 for ramp‐up events and ≤0.127 for ramp‐down events (both observed in Tohoku area). Higher CSI values were consistently seen in three wind farm areas, indicative of the improvement in detection probability for actual ramp events compared with benchmark.

Abstract Permafrost covers a wide area of the Northern Hemisphere, including high-altitude mountainous areas and even at mid-latitudes. There is concern that the thawing of mountain permafrost can cause slope instability and substantially impact alpine ecosystems, and because permafrost in mountainous areas is difficult to observe, detailed analyses have not been performed on its current distribution and future changes. Although previous studies have observed permafrost only at a limited number of points in Japan (e.g., Daisetsu Mountains, Mt. Fuji, and Mt. Tateyama in the Northern Japan Alps), we show that permafrost potentially exists in nine domains in Japan (Daisetsu Mountains, Mt. Fuji, Northern and Southern Japan Alps, Hidaka Mountains, Mt. Shiretokodake, Sharidake, Akandake, and Yotei). In the Daisetsu Mountains and Mt. Fuji, the environmental conditions required for maintaining at least some permafrost are projected to remain in the future if a decarbonized society is achieved (RCP2.6 or RCP4.5). However, if greenhouse gas emissions continue to increase (RCP8.5), the environmental conditions required for sustaining permafrost are projected to disappear in the second half of the twenty-first century. In other domains, the environmental conditions required for maintaining permafrost are either projected to disappear in the next ten years (Hidaka Mountains, Northern Japan Alps) or they have almost disappeared already (Southern Japan Alps, Mt. Shiretokodake, Sharidake, Akandake, and Yotei). Our projections show that climate change has a tremendous impact on Japan's mountain permafrost environment and suggests the importance of monitoring the mountain environment and considering measures for adapting to future climate change.

In this study, we assessed air quality (AQ) and urban climate during the mobility restrictions implemented in the Greater Tokyo Area, Japan, the world’s most populated region, in response to the COVID-19 pandemic. Observations from dense surface networks were analyzed using an interpretable machine learning approach. In parallel with a ∼50% reduction in mobility and an altered lifestyle of the population, we found limited reductions in nitrogen dioxide; decreases in fine particulate matter not entirely driven by local mobility; minor variations in ozone, with a positive (negative) tendency in areas with high (low) emissions; a decrease in air temperature consistent with mobility; and pollution levels and air temperature changes with well-defined, common spatiotemporal patterns. Specifically, cooling mainly occurred in urbanized areas with an improved AQ. Overall, although reductions in mobility were moderately effective in improving the typical indicators of urban AQ, including those known to negatively impact human health, the reductions in waste heat had a stronger impact on Tokyo’s urban heat island, suggestive of a strategy to minimize exposure to heat stress. These findings can help guide urban planning strategies and policies aimed at addressing climate change.

The Upper Chagres River Basin plays an important role in the stable operation of the Panama Canal. Previous studies have shown that there is a salient regional difference in the diurnal variation of precipitation in the basin. Precipitation during the rainy season peaks in the early afternoon throughout the basin, but precipitation is also observed in the morning at sites in the northern part of the basin. However, the cause of this is not clear due to limited ground observation. To address this issue, we conducted dynamical downscaling experiments with a horizontal grid spacing of 5 and 2 km and nested in a global atmospheric model with horizontal grid spacing of approximately 20 km. The results showed that the 2 km convection-permitting model successfully reproduced regional differences in observed diurnal variations. The downscaled results indicated that intensified low-level northeasterly winds over the southern Caribbean Sea triggered favorable conditions for morning rain with an orographic effect under the seaside coastal regime in the western Caribbean Sea. This is in contrast to precipitation peaks in the early afternoon under a landside coastal regime.

Abstract Climate scenario data set are indispensable for assessing future climate impacts. In this study, we developed statistically downscaled climate scenarios in Japan using modified bias correction method based on five general circulation models selected from the Coupled Model Intercomparison Project (CMIP) Phase 6 to facilitate impact assessments and adaptation strategies. Modification of time window of the original correction method results in successful agreement with the observed seasonal change of variables in each grid. The original CMIP6 models have a relatively small bias compared to CMIP5 models. The advantage of CMIP6‐based bias‐corrected scenarios is its availability for multiple global circulation models, which covered wide uncertainty in CMIP6 ensembles, with various emissions scenarios for representative concentration pathway (RCP) including RCP4.5, RCP2.6, and RCP8.5. Several temperature‐related indices derived from the CMIP6‐based climate scenarios agreed well with observations. The number of extremely hot days and nights increased nonlinearly in the future with additional global warming. An increase in the global warming level from 1 to 2°C above the early 1900s would increase the probability of the number of extremely hot days per year exceeding the 2018 case by 4.1 times. The development of bias‐corrected climate scenarios facilitates the study of various climate impacts on a CMIP6 basis.

