A high-performance polymer composite column for coronavirus nucleic acid purification.

Here, we report the development of a novel polymer composite (PC) purification column and kit. The performance of the PC columns was compared to conventional silica gel (SG) columns for the purification of nucleic acids from coronaviruses, including SARS-CoV-2, in 82 clinical samples. The results shows that PC-based purification outperforms silica gel (SG)-based purification by enabling a higher sensitivity (94%), accuracy (97%), and by eliminating false positives (100% specificity). The high specificity is critical for efficient patient triage and resource management during pandemics. Furthermore, PC-based purification exhibits three times higher analytical precision than a commonly used SG-based nucleic acid purification thereby enabling a more accurate quantification of viral loads and higher reproducibility.

Iridium oxide and cobalt hydroxide microfluidic-based potentiometric pH sensor.

Microliter volume pH determination is of great importance in the biomedical and industrial applications. The current available pH meter and measurement techniques are hard to reach the high demand of microliter volume pH determination in a repeatable, stable, and sensitivity manner. This work aims to fill the gap of microliter volume pH measurements while maintaining good sensing performance. The electrodeposited iridium oxide and cobalt hydroxide along with gold electrode served as working, counter, and reference electrode, respectively, for 10-12 µL volume pH measurements with Nernst constant of 55.9 ± 4.4 mV/pH. The electrodeposited thin film was further characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), Raman spectrometry, etc. to confirm its morphology and composition. The constructed pH sensor was used for human serum sample measurements to confirm the suitability of future applications. The results show that it has only 0.80% variation compared to a commercial pH meter with a limit of detection (LOD, or resolution) of ± 0.01 pH. It holds a great potential to be used in the future for microliter volume in situ pH measurements.

A High-Performance Polymer Composite Column for Coronavirus Nucleic Acid Purification.

Here, we report the development of a novel polymer composite (PC) purification column and kit. The performance of the PC columns was compared to conventional silica gel (SG) columns for the purification of nucleic acids from coronaviruses, including SARS-CoV-2, in 82 clinical samples. The results shows that PC-based purification outperforms silica gel (SG)-based purification by enabling a higher sensitivity (94%), accuracy (97%), and by eliminating false positives (100% selectivity). The high selectivity is critical for efficient patient triage and resource management during pandemics. Furthermore, PC-based purification exhibits three times higher analytical precision than a commonly used SG-based nucleic acid purification thereby enabling a more accurate quantification of viral loads and higher reproducibility.

High-fidelity profiling and modeling of heterogeneity in wastewater systems using milli-electrode array (MEA): Toward high-efficiency and energy-saving operation.

High energy consumption is a critical problem for wastewater treatment systems currently monitored using conventional "single point" probes and operated with manual or automatic open-loop control strategies, exhibiting significant time lag. This challenge is addressed in this study by profiling the variation of three critical water quality parameters (conductivity, temperature and pH) along the depth of a reactor at high spatiotemporal resolution in a real-time mode using flat thin milli-electrode array (MEA) sensors. The profiling accurately captured the heterogeneous status of the reactor under transient shocks (conductivity and pH) and slow lingering shock (temperature), providing an effective dataset to optimize the chemical dosage and energy requirement of wastewater treatment systems. Transient shock models were developed to validate the MEA profiles and calculate mass transfer coefficients. Monte Carlo simulation revealed high-resolution MEA profiling combined with fast closed-loop control strategies can save 59.50% of energy consumption (Temperature and oxygen consumption controls) and 45.29% of chemical dosage, and reach 16.28% performance improvement over the benchmark (defined with ideal conditions), compared with traditional "single-point" sensors that could only monitor the entire system through a single process state. This study demonstrated the capability of MEA sensors to profile reactor heterogeneity, visualize the variation of water quality at high resolution, provide complete datasets for accurate control, and ultimately lead to energy-saving operation with high resilience.

Towards high resolution monitoring of water flow velocity using flat flexible thin mm-sized resistance-typed sensor film (MRSF).

