The Prevalence of High-Risk Oral HPV Among Rural/Tribal Women In Mysore Who Are Chronic Smokeless Tobacco Users

Background: Human papillomavirus (HPV) is the most common sexually transmitted virus. Low-risk HPV can cause warts, and the virus often clears on its own. High-risk HPV can persist and can cause cervical, anogenital, and head and neck cancer, primarily of the oropharynx. Not all oral cancers are caused by HPV alone. Additional independent risk factors include alcohol, smoking, and smokeless tobacco consumption. Tobacco use, including cigarette smoking and tobacco chewing, is the primary cause of oral cancer worldwide. India has high prevalence of tobacco consumption and 60% of smokeless tobacco users are women. Tobacco users have a fifteen-fold increased risk of oral cancer compared to non-tobacco users. There may be interactions between smokeless tobacco use, oral HPV infection and oral cancer. Objective: The goal of this project is to examine the prevalence of oral HPV among women who are chronic tobacco users, and determine factors associated with HPV infection. The long-term objective is to inform oral cancer prevention strategies such as tobacco cessation and HPV vaccination within these communities. Materials and Methods: In June and July 2019, 50 chronic smokeless tobacco users and 50 non-users from rural areas of Mysore were consented to participate in this study. Interviewer administered questionnaires were used to assess demographic and socioeconomic characteristics, tobacco and alcohol use history, and oral health behaviors. Oral samples were collected using an all-collection swab to collect oral cells of the center of tongue, below the tongue, hard palate, buccal mucosa, and upper front gums and placed into a sterile Qiagen collection tube. The brush in the collection kit was used on these regions and placed in the same collection tube as the swab. Lastly, 10 mL saline was given to participants to gargle before spitting out into a separate sample container. The samples were tested with the digeneHC2 High Risk HPV DNA test kit (Qiagen) using microplate chemiluminescence for the qualitative detection of 13 high-risk types of HPV DNA. Results: The results show that the prevalence of high-risk oral HPV within these communities was very low (2%). Conclusions: This study shows that the prevalence of oral-HPV among chronic smokeless tobacco using women of the rural/tribal community in Mysore, India, is similar to the prevalence of oral-HPV for women in the United States.MHIRT Progra

