Creation of bladder assembloids mimicking tissue regeneration and cancer.

Current organoid models are limited by their inability to mimic mature organ architecture and associated tissue microenvironments1,2. Here we create multilayer bladder 'assembloids' by reconstituting tissue stem cells with stromal components to represent an organized architecture with an epithelium surrounding stroma and an outer muscle layer. These assembloids exhibit characteristics of mature adult bladders in cell composition and gene expression at the single-cell transcriptome level, and recapitulate in vivo tissue dynamics of regenerative responses to injury. We also develop malignant counterpart tumour assembloids to recapitulate the in vivo pathophysiological features of urothelial carcinoma. Using the genetically manipulated tumour-assembloid platform, we identify tumoural FOXA1, induced by stromal bone morphogenetic protein (BMP), as a master pioneer factor that drives enhancer reprogramming for the determination of tumour phenotype, suggesting the importance of the FOXA1-BMP-hedgehog signalling feedback axis between tumour and stroma in the control of tumour plasticity.

Forkhead box protein D2 suppresses colorectal cancer by reprogramming enhancer interactions.

Somatic stem cells contribute to normal tissue homeostasis, and their epigenomic features play an important role in regulating tissue identities or developing disease states. Enhancers are one of the key players controlling chromatin context-specific gene expression in a spatial and temporal manner while maintaining tissue homeostasis, and their dysregulation leads to tumorigenesis. Here, epigenomic and transcriptomic analyses reveal that forkhead box protein D2 (FOXD2) is a hub for the gene regulatory network exclusive to large intestinal stem cells, and its overexpression plays a significant role in colon cancer regression. FOXD2 is positioned at the closed chromatin and facilitates mixed-lineage leukemia protein-4 (MLL4/KMT2D) binding to deposit H3K4 monomethylation. De novo FOXD2-mediated chromatin interactions rewire the regulation of p53-responsive genes and induction of apoptosis. Taken together, our findings illustrate the novel mechanistic details of FOXD2 in suppressing colorectal cancer growth and suggest its function as a chromatin-tuning factor and a potential therapeutic target for colorectal cancer.

hnRNP K supports the maintenance of RORγ circadian rhythm through ERK signaling.

Retinoic acid-related orphan receptor γ (RORγ) maintains the circadian rhythms of its downstream genes. However, the mechanism behind the transcriptional activation of RORγ itself remains unclear. Here, we demonstrate that transcription of RORγ is activated by heterogeneous nuclear ribonucleoprotein K (hnRNP K) via the poly(C) motif within its proximal promoter. Interestingly, we confirmed the binding of endogenous hnRNP K within RORγ1 and RORγ2 promoter along with the recruitment of RNA polymerase 2 through chromatin immunoprecipitation (ChIP). Furthermore, an assay for transposase accessible chromatin (ATAC)-qPCR showed that hnRNP K induced higher chromatin accessibility within the RORγ1 and RORγ2 promoter. Then we found that the knockdown of hnRNP K lowers RORγ mRNA oscillation amplitude in both RORγ and RORγ-dependent metabolic genes. Moreover, we demonstrated that time-dependent extracellular signal-regulated kinase (ERK) activation controls mRNA oscillation of RORγ and RORγ-dependent metabolic genes through hnRNP K. Taken together, our results provide new insight into the regulation of RORγ by hnRNP K as a transcriptional activator, along with its physiological significance in metabolism.

Factors associated with quality of life among family caregivers of terminally ill cancer patients.

