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This paper discusses the instructional design experiences and processes shared by a multi-disciplinary group—including more than a dozen faculty, staff, and students—while developing a series of online courses on Model-based Systems Engineering (MBSE) for professional engineers, a project sponsored by the National Science Foundation. The team size, the complexity and uniqueness of the subject matter, the targeted learners, and the pre-determined research questions created a rare situation in which the team members collaborated and/or negotiated outside the realm of the traditional instructional design process. Over time the team went through two different types of instructional design processes, beginning with a waterfall-type process where the communication between the subject matter experts (SME) and the design team was somewhat limited and finally evolving to a collaborative process where the interaction between the two teams was more direct and immediate. The evolution of the design process and the dynamics between the SMEs and the design team resulted in several major design revisions implemented to improve the quality of the online courses.

Owing to the rapid climate change, a high-performance energy storage system (ESS) for efficient energy consumption has been receiving considerable attention. ESS, such as capacitors, usually has issues with the ion diffusion of electrode materials, resulting in a decrease in their capacitance. Notably, appropriate pore diameter and large specific surface area (SSA) may result in an effective ion diffusion. Therefore, graphene and multi-walled carbon nanotube (graphene@MWCNT) hybrid nanomaterials, with covalent bonds between the graphene and MWCNT, were prepared via an edge-chemistry reaction. The properties of these materials, such as high porosity, large SSA, and high electroconductivity, make them suitable to be used as electrode materials for capacitors. The optimal ratio of graphene to MWCNT can affect the electrochemical performance of the electrode material based on its physical and electrochemical properties. The supercapacitor using optimal graphene-based hybrid electrode material exhibited highest specific capacitance value as 158 F/g and excellent cycle stability.

Crohn's disease has a substantial negative impact on health-related quality of life (HRQoL).To examine the effects of risankizumab on HRQoL in Crohn's disease METHODS: We analysed data from patients with Crohn's disease from 12-week induction trials ADVANCE (N = 850) and MOTIVATE (N = 569) with risankizumab 600 mg or 1200 mg intravenous (IV) versus placebo IV and a 52-week maintenance trial FORTIFY (N = 462) with risankizumab 180 or 360 mg subcutaneous (SC) versus placebo SC. Outcomes included Inflammatory Bowel Disease Questionnaire (IBDQ), Functional Assessment of Chronic Illness Therapy-Fatigue (FACIT-F), 36-item Short Form Health Survey (SF-36), EuroQol 5-Dimension-5-Level (EQ-5D-5L) and work productivity. The mean change and percentages of patients achieving clinically meaningful improvement in all outcomes were determined at weeks 12 and 52.At week 12, more patients in the risankizumab 600 or 1200 mg groups achieved IBDQ response than with placebo (ADVANCE: 70.2%, 75.5% vs. 47.8%, p ≤ 0.001; MOTIVATE: 61.7%, 68.5% vs. 48.2%, p ≤ 0.01) and FACIT-F response (ADVANCE: 51.3%, 48.0% vs. 35.7%, p ≤ 0.01; MOTIVATE: 44.2%, 49.1% vs. 33.7%, p 0.05). These improvements persisted at week 52 with risankizumab maintenance treatment. Similar trends were observed for SF-36 physical and mental component summary scores, EQ-5D-5L and activity impairment within work productivity measures.Risankizumab induction therapy (600 or 1200 mg IV) led to clinically meaningful improvements in disease-specific and general patient-reported outcomes, including fatigue, in patients with moderate to severe Crohn's disease. These improvements were sustained after 52 weeks of risankizumab (180 or 360 mg SC) maintenance therapy.

