Shing, Ka Fai. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 158-188). / Abstracts in English and Chinese. / ABSTRACT --- p.i / 摘要 --- p.iv / ACKNOWLEDGEMENTS --- p.vi / TABLE OF CONTENTS --- p.vii / LIST OF TABLES --- p.x / LIST OF FIGURES --- p.xii / Chapter SECTION I: --- BACKGROUND --- p.1 / Chapter CHAPTER 1: --- CELL-FREE NUCLEIC ACIDS IN HUMAN BODILY FLUIDS --- p.2 / Chapter 1.1 --- Early studies on the presence of cell-free nucleic acids in human bodily fluids --- p.2 / Chapter 1.2 --- Circulating nucleic acids in plasma and serum --- p.2 / Chapter 1.2.1 --- Cancer Detection --- p.3 / Chapter 1.2.1.1 --- Circulating tumor-derived DNA --- p.3 / Chapter 1.2.1.2 --- Circulating tumor-derived RNA --- p.5 / Chapter 1.2.2 --- Prenatal diagnosis --- p.7 / Chapter 1.2.2.1 --- Circulating fetal DNA --- p.7 / Chapter 1.2.2.2 --- Circulating fetal messenger RNA --- p.11 / Chapter 1.2.2.3 --- Circulating placental microRNA --- p.13 / Chapter 1.3 --- Cell-free nucleic acids in urine --- p.14 / Chapter 1.3.1 --- Transrenal DNA (Tr-DNA) --- p.15 / Chapter 1.3.1.1 --- Biology of Tr-DNA --- p.15 / Chapter 1.3.1.2 --- Detection of fetal-derived Tr-DNA --- p.15 / Chapter 1.3.1.3 --- Potential problems associated with the detection of Tr-DNA --- p.16 / Chapter 1.3.2 --- Cell-free DNA in urine as released from the urinary tract --- p.17 / Chapter 1.4 --- Other bodily fluids with cell-free nucleic acids --- p.18 / Chapter 1.4.1 --- Amniotic fluid --- p.19 / Chapter 1.4.2 --- Cerebrospinal fluid (CSF) --- p.20 / Chapter 1.4.3 --- Peritoneal fluid --- p.20 / Chapter CHAPTER 2: --- MICRORNA IN HUMANS --- p.21 / Chapter 2.1 --- Introduction --- p.21 / Chapter 2.2 --- Biogenesis --- p.21 / Chapter 2.2.1 --- Transcription of microRNA genes --- p.21 / Chapter 2.2.2 --- Processing and maturation of microRNA precursors --- p.23 / Chapter 2.3 --- Mechanisms of gene regulation --- p.24 / Chapter 2.3.1 --- Cleavage of target mRNA --- p.24 / Chapter 2.3.2 --- Translational repression of mRNA --- p.25 / Chapter 2.4 --- Functional roles of microRNAs --- p.25 / Chapter 2.4.1 --- Oncogenesis --- p.25 / Chapter 2.4.2 --- Programmed cell death --- p.26 / Chapter 2.4.3 --- Cellular differentiation and development --- p.27 / Chapter 2.4.4 --- Regulation of physiological and cellular processes --- p.28 / Chapter 2.5 --- Aim of this thesis --- p.28 / Chapter SECTION II: --- MATERIALS AND METHODS --- p.30 / Chapter CHAPTER 3: --- QUANTITATIVE ANALYSIS OF CIRCULATING AND URINARY NUCLEIC ACIDS --- p.31 / Chapter 3.1 --- Preparation of samples --- p.31 / Chapter 3.1.1 --- Preparation of plasma --- p.31 / Chapter 3.1.2 --- Preparation of blood cells --- p.32 / Chapter 3.1.3 --- Preparation of placental tissue --- p.32 / Chapter 3.1.4 --- Preparation of urine and urine cell pellet --- p.32 / Chapter 3.2 --- Nucleic acid extraction --- p.33 / Chapter 3.2.1 --- "Extraction of small RNA-containing total RNA from plasma, blood cells and placental tissue" --- p.33 / Chapter 3.2.2 --- Extraction of DNA from urine --- p.37 / Chapter 3.3 --- Quantitative measurements of nucleic acids --- p.38 / Chapter 3.3.1 --- Principle of real-time quantitative PCR --- p.38 / Chapter 3.3.2 --- One-step QRT-PCR assays for mRNA quantification --- p.40 / Chapter 3.3.2.1 --- Principle --- p.40 / Chapter 3.3.2.2 --- Quantification of human placental lactogen （hPL) mRNA --- p.40 / Chapter 3.3.3 --- Two-step QRT-PCR assays for