Label-Free, Real-Time Monitoring of Cytochrome C Responses to Drugs in Microdissected Tumor Biopsies with a Multi-Well Aptasensor Platform.

Functional assays on intact tumor biopsies can potentially complement and extend genomics-based approaches for precision oncology, drug testing, and organs-on-chips cancer disease models by capturing key determinants of therapeutic response, such as tissue architecture, tumor heterogeneity, and the tumor microenvironment. Currently, most of these assays rely on fluorescent labeling, a semi-quantitative method best suited to be a single-time-point terminal assay or labor-intensive terminal immunostaining analysis. Here, we report integrated aptamer electrochemical sensors for on-chip, real-time monitoring of increases of cytochrome C, a cell death indicator, from intact microdissected tissues with high affinity and specificity. The platform features a multi-well sensor layout and a multiplexed electronic setup. The aptasensors measure increases in cytochrome C in the supernatant of mouse or human microdissected tumors after exposure to various drug treatments. Since the aptamer probe can be easily exchanged to recognize different targets, the platform could be adapted for multiplexed monitoring of various biomarkers, providing critical information on the tumor and its microenvironment. This approach could not only help develop more advanced cancer disease models but also apply to other complex in vitro disease models, such as organs-on-chips and organoids.

Fabrication and ex vivo evaluation of activated carbon-Pt microparticle based glutamate biosensor.

As one of the most abundant neurotransmitters in the brain and the spinal cord, glutamate plays many important roles in the nervous system. Precise information about the level of glutamate in the extracellular space of living brain tissue may provide new insights on fundamental understanding of the role of glutamate in neurological disorders as well as neurophysiological phenomena. Electrochemical sensor has emerged as a promising solution that can satisfy the requirement for highly reliable and continuous monitoring method with good spatiotemporal resolution for characterization of extracellular glutamate concentration. Recently, we published a method to create a simple printable glutamate biosensor using platinum nanoparticles. In this work, we introduce an even simpler and lower cost conductive polymer composite using commercially available activated carbon with platinum microparticles to easily fabricate highly sensitive glutamate biosensor using direct ink writing method. The fabricated biosensors are functionality superior than previously reported with the sensitivity of 5.73 ± 0.078 nA µM-1 mm-2, detection limit of 0.03 µM, response time less than or equal to 1 s, and a linear range from 1 µM up to 925 µM. In this study, we utilize astrocyte cell culture to demonstrate our biosensor's ability to monitor glutamate uptake process. We also demonstrate direct measurement of glutamate release from optogenetic stimulation in mouse primary visual cortex (V1) brain slices.

In Vivo Glutamate Sensing inside the Mouse Brain with Perovskite Nickelate-Nafion Heterostructures.

Glutamate, one of the main neurotransmitters in the brain, plays a critical role in communication between neurons, neuronal development, and various neurological disorders. Extracellular measurement of neurotransmitters such as glutamate in the brain is important for understanding these processes and developing a new generation of brain-machine interfaces. Here, we demonstrate the use of a perovskite nickelate-Nafion heterostructure as a promising glutamate sensor with a low detection limit of 16 nM and a response time of 1.2 s via amperometric sensing. We have designed and successfully tested novel perovskite nickelate-Nafion electrodes for recording of glutamate release ex vivo in electrically stimulated brain slices and in vivo from the primary visual cortex (V1) of awake mice exposed to visual stimuli. These results demonstrate the potential of perovskite nickelates as sensing media for brain-machine interfaces.

Printable Nonenzymatic Glucose Biosensors Using Carbon Nanotube-PtNP Nanocomposites Modified with AuRu for Improved Selectivity.

Nonenzymatic glucose biosensors have the potential for a more reliable in vivo functionality due to the reduced risk of biorecognition element degradation. However, these novel sensing mechanisms often are nanoparticle-based and have nonlinear responses, which makes it difficult to gauge their potential utility against more conventional enzymatic biosensors. Moreover, these nonenzymatic biosensors often suffer from poor selectivity that needs to be better addressed before being used in vivo. To address these problems, here we present an amperometric nonenzymatic glucose biosensor fabricated using one-step electrodeposition of Au and Ru nanoparticles on the surface of a carbon-nanotube-based platinum-nanoparticle hybrid in conductive polymer. Using benchtop evaluations, we demonstrate that the bimetallic catalyst of Au-Ru nanoparticles can enable the nonenzymatic detection of glucose with a superior performance and stability. Furthermore, our biosensor shows good selectivity against other interferents, with a nonlinear dynamic range of 1-19 mM glucose. The Au-Ru catalyst has a conventional linear range of 1-10 mM, with a sensitivity of 0.2347 nA/(µM mm2) ± 0.0198 (n = 3) and a limit of detection of 0.068 mM (signal-to-noise, S/N = 3). The biosensor also exhibits a good repeatability and stability at 37 °C over a 3 week incubation period. Finally, we use a modified Butler-Volmer nonlinear analytical model to evaluate the impact of geometrical and chemical design parameters on our nonenzymatic biosensor's performance, which may be used to help optimize the performance of this class of biosensors.

