Faculty Bibliography

BACKGROUND: More than 35 million people in developing countries are living with HIV infection. An enormous global effort is now underway to bring antiretroviral treatment to at least 3 million of those infected. While drug prices have dropped considerably, the cost and technical complexity of laboratory tests essential for the management of HIV disease, such as CD4 cell counts, remain prohibitive. New, simple, and affordable methods for measuring CD4 cells that can be implemented in resource-scarce settings are urgently needed. METHODS AND FINDINGS: Here we describe the development of a prototype for a simple, rapid, and affordable method for counting CD4 lymphocytes. Microliter volumes of blood without further sample preparation are stained with fluorescent antibodies, captured on a membrane within a miniaturized flow cell and imaged through microscope optics with the type of charge-coupled device developed for digital camera technology. An associated computer algorithm converts the raw digital image into absolute CD4 counts and CD4 percentages in real time. The accuracy of this prototype system was validated through testing in the United States and Botswana, and showed close agreement with standard flow cytometry (r = 0.95) over a range of absolute CD4 counts, and the ability to discriminate clinically relevant CD4 count thresholds with high sensitivity and specificity. CONCLUSION: Advances in the adaptation of new technologies to biomedical detection systems, such as the one described here, promise to make complex diagnostics for HIV and other infectious diseases a practical global reality.

We report here the adaptation of our electronic microchip technology towards the development of a new method for detecting and enumerating bacterial cells and spores. This new approach is based on the immuno-localization of bacterial spores captured on a membrane filter microchip placed within a flow cell. A combination of microfluidic, optical, and software components enables the integration of staining of the bacterial species with fully automated assays. The quantitation of the analyte signal is achieved through the measurement of a collective response or alternatively through the identification and counting of individual spores and particles. This new instrument displays outstanding analytical characteristics, and presents a limit of detection of approximately 500 spores when tested with Bacillus globigii (Bg), a commonly used simulant for Bacillus anthracis (Ba), with a total analysis time of only 5 min. Additionally, the system performed well when tested with real postal dust samples spiked with Bg in the presence of other common contaminants. This new approach is highly customizable towards a large number of relevant toxic chemicals, environmental factors, and analytes of relevance to clinical chemistry applications.

Although extensive amounts of research have been carried out on superconductor-normal metal-superconductor (SNS) electronic devices, the fabrication of superconductor SNS devices still remains difficult. Surface modification of high-temperature superconductors could be a way to control the interface of SNS electronic device fabrication. Here, we developed a cleaning method for thin films of high-temperature superconductor surface based on self-assembled monolayers. High-quality c-axis orientated YBa2Cu3O7-δ (i.e. , YBCO) and Y0.6Ca0.4Ba1.6La0.4Cu3O7-δ (i.e., TX-YBCO) thin films were deposited by standard laser ablation methods. YBCO/Au/YBCO and TX-YBCO/Au/TX-YBCO planar type junctions were fabricated by photolithography, focused-ion-beam milling, and ex situ sputter depositions. A 40-50 nm nanotrench was ion milled on the thin film by FIB, and a thin gold layer was deposited by an ex situ method on the nanotrench to connect the two separated high-temperature superconductor electrodes. SEM, AFM, and R vs T resistivity measurements were used to compare the corrosion layer formed in the interface of the SNS junctions with the SAM cleaned SNS junction. Evidence here suggests that the SAM cleaning method can be used to remove the degradation layer on the surface of cuprate superconductors. The obtained contact resistivity value, (10(-8) &UOmega; cm(2)) for a SNS junction with SAM treatment is comparable with that of SNS junctions fabricated by the in situ methods. (C) 2005 American Institute of Physics.