microRNA quantification --- p.45 / Chapter 3.3.3.1 --- Principle --- p.45 / Chapter 3.3.3.2 --- Advantages --- p.46 / Chapter 3.3.3.3 --- TaqMan® MicroRNA Assays --- p.47 / Chapter 3.3.4 --- QPCR assays for DNA quantification --- p.53 / Chapter 3.3.4.1 --- Principle --- p.53 / Chapter 3.3.4.2 --- Quantification of the leptin gene and the sex-determining region on Ychromosome gene --- p.53 / Chapter 3.4 --- Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) --- p.57 / Chapter 3.4.1 --- Principle --- p.57 / Chapter 3.4.2 --- Zinc finger protein gene assay for determining the fractional concentration of male DNA --- p.58 / Chapter 3.5 --- Statistical analyses --- p.65 / Chapter SECTION III: --- CIRCULATING PLACENTAL MICRORNAS IN MATERNAL PLASMA AS MARKERS FOR PRENATAL DIAGNOSIS　 --- p.66 / Chapter CHAPTER 4: --- THE EXISTENCE AND QUANTITATIVE DETECTION OF CELL-FREE MICRORNAS IN PLASMA --- p.67 / Chapter 4.1 --- Introduction --- p.67 / Chapter 4.2 --- Materials and methods --- p.69 / Chapter 4.2.1 --- Sample collection --- p.69 / Chapter 4.2.2 --- Experimental design --- p.69 / Chapter 4.2.3 --- RNA extraction and quantification --- p.72 / Chapter 4.3 --- Results --- p.75 / Chapter 4.3.1 --- Validation of two-step QRT-PCR system for miRNA quantification --- p.75 / Chapter 4.3.2 --- Detection of cell-free miRNA in maternal plasma --- p.82 / Chapter 4.4 --- Discussion --- p.82 / Chapter CHAPTER 5: --- SYSTEMATIC IDENTIFICATION AND CHARACTERIZATION OF PLACENTAL MICRORNAS IN MATERNAL PLASMA --- p.86 / Chapter 5.1 --- Introduction --- p.86 / Chapter 5.2 --- Materials and methods --- p.88 / Chapter 5.2.1 --- Sample collection --- p.88 / Chapter 5.2.2 --- Experimental design --- p.88 / Chapter 5.2.3 --- RNA extraction and miRNA quantification --- p.91 / Chapter 5.3 --- Results --- p.93 / Chapter 5.3.1 --- A systematic search for placental miRNAs in maternal plasma using two-step QRT-PCR assays --- p.93 / Chapter 5.3.2 --- Detection rate and clearance kinetics of placental miRNAs in maternal plasma --- p.97 / Chapter 5.3.3 --- Effects of filtering maternal plasma on the concentration of placental miRNA and mRNA --- p.99 / Chapter 5.3.5 --- Temporal profile of placental miRNA concentrations in maternal plasma across different trimesters of pregnancies --- p.103 / Chapter 5.4 --- Discussion --- p.115 / Chapter SECTION IV: --- DETECTION OF CELL-FREE DNA IN URINE --- p.119 / Chapter CHAPTER 6: --- HEMATOPOIETIC STEM CELL TRANSPLANTATION RECIPIENTS AS A MODEL TO STUDY CELL-FREE DNA IN URINE --- p.120 / Chapter 6.1 --- Introduction --- p.120 / Chapter 6.2 --- Materials and methods --- p.123 / Chapter 6.2.1 --- Sample collection --- p.123 / Chapter 6.2.2 --- Experimental design --- p.124 / Chapter 6.2.3 --- DNA extraction and quantification --- p.125 / Chapter 6.3 --- Results --- p.128 / Chapter 6.3.1 --- Validation of the zinc finger protein gene assay --- p.128 / Chapter 6.3.2 --- Fractional concentration of male DNA in blood cells and plasma of sex-mismatched HSCT patients --- p.129 / Chapter 6.3.3 --- Fractional concentration of male DNA in the urine and the urine cell pellets of sex-mismatched HSCT patients --- p.131 / Chapter 6.3.4 --- Size distribution of cell-free DNA in peripheral blood and urine samples of sex-mismatched HSCT patients --- p.132 / Amplicon size --- p.138 / Chapter 6.4 --- Discussion --- p.143 / Chapter SECTION V: --- CONCLUDING REMARKS --- p.147 / Chapter CHAPTER 7: --- CONCLUSION AND FUTURE PERSPECTIVES --- p.148 / Chapter 7.1 --- Circulating miRNA is a valuable resource for molecular analysis --- p.148 / Chapter 7.2 --- The presence of donor-derived DNA in the urine of HSCT recipients --- p.150 / Chapter 7.3 --- Prospects for future work --- p.152 / APPENDIX 1 --- p.154 / REFERENCES --- p.158

