International Research and Engineering Education: Impacts and Best Practices

In 2006, the IREE program funded 115 proposals from 82 U.S higher education institutions. Researchers and their faculty attended and presented their trip report at a 3-day conference held at Purdue University in November 2007. The first grantees conference was attended by 170 people, including 47 faculty members, 113 graduate students, 6 undergraduate students, and 6 NSF staff members. The 2007 IREE Grantees Conference was to provide a venue and facilitated opportunity for the IREE awardees, both students and faculty, to share experiences and what they gained from their time abroad under IREE. There are a set of 18 impacts of IREE that are categorized in three areas: Technical, professional, global/ trans-cultural. Based on these impacts, a set of best practices and recommendations are put forth to maximize learning and research outcomes of international research and engineering education

Best of Both Worlds: Foreign Language Preparation for Purdue University’s Undergraduate Global Engineering Education Program

Over the years, Purdue University has drastically increased the number of engineering students studying and interning abroad from less than 3% in 2000 to more than 10% in 2010. In order to increase the capacity of global engineering education curriculum, there is a need to create different study abroad programs to suit different student interests. Yet, the need of foreign language preparation remains in question. At Purdue University, researchers and administrators observed that students often self-select into study abroad programs of varying intensity according to the varying foreign language and GPA requirements. Case studies of student participants from four different Purdue education abroad programs will be demonstrated in this paper in the following order: (1) Global Engineering Alliance for Research and Education (GEARE), (2) International Research and Education for Engineering (IREE), (3) Global Internship, and (4) China Maymester Abroad Program. These case studies will be used to illustrate the importance of foreign language preparation and the varying needs. These results will also demonstrate that the achieved level of foreign language competency impacts technical outcomes and engineering professionalism

Cutting Edge: Induced Loss of Rasgrp1 in Peripheral CD4+ T Cells of Conditional Rasgrp1-Deficient Mice Reveals an Essential Role for Rasgrp1 in TCR/CD28-Induced Ras-MAPK Signaling.

Ras guanine nucleotide-releasing protein 1 (Rasgrp1) is a Ras guanine nucleotide exchange factor that participates in the activation of the Ras-ERK signaling pathway in developing T cells and is required for efficient thymic T cell positive selection. However, the role of Rasgrp1 in mature peripheral T cells has not been definitively addressed, in part because peripheral T cells from constitutive Rasgrp1-deficient mice show an abnormal activated phenotype. In this study, we generated an inducible Rasgrp1-deficient mouse model to allow acute disruption of Rasgrp1 in peripheral CD4+ T cells in the context of normal T cell development. TCR/CD28-mediated activation of Ras-ERK signaling was blocked in Rasgrp1-deficient peripheral CD4+ T cells. Furthermore, Rasgrp1-deficient CD4+ T cells were unable to synthesize IL-2 and the high-affinity IL-2R and were unable to proliferate in response to TCR/CD28 stimulation. These findings highlight an essential function for Rasgrp1 for TCR/CD28-induced Ras-ERK activation in peripheral CD4+ T cells

Gender differences in the relationship between loneliness and health-related behavioral risk factors among the Hakka elderly in Fujian, China

IntroductionTo explore gender differences in the relationship between loneliness and health-related behavioral risk factors (BRFs) among the Hakka elderly.MethodsLoneliness was measured by the UCLA Loneliness Scale Short-form (ULS-8). Seven BRFs were examined. Mann–Whitney U, Kruskal-Wallis, and post hoc tests were conducted to compare the differences in ULS-8 scores among the Hakka elderly with different BRFs. Generalized linear regression models were employed to examine the associations of specific BRF and its number with the ULS-8 scores among the Hakka elderly in male, female, and total samples.ResultsPhysical inactivity (B = 1.96, p < 0.001), insufficient leisure activities participation (B = 1.44, p < 0.001), unhealthy dietary behavior (B = 1.02, p < 0.001), and irregular sleep (B = 2.45, p < 0.001) were positively correlated with the ULS-8 scores, whereas drinking (B = −0.71, p < 0.01) was negatively associated with the ULS-8 scores in the total sample. In males, insufficient leisure activities participation (B = 2.35, p < 0.001), unhealthy dietary behavior (B = 1.39, p < 0.001), and irregular sleep (B = 2.07, p < 0.001) were positively associated with the ULS-8 scores. In females, physical inactivity (B = 2.69, p < 0.001) and irregular sleep (B = 2.91, p < 0.001) was positively correlated with the scores of ULS-8, while drinking (B = −0.98, p < 0.05) was negatively associated with the ULS-8 scores. More BRFs were significantly related to greater loneliness (p < 0.001).ConclusionThere are gender differences in the relationship between loneliness and BRFs among the Hakka elderly, and individuals with more BRFs were more likely to feel loneliness. Therefore, the co-occurrence of multiple BRFs requires more attention, and integrated behavioral intervention strategies should be adopted to reduce the loneliness of the elderly

