Development of micro immunosensors to study genomic and proteomic biomarkers related to cancer and Alzheimer's disease

A report from the National Institutes of Health defines a disease biomarker as a “characteristic that is objectively measured and evaluated as an indicator of normal biologic processes, pathogenic processes, or pharmacologic responses to a therapeutic intervention.” Early diagnosis is a crucial factor for incurable disease such as cancer and Alzheimer’s disease (AD). During the last decade researchers have discovered that biochemical changes caused by a disease can be detected considerably earlier as compared to physical manifestations/symptoms. In this dissertation electrochemical detection was utilized as the detection strategy as it offers high sensitivity/specificity, ease of operation, and capability of miniaturization and multiplexed detection. Electrochemical detection of biological analytes is an established field, and has matured at a rapid pace during the last 50 years and adapted itself to advances in micro/nanofabrication procedures. Carbon fiber microelectrodes were utilized as the platform sensor due to their high signal to noise ratio, ease and low-cost of fabrication, biocompatibility, and active carbon surface which allows conjugation with biorecognition moieties. This dissertation specifically focuses on the detection of 3 extensively validated biomarkers for cancer and AD. Firstly, vascular endothelial growth factor (VEGF) a cancer biomarker was detected using a one-step, reagentless immunosensing strategy. The immunosensing strategy allowed a rapid and sensitive means of VEGF detection with a detection limit of about 38 pg/mL with a linear dynamic range of 0–100 pg/mL. Direct detection of AD-related biomarker amyloid beta (Aβ) was achieved by exploiting its inherent electroactivity. The quantification of the ratio of Aβ1-40/42 (or Aβ ratio) has been established as a reliable test to diagnose AD through human clinical trials. Triple barrel carbon fiber microelectrodes were used to simultaneously detect Aβ1-40 and Aβ1-42 in cerebrospinal fluid from rats within a detection range of 100nM to 1.2μM and 400nM to 1μM respectively. In addition, the release of DNA damage/repair biomarker 8-hydroxydeoxyguanine (8-OHdG) under the influence of reactive oxidative stress from single lung endothelial cell was monitored using an activated carbon fiber microelectrode. The sensor was used to test the influence of nicotine, which is one of the most biologically active chemicals present in cigarette smoke and smokeless tobacco.

A study to identify the motivation compelling the business traveler to choose a specific airline in the Miami to Atlanta market and to develop a marketing strategy

The purpose of this study was to determine if the business traveler's behavior is influenced by brand loyalty. This brand loyalty, which became evident through the use of a survey, was then to be thoroughly evaluated. In order for this information to be best understood and utilized as the basis of future marketing strategies, much research was undertaken and its significance explained in relation to the airline industry as it exists at present. The results and conclusions of this study indicate that the airline industry is, for the most part, taking a successful approach in attracting business travelers. These travelers' business is highly valued due to the frequency with which they pay full-fare rates. The airlines view business travelers as a potential for great profit and their actions are in line with these philosophies.

Mindmapping as an adult learning notetaking strategy to increase the encoding of information in a corporate training course

A study was conducted to investigate the effectiveness, as measured by performance on course posttests, of mindmapping versus traditional notetaking in a corporate training class. The purpose of this study was to increase knowledge concerning the effectiveness of mindmapping as an information encoding tool to enhance the effectiveness of learning. Corporations invest billions of dollars, annually, in training programs. Given this increased demand for effective and efficient workplace learning, continual reliance on traditional notetaking is questionable for the high-speed and continual learning required on workers.^ An experimental, posttest-only control group design was used to test the following hypotheses: (1) there is no significant difference in posttest scores on an achievement test, administered immediately after the course, between adult learners using mindmapping versus traditional notetaking methods in a training lecture, and (2) there is no significant difference in posttest scores on an achievement test, administered 30 days after the course, between adult learners using mindmapping versus traditional notetaking methods in a training lecture. After a 1.5 hour instruction on mindmapping, the treatment group used mindmapping throughout the course. The control group used traditional notetaking. T-tests were used to determine if there were significant differences between mean posttest scores between the two groups. In addition, an attitudinal survey, brain hemisphere dominance survey, course dynamics observations, and course evaluations were used to investigate preference for mindmapping, its perceived effect on test performance, and the effectiveness of mindmapping instruction.^ This study's principal finding was that although the mindmapping group did not perform significantly higher on posttests administered immediately and 30 days after the course, than the traditional notetaking group, the mindmapping group did score higher on both posttests and reported higher ratings of the course on every evaluation criteria. Lower educated, right brain dominant learners reported a significantly positive learning experience. These results suggest that mindmapping enhances and reinforces the preconditions of learning. Recommendations for future study are provided. ^

Using a Bilingual Reading Strategy to Reduce SLD Numbers

Studies show that for Spanish speaking students, vocabulary expansion in Spanish increases fluency and comprehension in English. Therefore, creating a bilingual reading intervention program with a strong emphasis on vocabulary building in the early grades will help improve the students’ overall reading comprehension in English and avoid improper placement.