Novel flexible thin mm-sized resistance-typed sensor film (MRSF) fabricated using ink-jet printing technology (IPT) was developed in this study to monitor water flow rate in pipelines in real time in situ mode. The mechanism of MRSF is that the mm-sized interdigitated electrodes made by printing silver nanoparticles on an elastic polyimide film bend under different flow rates, leading to variation of the resistance of the sensor at different degrees of curvature. Continuous flow tests showed that MRSF possessed a high accuracy (0.2 m/s) and excellent sensitivity (0.1447/ms-1). A model of sensor resistance and flow velocity was established to unfold the correlation between the fundamentals of fluid mechanics and the mechanic flexibility of sensor materials. An analytical model yielded a high coefficient of determination (R2 > 0.93) for the relationship between the resistance increment of the MRSF and the square of the flow velocity at the velocity range of 0.25-2 m/s. Furthermore, a temperature-correction model was developed to quantify the effect of water temperature on the sensor resistance readings. MRSF exhibited a low temperature coefficient of resistance (TCR, 0.001) at the water temperature range of 20-60 °C. Computational fluid dynamics (CFD) simulations using the finite element method were conducted and confirmed both the underlying load assumptions and the deformation characteristics of the sensor film under various flow and material conditions. High-resolution monitoring of water flow rate using MRSF technology was expected to save at least 50% energy consumption for a given unit, especially under flow fluctuation. MRSF possesses a great potential to perform real-time in situ monitoring at high accuracy with ultralow cost, thus enabling the feedback control at high spatiotemporal resolution to reduce the overall energy consumption in water and wastewater systems.

Preparation, characterization and application of a protein hydrogel with rapid self-healing and unique autofluoresent multi-functionalities.

A smart hydrogel with dual self-healing and autofluoresent functionalities is presented. The protein hydrogel is fabricated by denaturing bovine serum albumin in a basic environment. Upon gelation, autofluorescence is induced and the protein hydrogel can be excited by a wide range of spectrum, ranging from 320 to 520 nm. It was also found that the as-prepared autofluorescent protein hydrogel possessed rapid and repetitive self-healing capability. Without any external stimulus, more than 90% recovery of the mechanical strength can be obtained within 10 min after destruction. Moreover, the as-prepared hydrogel exhibits excellent biocompatibility and cell attachment property after its pH adjustment to neutral pH, while both autofluorescence and self-healing properties were still retained. This study suggests a promising means to prepare multi-functional protein hydrogel with dual physicochemical functionalities, which holds great potential in biomedical related applications. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 81-91, 2019.

A Simple SERS-Based Trace Sensing Platform Enabled by AuNPs-Analyte/AuNPs Double-Decker Structure on Wax-Coated Hydrophobic Surface.

In this work, a simple and versatile SERS sensing platform enabled by AuNPs-analyte/AuNPs double-decker structure on wax-coated hydrophobic surface was developed using a portable Raman spectrometer. Wax-coated silicon wafer served as a hydrophobic surface to induce both aggregation and concentration of aqueous phase AuNPs mixed with analyte of interest. After drying, another layer of AuNPs was drop-cast onto the layer of AuNPs-analyte on the substrate to form double-decker structure, thus introducing more "hot spots" to further enhance the Raman signal. To validate the sensing platform, methyl parathion (pesticide), and melamine (a nitrogen-enrich compound illegally added to food products to increase their apparent protein content) were employed as two model compounds for trace sensing demonstration. The as-fabricated sensor showed high reproducibility and sensitivity toward both methyl parathion and melamine detection with the limit of detection at the nanomolar and sub-nanomolar concentration level, respectively. In addition, remarkable recoveries for methyl parathion spiked into lake water samples were obtained, while reasonably good recoveries for melamine spiked into milk samples were achieved. These results demonstrate that the as-developed SERS sensing platform holds great promise in detecting trace amount of hazardous chemicals for food safety and environment protection.

Digital, Rapid, Accurate, and Label-Free Enumeration of Viable Microorganisms Enabled by Custom-Built On-Glass-Slide Culturing Device and Microscopic Scanning.