OBJECTIVE: Limited research has examined the quality of life (QOL) and its correlates among family caregivers (FCs) during the final stage of terminal cancer. The purpose of this study was to investigate the determinants of overall QOL and its subdomains among Korean FCs at the very end of life. METHODS: For this cross-sectional study, we enrolled 299 FCs of terminal cancer patients from seven palliative care units. To assess FCs' QOL and its predictors, we used the Caregiver Quality Of Life Index-Cancer, which contains four domains. Possible determinants of caregiver QOL were categorized into patient, caregiver, and environmental factors. A multiple regression model was used to identify factors associated with FCs' QOL. RESULTS: Variance in each Caregiver Quality Of Life Index-Cancer domain was explained by different factors. FCs of younger patient felt more burden but were more likely to adapt positively. Emotional distress of FCs was strongly associated with total QOL, burdensomeness, and disruptiveness. Positive adaptation was related to more visits for care, FCs' religiousness, more social support, and satisfactory perceived quality of care. Financial concerns were more likely in married FCs, FCs with less social support, or low incomes. CONCLUSION: Emotional distress of FCs was the most important factor determining the overall and negative aspects of FCs' QOL, whereas various environmental factors were associated with positive coping. Appropriate support programs directed at these factors are needed to maintain and improve FCs' QOL.

Quality of life discordance between terminal cancer patients and family caregivers: a multicenter study.

BACKGROUND: Research studies on quality of life (QOL) discordance between cancer patients and family caregivers are limited, and the results are inconsistent. The objective of this study was to examine QOL discordance between patients and family caregivers in a hospice setting and to identify factors associated with the discordance. METHODS: We enrolled 178 patient-family caregiver pairs from six tertiary hospital hospice palliative care units in South Korea in this cross-sectional study. To establish groupings based on patient and family caregiver QOL levels, we measured the QOL of patient and family caregiver pairs using the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire Core 15 for Palliative Care and the Caregiver QOL Index-Cancer, respectively. Pairs were categorized into the following three groups: both good QOL pairs, only poor patient QOL, and only poor family caregiver QOL. Factors associated with only poor patient or only poor family caregiver QOL were compared to both good QOL pairs. A stepwise multivariate regression model was used to identify relevant factors. RESULTS: The QOL of family caregivers did not correlate significantly (P = 0.227) with QOL in terminally ill cancer patients. As well, poor emotional function in patients was the only significant factor associated with the only poor patient QOL group [adjusted odds ratio (aOR), 4.1; 95 % confidence interval (CI), 1.5-11.5]. However, emotionally distressed family caregivers (aOR, 10.2; 95 % CI, 2.8-37.5), family caregivers who professed a religion (aOR, 4.1; 95 % CI, 1.5-11.3), and family caregivers with low social support (aOR, 3.9; 95 % CI, 1.5-10.6) were independent predictors for the only poor family caregiver QOL group. CONCLUSIONS: Assessing the respective emotional status of both the patient and family caregiver is needed in hospice care to reduce the gap in QOL between the two groups. Further, more attention should be paid to the lack of social support for family caregivers.

Predictors for outcome of paper patch myringoplasty in patients with chronic tympanic membrane perforations.

The purpose of the present study is to evaluate the outcome of paper patch myringoplasty for chronic tympanic membrane (TM) perforations and to explore the predictive factors for a successful closure. A retrospective study was performed in a tertiary referral center. Data of the patients who met the inclusion criteria were analyzed: the treatment outcomes and the potential predictive factors including age, sex, the affected ear, hearing level, duration of perforation, causes, location and size of perforations, relationship between the perforation border and the malleus, status of TM surface, and the number of patch applications. Complete closure was achieved in 27 of the total 43 subjects. Among the 11 clinical and TM factors, only the perforation size remained significant as the predictor after multivariable logistic regression (p = 0.029, OR 4.4). The patients with perforation ≤ 5% of the TM showed higher closure rate (78.3%) than those with perforation >5% (45.0%). In conclusion, paper patch myringoplasty showed overall success rate of 62.8%. In patients with perforations smaller than 5% of the TM, the closure rate was 78.3%. The predictor of the treatment outcome was the perforation size. We can try paper patch myringoplasty first in patients who had dry chronic perforations smaller than 5% of the TM without middle ear disease.

Dietary magnesium intake and risk of cancer: a meta-analysis of epidemiologic studies.