Progress in immune checkpoint inhibitor-based combined regimens in the treatment of metastatic urothelial carcinoma

Metastatic urothelial carcinoma （mUC） yields poor prognosis and is difficult to treat. Immunotherapy strategies have developed rapidly in recent years， and clinical prognosis of mUC patients has been significantly improved. However， immune checkpoint inhibitors （ICIs） alone have limited efficacy， and some ICIs monotherapy indications for mUC have also been withdrawn in recent years. Therefore， combined therapy based on ICIs has become a research hotspot in the treatment of mUC. At present， multiple clinical studies of ICIs combined with chemotherapy， antibody-drug conjugates， and dual immunotherapy for multiple lines of treatment of mUC have proved that combined therapy possesses promising development prospect. In this article， latest progress in ICI-based combined treatment for mUC was reviewed， aiming to provide reference for clinicians

Seroprevalence Survey of Avian influenza A (H5) in wild migratory birds in Yunnan Province, Southwestern China

BACKGROUND: Highly pathogenic avian influenza virus (HPAIV) is a highly contagious disease which is a zoonotic pathogen of significant economic and public health concern. The outbreaks caused by HPAIV H5N1 of Asian origin have caused animal and human disease and mortality in several countries of Southeast Asia, such as Bangladesh, Cambodia, China, India, Indonesia, Laos, Myanmar, Thailand and Viet Nam. For the first time since 1961, this HPAIV has also caused extensive mortality in wild birds and has sparked debate of the role wild birds have played in the spread of this virus. Other than confirmed mortality events, little is known of this virus in wild birds. There is no report on the seroprevalence of avian influenza H5 infection in wild migratory birds in Yunnan Province. In this study we examined live wild birds in Yunnan Province for H5 specific antibody to better understand the occurrence of this disease in free living birds. METHODS: Sera from 440 wild birds were collected from in Kunming and Northern Ailaoshan of Yunnan Province, Southwestern China, and assayed for H5 antibodies using the hemagglutination inhibition (HI) assays. RESULTS: The investigation revealed that the seroprevalence of avian influenza H5 was as following: Ciconiiformes 2.6%, Strigiformes 13.04%, Passeriformes 20%, Cuculiformes 21.74%, Gruiformes 0%, Columbiformes 0%, Charadriiformes 0% and Coraciiformes 0%. Statistical analyses showed that there was a significant difference of prevalence between the orders (P < 0.01). Specific avian influenza H5 antibodies were detected in 23 of 440 (5.23%) sera. Mean HI titer 23 positive sera against H5 were 5.4 log(2). CONCLUSIONS: The results of the present survey indicated that the proportion of wild birds had previously infected AIV H5 at other times of the year. To our knowledge, this is the first seroprevalence report of avian influenza H5 infection in wild migratory birds in China’ s southwestern Yunnan Province. The results of the present survey have significant public health concerns

RHOA-regulated IGFBP2 promotes invasion and drives progression of BCR-ABL1 chronic myeloid leukemia

