Beta-defensin index: A functional biomarker for oral cancer detection.

There is an unmet clinical need for a non-invasive and cost-effective test for oral squamous cell carcinoma (OSCC) that informs clinicians when a biopsy is warranted. Human beta-defensin 3 (hBD-3), an epithelial cell-derived anti-microbial peptide, is pro-tumorigenic and overexpressed in early-stage OSCC compared to hBD-2. We validate this expression dichotomy in carcinoma in situ and OSCC lesions using immunofluorescence microscopy and flow cytometry. The proportion of hBD-3/hBD-2 levels in non-invasively collected lesional cells compared to contralateral normal cells, obtained by ELISA, generates the beta-defensin index (BDI). Proof-of-principle and blinded discovery studies demonstrate that BDI discriminates OSCC from benign lesions. A multi-center validation study shows sensitivity and specificity values of 98.2% (95% confidence interval [CI] 90.3-99.9) and 82.6% (95% CI 68.6-92.2), respectively. A proof-of-principle study shows that BDI is adaptable to a point-of-care assay using microfluidics. We propose that BDI may fulfill a major unmet need in low-socioeconomic countries where pathology services are lacking.

A Novel Approach for Glycosylated Hemoglobin Testing Using Microchip Affinity Electrophoresis.

OBJECTIVE: Effective management of diabetes largely benefits from early diagnosis followed by intensive long-term regulation of blood glucose. The levels of glycohemoglobin (HbA1 and HbA1c) have been used as standard biomarkers to assess long-term blood glucose concentrations for diabetes diagnosis and management. Gold standard laboratory methods for HbA1 and HbA1c testing are often costly and not widely available. Moreover, currently available point-of-care (POC) immunoassay-based glycohemoglobin tests may produce inaccurate test results for patients with co-existing diseases such as hemoglobin disorders and anemia. Here, we report a POC platform, HemeChip-GHb, for quantitative HbA1 detection leveraging paper-based affinity electrophoresis. METHODS: We describe the design and development of the HemeChip-GHb test. Feasibility and accuracy of the HemeChip-GHb system were demonstrated by testing blood samples collected from healthy donors, patients with prediabetes, and patients with diabetes. RESULTS: HbA1 levels measured with HemeChip-GHb show 0.96 correlation to the levels reported from the clinical standard HPLC tests, and with a bias of -0.72% based on Bland-Altman analysis. 99.6% of the HbA1 levels for paired HemeChip-GHb and HPLC fell within A and B zones of no difference in clinical outcome based on error grid analysis. CONCLUSION: Using HemeChip-GHb we achieved accurate diabetes status detection with sensitivity and specificity of 100%. SIGNIFICANCE: We presented a novel POC paper-based affinity electrophoresis platform that has the potential for accurately diagnosing diabetes, and addressing an unmet need for accurate and affordable diagnostics in resource-challenged environments.

Paper-based microchip electrophoresis for point-of-care hemoglobin testing.

Nearly 7% of the world's population live with a hemoglobin variant. Hemoglobins S, C, and E are the most common and significant hemoglobin variants worldwide. Sickle cell disease, caused by hemoglobin S, is highly prevalent in sub-Saharan Africa and in tribal populations of Central India. Hemoglobin C is common in West Africa, and hemoglobin E is common in Southeast Asia. Screening for significant hemoglobin disorders is not currently feasible in many low-income countries with the high disease burden. Lack of early diagnosis leads to preventable high morbidity and mortality in children born with hemoglobin variants in low-resource settings. Here, we describe HemeChip, the first miniaturized, paper-based, microchip electrophoresis platform for identifying the most common hemoglobin variants easily and affordably at the point-of-care in low-resource settings. HemeChip test works with a drop of blood. HemeChip system guides the user step-by-step through the test procedure with animated on-screen instructions. Hemoglobin identification and quantification is automatically performed, and hemoglobin types and percentages are displayed in an easily understandable, objective way. We show the feasibility and high accuracy of HemeChip via testing 768 subjects by clinical sites in the United States, Central India, sub-Saharan Africa, and Southeast Asia. Validation studies include hemoglobin E testing in Bangkok, Thailand, and hemoglobin S testing in Chhattisgarh, India, and in Kano, Nigeria, where the sickle cell disease burden is the highest in the world. Tests were performed by local users, including healthcare workers and clinical laboratory personnel. Study design, methods, and results are presented according to the Standards for Reporting Diagnostic Accuracy (STARD). HemeChip correctly identified all subjects with hemoglobin S, C, and E variants with 100% sensitivity, and displayed an overall diagnostic accuracy of 98.4% in comparison to reference standard methods. HemeChip is a versatile, mass-producible microchip electrophoresis platform that addresses a major unmet need of decentralized hemoglobin analysis in resource-limited settings.