植物合成多種發病機理相關蛋白以對抗病原體的侵襲。植物發病機理相關蛋白包括：核糖核酸酶；抗真菌蛋白；凝集素；胰蛋白酶抑制因子等。這些發病機理相關蛋白具有抗病毒，抗細菌，抗真菌，免疫調節及抗腫瘤等活性。從六種植物中提純了七個發病機理相關蛋白，包括三個凝集素，一個核糖核酸酶，兩個種抗真菌蛋白及一個胰蛋白酶抑制因子。 / 從西洋參須中提純了新的核糖核酸酶。核糖核酸酶分子量為26kDa，具有特异N末端氨基酸序列。此核糖核酸酶在 pH5 及 60℃ 條件下活性最高。它能抑制腫瘤細胞分裂及抑制人類後天免疫力缺乏症候群病毒逆轉錄酶活性。 / 從粉色菜豆及日本大花豆中提純了兩種凝集素。它們由兩個分子量為32kDa的亞基構成雙倍體。他們的活性穩定于0-60℃及3-12 pH。粉色菜豆凝集素的特异性糖基為木糖，日本大花豆凝集素的特异性糖基為半乳糖。從太子參中提純的凝集素分子量為33kDa，其活性穩定于0-60℃及2-5 pH。 這三種凝集素都具有抑制腫瘤細胞分裂及抑制人類後天免疫力缺乏症候群病毒逆轉錄酶活性。 / 提純的胰蛋白酶抑制因子分子量為21kDa。具有高耐熱及耐酸鹼性并表現出抑制腫瘤細胞分裂及抑制人類後天免疫力缺乏症候群病毒逆轉錄酶活性。從豇豆中提純的抗真菌肽分子量為5447Da，具有類防御素N末端氨基酸序列。 / Plants produce a diversity of proteins with antipathogenic activities. These proteins comprise among others, (i) ribonucleases, (ii) antifungal proteins, (iii) lectins and (iv) trypsin inhibitor with antiviral, antifungal and anticancer activities. The aim of this project was to isolate antipathogenic plant proteins including a ribonuclease from American ginseng branch roots, a trypsin inhibitor from rambutan seeds, defensin-like antifungal peptides from borlotti beans and king pole beans, and lectins from borlotti beans, Japanese large pinto beans and Pseudostellaria heterophylla. / The isolated 26-kDa ginseng branch root ribonuclease was monomeric with a novel N-terminal amino acid sequence. It exhibited maximal robonucleolytic activity toward yeast tRNA at pH 5 and 60℃. It inhibited proliferation of MCF7 human breast cancer cells and HepG2 human hepatoma cells. It also inhibited the activity of HIV-1 reverse transcriptase. / Both borlotti bean lectin and Japanese large pinto bean lectin were dimeric with a subunit molecular mass of 32-kDa. They were stable from 0℃ to 60℃ and from pH 3 to pH 12. Borlotti bean lectin was xylose-specific whereas Japanse large pinto bean lectin was galactose-specific. The 33-kDa Pseudostellaria heterophylla lectin could not be inhibited by the simple sugars tested. It was stable from 0℃ to 60℃ and from pH 2 to 5. All three isolated lectins suppressed proliferation of MCF7 and HepG2 cells and inhibited HIV-1 reverse transcriptase. / The isolated 21-kDa rambutan trypsin inhibitor has relatively high pH stability and thermostability, and exhibited HIV-1 reverse transcriptase inhibitory activity and antiproliferative activity toward a variety of tumor cells. The isolated 5447-Da king pole bean defensin-like peptide inhibited fungal growth. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Zhao, Yuan. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 202-222). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / Acknowledgements --- p.i / Abstract --- p.ii / 論文摘要 --- p.iv / Declaration --- p.v / Abbreviations --- p.vi / Table of Contents --- p.vii / List of Tables --- p.x / List of Figures --- p.xii / Chapter Chapter 1 --- Overview of Plant Defense-related Protein --- p.1 / Chapter 1.1 --- Overview of Lectins and hemagglutinins --- p.4 / Chapter 1.1.1 --- History and definition of lectins and hemagglutinins --- p.4 / Chapter 1.1.2 --- Occurrence and distribution of plant lectins --- p.6 / Chapter 1.1.3 --- Classification of lectins --- p.7 / Chapter 1.1.3.1 --- Classification of lectins on the basis of overall structure of lectin subunits --- p.7 / Chapter 1.1.3.2 --- Classification of lectins based on binding specificty to carbohydrates --- p.11 / Chapter 1.1.3.3 --- Classification of lectins according to families --- p.12 / Chapter 1.1.3.3.1 --- Legume lectins --- p.12 / Chapter 1.1.3.3.2 --- Monocot mannose-binding lectins --- p.13 / Chapter 1.1.3.3.3 --- Other lectins --- p.14 / Chapter 1.1.4 --- Defensive role of plant lectins --- p.15 / Chapter 1.1.5 --- Applications of plant lectins --- p.18 / Chapter 1.1.5.1 --- The antibacterial activity --- p.18 / Chapter 1.1.5.2 --- Anti-insect activity --- p.19 / Chapter 1.1.5.3 --- Antifungal activity --- p.21 / Chapter 1.1.5.4 --- The antiviral activity --- p.22 / Chapter 1.1.5.5 --- Lectin affinity chromatography --- p.23 / Chapter 1.1.5.6 --- Lectin microarray --- p.23 / Chapter 1.2 --- Overview of Ribonucleases --- p.26 / Chapter 1.2.1 --- History and definition of Ribonucleases --- p.26 / Chapter 1.2.2 --- Classification of Ribonucleases --- p.27 / Chapter 1.2.2.1 --- T1 Ribonucleases family --- p.27 / Chapter 1.2.2.2 --- RNase T2 family --- p.28 / Chapter 1.2.3 --- Biological activities of plant ribonucleases --- p.28 / Chapter 1.2.3.1 --- Phosphate remobilization --- p.28 / Chapter 1.2.3.2 --- Senescence --- p.29 / Chapter 1.2.3.3 --- Programmed cell death --- p.30 / Chapter 1.2.3.4 --- Plant defense --- p.31 / Chapter 1.2.3.5 --- RNA processing and decay --- p.32 / Chapter 1.2.3.6 --- Antitumor activities --- p.33 / Chapter 1.3 --- Other plant pathogen-related proteins --- p.34 / Chapter 1.3.1 --- Overview of chitinase --- p.34 / Chapter 1.3.1.1 --- Classification of chitinases --- p.35 / Chapter 1.3.1.2 --- Biological properties of chitinases --- p.38 / Chapter 1.3.2 --- Overview of plant ribosome-inactivating proteins (RIPs) --- p.41 / Chapter 1.3.2.1 --- Classification of RIPs --- p.42 / Chapter 1.3.2.2 --- Roles of RIPs in plants --- p.44 / Chapter 1.3.2.3 --- Possible application of RIPs --- p.46 / Chapter 1.3.3 --- Overview of thaumatin-like proteins (TLPs) --- p.50 / Chapter 1.3.3.1 --- Occurrence of TLPs --- p.51 / Chapter 1.3.3.2 --- Biological properties of TLPs --- p.52 / Chapter 1.4 --- Aim of this study --- p.54 / Chapter Chapter 2 --- Isolation of a lectin and an antifungal protein from Phaseolus vulgaris cv. Borlotti beans / Chapter 2.1 --- Introduction --- p.55 / Chapter 2.2 --- Materials and Methods --- p.55 / Chapter 2.3 --- Results --- p.64 / Chapter 2.4 --- Discussion --- p.79 / Chapter Chapter 3 --- Isolation of a lectin from Pinto beans (Phaseolus vulgaris pinto bean) / Chapter 3.1 --- Introduction --- p.82 / Chapter 3.2 --- Materials and Methods --- p.83 / Chapter 3.3 --- Results --- p.87 / Chapter 3.4 --- Discussion --- p.103 / Chapter Chapter 4 --- Isolation of a lectin from Pseudostellaria hetorophylla roots / Chapter 4.1 --- Introduction --- p.105 / Chapter 4.2 --- Materials and Methods --- p.107 / Chapter 4.3 --- Results --- p.110 / Chapter 4.4 --- Discussion --- p.122 / Chapter Chapter 5 --- Isolation of a ribonuclease from branch roots of American ginseng (Panax quinquefolium) / Chapter 5.1 --- Introduction --- p.124 / Chapter 5.2 --- Materials and Methods --- p.126 / Chapter 5.3 --- Results --- p.129 / Chapter 5.4 --- Discussion --- p.142 / Chapter Chapter 6 --- Isolation of a trypsin inhibitor in rambutan (Nephelium lappaceum L) seeds / Chapter 6.1 --- Introduction --- p.144 / Chapter 6.2 --- Materials and Methods --- p.147 / Chapter 6.3 --- Results --- p.152 / Chapter 6.4 --- Discussion --- p.163 / Chapter Chapter 7 --- Isoation of a defensin-like antifungal peptide from Phaseolus vulgaris cv. 