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.

Formation and investigation of nanostructured thin films

Small devices, in the range of nanometers, are playing a major role in today's technology. The field of nanotechnology is concerned with materials and systems whose structures and components exhibit novel and significantly improved physical, chemical and biological properties, phenomena and processes due to their small nanoscale size. Researches more and more are finding that structural features in the range of about 1 to 100 nanometers behave quite differently than isolated molecules (1 nanometer) or bulk materials. For comparison, a 10 nanometer structure is 1000 times smaller than the diameter of a human hair. The virtues of working in the nanodomain are increasingly recognized by the scientific community and discussed in the popular press. The use of such devices is expected to revolutionize our industries and lives. ^ This work mainly focuses on the fabrication, characterization and discovery of new nanostructured thin films. This research consists of the design of a new high-deposition rate nanoparticle machine for depositing nanostructured films from beams of nanoparticles and investigation film's unique optical and physical properties.^ A high-deposition rate nanoparticle machine was designed, built and successfully tested. Different nanostructured thin films were deposited from Copper, Gold, Iron and Zirconium targets with the grain size of between 1 to 20 nm under different conditions. Transmission Electron Microscopy (TEM), Atomic Force Microscopy (AFM), and x-ray diffraction (XRD) confirmed nanoscale grain size structures of deposited films. The optical properties of the nanostructured films deposited from copper, Iron and Zirconium targets were significantly different from optical properties of bulk and thin films. Zr, Cu and Fe films were transparent. Gold films revealed an epitaxial contact with the silicon substrate with interesting crystal structures. ^ The new high-deposition rate nanoparticle machine was able to deposit new nanostructured films with different properties from bulk and thin films reported in the literatures. ^

Capabilities of Police and Military Forces in Central America -- A Comparative Analysis of Guatemala, El Salvador, Honduras an d Nicaragua

A difficult transition to a new paradigm of Democratic Security and the subsequent process of military restructuring during the nineties led El Salvador, Honduras, Guatemala and Nicaragua to re-consider their old structures and functions of their armed forces and police agencies. This study compares the institutions in the four countries mentioned above to assess their current condition and response capacity in view of the contemporary security challenges in Central America. This report reveals that the original intention of limiting armies to defend and protect borders has been threatened by the increasing participation of armies in public security. While the strength of armies has been consolidated in terms of numbers, air and naval forces have failed to become strengthened or sufficiently developed to effectively combat organized crime and drug trafficking and are barely able to conduct air and sea operations. Honduras has been the only country that has maintained a proportional distribution of its armed forces. However, security has been in the hands of a Judicial Police, supervised by the Public Ministry. The Honduran Judicial Police has been limited to exercising preventive police duties, prohibited from carrying out criminal investigations. Nicaragua, meanwhile, possesses a successful police force, socially recognized for maintaining satisfactory levels of security surpassing the Guatemalan and El Salvadoran police, which have not achieved similar results despite of having set up a civilian police force separate from the military. El Salvador meanwhile, has excelled in promoting a Police Academy and career professional education, even while not having military attachés in other countries. Regarding budgetary issues, the four countries allocate almost twice the amount of funding on their security budgets in comparison to what is allocated to their defense budgets. However, spending in both areas is low when taking into account each country's GDP as well as their high crime rates. Regional security challenges must be accompanied by a professionalization of the regional armies focused on protecting and defending borders. Therefore, strong institutional frameworks to support the fight against crime and drug trafficking are required. It will require the strengthening of customs, greater control of illicit arms trafficking, investment in education initiatives, creating employment opportunities and facilitating significant improvements in the judicial system, as well as its accessibility to the average citizen.

Potential for Cell-Transplant Therapy with Human Neuronal Precursors to Treat Neuropathic Pain in Models of PNS and CNS Injury: Comparison of hNT2.17 and hNT2.19 Cell Lines

Effective treatment of sensory neuropathies in peripheral neuropathies and spinal cord injury (SCI) is one of the most difficult problems in modern clinical practice. Cell therapy to release antinociceptive agents near the injured spinal cord is a logical next step in the development of treatment modalities. But few clinical trials, especially for chronic pain, have tested the potential of transplant of cells to treat chronic pain. Cell lines derived from the human neuronal NT2 cell line parentage, the hNT2.17 and hNT2.19 lines, which synthesize and release the neurotransmitters gamma-aminobutyric acid (GABA) and serotonin (5HT), respectively, have been used to evaluate the potential of cell-based release of antinociceptive agents near the lumbar dorsal (horn) spinal sensory cell centers to relieve neuropathic pain after PNS (partial nerve and diabetes-related injury) and CNS (spinal cord injury) damage in rat models. Both cell lines transplants potently and permanently reverse behavioral hypersensitivity without inducing tumors or other complications after grafting. Functioning as cellular minipumps for antinociception, human neuronal precursors, like these NT2-derived cell lines, would likely provide a useful adjuvant or replacement for current pharmacological treatments for neuropathic pain.