Abstract Permafrost is known to occur in high mountainous areas such as the Daisetsu Mountains in Japan, which are located at the southernmost limit of the permafrost distribution in the world. In this study, areas with climatic conditions suitable for sustaining permafrost in the Daisetsu Mountains are projected using bias-corrected and downscaled climate model outputs and statistical relationships between surface air temperatures and permafrost areas. Using freezing and thawing indices, the size of the area in the Daisetsu Mountains where climatic conditions were suitable for permafrost were estimated to be approximately 150 km2 in 2010. Under the RCP8.5 scenario, this area is projected to decrease to about 30 km2 by 2050 and it is projected to disappear by around 2070. Under the RCP2.6 scenario, the area is projected to decrease to approximately 20 km2 by 2100. The degradation of mountain permafrost could potentially affect the stability of trekking trails due to slope displacement, and it may also have deleterious effects on current alpine ecosystems. It is therefore important to accurately monitor changes in the mountain ecosystem environment and to implement measures to adapt to an environment that is projected to change significantly in the future.

Assessing the impacts of climate change in multiple fields, such as energy, land and water resources, and human health and welfare is important to find effective strategies to adapt to a changing climate and to reduce greenhouse gases. Many phenomena influenced by climate change have diurnal fluctuations and are affected by simultaneous interactions among multiple meteorological factors. However, climate scenarios with detailed (at least hourly) resolutions are usually not available. To assess the impact of climate change on such phenomena while considering simultaneous interactions (e.g., synergies), climate scenarios with hourly fluctuations are indispensable. However, because meteorological indicators are not independent, the value of one indicator varies as a function of other indicators. Therefore, it is almost impossible to determine the functions that show all relationships among meteorological elements considering the geographical and temporal (both seasonal and time of a day) characteristics. Therefore, generating hourly scenarios that include possible combinations of meteorological indicators for each hourly observation unit is a challenging problem. In this study, we provide secondary future climate scenario datasets that have hourly fluctuations with reasonable combinations of meteorological indicator values that are likely to occur simultaneously, without losing the long-term climate change trend in the existing daily climate scenarios based on global climate models. Historical hourly weather datasets observed from 2017 to 2019 (the reference years) are used to retrieve short-term fluctuations. Bias-corrected daily future climate scenario datasets generated using four global climate models (GFDL CM3, HadGEM2-ES, MIROC5, and MRI-CGCM3) and two Representative Concentration Pathways (RCP8.5 and 2.6) are used to model long-term climate change. A total of 48 different types of hourly future scenario datasets for five meteorological indicators (temperature, solar radiation, humidity, rainfall, and wind speed) were acquired, targeting a projection period from 2020 to 2080, for 10 weather stations in Japan. The generated hourly climate scenario datasets can be used to project the quantitative impacts of climate change on targeted phenomena considering simultaneous interactions among multiple meteorological factors.

To investigate the cellular and molecular interactions between clear-cell renal cell carcinoma (ccRCC) and perinephric adipose tissue (PAT), perineoplasm PAT, PAT away from the neoplasm, renal sinus and subcutaneous adipose tissues were collected at the time of renal surgery for renal masses and conditioned medium (CM) was generated from 62 patients. Perineoplasm PAT CMs from 44 out of 62 (about 71%) of patients with ccRCC or benign renal diseases (e.g. oncocytomas, angiomyolipomas, multicystic kidney, interstitial fibrosis, etc.) enhanced the migration of CaKi-2 cells. Perineoplasm PAT CMs from ccRCC significantly increased migration of ACHN and CaKi-2 cells by ~8.2 and ~2.4 folds, respectively, relative to those from benign renal diseases, whereas there is no significant difference in migration between ccRCC and benign renal diseases in CMs collected from culturing PAT away from neoplasm, renal sinus and subcutaneous adipose tissues. High Fuhrman Grade was associated with increased migration of Caki-2 cells by perineoplasm PAT CMs. Perineoplasm PATs from pT3 RCCs overexpressed multiple WNTs and their CMs exhibited higher WNT/ß-catenin activity and increased the migration of Caki-2 cells compared to CMs from benign neoplasms. Addition of secreted WNT inhibitory factor-1 recombinant protein into perineoplasm PAT CMs completely blocked the cell migration. These results indicate that WNT related factors from perineoplasm PAT may promote progression of local ccRCC to locally advanced (pT3) disease by increasing ccRCC cell mobility.