'King Pole Bean' / Chapter 7.1 --- Introduction --- p.168 / Chapter 7.2 --- Materials and Methods --- p.170 / Chapter 7.3 --- Results --- p.173 / Chapter 7.4 --- Discussion --- p.181 / Chapter Chapter 8 --- Overall discussion --- p.183 / References --- p.186

A técnica de camada delgada em placas com meio de Middlebrook 7H11 foi comparada com a técnica padrão de cultura em meio de Stonebrink, a fim de se avaliar a sensibilidade e o tempo de detecção de micobactérias em amostras de leite, experimentalmente inoculadas com Mycobacterium bovis (estirpe AN5), em uma diluição 10-2, e submetidas a duas diferentes técnicas de processamento, utilizando-se da gordura (técnica 1)e do sedimento (técnica 2), descontaminadas pelo método de Petroff modificado (adicionado de Tween 80) e confrontadas com a técnica do leite total submetida ao método de Petroff e semeadas nos dois meios. Os resultados destas técnicas (1 e 2) foram comparados entre si pelo teste não paramétrico de Wilcoxon, e entre os resultados obtidos na técnica de leite total submetida ao método tradicional de Petroff, por meio do teste não paramétrico de Mann-Whitney. Posteriormente, amostras de leite nas diluições 10-3, 10-4 e 10-5 foram então submetidas às mesmas técnicas de processamento e descontaminação para averiguação de sensibilidade. Os resultados obtidos demonstraram que: 1) a técnica de cultivo de micobactérias em amostras de leite no meio de Middlebrook 7H11 se mostrou viável frente à tradicional; 2) a técnica de camada delgada permitiu a visualização precoce das micobactérias quando comparadas ao meio de Stonebrink; 3) as técnica 1 e 2 forneceram maior recuperação de micobactérias e maior proporção de cultivos positivos nos dois empregados; 4) a técnica de camada delgada em meio de Middlebrook 7H11 modificado pode ser usada como uma técnica complementar aos métodos tradicionais de diagnóstico da tuberculose bovina em amostras de leite para fins de vigilância epidemiológica. / The modified thin layer Middlebrook 7H11 cultivation technique was compared to the tradicional culture technique using the Stonebrink in order to evaluate the sensibility and the time for the detection of positive cultures of mycobacteria in milk samples, experimentally inoculated by Mycobacterium bovis (strain AN5), 10-2 dilution, were submitted by two types of procedures named technique 1 (milk fat) and 2 (milk sediment), descontamined by modified Petroff method, added with Tween 80, and compared with total milk samples submitted to tradicional Petroff method, in the same media types. The results of the two tecniques were compared within each other by the non-parametric Wilcoxon test, and within the results of standard milk submitted by the tradicional Petroff method, by the non-parametric Mann-Whitney. Milk samples were diluted to 10-3, 10-4 and 10-5 and submitted by the tecniques 1 and 2 descontaminated by the modified Petroff method, and total milk by tradicional Petroff method to averiguate the sensibility. The results found in this experiment demonstrated that 1) the modified thin layer technique of isolation of mycobacteria in milk samples