Hydrogen gas sensing using aluminum doped ZnO metasurfaces.

Hydrogen (H2) sensing is crucial in a wide variety of areas, such as industrial, environmental, energy and biomedical applications. However, engineering a practical, reliable, fast, sensitive and cost-effective hydrogen sensor is a persistent challenge. Here we demonstrate hydrogen sensing using aluminum-doped zinc oxide (AZO) metasurfaces based on optical read-out. The proposed sensing system consists of highly ordered AZO nanotubes (hollow pillars) standing on a SiO2 layer deposited on a Si wafer. Upon exposure to hydrogen gas, the AZO nanotube system shows a wavelength shift in the minimum reflectance by ∼13 nm within 10 minutes for a hydrogen concentration of 4%. These AZO nanotubes can also sense the presence of a low concentration (0.7%) of hydrogen gas within 10 minutes. Their rapid response time even for a low concentration, the possibility of large sensing area fabrication with good precision, and high sensitivity at room temperature make these highly ordered nanotube structures a promising miniaturized H2 gas sensor.

Emerging point-of-care technologies for sickle cell disease screening and monitoring.

INTRODUCTION: Sickle Cell Disease (SCD) affects 100,000 Americans and more than 14 million people globally, mostly in economically disadvantaged populations, and requires early diagnosis after birth and constant monitoring throughout the life-span of the patient. Areas covered: Early diagnosis of SCD still remains a challenge in preventing childhood mortality in the developing world due to requirements of skilled personnel and high-cost of currently available modalities. On the other hand, SCD monitoring presents insurmountable challenges due to heterogeneities among patient populations, as well as in the same individual longitudinally. Here, we describe emerging point-of-care micro/nano platform technologies for SCD screening and monitoring, and critically discuss current state of the art, potential challenges associated with these technologies, and future directions. Expert commentary: Recently developed microtechnologies offer simple, rapid, and affordable screening of SCD and have the potential to facilitate universal screening in resource-limited settings and developing countries. On the other hand, monitoring of SCD is more complicated compared to diagnosis and requires comprehensive validation of efficacy. Early use of novel microdevices for patient monitoring might come in especially handy in new clinical trial designs of emerging therapies.

A paper-based cantilever array sensor: Monitoring volatile organic compounds with naked eye.

Volatile organic compound (VOC) detection is critical for controlling industrial and commercial emissions, environmental monitoring, and public health. Simple, portable, rapid and low-cost VOC sensing platforms offer the benefits of on-site and real-time monitoring anytime and anywhere. The best and most practically useful approaches to monitoring would include equipment-free and power-free detection by the naked eye. In this work, we created a novel, paper-based cantilever sensor array that allows simple and rapid naked-eye VOC detection without the need for power, electronics or readout interface/equipment. This simple VOC detection method was achieved using (i) low-cost paper materials as a substrate and (ii) swellable thin polymers adhered to the paper. Upon exposure to VOCs, the polymer swelling adhered to the paper-based cantilever, inducing mechanical deflection that generated a distinctive composite pattern of the deflection angles for a specific VOC. The angle is directly measured by the naked eye on a 3-D protractor printed on a paper facing the cantilevers. The generated angle patterns are subjected to statistical algorithms (linear discriminant analysis (LDA)) to classify each VOC sample and selectively detect a VOC. We classified four VOC samples with 100% accuracy using LDA.

A disposable power source in resource-limited environments: A paper-based biobattery generating electricity from wastewater.

We report a novel paper-based biobattery which generates power from microorganism-containing liquid derived from renewable and sustainable wastewater which is readily accessible in the local environment. The device fuses the art of origami and the technology of microbial fuel cells (MFCs) and has the potential to shift the paradigm for flexible and stackable paper-based batteries by enabling exceptional electrical characteristics and functionalities. 3D, modular, and retractable battery stack is created from (i) 2D paper sheets through high degrees of folding and (ii) multifunctional layers sandwiched for MFC device configuration. The stack is based on ninja star-shaped origami design formed by eight MFC modular blades, which is retractable from sharp shuriken (closed) to round frisbee (opened). The microorganism-containing wastewater is added into an inlet of the closed battery stack and it is transported into each MFC module through patterned fluidic pathways in the paper layers. During operation, the battery stack is transformed into the round frisbee to connect eight MFC modules in series for improving the power output and simultaneously expose all air-cathodes to the air for their cathodic reactions. The device generates desired values of electrical current and potential for powering an LED for more than 20min.