The study aimed to identify the factors associated with the 6-min walk distance (6MWD) and to provide reference values for the 6MWD in individuals with schizophrenia (SCZ) in Taiwan.A proportional stratified sampling method was utilized based on distribution of gender, age and body mass index (BMI) at the study hospital. The 6-minute walk test was conducted according to the American Thoracic Society protocol.A total of 237 patients with SCZ completed the 6-minute walk test. The 6MWD was significantly associated with age, height, weight, and length of the onset of SCZ. Stepwise linear regression revealed that height and age were significant determinants of 6MWD. The reference values for males and females at different age groups were determined. Notably, females over 60 walked substantially shorter than the age younger than 60.Height and age were the main predictors for 6MWD among people with SCZ in Taiwan. The established reference values can be used to identify those at risk of poor cardiorespiratory fitness and as a target outcome during exercise programs in psychiatric rehabilitation. Our results highlight that older females with SCZ may be a priority group to target with exercise interventions to mitigate the faster decline in cardiorespiratory fitness.IMPLICATIONS FOR REHABILITATIONHeight and age were predictors of 6-min walk distance (6MWD) in schizophrenia (SCZ).The established age- and gender reference values for the 6MWD can be used to identify those at risk of poor cardiorespiratory fitness.Females with SCZ over age 60 may be a priority group to target with exercise interventions to mitigate the faster decline in cardiorespiratory fitness.

PurposeGiven the importance of third-party endorsements as external cues during purchase evaluations, the manipulation of endorsements may raise consumers’ suspicion and thereby reduce the effectiveness of marketing messages. Consumers find it increasingly difficult to distinguish between authentic and biased brand reviews. This research conducts two experiments regarding brand rankings of third-parties in four industries. Study 1 aims to probes the interaction effect of persuasion knowledge and fairness of rating method (fair vs. unfair) on brand rankings’ credibility and brand image enhancement. Study 2 examines the interplay of dispositional persuasion knowledge and reputation of third parties (high vs. low).Design/methodology/approachThis research builds on an analysis of data (Study 1, N = 122; Study 2, N = 123) from Taiwan, where brand rankings held by third-party organizations are regular. The hypotheses were tested using SPSS’s PROCESS macro.FindingsDrawing on previous research of persuasion knowledge, the authors find that a fair rating method and a high reputation of third-party organization can mitigate the negative effect of persuasion knowledge.Practical implicationsThe results of this research suggest that marketers need to manage third-party endorsements carefully. Consumer skepticism toward these endorsements (e.g. third-party’s brand rankings) can be weakened when persuasion attempt is considered appropriate.Originality/valueThis is one of the first efforts to empirically explore the influences of persuasion knowledge on third-party endorsements. The importance of this work is underscored by the fact that a growing number of third-party endorsements are sponsored or even manipulated by brand owners.

Low molecular weight heparin (LMWH) has been given to reproductive-age women with various indications. This study aims to assess the benefits and risks of such use.We retrospectively reviewed data (n = 204) between Jan 2016 and May 2019. Logistic regression analysis was conducted to evaluate the correlation between indications and reproductive outcomes.LMWH use had higher odds of live birth in women less than 30 years of age (OR: 4.98; 95% CI = 1.13-21.98; p = 0.034) and with protein S deficiency (OR: 3.90; 95% CI = 1.77-8.59; p = 0.001). For the subgroup of recurrent pregnant loss, LMWH use was only advantageous to women with protein S deficiency (OR: 2.45; 95%:1.01-5.97; p = 0.048). Risks such as preterm delivery, small-for-gestational-age, placental abruption, antepartum/postpartum hemorrhage were not significantly increased among subgroups. Women treated with LMWH and who had successful live births (n = 171) had a slightly increased risk of postpartum hemorrhage compared to controls (n = 8058) during this period in our institution (2.9% vs 1.2%, p 0.001).LMWH administration produces a higher chance of live-birth to women younger than 30 years of age or with protein S deficiency. However, risk of postpartum hemorrhage is increased.

Adult T-cell leukemia/lymphoma (ATL) is a genetically complex hematological malignancy derived from mature T cells. Using an integrative approach, we previously identified genes recurrently associated with super-enhancers in ATL. One of those genes was TP73, a TP53 family gene; however, the roles and function of TP73 and its super-enhancer in ATL pathogenesis are poorly understood. Our study demonstrates that TP73 is highly activated under the control of a super-enhancer in ATL cells but not in normal T cells or other hematological malignancies examined. Full-length TP73 is required for ATL cell maintenance in vitro and in vivo via the regulation of cell proliferation and DNA damage response pathways. Notably, recurrent deletions of TP73 exons 2-3 were observed in a fraction of primary ATL cases that harbored the super-enhancer, while induction of this deletion in cell lines further increased proliferation and mutational burden. Our study suggests that formation of the TP73 intragenic super-enhancer and genetic deletion are likely sequentially acquired in relation to intracellular state of ATL cells, which leads to functional alteration of TP73 that confers additional clonal advantage.