The aim of this study was to investigate the association between dietary magnesium and the risk of overall cancer using a meta-analysis. We searched PubMed, SCOPUS, and the Cochrane Review through November 2012. All the articles searched were independently reviewed by 3 authors based on predetermined selection criterion. A total of 13 epidemiologic studies, 6 case-control studies, and 7 prospective cohort studies involving 1,236,004 participants were included in the final analysis. When all studies were pooled, the relative risk (RR) of overall cancer for the highest level of dietary magnesium intake was 0.801 [95% confidence interval (CI): 0.664-0.966) compared with the lowest level of dietary magnesium intake. In subgroup meta-analyses by study design, there was a significant inverse association between dietary magnesium and the risk of cancer in case-control studies (RR = 0.663, 95% CI: 0.475-0.925), whereas there was no significant association in prospective cohort studies (RR = 0.888, 95% CI: 0.745-1.060). Furthermore, there was a significant preventive effect of dietary magnesium for colorectal cancer (RR = 0.775, 95% CI: 0.655-0.919), but not for other cancer. Our meta-analysis showed that higher dietary magnesium intake seems to have a protective effect for cancer, especially colorectal cancer and in females.

The survival time of terminal cancer patients: prediction based on clinical parameters and simple prognostic scores.

AIM: By examining clinical parameters associated with survival time and analyzing patients' survival times using prognostic scores, this study aimed to provide helpful information related to the treatment of terminal cancer patients. METHODS: We retrospectively reviewed the medical records of 415 inpatients who died in the hospices of two hospitals from March 2009 to August 2011 then analyzed differences in survival times and relative risk for clinical parameters and prognostic scores. RESULTS: There were 15 parameters associated with survival time. Performance decline was the most influential factor. The optimal scores for predicting four-week survival were over 4.5 on the Palliative Prognostic Index (PPI), over 10 on the Palliative Prognostic (PaP) Score, and 30 or under on the Palliative Performance Scale (PPS). CONCLUSION: Performance decline is a major factor affecting survival time. The PaP is the most useful tool for predicting four-week survival, with an optimal value of over 10.

The effects of fucoidan-rich polysaccharides extracted from Sargassum horneri on enhancing collagen-related skin barrier function as a potential cosmetic product.

BACKGROUND: Sargassum horneri came ashore after flowing from the South China Sea to Jeju Island a few years ago. This caused a significant environmental impact on coastal areas where S. horneri has accumulated because of decomposition and the release of toxic substances, such as hydrogen sulfide. AIMS: In this study, we evaluated a biological ingredient prepared from fucoidan-rich S. horneri and demonstrated its antiwrinkle effects on ultraviolet B (UVB)-induced fibroblast cells. MATERIALS AND METHODS: Fucoidan samples from S. horneri were prepared according to a previously published process with modifications. The compositional analysis of S. horneri fucoidan extract (SHFE) as well as its effects on antiaging were examined to determine its utility as a functional material. RESULTS: SHFE exhibited antioxidant properties using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay. Treatment of UVB-induced fibroblasts with SHFE significantly increased the synthesis of procollagen compared with adenosine treatment and inhibited MMP-1 and MMP-3 expression. In a clinical study, SHFE lotion improved skin barrier effects in forearms and transepidermal water loss (TEWL) values were reduced after 3 weeks of use compared with a placebo. CONCLUSION: SHFE has utility as an additive with functional antiaging effects for a range of cosmetic products as it restores skin hydration in the epidermal barrier.

Glycation-mediated tissue-level remodeling of brain meningeal membrane by aging.