The Philadelphia 9;22 chromosome translocation has two common isoforms that are preferentially associated with distinct subtypes of leukemia. The p210 variant is the hallmark of chronic myeloid leukemia (CML) whereas p190 is frequently associated with B-cell acute lymphoblastic leukemia. The only sequence difference between the two isoforms is the guanidine exchange factor domain. This guanidine exchange factor is reported to activate RHO family GTPases in response to diverse extracellular stimuli. It is not clear whether and, if so, how RHOA contributes to progression of p210 CML. Here we show that knockout of RHOA in the K562 and KU812, p210-expressing cell lines leads to suppression of leukemogenesis in animal models in vivo. RNA-sequencing analysis of the mock control and null cells demonstrated a distinct change in the gene expression profile as a result of RHOA deletion, with significant downregulation of genes involved in cell activation and cell adhesion. Cellular analysis revealed that RHOA knockout leads to impaired cell adhesion and migration and, most importantly, the homing ability of leukemia cells to the bone marrow, which may be responsible for the attenuated leukemia progression. We also identified IGFBP2 as an important downstream target of RHOA. Further mechanistic investigation showed that RHOA activation leads to relocation of the serum response factor (SRF) into the nucleus, where it directly activates IGFBP2. Knockout of IGFBP2 in CML cells suppressed cell adhesion/invasion, as well as leukemogenesis in vivo. This elevated IGFBP2 expression was confirmed in primary CML samples. Thus, we demonstrate one mechanism whereby the RHOA-SRF-IGFBP2 signaling axis contributes to the development of leukemia in cells expressing the p210 BCR-ABL1 fusion kinase

Intratumoral Delivery of a PD-1-blocking scFv encoded in Oncolytic HSV-1 Promotes Antitumor Immunity and Synergizes with TIGIT Blockade

恶性肿瘤已严重威胁人类健康和生命，现有的治疗手段远远未能满足临床需求。厦门大学研究团队联合浙江养生堂生物科技有限公司、养生堂有限公司进行协同攻关，研制出新一代肿瘤免疫治疗药物——“注射用重组人PD-1抗体单纯疱疹病毒”。研究发现，重组表达PD-1单链抗体的溶瘤病毒具有“双药合一”抗肿瘤的突出优势，提高肿瘤治愈率。相关成果于2020年3月3日以研究论文形式在线发表于Cancer Immunology Research（《癌症免疫学研究》）期刊，为指导新型溶瘤病毒的升级改造和突破肿瘤免疫耐受提供了新的思路，为重组表达PD-1单链抗体的溶瘤病毒药物运用于肿瘤临床治疗奠定了坚实的理论基础。 厦门大学公共卫生学院夏宁邵教授和黄承浩助理教授为该论文的共同通讯作者，博士生林超龙和任文峰为该论文的共同第一作者。【Abstract】Oncolytic virotherapy can lead to systemic antitumor immunity, but the therapeutic potential of oncolytic viruses (OVs) in humans is limited due to their insufficient ability to overcome the immunosuppressive tumor microenvironment (TME). Here, we showed that locoregional oncolytic virotherapy upregulated the expression of PD-L1 in the TME, which was mediated by virus-induced type I and type II interferons (IFNs). To explore PD-1/PD-L1 signaling as a direct target in tumor tissue, we developed a novel immunotherapeutic herpes simplex virus (HSV), OVH-aMPD-1, that expressed a single-chain variable fragment (scFv) against PD-1 (aMPD-1 scFv). The virus was designed to locally deliver aMPD-1 scFv in the TME to achieve enhanced antitumor effects. This virus effectively modified the TME by releasing damage associated molecular patterns (DAMPs), promoting antigen cross-presentation by dendritic cells, and enhancing the infiltration of activated T cells; these alterations resulted antitumor T cell activity which led to reduced tumor burdens in a liver cancer model. Compared with OVH, OVH-aMPD-1 promoted the infiltration of myeloid-derived suppressor cells (MDSCs),resulting in significantly higher percentages of CD155+ G-MDSCs and M-MDSCs in tumors. In combination with TIGIT blockade, this virus enhanced tumor-specific immune responses in mice with implanted subcutaneous tumors or invasive tumors. These findings highlighted that intratumoral immunomodulation with an OV expressing aMPD-1 scFv could be an effective standalone strategy to treat cancers or drive maximal efficacy of a combination therapy with other immune checkpoint inhibitors.This work was supported by grant 2018ZX10301404-001-002 from the National Science and Technology Major Project and grant 81571990 from the National Natural Science Foundation of China.该研究获得了国家自然科学基金、国家科技重大专项的资助

Identificatioh of amino acids involved in SAM binding and methyl group transfer of BaMV capping enzyme