인간 유래 ChaC 단백질의 glutathione 대사에서 구조밎 기능이 대한 연구

학위논문(박사)--서울대학교 대학원 :약학대학 약학과,2020. 2. Byung Woo Han.Glutathione (GSH) degradation plays an essential role not only in the GSH homeostasis but also in housekeeping functions, which regulate cell survival especially in cancer cells. Among human GSH degradation enzymes, cytosolic ChaC1 and ChaC2 enzymes act on GSH to form 5-L-oxoproline and Cys-Gly specifically in cytosol and share 60% sequence similarities. Here, I report the crystal structures of ChaC2 in two different conformations and compare structural features with other known γ-glutamylcyclotransferase enzymes. The unique domain-swapped loop of ChaC2 seems to function as a gate to achieve specificity for GSH binding and regulate the constant GSH degradation rate. Intensive structural and biochemical analyses of ChaC2 revealed that Glu74 and Glu83 play crucial roles in directing the assembly of the swapping dimer and in modulating the enzyme activity. Based on the docking study of GSH to ChaC2 and binding assay, I propose a substrate binding mode and catalytic mechanism. I also found that overexpression of ChaC2, but not ChaC2 E74Q or ChaC2 E83Q, significantly promoted breast cancer cell line proliferation, suggesting that the GSH degradation role of ChaC protein plays a role in breast cancer cell growth. My structural and functional analyses of ChaC2 will contribute to the development of modulators in the ChaC family, which could effectively regulate the progression of GSH degradation-related diseases and cancers.Glutathione(GSH)의 분해는 GSH의 항상성을 유지할 뿐만 아니라, 암 세포에서 세포 생존에 중요한 역할을 한다고 알려져 있다. 특히 GSH 분해를 담당하는 효소들 중 ChaC1과 ChaC2는 공통적으로 GSH를 분해하여 5-L-oxoproline 과 Cys-Gly 펩타이드를 만드는 역할을 하는데, 서로의 아미노산 서열은 60% 정도 유사하다고 알려져 있다. 본 연구에서는 ChaC2의 각각 다른 두 가지 형태의 결정 구조를 밝혔고, 이 구조들을 기존에 알려져 있던 다른 γ-glutamylcyclotransferase 계열의 효소들과 비교하였다. ChaC2 결정 구조에서 발견되는 domain-교환 loop은 GSH와의 결합 특이성을 부여하는 부분으로, GSH 분해 속도를 조절하는 데 중요한 역할을 하는 것으로 보인다. 여러 생화학적인 실험 결과와 단백질 구조 분석을 통하여 Glu74와 Glu83이 이합체의 형성과 단백질 활성의 조절에 중요한 역할을 한다는 것을 알 수 있었고, GSH와 ChaC2 사이의 docking 연구와 결합 실험을 통해 GSH와 ChaC2의 사이의 결합 방식과 ChaC2의 반응 메커니즘을 제시하였다. 뿐만 아니라, ChaC2의 과발현이 E74Q와 E83Q 변이 단백질들에 비해 유방암 세포의 생존을 증가시키는 실험 결과를 토대로 ChaC2에 의한 GSH의 분해가 유방암의 발달에 특정 역할을 할 것이라 제시할 수 있었다. 결과적으로, ChaC2의 구조와 기능에 대한 본 연구는 ChaC 계열 단백질들을 조절할 수 있는 분자의 개발에 기여하고, 더 나아가 GSH 분해와 관련된 질병과 암을 효율적으로 조절할 수 있는 치료의 기반을 제공한다.Abstract i Table of Contents ii List of Tables iv List of Figures v Abbreviations vi I. Introduction １ 1. Study Background １ 2. Purpose of Research ６ II. Materials and methods ８ 1. Bioinformatics analysis ８ 2. Cloning and protein production ８ 2.1. Cloning and protein production of ChaC2 ８ 2.2. Cloning and protein production of ChaC1 10 3. Mutagenesis 13 4. Crystallization 13 4.1. Crystallization of ChaC2 13 4.2. Crystallization of ChaC1 15 5. X-ray diffraction data collection, structure determination 18 6. Preparation of ChaC2-overexpressing cells 20 7. GSH degradation activity of ChaC enzymes by LC-MS 21 8. GSH assays 22 9. Viability assay (MTT assay) 22 10. Colony-forming assay 23 11. Immunoblotting 23 12. Docking study 23 13. Surface plasmon resonance experiment 23 14. Statistical analysis 24 15. Accession numbers 23 III. Results 25 1. ChaC2 structure was determined through rational Met 25 2. The overall structure of ChaC2 adopts the GGCT fold 31 3. Domain-swapping structure of the ChaC2 homodimer. 33 4. Single mutations of E74Q and E83Q induced conformational 36 5. Structural comparison of human ChaC2 and other GGCT 47 6. Purified ChaC proteins have GGCT activity in vitro 51 7. ChaC2 E74Q and ChaC2 E83Q mutation significantly reduced 53 8. ChaC2 overexpression promotes cell proliferation in MCF-7 55 IV. Discussion 58 1. Role of domain-swapped ChaC2 homodimer conformation 58 2. Proposed mechanism of substrate recognition and GSH... 61 3. Correlation of ChaC2, GSH degradation, and breast cancer 68 Reference 70 Abstract in Korean 73 Acknowledgements 74Docto

Vietnamese American women’s beliefs and perceptions on cervical cancer, cervical cancer screening, and cancer prevention vaccines: A community-based participatory study

Cervical cancer remains commonly diagnosed in Vietnamese American women. Despite efforts to increase cervical cancer screening among Vietnamese American women, participation rates are persistently lower than the national goal. The objective of this study is to explore beliefs of Vietnamese American women about cervical cancer, cervical cancer screening, and cancer prevention vaccines. A qualitative descriptive investigation captured group perceptions about meaning and beliefs of cervical cancer, screening, and cancer prevention vaccines, and participants’ stories using a community-based participatory research approach. Forty Vietnamese American women were recruited from the Portland, Oregon metropolitan area into four focus groups. Using a process of directed content analysis, focus group transcripts were coded for themes. We found that cervical cancer continues to be a difficult topic to discuss, and Vietnamese American women may not bring the topic up themselves to their health care providers. Some women experienced intense emotions of fear or shame of having their cervix examined. Women delayed seeking cervical cancer screening and needed to have early warning signs, which guided them as to when to seek health care. Women focused on cleanliness through vaginal and/or perineal washing as primary prevention for cervical cancer. There were limited awareness and knowledge about cancer prevention vaccines, specifically the human papillomavirus. Some women relied heavily on their informal social networks of family, friends, or community for health knowledge. Fear and misunderstanding dominated the beliefs of Vietnamese American women about cervical cancer screening and prevention. These findings underscored the importance of having culturally-specific findings, which will inform a multicomponent intervention to promote cervical cancer screening and cancer prevention vaccine uptake within this population

ESTIMATION OF VARIABILITY ANALYSIS PARAMETERS FOR MAJOR GROWTH AND FLOWERING TRAITS OF Lilium leichtlinii var. maximowiczii GERMPLASM