The Use of Literature to Combat Bullying

Bullying is a pervasive phenomenon. This study examined what teachers think encourages bullying among young people, and what effects teachers believe reader response strategies would have on their students. The study found teachers implementing reader response strategies in discussing literature were able to influence behavior in students and reduce bullying.

Near Term Computer Management Strategy For Hospitality Managers and Computer System Vendors

In his dialogue - Near Term Computer Management Strategy For Hospitality Managers and Computer System Vendors - by William O'Brien, Associate Professor, School of Hospitality Management at Florida International University, Associate Professor O’Brien initially states: “The computer revolution has only just begun. Rapid improvement in hardware will continue into the foreseeable future; over the last five years it has set the stage for more significant improvements in software technology still to come. John Naisbitt's information electronics economy¹ based on the creation and distribution of information has already arrived and as computer devices improve, hospitality managers will increasingly do at least a portion of their work with software tools.” At the time of this writing Assistant Professor O’Brien will have you know, contrary to what some people might think, the computer revolution is not over, it’s just beginning; it’s just an embryo. Computer technology will only continue to develop and expand, says O’Brien with citation. “A complacent few of us who feel “we have survived the computer revolution” will miss opportunities as a new wave of technology moves through the hospitality industry,” says ‘Professor O’Brien. “Both managers who buy technology and vendors who sell it can profit from strategy based on understanding the wave of technological innovation,” is his informed opinion. Property managers who embrace rather than eschew innovation, in this case computer technology, will benefit greatly from this new science in hospitality management, O’Brien says. “The manager who is not alert to or misunderstands the nature of this wave of innovation will be the constant victim of technology,” he advises. On the vendor side of the equation, O’Brien observes, “Computer-wise hospitality managers want systems which are easier and more profitable to operate. Some view their own industry as being somewhat behind the times… They plan to pay significantly less for better computer devices. Their high expectations are fed by vendor marketing efforts…” he says. O’Brien warns against taking a gamble on a risky computer system by falling victim to un-substantiated claims and pie-in-the-sky promises. He recommends affiliating with turn-key vendors who provide hardware, software, and training, or soliciting the help of large mainstream vendors such as IBM, NCR, or Apple. Many experts agree that the computer revolution has merely and genuinely morphed into the software revolution, informs O’Brien; “…recognizing that a computer is nothing but a box in which programs run.” Yes, some of the empirical data in this article is dated by now, but the core philosophy of advancing technology, and properties continually tapping current knowledge is sound.

Specific Increase in MDR1 Mediated Drug-Efflux in Human Brain Endothelial Cells following Co-Exposure to HIV-1 and Saquinavir

Persistence of HIV-1 reservoirs within the Central Nervous System (CNS) remains a significant challenge to the efficacy of potent anti-HIV-1 drugs. The primary human Brain Microvascular Endothelial Cells (HBMVEC) constitutes the Blood Brain Barrier (BBB) which interferes with anti-HIV drug delivery into the CNS. The ATP binding cassette (ABC) transporters expressed on HBMVEC can efflux HIV-1 protease inhibitors (HPI), enabling the persistence of HIV-1 in CNS. Constitutive low level expression of several ABC-transporters, such as MDR1 (a.k.a. P-gp) and MRPs are documented in HBMVEC. Although it is recognized that inflammatory cytokines and exposure to xenobiotic drug substrates (e.g HPI) can augment the expression of these transporters, it is not known whether concomitant exposure to virus and anti-retroviral drugs can increase drug-efflux functions in HBMVEC. Our in vitro studies showed that exposure of HBMVEC to HIV-1 significantly up-regulates both MDR1 gene expression and protein levels; however, no significant increases in either MRP-1 or MRP-2 were observed. Furthermore, calcein-AM dye-efflux assays using HBMVEC showed that, compared to virus exposure alone, the MDR1 mediated drug-efflux function was significantly induced following concomitant exposure to both HIV-1 and saquinavir (SQV). This increase in MDR1 mediated drug-efflux was further substantiated via increased intracellular retention of radiolabeled [3H-] SQV. The crucial role of MDR1 in 3H-SQV efflux from HBMVEC was further confirmed by using both a MDR1 specific blocker (PSC-833) and MDR1 specific siRNAs. Therefore, MDR1 specific drug-efflux function increases in HBMVEC following co-exposure to HIV-1 and SQV which can reduce the penetration of HPIs into the infected brain reservoirs of HIV-1. A targeted suppression of MDR1 in the BBB may thus provide a novel strategy to suppress residual viral replication in the CNS, by augmenting the therapeutic efficacy of HAART drugs.