Accurately measuring the number of viable microorganisms plays an essential role in microbiological studies. Since the conventional agar method of enumerating visible colonies is time-consuming and not accurate, efforts have been made towards overcoming these limitations by counting the invisible micro-colonies. However, none of studies on micro-colony counting was able to save significant time or provide accurate results. Herein, we developed an on-glass-slide cell culture device that enables rapid formation of micro-colonies on a 0.38 mm-thick gel film without suffering from nutrient and oxygen deprivation during bacteria culturing. Employing a phase contrast imaging setup, we achieved rapid microscopic scanning of micro-colonies within a large sample area on the thin film without the need of fluorescent staining. Using Escherichia coli (E. coli) as a demonstration, our technique was able to shorten the culturing time to within 5 h and automatically enumerate the micro-colonies from the phase contrast images. Moreover, this method delivered more accurate counts than the conventional visible colony counting methods. Due to these advantages, this imaging-based micro-colony enumeration technique provides a new platform for the quantification of viable microorganisms.

Sensitive and Selective NH3 Monitoring at Room Temperature Using ZnO Ceramic Nanofibers Decorated with Poly(styrene sulfonate).

Ammonia (NH3) gas is a prominent air pollutant that is frequently found in industrial and livestock production environments. Due to the importance in controlling pollution and protecting public health, the development of new platforms for sensing NH3 at room temperature has attracted great attention. In this study, a sensitive NH3 gas device with enhanced selectivity is developed based on zinc oxide nanofibers (ZnO NFs) decorated with poly(styrene sulfonate) (PSS) and operated at room temperature. ZnO NFs were prepared by electrospinning followed by calcination at 500 °C for 3 h. The electrospun ZnO NFs are characterized to evaluate the properties of the as-prepared sensing materials. The loading of PSS to prepare ZnO NFs/PSS composite is also optimized based on the best sensing performance. Under the optimal composition, ZnO NFs/PSS displays rapid, reversible, and sensitive response upon NH3 exposure at room temperature. The device shows a dynamic linear range up to 100 ppm and a limit of detection of 3.22 ppm and enhanced selectivity toward NH3 in synthetic air, against NO2 and CO, compared to pure ZnO NFs. Additionally, a sensing mechanism is proposed to illustrate the sensing performance using ZnO NFs/PSS composite. Therefore, this study provides a simple methodology to design a sensitive platform for NH3 monitoring at room temperature.

Dual functional rhodium oxide nanocorals enabled sensor for both non-enzymatic glucose and solid-state pH sensing.

Both pH-sensitive and glucose-responsive rhodium oxide nanocorals (Rh2O3 NCs) were synthesized through electrospinning followed by high-temperature calcination. The as-prepared Rh2O3 NCs were systematically characterized using various advanced techniques including scanning electron microscopy, X-ray powder diffraction and Raman spectroscopy, and then employed as a dual functional nanomaterial to fabricate a dual sensor for both non-enzymatic glucose sensing and solid-state pH monitoring. The sensing performance of the Rh2O3 NCs based dual sensor toward pH and glucose was evaluated using open circuit potential, cyclic voltammetry and amperometric techniques, respectively. The results show that the as-prepared Rh2O3 NCs not only maintain accurate and reversible pH sensitivity of Rh2O3, but also demonstrate a good electrocatalytic activity toward glucose oxidation in alkaline medium with a sensitivity of 11.46 µA mM-1 cm-2, a limit of detection of 3.1 µM (S/N = 3), and a reasonable selectivity against various interferents in non-enzymatic glucose detection. Its accuracy in determining glucose in human serum samples was further demonstrated. These features indicate that the as-prepared Rh2O3 NCs hold great promise as a dual-functional sensing material in the development of a high-performance sensor forManjakkal both solid-state pH and non-enzymatic glucose sensing.

A high-performance electrochemical sensor for biologically meaningful l-cysteine based on a new nanostructured l-cysteine electrocatalyst.

