Grace M. Hwang

Spectral sensitivity of two-dimensional nanohole array surface plasmon polariton resonance sensor

An analytical expression of spectral sensitivity derived from a surface plasmon polariton dispersion relation for a two-dimensional nanohole array surface plasmon polariton resonance sensor is presented. The sensitivity of the nanohole array sensor depends on the periodicity of the array and the order of the excited surface plasmon polariton modes. The analytical expression is further confirmed by rigorous electromagnetic simulation and validated by experimental results. Real-time monitoring of protein-protein specific bonding is performed to demonstrate the integrated microfluidic nanohole array surface plasmon resonance biosensor.

Recent advances in peptide probe-based biosensors for detection of infectious agents

Recent biological terrorism threats and outbreaks of microbial pathogens clearly emphasize the need for biosensors that can quickly and accurately identify infectious agents. The majority of rapid biosensors generate detectable signals when a molecular probe in the detector interacts with an analyte of interest. Analytes may be whole bacterial or fungal cells, virus particles, or specific molecules, such as chemicals or protein toxins, produced by the infectious agent. Peptides and nucleic acids are most commonly used as probes in biosensors because of their versatility in forming various tertiary structures. The interaction between the probe and the analyte can be detected by various sensor platforms, including quartz crystal microbalances, surface acoustical waves, surface plasmon resonance, amperometrics, and magnetoelastics. The field of biosensors is constantly evolving to develop devices that have higher sensitivity and specificity, and are smaller, portable, and cost-effective. This mini review discusses recent advances in peptide-dependent rapid biosensors and their applications as well as relative advantages and disadvantages of each technology.

Plasmonic Sensing of Biological Analytes Through Nanoholes

A transmission-based surface plasmon resonance (SPR) sensor for label-free detection of protein-carbohydrate and protein-protein binding proximate to a perforated gold surface is demonstrated. An SPR instrument makes real-time measurements of the resonant wavelength and/or the resonant angle of incidence of transmitted light; both are influenced by the presence of proteins at the gold surface-liquid interface. Ethylene glycol solutions with known refractive indexes were used to calibrate the instrument. A paired polarization-sensitive detector achieved an overall detection resolution of ~ 6.6 times 10-5 refractive index units (RIU). Proof of principle experiments was performed with concanavalin A (Con A) binding to gold-adsorbed ovomucoid and anti-bovine serum albumin (BSA) binding to gold-adsorbed BSA.

Entamoeba histolytica Induces Intestinal Cathelicidins but Is Resistant to Cathelicidin-Mediated Killing

ABSTRACT The enteric protozoan parasite Entamoeba histolytica is the cause of potentially fatal amebic colitis and liver abscesses. E. histolytica trophozoites colonize the colon, where they induce inflammation, penetrate the mucosa, and disrupt the host immune system. The early establishment of E. histolytica in the colon occurs in the presence of antimicrobial human (LL-37) and murine (CRAMP [cathelin-related antimicrobial peptide]) cathelicidins, essential components of the mammalian innate defense system in the intestine. Studying this early step in the pathogenesis of amebic colitis, we demonstrate that E. histolytica trophozoites or their released proteinases, including cysteine proteinase 1 (EhCP1), induce intestinal cathelicidins in human intestinal epithelial cell lines and in a mouse model of amebic colitis. Despite induction, E. histolytica trophozoites were found to be resistant to killing by these antimicrobial peptides, and LL-37 and CRAMP were rapidly cleaved by released amebic cysteine proteases. The cathelicidin fragments however, did maintain their antimicrobial activity against bacteria. Degradation of intestinal cathelicidins is a novel function of E. histolytica cysteine proteinases in the evasion of the innate immune system in the bowel. Thus, early intestinal epithelial colonization of invasive trophozoites involves a complex interplay in which the ultimate outcome of infection depends in part on the balance between degradation of cathelicidins by amebic released cysteine proteinases and upregulation of proinflammatory mediators which trigger the inflammatory response.