was practicable against the tradicional one; 2) the time needed for the detection of mycobacteria colonies was slightly less with the thin layer technique than the tradicional culture method; 3) techniques 1 and 2 gave more number of colonies and more proportion of positive culture than the tradicional one in all media used; 4) this technique should be used as a complementary method to the tradicional one in the diagnostic of bovine tuberculosis from milk samples in orther to improve epidemiologic vigilance.

O presente trabalho descreve o desenvolvimento de uma técnica de nested-PCR para a detecção de ADN do vírus da laringotraqueíte infecciosa (VLTI) utilizando primers que amplificam uma região do gene que codifica a glicoproteína E viral. A técnica padronizada amplificou amostras isoladas e de campo, mostrando alta sensibilidade e especificidade tomando o isolamento em ovos embrionados SPF como técnica de referência. O seqüenciamento de uma amostra positiva por nested-PCR confirmou a identidade do produto amplificado. Foram submetidas a nested-PCR padronizada amostras de traquéia, pulmão e conjuntiva de 51 granjas de poedeiras comerciais procedentes da região de Bastos, Estado de São Paulo, que apresentou um surto caracterizado por sinais respiratórios, queda na produção de ovos e aumento da mortalidade. Vinte e três granjas foram positivas a nested-PCR e vinte e duas amostras foram isoladas. Considerando os resultados das duas técnicas, vinte e quatro granjas resultaram positivas. Não foram detectados os vírus das doenças de Newcastle, pneumovirose aviaria, nem Mycoplasma gallisepticum. O vírus da bronquite infecciosa das galinhas foi detectado em uma granja, e Mycoplasma synoviae em oito. A alta freqüência de ocorrência do VLTI, a alta concordância entre o quadro clínico observado e detecção do VLTI e o resultados do diagnóstico diferencial demonstram que o VLTI foi o agente etiológico causador de um surto de doença respiratória observado na região de Bastos, Estado de São Paulo. / The present work describes the development of a nested-PCR technique for the detection of Infectious laryngotracheitis virus (ILTV) DNA using primers to amplify a region that encodes the glycoprotein E. The standardized technique amplified isolated and from clinical strains, offering high sensitivity and specificity, using the isolation in SPF chicken embryos such as reference test. The identity of the amplified product of the nested-PCR was confirmed by DNA sequencing. Trachea, lung and conjunctiva samples from fifty-one commercial layer farms from Bastos region, São Paulo State, which showed an outbreak characterized by respiratory signs, decreased egg production and increased mortality, were tested by the standardized nested-PCR. Twenty-three farms were positive by nested-PCR and samples of twenty-two farms were isolated. Twenty-four farms were positives when the results of both techniques were considered. Newcastle disease virus, Avian Pneumovirus and Mycoplasma gallisepticum were not detected. Infectious bronchitis virus was detected in one farm and Mycoplasma synoviae was detected in eight farms. The high frequency of occurrence of ILTV, the high agreement between clinical signs observed and ILTV detection and the results of differential diagnosis demonstrate that ILTV was the etiological agent of an outbreak respiratory disease observed in Bastos region, São Paulo State.