Social jetlag, the discrepancy between social and biological timing, has been suggested to disturb metabolic functions. However, the relationship between social jetlag and obesity has been inconsistent in other studies. In this study we examined the association between social jetlag and obesity among day and shift workers. We invited 2508 day workers and 1383 shift workers from a hospital worker health cohort to participate in a 2018-2019 survey on their sleep behaviors. Shift-specific social jetlag was quantified using the Munich ChronoType Questionnaire, and body mass index was measured during annual physical examinations. The distributions of shift-specific social jetlag were illustrated, and logistic regression analysis was used to examine the association between social jetlag and obesity. We found that high level of social jetlag (odds ratio [OR] = 1.26, 95% confidence interval [CI] = 1.08-1.47) and positive social jetlag (OR = 2.25, 95% CI = 1.30-3.90) during evening shifts were associated with obesity after adjustment for age, sex, health behaviors, and sleep quality. During night shift periods, sleep time varied greatly on free days, but the participants slept at similar times, namely 16:00, on workdays. In conclusion, phase advance on workdays and high levels of social jetlag were associated with obesity. Sleep timing should therefore be recommended according to the relative phase of individuals' preferred sleep time and work time.

To investigate the relationship between abnormal glucose tolerance during pregnancy and maternal-fetal outcomes, after categorizing women into groups with different levels of gestational abnormal glucose tolerance.A total of 1858 pregnant women who received two-step screening for gestational diabetes mellitus (GDM) at Fooyin University Hospital were categorized into four groups, according to their glucose abnormalities, and their maternal-fetal outcomes were investigated from October 2015 to June 2020.Among the groups having different levels of abnormal glucose tolerance, there were significant differences and trends in mother's age, currently married status, and prepregnancy overweight or obesity, incidences of cesarean section, preterm, and gestational hypertension or preeclampsia; and with respect to neonatal incidence of large for gestational age (LGA), average weight, and average height (p 0.05). After adjusting for potential factors, there were higher incidences of cesarean section (AOR = 1.71; 95% confidence interval [CI]: 1.12-2.61), preterm (AOR = 2.20; 95% CI: 1.23-3.91), neonatal LGA (AOR = 4.94; 95% CI: 2.87-8.51), and neonatal intensive care unit (NICU) admission (AOR = 2.66; 95% CI: 1.14-6.24) in the GDM group, relative to the control group. Furthermore, the women in the oral glucose tolerance test (OGTT)-1 group had a higher incidence of neonatal LGA when compared with the women in the normal group (AOR = 2.31; 95% CI: 1.02-5.33).We found higher incidences of cesarean section, preterm, and neonatal LGA and NICU admission in the GDM women, and a higher incidence of neonatal LGA in the OGTT-1 group, relative to control group.

PurposeFood packaging pictures are one of the most important extrinsic cues for consumers to evaluate food products before purchasing. Over the past decades, marketers have used exaggerated pictures to attract consumers' attention, enhance their attitude toward a product and increase their purchase intention. This study examined the interplay of “puff-up” product picture, food type and picture type in influencing consumers' responses via persuasion knowledge.Design/methodology/approachThe research comprises two 2 × 2 factorial experiments. Study 1 tested the interaction effect of puffery (high vs low) and food type (utilitarian vs. hedonic) on consumers' responses using two fictitious brands of prepared food, whereas Study 2 tested the interaction effect of puffery (high vs. low) and picture type (ingredients vs. cooked food) using a fictitious brand of Chinese delicacy.FindingsResults demonstrated that the degree of picture puffery did not influence consumers' responses to utilitarian food and ingredient image. Conversely, consumers were sensitive to puffery when they see hedonic food and cooked-food image. Our findings also suggested that consumers' persuasion knowledge mediates the relationship between puffery and their responses.Practical implicationsThe presented findings facilitate marketers to know consumers' attitude about food puffery pictures.Originality/valueThis research is one of the first efforts to empirically explore the influences of persuasion knowledge on food puffery pictures. The importance of this work is underscored by the fact that a growing number of visual exaggerations are adopted on food packaging.