As a new class of l-cysteine electrocatalyst explored in this study, Au/CeO2 composite nanofibers (CNFs) were employed to modify the screen printed carbon electrode (SPCE) to fabricate a novel l-cysteine (CySH) electrochemical sensor with high performance. Its electrochemical behavior and the roles of Au and CeO2 in the composite toward electro-oxidation of CySH were elucidated and demonstrated using cyclic voltammetry and amperometry techniques for the first time through the comparison with pure CeO2 NFs. More specifically, the Au/CeO2 CNFs modified SPCE possessed greatly enhanced electrocatalytic activity toward CySH oxidation. An ultra high sensitivity of 321 µA mM-1cm-2 was obtained, which is almost 2.7 times higher than that of pure CeO2 NFs, revealing that the presence of Au imposed an important influence on the electrocatalytic activity toward CySH. The detailed reasons on such high performance were also discussed. In addition, the as-prepared sensor showed a low detection limit of 10 nM (signal to noise ratio of 3), a wide linear range up to 200 µM for the determination of CySH, an outstanding reproducibility and good long-term stability, as well as an excellent selectivity against common interferents such as tryptophan, tyrosine, methionine, ascorbic acid and uric acid. All these features indicate that the Au/CeO2 composite nanofiber is a promising candidate as a new class of l-cysteine electrocatalyst in the development of highly sensitive and selective CySH electrochemical sensor.

Repetitive Biomimetic Self-healing of Ca(2+)-Induced Nanocomposite Protein Hydrogels.

Self-healing is a capacity observed in most biological systems in which the healing processes are autonomously triggered after the damage. Inspired by this natural behavior, researchers believed that a synthetic material possessing similar self-recovery capability could also be developed. Albeit various intrinsic self-healing systems have been developed over the past few decades, restriction on the biocompatibility due to the required synthetic conditions under extreme pH and with poisonous cross-linker significantly limits their application in biomedical field. In this study, a highly biocompatible nanocomposite protein hydrogel with excellent biomimetic self-healing property is presented. The self-healing protein gel is made by inducing calcium ions into the mixture of heat-induced BSA nano-aggregates and pristine BSA molecules at room temperature and under physiological pH due to the ion-mediated protein-protein association and the bridging effect of divalent Ca(2+) ions. The as-prepared protein hydrogel shows excellent repetitive self-healing properties without using any external stimuli at ambient condition. Such outstanding self-recovery performance was quantitatively evaluated/validated by both dynamic and oscillatory rheological analysis. Moreover, with the presence of calcium ions, the self-healing behavior can be significantly facilitated/enhanced. Finally, the superior biocompatibility demonstrated by in vitro cytotoxicity analysis suggests that it is a promising self-healing material well-suited for biomedical applications.

ELP-OPH/BSA/TiO2 nanofibers/c-MWCNTs based biosensor for sensitive and selective determination of p-nitrophenyl substituted organophosphate pesticides in aqueous system.

A novel biosensor for rapid, sensitive and selective monitoring of p-nitrophenyl substituted organophosphate pesticides (OPs) in aqueous system was developed using a functional nanocomposite which consists of elastin-like-polypeptide-organophosphate hydrolase (ELP-OPH), bovine serum albumin (BSA), titanium dioxide nanofibers (TiO2NFs) and carboxylic acid functionalized multi-walled carbon nanotubes (c-MWCNTs). ELP-OPH was simply purified from genetically engineered Escherichia coli based on the unique phase transition of ELP and thus served as biocatalyst for OPs, while BSA was used to stabilize OPH activity in the nanocomposite. TiO2NFs was employed to enrich organophosphates in the nanocomposite due to its strong affinity with phosphoric group in OPs, while c-MWCNTs was used to enhance the electron transfer in the amperometric detection as well as for covalent immobilization of ELP-OPH. ELP-OPH/BSA/TiO2NFs/c-MWCNTs nanocomposite were systematically characterized using field emission scanning electron microscopy (SEM), Raman spectra, Fourier Transform infrared spectroscopy (FTIR) and X-ray Diffraction (XRD). Under the optimized operating conditions, the ELP-OPH/BSA/TiO2NFs/c-MWCNTs based biosensor for OPs shows a wide linear range, a fast response (less than 5s) and limits of detection (S/N=3) as low as 12nM and 10nM for methyl parathion and parathion, respectively. Such excellent sensing performance can be attributed to the synergistic effects of the individual components in the nanocomposite. Its further application for selectively monitoring OPs compounds spiked in lake water samples was also demonstrated with good accuracy. These features indicate that the developed nanocomposite offers an excellent biosensing platform for rapid, sensitive and selective detection of organophosphates compounds.