This experiment was carried out to evaluate the Lilium leichtlinii var maximowiczii germplasm collected from the different natural habitats from all over Korea. In total 30 accessions were studied for nine traits viz. plant height, leaf length, leaf width, the numbers of flowers, flower diameter, length of outer tepal, width of outer tepal, the number of leaf burn, and days to flowering in randomized block design with three replications. The ANOVA revealed highly significant variability prevailing among the investigated genotypes for almost all studied traits (except leaf width). The higher estimated value of GCV, PCV, heritability (H2), and genetic advance as percent of the mean was obtained for the number of flowers and leaf burn. The moderate to high GCV and PCV coupled with higher heritability estimates (H2) and GAM were found for plant height and flower diameter. Progeny selection would be effective as the prevalence of additive gene effect for these traits. Besides, leaf width, leaf length, length of outer tepal, the width of outer tepal, and days to flowering traits were possessed moderate to low GCV and PCV value coupled with the moderate value of heritability estimate with the low level of GAM proved to be the prevalence of non-additive genetic effect thereby indicating the necessity of alternative breeding approach for these traits improvement and breeding scheme. For the former group of traits breeding hybridization and selection would be an effective method, and primarily mean performance of these traits would be very handy for the decision of selection

Evaluating a First-Year Veterans Affairs Nurse Practitioner Residency Program: Analysis for Change

Background: The Veteran Affairs Portland Healthcare System (VAPORHCS) is experiencing a shortage of primary care physicians. To help meet this demand for primary care providers in outpatient clinics, VAPORHCS turned to nurse practitioners (NPs). A primary care nurse practitioner residency (PC-NPR) program was developed to support novice NP’s transition to practice. Purpose: To describe the development of evaluation tools and an evaluation of the effectiveness and efficiency of the PC-NPR program’s curriculum in supporting NP residents’ progression through the program and transition to practice utilizing accreditation standards. Methods: The development of evaluation tools using a combined approach guided by Meleis’ Transition Theory and the Centers for Disease Control and Prevention Program Evaluation Framework. Evaluation tools included a 12-item curriculum questionnaire developed from national accreditation pre-publication standards and a focus group interview. Results: There was a 54% (n=13) response rate. The questionnaire had some negative responses to three statements. The remaining responses were positive. The two main themes of the focus group were transition to practice support and curricular improvement. Conclusion: This evaluation contributed to a comprehensive program evaluation. Results are being used to make timely improvements to the program objectives and curriculum in preparation for seeking the Commission on Collegiate Nursing Education national accreditation

Reproductive Biology of Moonlight Gourami (Trichopodus microlepis) in U Minh Thuong and U Minh Ha National Parks

This study aimed to provide information about the reproductive biology of moonlight gourami (Trichopodus microlepsis) distributed in U Minh National Parks. Fish samples were collected monthly from January to October 2019 at two locations: U Minh Ha (UMH, Ca Mau, n=462) and U Minh Thuong (UMT, Kien Giang, n=635). Results showed that the maturation-stage ratios of females were similar between the two fish populations. Mature females (with stage IV-ovaries) and the increase in their gonadal somatic index (GIS) were found from June to October, indicating that this period is their spawning season. GSI reached the highest values in August with 8.04% in UMH and 6.44% in UMT. Female fish of the UMH population showed higher average fecundity (7,483±3,008 eggs/female) and larger egg diameter (762±55 µm) than those of UMT (6,898±2,952 eggs/female and 754±43 µm, respectively), but these differences were not significant (p>0.05)

Effects of maturity stages and fermentation of cocoa beans on total phenolic contents and antioxidant capacities in raw cocoa powder

Consumption of cocoa (Theobroma cacao L.) and cocoa products is associated with numerous health benefits due to their high levels of polyphenols and antioxidant capacities. In this study, changes of total phenolic contents (TPC) and antioxidant capacities (AC) of raw cocoa powder at four maturity stages, under different fermentation methods and fermentation duration of cocoa beans were investigated. The TPC and AC were measured using Folin–Ciocalteu and Trolox equivalent antioxidant capacity (TEAC)/ABTS assay, respectively. In term of maturity stages, the powder of unfermented cocoa beans harvested from the stage one contained significantly lower levels of TPC (6.39 ± 0.02 g CE/100 g DM) and AC (26.82 ± 0.13 mol TE/100 g DM) than those from the beans harvested from the stage two, three and four. For all maturity stages studied, fermenting cocoa beans increased antioxidant capacities of the raw cocoa powder. Cocoa fermentation using the commercial enzyme Pectinex® Ultra SP-L resulted in lower TPC but higher AC in comparison to those treated without enzymes, however prolonged fermentation time in cocoa beans significantly reduced TPC and AC in the powder. Strong correlations between TPC and AC of fermented samples with (R = 0.923) and without enzyme supplement (R = 0.942) were obtained. Two-way anova analyses showed that changes of TPC and AC of cocoa beans were dependent on maturity stages, fermentation methods and fermentation duration. As a conclusion, fermentation of cocoa beans harvested at the maturity stage two and three was found to be optimum for the high levels of AC and TPC attainment; fermented beans with commercial enzyme could be utilized to reduce labor cost by shortening the fermentation duration