竹嵌紋病毒(bamboo mosaic virus)基因體其上open reading frame 1 (ORF1)長約4.1 kb，可製造出一個分子量約155 kDa的複製酵素，此酵素由N端至C端分別為戴帽酵素(capping enzyme)、似解旋酵素區段(helicase-like domain)、RNA聚合酵素(RNA-dependent RNA polymerase, RdRp)。之前的研究結果顯示由酵母菌(Saccharomyces cerevisiae)生產之病毒戴帽酵素同時具有利用SAM及GTP這兩個受質的能力，也就是同時具有轉甲基酵素和guanylyltransferase之活性。其作用機制為：轉甲基酵素先利用SAM進行GTP的甲基化，形成m7GTP分子，guanylyltransferase 再將m7GTP轉換成m7GMP並以共價鍵型式結合在酵素之活化中心胺基酸。在此研究中，我們試著找出病毒戴帽酵素活化中心與受質之結合位氨基酸。結果顯示H66、K121、D122、R125、Y213、D310、K344、W406這幾個氨基酸的突變會嚴重破壞轉甲基酵素的活性，藉由UV cross-linking得知H66、K121、D122、R125、Y213是結合AdoMet必須的氨基酸。而H68的突變會增強轉甲基酵素的活性，但是卻喪失了guanylyltransferase的活性，推測此氨基酸為m7GMP之共價鍵結位置。我們也嘗試利用UV cross-linking的方法來探討酵素與GTP結合的關係，可惜一直無法如願，期待將來可利用其他方法來研究。另一方面，為了想進一步瞭解竹嵌紋病毒複製酵素之複製機制，我們嘗試建立一個以酵母菌為寄主的生體內分析系統。我們將兩個質體同時送入酵母菌中，pYEB1含有完整的ORF1基因，能在酵母菌體內製造出完整的竹嵌紋病毒複製酵素，而另一個質體則含修改過的竹嵌紋病毒CDNA。假定在酵母Gal promoter的驅動下，會轉錄出修改過的病毒正股RNA，同時，由pYEB1表達之複製酵素以修改過的病毒正股RNA為模版，複製出相對應的負股RNA，再以之為模版複製出次基因體RNA。如此一來，次基因體RNA就可被核醣體轉譯，產生移動蛋白與外鞘蛋白。實驗結果顯示，我們無法偵測到移動蛋白及外鞘蛋白的存在。這樣的結果使我們不禁猜測，也許竹嵌紋病毒複製酵素不適用於in-trans的系統。因此，我們改變策略，只將一含有竹嵌紋病毒全長CDNA的質體送入酵母菌中。但是，很遺憾的，我們仍然無法偵測到複製酵素的活性。由以上的結果，我們推測或許由酵母Gal promoter驅動表達出的病毒正股RNA無法被複製酵素辨識，抑或許酵母菌體內缺乏竹嵌紋病毒複製酵素C端聚合酵素活性必須的環境及因子，致使此酵母菌生體內的分析系統無法成功。Open reading frame 1 (ORF1) of Bamboo mosaic virus (BaMV) encodes a 155-kDa replicase that contains capping enzyme, helicase-like domain, and RNA dependent RNA polymerase (RdRp) on the order of from N to C termini. The viral capping enzyme expressed in yeast uses both SAM and GTP as substrates for methyltransferase (MT) and guanylyltransferase (GT) activity. The MT activity transfers the methyl group from SAM to GTP; subsequently, the GT activity catalyzes transguanylation via the intermediate m7GMP-enzyme complex. In this study, we tried to locate the residues, which are involved in binding SAM and GTP. The results showed that mutations of H66, K121, D122, R125, Y213, D310, K344, and W406 destroyed or greatly reduced the MT activity, and mutation of H68 prevented the GT reaction but increased MT activity. The data from UV cross-linking suggest that residues H66, K121, D122, R125 and Y213 are necessary for SAM binding, and H68 might be the residue for forming the covalent bond to m7GMP in m7GMP-enzyme complex. UV cross-linking was also used to find the effect of mutations for GTP binding; however, no GTP-enzyme complex could be detected after UV cross-linking reaction. In the other project, we tried to set up an in vivo replication system using yeast as a host to study BaMV replicase activity. We transferred two plasmids into yeast so that the BaMV replicase can be expressed from one plasmid and a modified BaMV positive-stranded RNA can be transcribed from the other by galactose-inducible yeast GAL promoter. We expected that BaMV replicase expressed in yeast can lead to the synthesis of negative-stranded RNA by using the modified BaMV positive-stranded RNA as template; subsequently, the subgenomic mRNA coding for coat protein and movement protein can be produced. However, neither coat protein nor movement protein could be detected in the recombinant yeast. Neither was the reporter protein expressed when its coding RNA was continuously transcribed downstream the coding sequence of BaMV replicase. These results suggest that the BaMV positive-stranded RNA driven by yeast GAL promoter can't be used by BaMV replicase, or yeast cell lacks the necessary factors for BaMV RdRp activity.