A Biocompatible and Biodegradable Protein Hydrogel with Green and Red Autofluorescence: Preparation, Characterization and In Vivo Biodegradation Tracking and Modeling.

Because of its good biocompatibility and biodegradability, albumins such as bovine serum albumin (BSA) and human serum albumin (HSA) have found a wide range of biomedical applications. Herein, we report that glutaraldehyde cross-linked BSA (or HSA) forms a novel fluorescent biological hydrogel, exhibiting new green and red autofluorescence in vitro and in vivo without the use of any additional fluorescent labels. UV-vis spectra studies, in conjunction with the fluorescence spectra studies including emission, excitation and synchronous scans, indicated that three classes of fluorescent compounds are presumably formed during the gelation process. SEM, FTIR and mechanical tests were further employed to investigate the morphology, the specific chemical structures and the mechanical strength of the as-prepared autofluorescent hydrogel, respectively. Its biocompatibility and biodegradability were also demonstrated through extensive in vitro and in vivo studies. More interestingly, the strong red autofluorescence of the as-prepared hydrogel allows for conveniently and non-invasively tracking and modeling its in vivo degradation based on the time-dependent fluorescent images of mice. A mathematical model was proposed and was in good agreement with the experimental results. The developed facile strategy to prepare novel biocompatible and biodegradable autofluorescent protein hydrogels could significantly expand the scope of protein hydrogels in biomedical applications.

Protein Microspheres with Unique Green and Red Autofluorescence for Noninvasively Tracking and Modeling Their in Vivo Biodegradation.

Bovine serum albumin (BSA) microspheres were prepared through a facile and low-cost route including a high-speed dispersion of BSA in cross-linking solution followed by spray drying. Interestingly the as-prepared BSA microspheres possess unique blue-green, green, green-yellow, and red fluorescence when excited by specific wavelengths of laser or LED light. The studies of UV-visible reflectance spectra and fluorescence emission spectra indicated that four classes of fluorescent compounds are presumably formed during the fabrication processes. The formation and the potential contributors for the unique green and red autofluorescence were also discussed and proposed though the exact structures of the fluorophores formed remain elusive due to the complexity of the protein system. The effect of spray-drying conditions on the morphology of spray-dried samples was investigated and optimized. FTIR was further employed to characterize the formation of the functional groups in the as-prepared autofluorescent microspheres. Good in vitro and in vivo biocompatibility was demonstrated by the cytotoxicity test on the A549 cancer cells and tissue histological analysis, respectively. The autofluorescent BSA microspheres themselves were then applied as a novel tracer for convenient tracking/modeling of the biodegradation of autofluorescent BSA microspheres injected into mouse model based on noninvasive, time-dependent fluorescence images of the mice, in which experimental data are in good agreement with the proposed mathematical model. All these studies indicate that the as-developed protein microspheres exhibiting good biocompatibility, biodegradability, and unique autofluorescence, can significantly broaden biomedical applications of fluorescent protein particles.

Flat microliter membrane-based microbial fuel cell as "on-line sticker sensor" for self-supported in situ monitoring of wastewater shocks.

Novel flat membrane-based microbial fuel cell (MMFC) sensors were developed by compacting two filter membranes coated with carbon ink. High micro-porosity and hydrophilicity of membranes offered the distinct advantages of short acclimation period (couple hours), simple compact configuration with microliter size, and high sensitivity and stability. MMFC sensors were examined at two toxic shocks (chromium and nickel) in a batch-mode test chamber, and rapidly responded to shock types and concentrations. The variation of voltage output was correlated with open circuit potential (OCP). Filter membranes facilitated bacterial attachment and shortened acclimation. The MMFC sensors showed good reusability and recovered several days after toxic shocks. The robustness of MMFC sensors was validated through 1-month tests. The stability of sensor signals was examined with coefficient of variance (CV) statistical analysis. The flat microliter MMFC has a great potential as "on-line sticker sensor" for real time in situ monitoring of wastewater quality.