Collagen is a prominent target of nonenzymatic glycation, which is a hallmark of aging and causes functional alteration of the matrix. Here, we uncover glycation-mediated structural and functional changes in the collagen-enriched meningeal membrane of the human and mouse brain. Using an in vitro culture platform mimicking the meningeal membrane composed of fibrillar collagen, we showed that the accumulation of advanced glycation end products (AGEs) in the collagen membrane is responsible for glycation-mediated matrix remodeling. These changes influence fibroblast-matrix interactions, inducing cell-mediated ECM remodeling. The adherence of meningeal fibroblasts to the glycated collagen membrane was mediated by the discoidin domain-containing receptor 2 (DDR2), whereas integrin-mediated adhesion was inhibited. A-kinase anchoring protein 12 (AKAP12)-positive meningeal fibroblasts in the meningeal membrane of aged mice exhibited substantially increased expression of DDR2 and depletion of integrin beta-1 (ITGB1). In the glycated collagen membrane, meningeal fibroblasts increased the expression of matrix metalloproteinase 14 (MMP14) and less tissue inhibitor of metalloproteinase-1 (TIMP1). In contrast, the cells exhibited decreased expression of type I collagen (COL1A1). These results suggest that glycation modification by meningeal fibroblasts is intimately linked to aging-related structural and functional alterations in the meningeal membrane.

Immature Citrus sunki peel extract exhibits antiobesity effects by ß-oxidation and lipolysis in high-fat diet-induced obese mice.

The peel of Citrus sunki HORT. ex TANAKA has been widely used in traditional Asian medicine for the treatment of many diseases, including indigestion and bronchial asthma. In this study, we investigated the antiobesity activity of immature C. sunki peel extract (designated CSE) using high-fat diet (HFD)-induced obese C57BL/6 mice and mature 3T3-L1 adipocytes. In the animal study, body weight gain, adipose tissue weight, serum total cholesterol, and triglyceride in the CSE-administered group decreased significantly compared to the HFD group. Also, CSE supplementation reduced serum levels of glutamic pyruvic transaminase, glutamic oxaloacetic transaminase, and lactate dehydrogenase. Moreover, it significantly decreased the accumulation of fatty droplets in liver tissue, suggesting a protective effect against HFD-induced hepatic steatosis. Dietary supplementation with CSE reversed the HFD-induced decrease in the phosphorylation levels of AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC), which are related to fatty acid ß-oxidation, in the epididymal adipose tissue. Also, CSE increased AMPK and ACC phosphorylation in mature 3T3-L1 adipocytes. CSE also enhanced lipolysis by phosphorylation of cAMP-dependent protein kinase (PKA) and hormone-sensitive lipase (HSL) in mature 3T3-L1 adipocytes. These results suggest that CSE had an antiobesity effect via elevated ß-oxidation and lipolysis in adipose tissue.

Anti-obesity properties of a Sasa quelpaertensis extract in high-fat diet-induced obese mice.

This study explores the anti-obesity properties of a Sasa quelpaertensis leaf extract (SQE) in high-fat diet (HFD)-induced obese C57BL/6 mice and mature 3T3-L1 adipocytes. SQE administration with HFD for 70 d significantly decreased the body weight gain, adipose tissue weight, and serum total cholesterol and triglyceride levels in comparison with the HFD group. SQE administration also reduced the serum levels of glutamic oxaloacetic transaminase, glutamic pyruvic transaminase and lactate dehydrogenase, and the accumulation of lipid droplets in the liver, suggesting a protective effect against HFD-induced hepatic steatosis. SQE administration restored the HFD-induced decreases with phosphorylation of AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC) in epididymal adipose tissue. SQE also induced AMPK phosphorylation in mature 3T3-L1 adipocytes. These results suggest that SQE exerted an anti-obesity effect on HFD-induced obese mice by activating AMPK in adipose tissue and reducing lipid droplet accumulation in the liver.

Fucoxanthin exerts differing effects on 3T3-L1 cells according to differentiation stage and inhibits glucose uptake in mature adipocytes.