PurposeIn recent years, there is a steady increase of studies documenting the emergence of “contracting back-in” in many countries around the world, that is, governments bringing previously contracted services back in-house and once again relying on public employees to deliver these services. Through a survey of the academic discourse on contracting back-in, this manuscript describes the phenomenon of contracting back-in and discusses its implications for “bringing the state back in”.Design/methodology/approachThe manuscript conducts a systematic review of existing literature on contracting back-in with a focus on the scope and rationale of contracting back-in.FindingsThe existing literature demonstrates that contracting back-in is often as common as contracting out and can be found in a variety of countries and across different service areas. Existing studies also suggest that contracting back-in is driven by a mix of managerial, political, and environmental factors. The rise of contracting back-in could imply a revival of the administrative state in public governance and a more dynamic, potentially more complicated, system of public service delivery.Originality/valueThe manuscript contributes to the special issue on “bringing the state back in” through the lens of government contracting. It adds a number of implications to the discussion on the emergence of the “new” administrative state and strategies to reinvent it.

Obstructive sleep apnea (OSA) is a common disorder characterized by recurrent upper airway obstruction during sleep. Despite the availability of continuous positive airway pressure (CPAP) as the gold standard treatment, it is not well tolerated by all patients. Accordingly, research has increasingly focused on developing methods for OSA endotyping, which aims to identify underlying pathophysiological mechanisms of the disorder to help guide treatment for CPAP-intolerant individuals. Four key endotypic traits have been identified, namely: collapsibility, upper airway muscle compensation, arousal threshold and loop gain. However, most methods for extracting these traits require specialized training and equipment not available in a standard sleep clinic, which has hampered the ability to assess the full impact of these traits on OSA outcomes. This paper aims to provide an overview of current methods for OSA endotyping, focusing on the Endo-Phenotyping Using Polysomnography (PUP) method and its cloud-based extension, PUPpy, which offer scalable and accessible ways to estimate endotypic traits from standard polysomnography. We discuss the potential for these methods to facilitate precision medicine for OSA patients and the challenges that need to be addressed for their translation into clinical practice.

INTRODUCTION: Dose escalation is one of the treatment approaches studied and suggested in advanced therapies for Crohn's disease (CD) and ulcerative colitis (UC). This study aimed to identify and characterize the dosing escalation patterns of advanced therapies in CD and UC. METHODS: Two systematic literature reviews (SLRs) were conducted in accordance with Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. MEDLINE®, Embase®, and Cochrane Library were searched for articles published between January 2011 and October 2021 and limited to non-interventional studies in English language. Congress and bibliographic searches were also conducted. Articles were screened by two independent researchers. Dose escalation patterns were described and summarized considering the regional regulatory label recommendation (in North America [NA] or outside of North America [ONA]). RESULTS: Among 3190 CD and 2116 UC articles identified in the Ovid searches, 100 CD and 54 UC studies were included in the SLR, with more studies conducted ONA. Most studies reported an initial maintenance dose pattern aligned with the lower starting dose per local regulatory label; however, several ONA studies (n = 13 out of 14) reported ustekinumab every 8 weeks as starting maintenance pattern in CD. In ONA studies, the median within-guideline escalation rates in CD and UC were 43% in ustekinumab (CD only), 33% and 32% for vedolizumab; 29% and 39% for adalimumab; and 14% and 10% for infliximab. Evidence regarding dose escalation patterns for tofacitinib, certolizumab pegol, and golimumab was limited. Some dose escalation patterns outside of label recommendations were observed including ustekinumab every 8 weeks to every 4 weeks and vedolizumab every 8 weeks to every 6 weeks. CONCLUSION: Dose escalation strategies are widely documented in the literature. The reported dose escalation patterns and escalation rates vary by region and by CD and UC. Most escalation patterns reported were aligned with regulatory recommendations while some reported more diverse or aggressive dose escalation. PROSPERO REGISTRATION: CRD42021289251.

In recent years, sustainability issues have been widely recognized by various sectors worldwide; therefore, this concept should be extensively integrated into university education. This study takes the integration of the Sustainable Development Goals (SDGs) and architectural design courses as an example to provide possibilities for the reuse of discarded concrete on campus, seeking opportunities to fulfill sustainable development on campus. Results showed that: (1) The course planning, operation, communication, SDG integration, and overall satisfaction were all well-received by the students. (2) By integrating the SDGs into the architecture design course and inter-school exchange activities, students quickly gained a better understanding of the SDGs and were able to contribute to promoting a sustainable campus. (3) The guidance provided by the teacher is crucial. To focus on the discussion topics, it is recommended to concentrate on one or two topics at a time to have in-depth discussions. (4) It is better to operate the course at a real site to allow students to experience and address real issues. In this case, the proposed solutions can have close ties with the environment of the site to truly embody the spirit of the SDGs.