Hsp90 Inhibitors as New Leads to Target Parasitic Diarrheal Diseases

ABSTRACT Entamoeba histolytica and Giardia lamblia are anaerobic protozoan parasites that cause amebiasis and giardiasis, two of the most common diarrheal diseases worldwide. Current therapy relies on metronidazole, but resistance has been reported and the drug has significant adverse effects. Therefore, it is critical to search for effective, better-tolerated antiamebic and antigiardial drugs. We synthesized several examples of a recently reported class of Hsp90 inhibitors and evaluated these compounds as potential leads for antiparasitic chemotherapy. Several of these inhibitors showed strong in vitro activity against both E. histolytica and G. lamblia trophozoites. The inhibitors were rescreened to discriminate between amebicidal and giardicidal activity and general cytotoxicity toward a mammalian cell line. No mammalian cytotoxicity was found at >100 μM for 48 h for any of the inhibitors. To understand the mechanism of action, a competitive binding assay was performed using the fluorescent ATP analogue bis-ANS (4,4′-dianilino-1,1′-binaphthyl-5,5′-disulfonic acid dipotassium salt) and recombinant E. histolytica Hsp90 preincubated in both the presence and absence of Hsp90 inhibitors. There was significant reduction in fluorescence compared to the level in the control, suggesting that E. histolytica Hsp90 is a selective target. The in vivo efficacy and safety of one Hsp90 inhibitor in a mouse model of amebic colitis and giardiasis was demonstrated by significant inhibition of parasite growth at a single oral dose of 5 mg/kg of body weight/day for 7 days and 10 mg/kg/day for 3 days. Considering the results for in vitro activity and in vivo efficacy, Hsp90 inhibitors represent a promising therapeutic option for amebiasis and giardiasis.

A model-based tool to predict the propagation of infectious disease via airports.

Summary Epidemics of novel or re-emerging infectious diseases have quickly spread globally via air travel, as highlighted by pandemic H1N1 influenza in 2009 (pH1N1). Federal, state, and local public health responders must be able to plan for and respond to these events at aviation points of entry. The emergence of a novel influenza virus and its spread to the United States were simulated for February 2009 from 55 international metropolitan areas using three basic reproduction numbers (R 0): 1.53, 1.70, and 1.90. Empirical data from the pH1N1 virus were used to validate our SEIR model. Time to entry to the U.S. during the early stages of a prototypical novel communicable disease was predicted based on the aviation network patterns and the epidemiology of the disease. For example, approximately 96% of origins (R 0 of 1.53) propagated a disease into the U.S. in under 75 days, 90% of these origins propagated a disease in under 50 days. An R 0 of 1.53 reproduced the pH1NI observations. The ability to anticipate the rate and location of disease introduction into the U.S. provides greater opportunity to plan responses based on the scenario as it is unfolding. This simulation tool can aid public health officials to assess risk and leverage resources efficiently.

Detection of respiratory viruses on air filters from aircraft

Aims:  To evaluate the feasibility of identifying viruses from aircraft cabin air, we evaluated whether respiratory viruses trapped by commercial aircraft air filters can be extracted and detected using a multiplex PCR, bead‐based assay.

An analysis on the detection of biological contaminants aboard aircraft.