Progression of 3T3-L1 preadipocyte differentiation is divided into early (days 0-2, D0-D2), intermediate (days 2-4, D2-D4), and late stages (day 4 onwards, D4-). In this study, we investigated the effects of fucoxanthin, isolated from the edible brown seaweed Petalonia binghamiae, on adipogenesis during the three differentiation stages of 3T3-L1 preadipocytes. When fucoxanthin was applied during the early stage of differentiation (D0-D2), it promoted 3T3-L1 adipocyte differentiation, as evidenced by increased triglyceride accumulation. At the molecular level, fucoxanthin increased protein expression of peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer-binding protein α (C/EBPα), sterol regulatory element-binding protein 1c (SREBP1c), and aP2, and adiponectin mRNA expression, in a dose-dependent manner. However, it reduced the expression of PPARγ, C/EBPα, and SREBP1c during the intermediate (D2-D4) and late stages (D4-D7) of differentiation. It also inhibited the uptake of glucose in mature 3T3-L1 adipocytes by reducing the phosphorylation of insulin receptor substrate 1 (IRS-1). These results suggest that fucoxanthin exerts differing effects on 3T3-L1 cells of different differentiation stages and inhibits glucose uptake in mature adipocytes.

Stability of Quantum-Dot Light Emitting Diodes with Alkali Metal Carbonates Blending in Mg Doped ZnO Electron Transport Layer.

We report here the fabrication of highly efficient and long-lasting quantum-dot light emitting diodes (QLEDs) by blending various alkali metal carbonate in magnesium (Mg) doped zinc oxide (ZnO) (MZO) electron transport layer (ETL). Alkali metal carbonates blending in MZO, X2CO3:MZO, control the band-gap, electrical properties, and thermal stability. This can therefore enhance the operational lifetime of QLEDs. It is found that the conductivity of X2CO3:MZO film can be controlled and the thermal stability of ETLs could be improved by X2CO3 blending in MZO. The inverted red QLEDs (R-QLEDs) with Cs2CO3:MZO, Rb2CO3:MZO, and K2CO3:MZO ETLs exhibited the operational lifetime of 407 h for the R-QLEDs with Cs2CO3:MZO, 620 h with Rb2CO3:MZO and 94 h with K2CO3:MZO ETLs at T95 with the initial luminance of 1000 cd/m2. Note that all red QLEDs showed the high brightness over 150,000 cd/m2. But the R-QLEDs with Na2CO3:MZO and Li2CO3:MZO ETLs exhibited shorter operational lifetime and poor brightness than the R-QLED with pristine MZO ETL.

Fluid-Matrix Interface Triggers a Heterogeneous Activation of Macrophages.

Spontaneous activation of macrophages in response to inflammation is a key part of innate immunity and host defense. Macrophages represent a heterogeneous population of cells with different phenotypic profiles performing distinct functions in host defense. Although a spectrum of macrophage activation stages exists in an inflamed region, the effect of local physical conditions on the heterotypic activation of macrophages is unknown. Here, we introduce an in vivo fluid-matrix interface analogous culture platform, an asymmetric microenvironment, facilitating the formation of macrophage aggregates (MAs). Macrophages were self-assembled to form MAs of ∼100 µm diameter at the collagen matrix-medium interface upon phorbol-12-myristate-13-acetate treatment. The macrophages within the half-embedded MAs into the matrix were heterogeneously activated, resulting in inhomogeneous cell-cell and cell-matrix interactions within the aggregates. Our demonstration may aid in a better understanding of the acquisition of macrophage heterogeneity in response to tissue-specific microenvironments.

Astrocyte-Encapsulated Hydrogel Microfibers Enhance Neuronal Circuit Generation.

Astrocytes, the most representative glial cells in the brain, play a multitude of crucial functions for proper neuronal development and synaptic-network formation, including neuroprotection as well as physical and chemical support. However, little attention has been paid, in the neuroregenerative medicine and related fields, to the cytoprotective incorporation of astrocytes into neuron-culture scaffolds and full-fledged functional utilization of encapsulated astrocytes for controlled neuronal development. In this article, a 3D neurosupportive culture system for enhanced induction of neuronal circuit generation is reported, where astrocytes are confined in hydrogel microfibers and protected from the outside. The astrocyte-encapsulated microfibers significantly accelerate the neurite outgrowth and guide its directionality, and enhance the synaptic formation, without any physical contact with the neurons. This astrocyte-laden system provides a pivotal culture scaffold for advanced development of cell-based therapeutics for neural injuries, such as spinal cord injury.

hnRNP K Supports High-Amplitude D Site-Binding Protein mRNA (Dbp mRNA) Oscillation To Sustain Circadian Rhythms.