Background This retrospective study gathered medical/pharmacy claims data on patients with IBD between 01/01/2000 and 03/31/2019 from the IBM® MarketScan® commercial claims database to assess the real-world impact of fatigue on healthcare costs in patients newly diagnosed with inflammatory bowel disease (IBD). Methods Eligible participants were ≥18 years, newly diagnosed with IBD (≥2 separate claims) and had ≥12 months of continuous database enrollment before and after fatigue diagnosis. The date of fatigue diagnosis was the index date; participants were followed for 12 months post-index. Patients with (cases) or without (controls) fatigue were matched 1:1 by propensity score matching. Patients with evidence of prior IBD diagnosis/treatment, or those with a chronic disease other than IBD wherein fatigue is the primary symptom, were excluded. Healthcare resource utilization (HCRU), including hospitalizations, inpatient and outpatient visits, and associated costs was compared between cases and controls. Results Matched IBD cohorts (21,321 cases/21,321 controls) were identified (42% CD and 58% UC) with similar baseline characteristics (average age: 46 years; 60% female). Cases versus controls had significantly more all-cause outpatient visits (incidence rate ratio [IRR], 95% confidence intervals [95% CI]: 1.64 [1.61, 1.67], P Conclusions Presence of fatigue is associated with an increase in HCRU and total medical costs among patients newly diagnosed with IBD.

Background Stroke may cause debilitating neurological deficiencies that result in motor, sensory, and cognitive deficits and poorer psychosocial functioning. Prior studies have provided some initial evidence for the significant roles of health literacy and poor oral health for old people. However, few studies have focused on the health literacy of individuals who had a stroke; therefore, the relationships between the health literacy and oral health-related quality of life (OHRQoL) among middle-aged and older adults who had a stroke are unknown. We aimed to assess the relationships between stroke prevalence, health literacy status, and OHRQoL in middle-aged and older adults. Methods We retrieved the data from The Taiwan Longitudinal Study on Aging, a population-based survey. For each eligible subject, we gathered data in 2015 on age, sex, level of education, marital status, health literacy, the activity daily living (ADL), stroke history and OHRQoL. We evaluated the respondents’ health literacy by using a nine-item health literacy scale and categorized their health literacy level as low, medium, or high. OHRQoL was identified based on the Taiwan version of the Oral Health Impact Profile (OHIP-7T). Results The final study contained 7702 community-based dwelling elderly people (3630 male and 4072 female) were analysis in our study. Stroke history was reported in 4.3% of participants, 25.3% reported low health literacy, and 41.9% had at least one ADL disability. Furthermore, 11.3% of participants had depression, 8.3% had cognitive impairment, and 3.4% had poor OHRQoL. Age, health literacy, ADL disability, stroke history, and depression status were significantly associated with poor OHRQoL after sex and marital status was adjusted. Medium (odds ratio [OR] = 1.784, 95% confidence interval [CI] = 1.177, 2.702) to low health literacy (OR = 2.496, 95% CI = 1.628, 3.828) was significantly associated with poor OHRQoL. Conclusions Base our study results, people with stroke history had poor OHRQoL. Lower health literacy and ADL disability were associated with worse QHRQoL. Further studies are necessary to define practical strategies for reducing the risk of stroke and oral health with constantly lower health literacy, thereby improving the quality of life and providing health care of older people.