In the last decade, saliva has been advocated as a non-invasive alternative to blood as a diagnostic fluid. However, use of saliva has been hindered by the inadequate sensitivity of current methods to detect the lower salivary concentrations of many constituents compared to serum. Furthermore, developments in the areas related to lab-on-a-chip systems for saliva-based point of care diagnostics are complicated by the high viscosity and heterogeneous properties associated with this diagnostic fluid. The biomarker C-reactive protein (CRP) is an acute phase reactant and a well-accepted indicator of inflammation. Numerous clinical studies have established elevated serum CRP as a strong, independent risk factor for the development of cardiovascular disease (CVD). CVD has also been associated with oral infections (i.e. periodontal diseases) and there is evidence that systemic CRP may be a link between the two. Clinical measurements of CRP in serum are currently performed with "high sensitivity" CRP (hsCRP) enzyme-linked immunosorbent assay (ELISA) tests that lack the sensitivity for the detection of this important biomarker in saliva. Because measurement of salivary CRP may represent a novel approach for diagnosing and monitoring chronic inflammatory disease, including CVD and periodontal diseases, the objective of this study was to apply an ultra-sensitive microchip assay system for the measurement of CRP in human saliva. Here, we describe this novel lab-on-a-chip system in its first application for the measurement of CRP in saliva and demonstrate its advantages over the traditional ELISA method. The increased sensitivity of the microchip system (10 pg ml(-1) of CRP with 1000-fold dilution of saliva sample) is attributed to its inherent increased signal to noise ratio, resulting from the higher bead surface area available for antigen/antibody interactions and the high stringency washes associated with this approach. Finally, the microchip assay system was utilized in this study to provide direct experimental evidence that chronic periodontal disease may be associated with higher levels of salivary CRP.

We demonstrate in this paper that two-dimensional (2-D) layered ceramics, materials that are highly anisotropic in terms of structure and properties can be used to induce the formation of polymer-covered metal nanorods. The procedure took advantage of the intrinsic planar, layered ordering of the metal cations suitable to be reduced and can be further used to engineer one-dimensional (1-D) metal alloy nanostructures by appropriate doping of the initial layered ceramic lattice with suitable cationic species. The procedure involved the formation in an intermediate step of a polymer-layered ceramic nanocomposite, highly porous to the diffusion of the reducing agent. Two structurally similar layered bismuthates, Bi2Sr2CaCu2O8+delta and Bi6Sr2CaO12 were used as the precursor layered ceramics and the redox-active metal cations were Cu2+ and Bi3+.

Molybdenum dioxide and vanadium-doped molybdenum dioxide microcrystals were synthesized by a totally new method involving a redox-active monomer route. The two related dioxides had virtually identical crystal structures and the three precursors used in the process could be subsequently divided in two subgroups of two members each, based on their strong similarities in composition and crystal structure, respectively. Molybdenum trioxide (orthorhombic and hexagonal forms) and the vanadium-doped (10%, molar) molybdenum trioxide (hexagonal form) were used in the redox process with excess pyrrole, and at relatively high temperature (325 degrees C) and long reaction time (12 days), formed a heterogeneous new type of organic-inorganic microcomposite. The new material could be described as a molybdenum dioxide crystal in a polymer box, with the size of the crystallite grain depending on the dimensions of the initial molybdenum trioxide grain and the ratio between the unit cell volumes of the initial and final oxide. The procedure took advantage of the relatively high reduction potentials of the Mo6+ and V5+ oxide surface centers towards the oxidative polymerization of pyrrole at intermediate temperatures, and the high catalytic activity of the metal cations in the molybdenum trioxide-based materials towards the total oxidation of the carbon atoms in the organic moiety, at elevated temperatures. This new synthetic route offers good perspectives as precursor composites for nanocrystal engineering in practical applications like heterogeneous catalysis, chemical sensing, nanostructured metallic catalysts, etc.

Although the superconductor/normal metal (SN) interface is one of the most important controlling factors in determining the performance of superconductor/normal metal/superconductor (SNS) junctions, controlling the SN interface still remains difficult. The in situ deposit techniques have been widely used to cope with this problem, but they limit the types of SNS junctions that can be explored. Soft Chemistry Etching methods were developed to fabricate the YBCO/Au/YBCO and TX-YBCO/Au/TX-YBCO SNS junction nano-devices with ex situ deposit. The planar SNS junctions controlled by such methods exhibited good transport properties above 80 K with less than 10(-8) Omega-cm(2) contact resistivity. The current state of the suitable fabrication methods of SNS nano-devices will be described with experiment results.