Recent evidence has strongly supported that the rate of aging is controlled, at least to some extent, by evolutionarily conserved nutrient sensing pathways (e.g. the insulin/IGF-1-signaling, mTOR, AMPK, and sirtuins) from worms to humans. These pathways are also commonly involved in carcinogenesis and cancer metabolism. Agents (e.g. metformin, resveratrol, and Rhodiola) that target these nutrient sensing pathways often have both anti-aging and anti-cancer efficacy. These agents not only reprogram energy metabolism of malignant cells, but also target normal postmitotic cells by suppressing their conversion into senescent cells, which confers systematic metabolism benefits. These agents are fundamentally different from chemotherapy (e.g. paclitaxel and doxorubicin) or radiation therapy that causes molecular damage (e.g. DNA and protein damages) and thereby no selection resistance may be expected. By reviewing molecular mechanisms of action, epidemiological evidence, experimental data in tumor models, and early clinical study results, this review provides information supporting the promising use of agents with both anti-aging and anti-cancer efficacy for cancer chemoprevention.

S phase kinase-associated protein 2 (Skp2) has been shown to be required for spontaneous tumor development that occurs in the retinoblastoma protein (pRb) deficient mice. Here we have demonstrated that flavokawain A (FKA), a novel chalcone from the kava plant, selectively inhibited the growth of pRb deficient cell lines and resulted in a proteasome-dependent and ubiquitination-mediated Skp2 degradation. Degradation of Skp2 by FKA was found to be involved in a functional Cullin1, but independent of Cdh1 expression. Further studies have demonstrated that FKA docked into the ATP binding pocket of the precursor cell-expressed developmentally down-regulated 8 (NEDD8)-activating enzyme (NAE) complex, inhibited NEDD8 conjugations to both Cullin1 and Ubc12 in PC3 cells and Ubc12 NEDDylation in an in vitro assay. Finally, dietary feeding of the autochthonous transgenic adenocarcinoma of the mouse prostate (TRAMP) mice with FKA inhibited the formation of high-grade prostatic intra-epithelial neoplasia lesions (HG-PIN) and prostate adenocarcinomas, reduced the tumor burden and completely abolished distant organ metastasis. Immunohistochemistry studies revealed that dietary FKA feeding resulted in marked anti-proliferative and apoptotic effects via down-regulation of Skp2 and NEDD8 and up-regulation of p27/Kip1 in the prostate of TRAMP mice. Our findings therefore provide evidence that FKA is a promising NEDDylation inhibitor for targeting Skp2 degradation in prostate cancer prevention and treatment.

Increasing evidence has indicated that Wnt signaling plays complex roles in castration resistant prostate cancer (CRPC). Although not all data were consistent, β-catenin nuclear localization and its co-localization with androgen receptor (AR) were more frequently observed in CRPC compared to hormone naïve prostate cancer. This direct interaction between AR and β-catenin seemed to elicit a specific expression of a set of target genes in low androgen conditions in CRPC. Paracrine Wnt signaling also was shown to aid resistance to chemotherapy and androgen deprivation therapy. Results from the next generation sequencing studies (i.e. RNA-seq and whole exosome sequcing) of CRPC specimens have identified the Wnt pathway as one of the top signaling pathways with significant genomic alterations in CRPC, whereas, Wnt pathway alterations were virtually absent in hormone naïve primary prostate cancer. Furthermore, Wnt signaling has been suggested to play an important role in cancer stem cell functions in prostate cancer recurrence and resistance to androgen deprivation therapy. Therefore, in this review we have summarized existing knowledge regarding potential roles of Wnt signaling in CRPC and underline Wnt signaling as a potential therapeutic target for CRPC. Further understanding of Wnt signaling in castration resistance may eventually contribute new insights into possible treatment options for this incurable disease.