PDF file - 76K, Summary statistics for Fig. 1C and Fig.2A

PDF file - 410K, Calcium influx in myeloid cells induced by CCL1 and SDF1

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PDF file - 70K, CCR8+ myeloid cells induce FoxP3 expression in T cells

In response to chemical stimuli from cancer cells, mesenchymal stem cells (MSC) can differentiate into cancer-associated fibroblasts (CAF) and promote tumor progression. How mechanical stimuli such as stiffness of the extracellular matrix (ECM) contribute to MSC phenotype in cancer remains poorly understood. Here, we show that ECM stiffness leads to mechano-signal transduction in MSC, which promotes mammary tumor growth in part through secretion of the signaling protein prosaposin. On a stiff matrix, MSC cultured with conditioned media from mammary cancer cells expressed increased levels of α-smooth muscle actin, a marker of CAF, compared with MSC cultured on a soft matrix. By contrast, MSC cultured on a stiff matrix secreted prosaposin that promoted proliferation and survival of mammary carcinoma cells but inhibited metastasis. Our findings suggest that in addition to chemical stimuli, increased stiffness of the ECM in the tumor microenvironment induces differentiation of MSC to CAF, triggering enhanced proliferation and survival of mammary cancer cells. Cancer Res; 77(22); 6179–89. ©2017 AACR.

This file contains supplementary figures S1-S10 and supplementary materials and methods. Figure S1 shows experimental design of mesenchymal stem cells in this article. Figure S2 shows differentiation of transgenic mesenchymal stem cells. Figure S3 shows differentiation and gel contraction of mesenchymal stem cells. Figure S4 shows YAP activity in mesenchymal stem cells on a stiff matrix. Figure S5 shows expression and activation of YAP-related signaling molecules in mesenchymal stem cells. Figure S6 shows contribution of mesenchymal stem cells to tumor progression. Figure S7 shows prosaposin detection in mammary tumors. Figure S8 shows the role of prosaposin for ERK and caspase 3. Figure S9 shows contribution of prosaposin to late tumor growth and progression. Figure S10 shows the role of stiff matrix for mesenchymal stem cells from B6 mice.

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PDF file - 51K, CCL1-induced IL6 production

PDF file - 169K, CCR8 expression in myeloid cells isolated from blood of cancer patients

Purpose: Chemokines are involved in cancer-related inflammation and malignant progression. In this study, we evaluated expression of CCR8 and its natural cognate ligand CCL1 in patients with urothelial carcinomas of bladder and renal cell carcinomas.Experimental Design: We examined CCR8 expression in peripheral blood and tumor tissues from patients with bladder and renal carcinomas. CCR8-positive myeloid cells were isolated from cancer tissues with magnetic beads and tested in vitro for cytokine production and ability to modulate T-cell function.Results: We show that monocytic and granulocytic myeloid cell subsets in peripheral blood of patients with cancer with urothelial and renal carcinomas display increased expression of chemokine receptor CCR8. Upregulated expression of CCR8 is also detected within human cancer tissues and primarily limited to tumor-associated macrophages. When isolated, CD11b+CCR8+ cell subset produces the highest levels of proinflammatory and proangiogenic factors among intratumoral CD11b myeloid cells. Tumor-infiltrating CD11b+CCR8+ cells selectively display activated Stat3 and are capable of inducing FoxP3 expression in autologous T lymphocytes. Primary human tumors produce substantial amounts of the natural CCR8 ligand CCL1.Conclusions: This study provides the first evidence that CCR8+ myeloid cell subset is expanded in patients with cancer. Elevated secretion of CCL1 by tumors and increased presence of CCR8+ myeloid cells in peripheral blood and cancer tissues indicate that CCL1/CCR8 axis is a component of cancer-related inflammation and may contribute to immune evasion. Obtained results also implicate that blockade of CCR8 signals may provide an attractive strategy for therapeutic intervention in human urothelial and renal cancers. Clin Cancer Res; 19(7); 1670–80. ©2013 AACR.

PDF file - 31K, Figure S1. GRK5 phosphorylates non-GPCR substrates. Figure S2. Moesin regulates migration and invasion of PCa cells.