Circadian gene expression is defined by the gene-specific phase and amplitude of daily oscillations in mRNA and protein levels. D site-binding protein mRNA (Dbp mRNA) shows high-amplitude oscillation; however, the underlying mechanism remains elusive. Here, we demonstrate that heterogeneous nuclear ribonucleoprotein K (hnRNP K) is a key regulator that activates Dbp transcription via the poly(C) motif within its proximal promoter. Biochemical analyses identified hnRNP K as a specific protein that directly associates with the poly(C) motif in vitro Interestingly, we further confirmed the rhythmic binding of endogenous hnRNP K within the Dbp promoter through chromatin immunoprecipitation as well as the cycling expression of hnRNP K. Finally, knockdown of hnRNP K decreased mRNA oscillation in both Dbp and Dbp-dependent clock genes. Taken together, our results show rhythmic protein expression of hnRNP K and provide new insights into its function as a transcriptional amplifier of Dbp.

The Poly(C) Motif in the Proximal Promoter Region of the D Site-Binding Protein Gene (Dbp) Drives Its High-Amplitude Oscillation.

The D site-binding protein (Dbp) supports the rhythmic transcription of downstream genes, in part by displaying high-amplitude cycling of its own transcripts compared to other circadian-clock genes. However, the underlying mechanism remains elusive. Here, we demonstrated that the poly(C) motif within the Dbp proximal promoter, in addition to an E-box element, provoked transcriptional activation. Furthermore, we generated a cell line with poly(C) deleted to demonstrate the endogenous effect of the poly(C) motif within the Dbp promoter. We investigated whether RNA polymerase 2 (Pol2) recruitment on the Dbp promoter was decreased in the cell line with poly(C) deleted. Next, assay for transposase-accessible chromatin (ATAC)-quantitative PCR (qPCR) showed that the poly(C) motif induced greater chromatin accessibility within the region of the Dbp promoter. Finally, we determined that the oscillation amplitude of endogenous Dbp mRNA of the cell line with poly(C) deleted was decreased, which affected the oscillation of other clock genes that are controlled by Dbp Taken together, our results provide new insights into the function of the poly(C) motif as a novel cis-acting element of Dbp, along with its significance in the regulation of circadian rhythms.

Beneficial Effects of Appropriate Sleep Duration on Depressive Symptoms and Perceived Stress Severity in a Healthy Population in Korea.

BACKGROUND: Recently, several institutions, including the American Academy of Sleep Medicine, the Sleep Research Society, and the National Sleep Foundation, have made consensus recommendations concerning appropriate sleep duration for adults. Although numerous studies conducted in Western populations have provided evidence of the harmful effects of short or long sleep duration on mental health, it is still unclear whether these consensus recommendations are appropriate in Korean culture. METHODS: Data from 1,892 subjects with no history of medical or psychiatric diagnoses were selected from the Korea National Health and Nutrition Examination Survey of 2014. Subjects were divided into seven groups based on their sleep duration (≤4, 5, 6, 7, 8, 9, and ≥10 hours). Depressive symptoms were measured using the Patient Health Questionnaire-9 and perceived stress severity was evaluated using a Likert-type scale. Group differences in depressive symptoms and severity of stress were analyzed using an analysis of covariance. RESULTS: Depressive symptoms in subjects with sleep duration of ≤4 hours (5.7±5.9) or 5 hours (3.4±3.9) were higher than in subjects with a sleep duration of 7 (2.2±2.9) or 8 hours (2.2±2.9) (corrected P<0.05). Furthermore, subjects with a short sleep duration (5 hours or below) had greater perceived stress severity than subjects with a sleep duration of 7 or 8 hours (corrected P<0.05). CONCLUSION: Our results suggest that maintaining an appropriate sleep duration as found in the recent consensus recommendation is important for mental health, even in healthy subjects without any medical or psychiatric illnesses, in Korea.