Abstract Background Lysophosphatidic acid (LPA), generated by autotaxin (ATX), is a bioactive lipid mediator that binds to the receptors (LPA1–6), and serves as an important mediator in inflammation. Previous studies have demonstrated that LPA-LPA1 cascade contributes to arthritis and skin sclerosis. In this study, we examined the role of LPA signals in murine Candida albicans water-soluble fraction (CAWS)-induced vasculitis. Methods ATX and LPA receptor expressions were analyzed by immunohistochemistry and quantitative reverse transcription-polymerase chain reaction. Effects of LPA1 inhibition on CAWS-induced vasculitis were evaluated in LPA1-deficient mice or using an LPA1 antagonist, LA-01. Migration activity was assessed using a chemotaxis chamber. The number of migrated fluorescently labeled neutrophils, which were transferred into the vasculitis mice, was counted in the aortic wall. CXCL1 and IL-8 concentrations were determined by enzyme-linked immunosorbent assay. Results ATX and LPA1 were highly expressed in the inflamed region of CAWS-induced vasculitis. Severity of the vasculitis in LPA1-deficient mice was suppressed. The LPA1 antagonist, LA-01, also ameliorated the CAWS-induced vasculitis. LPA induced neutrophil migration, which was inhibited by LA-01 in vitro. Infiltration of transferred neutrophils from LPA1-deficient mice into the coronary arteries was suppressed. LA-01 also inhibited the infiltration of wild-type neutrophils. Expression of CXCL1 and IL-8 in human endothelial cells was enhanced by LPA, but was inhibited by LA-01. ATX and LPA1 expression levels were higher in the affected skin region of vasculitis patients than in healthy controls. Conclusions These results suggest that LPA-LPA1 signaling contributes to the development of vasculitis via chemoattractant production from endothelial cells followed by neutrophil recruitment. Thus, LPA1 has potential as a novel target for vasculitis therapies.

Diabetes is a major cause of kidney disease. Here Kikuchi et al. show that phenol sulfate, a gut microbiota-derived metabolite, is increased in diabetic kidney disease and contributes to the pathology by promoting kidney injury, suggesting phenyl sulfate could be used a marker and therapeutic target for the treatment of diabetic kidney disease.

Introduction Kawasaki disease (KD), first described as an acute, febrile, mucocutaneous lymph node syndrome in 1967 (1), is the most common systemic vasculitis of children and infants. KD is recognized [...], Environmental triggers, including those from pathogens, are thought to play an important role in triggering autoimmune diseases, such as vasculitis, in genetically susceptible individuals. The mechanism by which activation of the innate immune system contributes to vessel-specific autoimmunity in vasculitis is not known. Systemic administration of Candida albicans water-soluble extract (CAWS) induces vasculitis in the aortic root and coronary arteries of mice that mimics human Kawasaki disease. We found that Dectin-2 signaling in macrophages resident in the aortic root of the heart induced early CCL2 production and the initial recruitment of [CCR2.sup.+] inflammatory monocytes (iMos) into the aortic root and coronary arteries. iMos differentiated into monocyte-derived dendritic cells (Mo-DCs) in the vessel wall and were induced to release IL-1[beta] in a Dectin-2/Syk/NLRP3 inflammasome-dependent pathway. IL-1[beta] then activated cardiac endothelial cells to express CXCL1 and CCL2 and adhesion molecules that induced neutrophil and further iMo recruitment and accumulation in the aortic root and coronary arteries. Our findings demonstrate that Dectin-2-mediated induction of CCL2 production by macrophages resident in the aortic root and coronary arteries initiates vascular inflammation in a model of Kawasaki disease, suggesting an important role for the innate immune system in initiating vasculitis.