序…………………………………………………………………………………...... 1 第一章：竹嵌紋病毒戴帽酵素中參與受質SAM結合與催化 甲基轉移能力之氨基酸探討…………..…...……….………...…….. 5 第一節、緒言……………………………………………………………………….. 8 第二節、材料與方法…………………………………………………………...... 11 第三節、結果 壹、竹嵌紋病毒戴帽酵素之轉甲基酵素活性測試……………….....18 貳、突變酵素之轉甲基酵素活性測試……………………………….…20 參、突變酵素之SAM結合能力測試…………………………………..21 肆、尋找以UV光能量使戴帽酵素與GTP形成共價鍵結 之條件………………………………………………...………………….22 第四節、討論………………………………………………………………………25 第二章：建立竹嵌紋病毒於酵母菌中之完整複製系統……........…….…44 第一節、緒言………………………………………………………………………47 第二節、材料與方法………………………………………………………..……50 第三節、結果 壹、in-trans分析系統………………………………………………………56 貳、in-cis分析系統…………………………………………………………57 第四節、討論………………………………………………………………………59 參考文獻…………………………………………………………………………….75 表目錄 表 1-1、SC + KIWI 培養基……………………………………………………70 表 1-2、放射性標定 GTP 與竹嵌紋病毒戴帽酵素之 UV cross-linking條件……………………………………………..………..71 表 2-1、PCR 所使用引子之序列…………………………………….……….72 表 2-2、含不同表現質體之酵母菌與其對應之培養基…………………..73 圖目錄 圖1-1、竹嵌紋病毒基因體RNA圖譜及其基因產物蛋白質簡圖。.....29 圖1-2、竹嵌紋病毒戴帽酵素氨基酸序列及定點突變之位置。.............30 圖1-3、竹嵌紋病毒戴帽酵素之定量分析。..................................................31 圖1-4、竹嵌紋病毒戴帽酵素之轉甲基酵素活性分析。...........................32 圖1-5、不同甲基接受者對竹嵌紋病毒戴帽酵素之轉甲基酵素活 性影響。......................................................................................................33 圖1-6、鎂離子濃度對竹嵌紋病毒戴帽酵素之轉甲基酵素活性影 響。...............................................................................................................34 圖1-7、竹嵌紋病毒戴帽酵素丙氨酸置換突變酵素之轉甲基酵素 活性分析。................................................................................................35 圖1-8、竹嵌紋病毒戴帽酵素WT與H68A之轉甲基酵素活性對 甲基接受者之專一性分析。................................................................36 圖1-9、竹嵌紋病毒戴帽酵素WT與H68A之轉甲基酵素活性時 間曲線。......................................................................................................37 圖1-10、竹嵌紋病毒戴帽酵素丙氨酸置換突變酵素之戴帽活性以 及SAM結合能力。...............................................................................38 圖1-11、利用UV cross-linking方式偵測竹嵌紋病毒戴帽酵素與 GTP之結合能力。.................................................................................39 圖1-12、不同核種對UV cross-linking之影響。.........................................40 圖1-13、酵母菌細胞膜上竹嵌紋病毒戴帽酵素與GTP結合之 UV cross-linking結果。....................................................................41 圖1-14、酵母菌細胞膜上病毒融合蛋白B3GFP之戴帽活性以及 UV cross-linking結果。.....................................................................42 圖1-15、竹嵌紋病毒戴帽酵素之戴帽反應示意圖。………….............….43 圖2-1、竹嵌紋病毒複製循環。..........................................................................62 圖2-2、酵母菌生體內in-trans分析系統圖解。...........................................63 圖2-3、質體p424-BGFP、p424-BGA以及p424-BV之構築流程。....64 圖2-4、質體p424-B5+以及p424-B5-之構築流程。...................................65 圖2-5、質體pLex-BGFP以及pHyb-BGFP之構築流程。......................66 圖2-6、質體pYECB以及pYECBde之構築流程。..................................67 圖2-7、竹嵌紋病毒複製酵素活性。................................................................68 圖2-8、竹嵌紋病毒複製酵素活性。................................................................69 附錄目錄 縮寫表………………………………………………………………附錄