The Central European initiative : an approach to regional stabilization and democratic consolidation

This thesis examines how multilateral institutions can contribute to democratization and regional stability. It is a case study of the Central European Initiative (CEI), a regional intergovernmental organization founded by Italy and Austria in 1989 to undertake a regional integration process, and of its role in stabilizing post-communist democracies. Documents were collected at the CEI offices in Trieste, Italy, and interviews conducted with CEI officers, and data obtained from the websites of related organizations and of CEI member countries. The thesis probes the relevance of concepts derived from theories of international regimes and social constructivism. It shows that the CEI diffuses norms and institutional rules conducive to consolidating democracies, including the development of a free civil society, a relatively autonomous political society, rule of law, state bureaucracies that are usable by the new democratic governments, and functioning free market economies, addition, and fosters habits of dialogue, socializes participants, and supports the creation of supra-national identities.

Screening for AmpR-Specific Inhibitors to Combat P. aeruginosa Infections

Pseudomonas aeruginosa, a Gram-negative bacterium, an opportunistic pathogen that infects individuals suffering from reduced immunity or damaged tissue. The treatment of these infections has become a major problem due to its increasing antibiotic resistance. Many multi-drug resistant isolates of P. aeruginosa can thwart most antibiotic classes including ?- lactams, fluoroquinolones, and aminoglycosides. Its ability to combat ?-lactams is in part due to expression of AmpC, a major chromosomally encoded ?-lactamase. The expression of ampC is positively regulated by AmpR. Besides antibiotic resistance, AmpR is an important regulator of various factors that are required for establishing acute and chronic infections. Loss of ampR makes P. aeruginosa susceptible to ?-lactams and less virulent than the wild type. We hypothesize that AmpR is a potential therapeutic target. In the absence of new drugs in the pipeline, the aim of this study is to find an AmpR-specific inhibitor to assist and improve the use of currently available ?- lactam treatment. A small-molecule library from Torrey Pines Institute will be used in this study. Two reporter systems, lux and lacZ, fused to a PampC promotor will be used to assess AmpR activity. Positive hits will be those that inhibit 50% PampC activity in the presence of sub inhibitory concentration of imipenem, a ?- lactam. The top positive hits will be screened for their ability to cause human cell-cytotoxicity. The non-cytotoxic hits will be assessed for their ability to affect P. aeruginosa virulence and antibiotic resistance using various in vitro assays. Determination of potential AmpR inhibitors will prove to be useful in fighting off infections and may save countless patients suffering from these infections.

Left to right: Oliva and Damon at TS/Ph-1b, Taylor Slough

This material is based upon work supported by the National Science Foundation through the Florida Coastal Everglades Long-Term Ecological Research program under Cooperative Agreements #DBI-0620409 and #DEB-9910514. This image is made available for non-commercial or educational use only.

Left to right: Oliva and Damon at TS/Ph-1b, Taylor Slough

This material is based upon work supported by the National Science Foundation through the Florida Coastal Everglades Long-Term Ecological Research program under Cooperative Agreements #DBI-0620409 and #DEB-9910514. This image is made available for non-commercial or educational use only.

Left to right: Laura and Damon collecting periphyton at TS/Ph-2, Taylor Slough

This material is based upon work supported by the National Science Foundation through the Florida Coastal Everglades Long-Term Ecological Research program under Cooperative Agreements #DBI-0620409 and #DEB-9910514. This image is made available for non-commercial or educational use only.

Edward Castaneda retrieving a soil core to assess sedimentation and nutrient accumulation rates in a dwarf mangrove forest, Taylor Slough

This material is based upon work supported by the National Science Foundation through the Florida Coastal Everglades Long-Term Ecological Research program under Cooperative Agreements #DBI-0620409 and #DEB-9910514. This image is made available for non-commercial or educational use only.