Lymphoid enhancer factor 1 (LEF-1) mediates Wnt signaling via recruitment of β-catenin to target genes. The LEF1 gene is aberrantly transcribed in colon cancers because promoter 1 (P1) is a Wnt target gene and is activated by TCF–β-catenin complexes. A second promoter in intron 2 (P2) produces dominant negative LEF-1 isoforms (dnLEF-1), but P2 is silent because it is repressed by an upstream distal repressor element. In this study we identify Yin Yang 1 (YY1) transcription factor as the P2-specific factor necessary for repression. Site-directed mutagenesis and EMSA were used to identify a YY1-binding site at +25 in P2, and chromatin immunoprecipitation assays detected YY1 binding to endogenous LEF1 P2. Mutation of this site relieves P2 repression in transient transfections, and knockdown of endogenous YY1 relieves repression of integrated P2 reporter constructs and decreases the H3K9me3 epigenetic marks. YY1 is responsible for repressor specificity because introduction of a single YY1-binding site into the P1 promoter makes it sensitive to the distal repressor. We also show that induced expression of dnLEF-1 in colon cancer cells slows their rate of proliferation. We propose that YY1 plays an important role in preventing dnLEF-1 expression and growth inhibition in colon cancer.

Wnt regulation of gene expression requires binding of LEF/T-cell factor (LEF/TCF) transcription factors to Wnt response elements (WREs) and recruitment of the activator beta-catenin. There are significant differences in the abilities of LEF/TCF family members to regulate Wnt target genes. For example, alternatively spliced isoforms of TCF-1 and TCF-4 with a C-terminal "E" tail are uniquely potent in their activation of LEF1 and CDX1. Here we report that the mechanism responsible for this unique activity is an auxiliary 30-amino-acid DNA interaction motif referred to here as the "cysteine clamp" (or C-clamp). The C-clamp contains invariant cysteine, aromatic, and basic residues, and surface plasmon resonance (SPR) studies with recombinant C-clamp protein showed that it binds double-stranded DNA but not single-stranded DNA or RNA (equilibrium dissociation constant = 16 nM). CASTing (Cyclic Amplification and Selection of Targets) experiments were used to test whether this motif influences WRE recognition. Full-length LEF-1, TCF-1E, and TCF-1E with a mutated C-clamp all bind nearly identical WREs (TYYCTTTGATSTT), showing that the C-clamp does not alter WRE specificity. However, a GC element downstream of the WRE (RCCG) is enriched in wild-type TCF-1E binding sites but not in mutant TCF-1E binding sites. We conclude that the C-clamp is a sequence-specific DNA binding motif. C-clamp mutations destroy the ability of beta-catenin to regulate the LEF1 promoter, and they severely impair the ability of TCF-1 to regulate growth in colon cancer cells. Thus, E-tail isoforms of TCFs utilize two DNA binding activities to access a subset of Wnt targets important for cell growth.

To simulate the current climate, a 20-year integration of a non-hydrostatic regional climate model (NHRCM) with grid spacing of 5 and 2 km (NHRCM05 and NHRCM02, respectively) was nested within the AGCM. The three models did a similarly good job of simulating surface air temperature, and the spatial horizontal resolution did not affect these statistics. NHRCM02 did a good job of reproducing seasonal variations in surface air temperature. NHRCM05 overestimated annual mean precipitation in the western part of Panama and eastern part of the Pacific Ocean. NHRCM05 is responsible for this overestimation because it is not seen in MRI-AGCM. NHRCM02 simulated annual mean precipitation better than NHRCM05, probably due to a convection-permitting model without a convection scheme, such as the Kain and Fritsch scheme. Therefore, the finer horizontal resolution of NHRCM02 did a better job of replicating the current climatological mean geographical distributions and seasonal changes of surface air temperature and precipitation.

The antitumour activity and immunomodulating activity of synthesized lignin-like polyphenols in mice were determined. Polymerized polyphenols were prepared using caffeic acid as the precursor. The intake of both polymeric and monomeric caffeic acid inhibited the growth of B7-P815 tumours in mice, but we found that only the monomers inhibited tumour growth in vitro. Oral administration of polymerized caffeic acid, but not monomeric caffeic acid, substantially increased interferon-γ production in splenocytes stimulated with phorbol 12-myristate 13-acetate and ionomycin. Moreover, natural killer activity was markedly upregulated in mice that were administered polymerized caffeic acid compared with mice administered water or caffeic acid monomers. Lastly, we determined that caffeic acid monomers were absorbed more easily than polymers, but ingested polymers were less susceptible to degradation into monomers by digesting enzymes. Taken together, our results suggest that polymerized caffeic acid administered orally can upregulate systemic immunity and inhibit tumour cell growth.