Cystatin M is a secreted inhibitor of lysosomal cysteine proteases. Several lines of evidence indicate that cystatin M is a tumor suppressor important in breast malignancy; however, the mechanism(s) that leads to inactivation of cystatin M during cancer progression is unknown. Inspection of the human cystatin M locus uncovered a large and dense CpG island within the 5′ region of this gene (termed CST6). Analysis of cultured human breast tumor lines indicated that cystatin M expression is either undetectable or in low abundance in several lines; however, enhanced gene expression was measured in cells cultured on the DNA demethylating agent 5-aza-2′-deoxycytidine (5-aza-dC). Increased cystatin M expression does not correlate with a cytotoxic response to 5-aza-dC; rather, various molecular approaches indicated that the CST6 gene was aberrantly methylated in these tumor lines as well as in primary breast tumors. Moreover, 60% (12 of 20) of primary tumors analyzed displayed CST6 hypermethylation, indicating that this aberrant characteristic is common in breast malignancies. Finally, preinvasive and invasive breast tumor cells were microdissected from nine archival breast cancer specimens. Of the five tumors displaying CST6 gene methylation, four tumors displayed methylation in both ductal carcinoma in situ and invasive breast carcinoma lesions and reduced expression of cystatin M in these tumors was confirmed by immunohistochemistry. In summary, this study establishes that the tumor suppressor cystatin M is a novel target for epigenetic silencing during mammary tumorigenesis and that this aberrant event can occur before development of invasive breast cancer. (Cancer Res 2006; 66(16): 7899-909)

Table S1: Oligonucleotide primer sequences. Table S2: Data obtained from the Cox model. Figure S1: Detailed structure of the human TRIM29 gene locus. Figure S2: Additional TRIM29 gene methylation and transcript abundance analysis. Figure S3: Analysis of gene expression in breast cancer cells with TRIM29 knockdown. Figure S4: qRT-PCR analysis of TWIST1 and N-cadherin mRNA abundance in TRIM29 knockdown and control SK-BR-3 cells. Figure S5: Results of multivariate survival analysis examining TRIM29 expression vs patient age (A), lymph node status (B), size (C), Grade (D), and ER status (E).

Supplementary Figure 3 from Epigenetic Silencing of the Tumor Suppressor Cystatin M Occurs during Breast Cancer Progression

Supplementary Figure 1 from Epigenetic Silencing of the Tumor Suppressor Cystatin M Occurs during Breast Cancer Progression

G protein–coupled receptor kinases (GRK) regulate diverse cellular functions ranging from metabolism to growth and locomotion. Here, we report an important contributory role for GRK5 in human prostate cancer. Inhibition of GRK5 kinase activity attenuated the migration and invasion of prostate cancer cells and, concordantly, increased cell attachment and focal adhesion formation. Mass spectrometric analysis of the phosphoproteome revealed the cytoskeletal-membrane attachment protein moesin as a putative GRK5 substrate. GRK5 regulated the subcellular distribution of moesin and colocalized with moesin at the cell periphery. We identified amino acid T66 of moesin as a principal GRK5 phosphorylation site and showed that enforcing the expression of a T66-mutated moesin reduced cell spreading. In a xenograft model of human prostate cancer, GRK5 silencing reduced tumor growth, invasion, and metastasis. Taken together, our results established GRK5 as a key contributor to the growth and metastasis of prostate cancer. Cancer Res; 74(13); 3489–500. ©2014 AACR.

Human noroviruses (HuNVs) are the leading cause of gastroenteritis worldwide. NS1.2 is critical for HuNV pathogenesis, but the function is still unclear. The GII NS1.2 of HuNVs, unlike GI NS1.2, was localized to the endoplasmic reticulum (ER) and lipid droplets (LDs) and is accompanied by a distorted-filamentous ER morphology and aggregated-enlarged LDs. LC3 was recruited to the NS1.2-localized membrane through an autophagy-independent pathway. NS1.2, expressed from a cDNA clone of GII.4 norovirus, formed complexes with NTPase and NS4, which exhibited aggregated vesicle-like structures that were also colocalized with LC3 and LDs. NS1.2 is structurally divided into three domains from the N terminus: an inherently disordered region (IDR), a region that contains a putative hydrolase with the H-box/NC catalytic center (H-box/NC), and a C-terminal 251–330 a.a. region containing membrane-targeting domain. All three functional domains of NS1.2 were required for the induction of the filamentous ER. The IDR was essential for LC3 recruitment by NS1.2. Both the H-Box/NC and membrane-targeting domains are required for the induction of aggregated-enlarged LDs, NS1.2 self-assembly, and interaction with NTPase. The membrane-targeting domain was sufficient to interact with NS4. The study characterized the NS1.2 domain required for membrane targeting and protein–protein interactions, which are crucial for forming a viral replication complex.

Skin fibroblasts are the primary cells of DENV infection through mosquito bites. Establishing a successful infection in dermal fibroblasts might be critical for dengue disease.