A stackable, two-chambered, paper-based microbial fuel cell.

We developed a stackable and integrable paper-based microbial fuel cell (MFC) for potentially powering on-chip paper-based devices. Four MFCs were prepared on a T-shaped filter paper which was eventually folded three times to connect these MFCs in series. Each MFC was fabricated by sandwiching multifunctional paper layers for two-chambered fuel cell configuration. One drop of bacteria-containing anolyte into the anodic inlet and another drop of potassium ferricyanide for cathodic reaction flowed through patterned fluidic pathways within the paper matrix, both vertically and horizontally, reaching each of the four MFCs and filling the reservoir of each device. Bacterial respiration then transferred electrons to the anode, which traveled across an external load to the cathode where they combined with protons. The MFC stack connected in series generated a high power density (1.2µW/cm(2)), which is two orders of magnitude higher than the previous report on the paper-based MFC stack. This work will represent the fusion of the art of origami and paper-based MFC technology, which could provide a paradigm shift for the architecture and design of paper-based batteries.

A 3D paper-based enzymatic fuel cell for self-powered, low-cost glucose monitoring.

In this work, we demonstrate a novel low-cost, self-powered paper-based biosensor for glucose monitoring. The device operating mechanism is based on a glucose/oxygen enzymatic fuel cell using an electrochemical energy conversion as a transducing element for glucose monitoring. The self-powered glucose biosensor features (i) a 3D origami paper-based structure for easy system integration onto paper, (ii) an air-cathode on paper for low-cost production and easy operation, and (iii) a screen printed chitosan/glucose oxidase anode for stable current generation as an analytical signal for glucose monitoring. The sensor showed a linear range of output current at 1-5mM glucose (R(2)=0.996) with a sensitivity of 0.02 µA mM(-1). The advantages offered by such a device, including a low cost, lack of external power sources/sophisticated external transducers, and the capacity to rapidly generate reliable results, are well suited for the clinical and social settings of the developing world.

Bacteria-powered battery on paper.

Paper-based devices have recently emerged as simple and low-cost paradigms for fluid manipulation and analytical/clinical testing. However, there are significant challenges in developing paper-based devices at the system level, which contain integrated paper-based power sources. Here, we report a microfabricated paper-based bacteria-powered battery that is capable of generating power from microbial metabolism. The battery on paper showed a very short start-up time relative to conventional microbial fuel cells (MFCs); paper substrates eliminated the time traditional MFCs required to accumulate and acclimate bacteria on the anode. Only four batteries connected in series provided desired values of current and potential to power an LED for more than 30 minutes. The battery featured (i) a low-cost paper-based proton exchange membrane directly patterned on commercially available parchment paper and (ii) paper reservoirs for holding the anolyte and the catholyte for an extended period of time. Based on this concept, we also demonstrate the use of paper-based test platforms for the rapid characterization of electricity-generating bacteria. This paper-based microbial screening tool does not require external pumps/tubings and represents the most rapid test platform (<50 min) compared with the time needed by using traditional screening tools (up to 103 days) and even recently proposed MEMS arrays (< 2 days).

Paper-based batteries: a review.

There is an extensively growing interest in using paper or paper-like substrates for batteries and other energy storage devices. Due to their intrinsic characteristics, paper (or paper-like) batteries show outstanding performance while retaining low cost, multifunctionality, versatility, flexibility and disposability. In this overview, we review recent achievements in paper (or paper-like) batteries as well as their applications. Various types of paper power devices are discussed including electrochemical batteries, biofuel cells, lithium-ion batteries, supercapacitors, and nanogenerators. Further scientific and technological challenges in this field are also discussed.

A paper-based microbial fuel cell: instant battery for disposable diagnostic devices.

We present a microfabricated paper-based microbial fuel cell (MFC) generating a maximum power of 5.5 µW/cm(2). The MFC features (1) a paper-based proton exchange membrane by infiltrating sulfonated sodium polystyrene sulfonate and (2) micro-fabricated paper chambers by patterning hydrophobic barriers of photoresist. Once inoculum and catholyte were added to the MFC, a current of 74 µA was generated immediately. This paper-based MFC has the advantages of ease of use, low production cost, and high portability. The voltage produced was increased by 1.9 × when two MFC devices were stacked in series, while operating lifetime was significantly enhanced in parallel.