Transmissions based precautions are measures implemented in various clinical health care settings as a means to prevent the transmission of infectious diseases and decrease instances of healthcare acquired infections (HAI). HAI’s result in increased cost to hospitals, longer hospitalization for patients, increased patient suffering, and fatal patient outcomes. While staff member adherence to transmissions based precautions are mandated through various organizations and hospital policies, a review of literature indicates little research has been conducted regarding visitor compliance with transmission-based precautions. The potential implications in healthcare from visitor non-adherence acquired infections are unknown; revealing a gap in literature and supporting the need for further research to describe the phenomenon. Through utilization of a descriptive online survey instrument, the purpose of this descriptive study is to gain insight into why nurses believe visitors may or may not be compliant with transmission-based precautions. To collect the data, an online descriptive survey instrument was developed and distributed via email to all graduate students’ enrolled East Tennessee State University’s College of Nursing. Only ten participants met the eligibility requirements to participate in this study. Data was analyzed though a predictive analytics software and grouping responses into themes. Responses suggest that nurses feel visitors are not complying with transmission-based precautions because of a lack in education, not perceiving the infection as a threat, prior exposure to loved one at home, and inconvenience.

abstract: Advances in health care have resulted in an increase in life expectancy causing a rapid growth in the number of older adults at a global level. At the same time, socioeconomic development is transitioning family structures and social relationships. With reduced family engagement, many older adults are more at risk for physical and psychological health issues including loneliness, which is considered a public health issue affecting their quality of life and well-being. This descriptive, exploratory study aims to describe the significance of loneliness in three northern regions of the Sultanate of Oman. The purpose of this study is to examine the prevalence and correlates of loneliness and the relationship of loneliness to health statuses among older Omani adults aged 60 years and above. A demographic data questionnaire, the UCLA loneliness scale, and SF-12-v-1 health status instruments were used for data collection. The sample includes 113 Omani older adults, male (n = 36) and female (n = 77), who experienced a mixture from low to high and severe levels of loneliness. Among these older adults, 34.5% perceived low level, 34.5% moderate level, 22.1% high, and 8.8% were severely lonely. The main demographic factors that were associated with the older adults level of loneliness were female gender, older age 80 years and above, living with others who were not a family member, and being unemployed. When controlling for demographic and environmental factors loneliness was a significant predictor (p < .001) for lower mental health status but not for physical health status (p > .05). / Dissertation/Thesis / Doctoral Dissertation Nursing 2019

No description available.