The spread of infectious disease via commercial airliner travel is a significant and realistic threat. To shed some light on the feasibility of detecting airborne pathogens, a sensor integration study has been conducted and computational investigations of contaminant transport in an aircraft cabin have been performed. Our study took into consideration sensor sensitivity as well as the time-to-answer, size, weight and the power of best available commercial off-the-shelf (COTS) devices. We conducted computational fluid dynamics simulations to investigate three types of scenarios: (1) nominal breathing (up to 20 breaths per minute) and coughing (20 times per hour); (2) nominal breathing and sneezing (4 times per hour); and (3) nominal breathing only. Each scenario was implemented with one or seven infectious passengers expelling air and sneezes or coughs at the stated frequencies. Scenario 2 was implemented with two additional cases in which one infectious passenger expelled 20 and 50 sneezes per hour, respectively. All computations were based on 90 minutes of sampling using specifications from a COTS aerosol collector and biosensor. Only biosensors that could provide an answer in under 20 minutes without any manual preparation steps were included. The principal finding was that the steady-state bacteria concentrations in aircraft would be high enough to be detected in the case where seven infectious passengers are exhaling under scenarios 1 and 2 and where one infectious passenger is actively exhaling in scenario 2. Breathing alone failed to generate sufficient bacterial particles for detection, and none of the scenarios generated sufficient viral particles for detection to be feasible. These results suggest that more sensitive sensors than the COTS devices currently available and/or sampling of individual passengers would be needed for the detection of bacteria and viruses in aircraft.

Collection of Aerosolized Human Cytokines Using Teflon® Filters

Background Collection of exhaled breath samples for the analysis of inflammatory biomarkers is an important area of research aimed at improving our ability to diagnose, treat and understand the mechanisms of chronic pulmonary disease. Current collection methods based on condensation of water vapor from exhaled breath yield biomarker levels at or near the detection limits of immunoassays contributing to problems with reproducibility and validity of biomarker measurements. In this study, we compare the collection efficiency of two aerosol-to-liquid sampling devices to a filter-based collection method for recovery of dilute laboratory generated aerosols of human cytokines so as to identify potential alternatives to exhaled breath condensate collection. Methodology/Principal Findings Two aerosol-to-liquid sampling devices, the SKC® Biosampler and Omni 3000™, as well as Teflon® filters were used to collect aerosols of human cytokines generated using a HEART nebulizer and single-pass aerosol chamber setup in order to compare the collection efficiencies of these sampling methods. Additionally, methods for the use of Teflon® filters to collect and measure cytokines recovered from aerosols were developed and evaluated through use of a high-sensitivity multiplex immunoassay. Our results show successful collection of cytokines from pg/m3 aerosol concentrations using Teflon® filters and measurement of cytokine levels in the sub-picogram/mL concentration range using a multiplex immunoassay with sampling times less than 30 minutes. Significant degradation of cytokines was observed due to storage of cytokines in concentrated filter extract solutions as compared to storage of dry filters. Conclusions Use of filter collection methods resulted in significantly higher efficiency of collection than the two aerosol-to-liquid samplers evaluated in our study. The results of this study provide the foundation for a potential new technique to evaluate biomarkers of inflammation in exhaled breath samples.

Detecting infectious and biological contaminants aboard aircraft - Is it feasible?

The spread of disease via commercial airliner travel is a significant and realistic threat. To shed some light on the feasibility of detecting airborne pathogens a sensor integration study has been conducted in this work, in addition to computational investigations of the contaminant transport in an aircraft cabin. Our sensor study took into consideration sensor sensitivity as well as the size, weight and power of best available commercial off-the-shelf devices. Computational fluid dynamic simulations were made to investigate extreme coughing and sneezing scenarios. The principle finding was that contaminant concentrations in aircraft are below the sensitivity thresholds of current state of the art devices. Therefore, this study proposes the development of a novel bio-detector to achieve single particle detection limits for pathogen sensing in airline cabins.The spread of disease via commercial airliner travel is a significant and realistic threat. To shed some light on the feasibility of detecting airborne pathogens a sensor integration study has been conducted in this work in addition to computational investigations of the contaminant transport in an aircraft cabin. Our sensor study took into consideration the sensor sensitivity as well as the size, weight and power of commercial off the shelf devices. The computational fluid dynamic simulations were implemented for investigating extreme coughing and sneezing scenarios. The principle finding was that the contaminant concentration is relatively low when compared to the sensitivity available in current state of the art devices. Therefore, this study proposes a novel device/method to detect existence of pathogens. Moreover, it concludes that a collector is also required.