Ecohydrology, evapotranspiration and hydrogeochemistry of carbonate mangrove wetlands

Coastal environments can be highly susceptible to environmental changes caused by anthropogenic pressures and natural events. Both anthropogenic and natural perturbations may directly affect the amount and the quality of water flowing through the ecosystem, both in the surface and subsurface and can subsequently, alter ecological communities and functions. The Florida Everglades and the Sian Ka'an Biosphere Reserve (Mexico) are two large ecosystems with an extensive coastal mangrove ecotone that represent a historically altered and pristine environment, respectively. Rising sea levels, climate change, increased water demand, and salt water intrusion are growing concerns in these regions and underlies the need for a better understanding of the present conditions. The goal of my research was to better understand various ecohydrological, environmental, and hydrogeochemical interactions and relationships in carbonate mangrove wetlands. A combination of aqueous geochemical analyses and visible and near-infrared reflectance data were employed to explore relationships between surface and subsurface water chemistry and spectral biophysical stress in mangroves. Optical satellite imagery and field collected meteorological data were used to estimate surface energy and evapotranspiration and measure variability associated with hurricanes and restoration efforts. Furthermore, major ionic and nutrient concentrations, and stable isotopes of hydrogen and oxygen were used to distinguish water sources and infer coastal groundwater discharge by applying the data to a combined principal component analysis-end member mixing model. Spectral reflectance measured at the field and satellite scales were successfully used to estimate surface and subsurface water chemistry and model chloride concentrations along the southern Everglades. Satellite imagery indicated that mangrove sites that have less tidal flushing and hydrogeomorphic heterogeneity tend to have more variable evapotranspiration and soil heat flux in response to storms and restoration. Lastly, water chemistry and multivariate analyses indicated two distinct fresh groundwater sources that discharge to the phosphorus-limited estuaries and bays of the Sian Ka'an Biopshere Reserve; and that coastal groundwater discharge was an important source for phosphorus. The results of the study give us a better understanding of the ecohydrological and hydrogeological processes in carbonate mangrove environments that can be then be extrapolated to similar coastal ecosystems in the Caribbean.

Labor turnover : its causes, consequences, and control strategy in five selected restaurants in the Miami area

The purpose of this study is to identify and analyze the basic causes of food service employee turnover in five selected restaurants in the Miami area. The withdrawal behavior in this study is treated in terms of controllable turnover, for the purpose of management, learning more about what action to take to solve this problem which has eaten into the fabric of the hospitality industry. The aim is to find out from the food service employees and management view of work for the purpose of identifying the variables which cause an employee to voluntarily leave a job. The objective is therefore, to analyze and describe the problem of labor turnover in these selected restaurants. Such description must precede efforts to arrive at solutions to the problem if these efforts are ever to be more than haphazard and superficial. Sigmund Freud once stated: "The true beginning of scientific activity consists in describing phenomena and only then in proceeding to group, classify and correlate them."1 The nature of the study is basically descriptive survey. Data is collected by the use of management questionnaire, food service employee questionnaire and finally employees job description index. The survey consisted of a series of well defined questions with open and closed endings dealing with employee with employee turnover. As Robert Ferber and P. J. Verdoom state in their book titled Research Method in Economics of Business: "Structured questionnaires, by supplying question formulations in very specific terms as well as the different possible answers are easier for the sample members to answer and also serve to reduce the danger of interviewer bias."2 The answers to the prepared questionnaire by sample members were then recorded. The results of the questionnaire responses were then compiled for presentation and analysis. 1 Julian Simon, Basic Research Methods in Social Science. Random House, New York, 1969, p.53. 2 Robert J. Ferber and P.J. Verdoon, Research Methods in Economics and Business, The McMillan Company, 1962, p. 20 9 .