This qualitative study explored the perceptions of evangelical freshmen students attending the University of Texas at Arlington and the University of Texas at Dallas during the spring semester of 2006 in the context of student alienation. The purpose of this study was to explore the possibility that evangelicals attending secular universities were perceiving alienation through their interactions with their universities. It was hypothesized that the modern university, having evolved into its present naturalistic worldview condition, might prove alienating to evangelicals from a worldview standpoint. Assuming the possibility that alienation might prove to be a reality for evangelicals, the subordinate purposes were intended to discover the types and sources of alienation, the possible evangelical coping strategies, and their perceptions of the university's reaction to them as evangelicals. During the spring semester of 2006, I conducted two live interviews with twenty participants. The first interview included a questionnaire which was administered for the purpose of providing insight into each participant's religiosity or evangelical commitment. The first interview (conducted prior to spring break) asked the students to reflect back upon their first semester experience (the fall of 2005). The second interview, conducted towards the end of the spring semester, was oriented towards the second semester experience. I found that all evangelicals but one had successfully assimilated socially and academically into their respective university. Their academic assimilation was primarily manifested by their relatively high academic achievement. Although they did experience worldview related incongruence, it was not severe enough to manifest any related attrition. I found the most severe incongruence to be related to the perceptions of a negative university moral ethos combined with the prevailing naturalistic monism of the university that relegated the Christian worldview to marginalization or irrelevance. I also found that the high level of social integration was primarily related to participant affiliation with various evangelical entities independent of the university. The data revealed that zero participants lost or abandoned their evangelical faith during their freshman year, and the students' perceived that they had actually experienced positive growth in their spiritual lives as a result of the overall college freshman experience.

The oral pathogen Streptococcus mutans is a known etiological agent for dental root decay and coronal caries. It has been hypothesized, but not yet proven, that S. mutans expression of virulence genes in dental plaque may be influenced by its interaction with co-aggregating partners, notably Fusobacterium nucleatum and Actinomyces viscosus. Investigation of the suitability of mixed cultures of S. mutans with F. nucleatum versus S. mutans with A. viscosus proved that A. viscosus was a better target in the present laboratory setting. Furthermore, A. viscosus, a causative agent of mandible osteomyelitis and endocarditis, has been shown to have direct interaction ability with S. mutans. DNA microarray analysis was used in the present study to investigate the influence of co-aggregation with A. viscosus on the expression of S. mutans genes. Microarrays have been used successfully in the analysis of differential gene expression in S. mutans as a function of culture conditions, such as in biofilms versus planktonic states. This technology however, has not yet been applied to the analysis of homologous versus heterologous cultures. The present study was conducted in order to identify potential problems associated with the application of microarray analysis to mixed cultures. The data obtained encourage the further testing of microarrays for the analysis of heterologous cultures of oral bacteria.

Surface functionalization of living cells for cell therapeutics has gained substantial momentum in the last two decades. From encapsulating islets of Langerhans, to cell laden gels for tissue scaffolds, to individual cell encapsulation in thin hydrogels, to surface adhesives and inert surface camouflage, modification of living cell surfaces has a wide array of important applications. Here we use hydrogel encapsulation of individual cells as a mode of protection from mechanical forces for high throughput cell printing, and chemical stimuli for the isolation of rare cells in blood. In the first study, we review methods of surface functionalization and establish a metric of potential target biomarkers for circulating tumor cell (CTC) isolation. For extended applications in cancer detection through a fluid biopsy, common surface antigen densities were quantitatively assessed in relation to peripheral blood mononuclear cells (PBMCs) for potential targets of cell specific encapsulation. We then look to commercialization of our process after considering biopsy volumes and cell therapy dose sizes. Undesired batch-to-batch variation in our in-house synthesized photo-initiator could be eliminated by the use of fluorescein, a commercial fluorochrome of similar initiating power to our current eosin initiating system. Fluorescence and hydrogel generation were compared indicating a fluorescein conjugate has comparable power to that of our in-house conjugated eosin. Parameters involving the number of cells and fluid volumes processed were then analyzed systematically. Key parameters were studied to determine optimal equipment and protocol for clinically relevant batch sizes. The final study looks at the mechanical protection provided by thin hydrogel encapsulation. With growing interests in 3D bioprinting and goals of viable whole organ printing for transplant, high resolution and high throughput printing is a growing need. 3D bioprinting presents intense mechanical stimuli in the process that cells must endure. Here we analyze how hydrogel encapsulation reinforces the cellular membrane allowing cells to withstand the damaging forces associated with bioprinting.