Simple Fabrication of Flexible Biosensor Arrays Using Direct Writing for Multianalyte Measurement from Human Astrocytes.

Simultaneous measurements of glucose, lactate, and neurotransmitters (e.g., glutamate) in cell culture over hours and days can provide a more dynamic and longitudinal perspective on ways neural cells respond to various drugs and environmental cues. Compared with conventional microfabrication techniques, direct writing of conductive ink is cheaper, faster, and customizable, which allows rapid iteration for different applications. Using a simple direct writing technique, we printed biosensor arrays onto cell culture dishes, flexible laminate, and glass to enable multianalyte monitoring. The ink was a composite of PEDOT:PSS conductive polymer, silicone, activated carbon, and Pt microparticles. We applied 0.5% Nafion to the biosensors for selectivity and functionalized them with oxidase enzymes. We characterized biosensors in phosphate-buffered saline and in cell culture medium supplemented with fetal bovine serum. The biosensor arrays measured glucose, lactate, and glutamate simultaneously and continued to function after incubation in cell culture at 37 °C for up to 2 days. We cultured primary human astrocytes on top of the biosensor arrays and placed arrays into astrocyte cultures. The biosensors simultaneously measured glucose, glutamate, and lactate from astrocyte cultures. Direct writing can be integrated with microfluidic organ-on-a-chip platforms or as part of a smart culture dish system. Because we print extrudable and flexible components, sensing elements can be printed on any 3D or flexible substrate.

Ex vivo electrochemical measurement of glutamate release during spinal cord injury.

Excessive glutamate release following traumatic spinal cord injury (SCI) has been associated with exacerbating the extent of SCI. However, the mechanism behind sustained high levels of extracellular glutamate is unclear. Spinal cord segments mounted in a sucrose double gap recording chamber are an established model for traumatic spinal cord injury. We have developed a method to record, with micro-scale printed glutamate biosensors, glutamate release from ex vivo rat spinal cord segments following injury. This protocol would work equally well for similar glutamate biosensors.

Comments on "Antibiotic pollution in surface fresh waters: Occurrence and effects", Science of the Total Environment, 664, 793-804 (2019).

This article found mistakes and misunderstandings regarding the collection of occurrence data of antibiotics in surface freshwater from the previous literature. These mistakes and misunderstandings were corrected to avoid the propagation of inaccurate information in the scientific literature.

Facile fabrication of flexible glutamate biosensor using direct writing of platinum nanoparticle-based nanocomposite ink.

Glutamate excitotoxicity is a pathology in which excessive glutamate can cause neuronal damage and degeneration. It has also been linked to secondary injury mechanisms in traumatic spinal cord injury. Conventional bioanalytical techniques used to characterize glutamate levels in vivo, such as microdialysis, have low spatiotemporal resolution, which has impeded our understanding of this dynamic event. In this study, we present an amperometric biosensor fabricated using a simple direct ink writing technique for the purpose of in vivo glutamate monitoring. The biosensor is fabricated by immobilizing glutamate oxidase on nanocomposite electrodes made of platinum nanoparticles, multi-walled carbon nanotubes, and a conductive polymer on a flexible substrate. The sensor is designed to measure extracellular dynamics of glutamate and other potential biomarkers during a traumatic spinal cord injury event. Here we demonstrate good sensitivity and selectivity of these rapidly prototyped implantable biosensors that can be inserted into a spinal cord and measure extracellular glutamate concentration. We show that our biosensors exhibit good flexibility, linear range, repeatability, and stability that are suitable for future in vivo evaluation.

An electrically injected AlGaN nanowire defect-free photonic crystal ultraviolet laser.

We report on the demonstration of an electrically injected AlGaN nanowire photonic crystal laser that can operate in the ultraviolet spectral range. The nanowire heterostructures were grown on sapphire substrate using a site-controlled selective area growth process. By exploiting the topological high-Q resonance of a defect-free nanowire photonic crystal, we have demonstrated electrically pumped lasers that can operate at 369.5 nm with a relatively low threshold current density of ~2.1 kA/cm2 under continuous wave operation at room-temperature. This work provides a promising approach for achieving low threshold semiconductor laser diodes operating in the UV spectral range that were previously difficult.

Four-year safety follow-up of the tetravalent dengue vaccine efficacy randomized controlled trials in Asia and Latin America.

OBJECTIVE: Our objective was to describe the risk of hospital admission for virologically confirmed dengue (VCD) and the risk of clinically severe hospitalized VCD occurring up to 4 years after the first dose (years 1 to 4) in three randomized clinical trials comparing tetravalent dengue vaccine with placebo. METHODS: The relative risks (RR) for hospitalized VCD from first dose to year 4 were estimated by year and age-group in individual and combined studies. RESULTS: Overall, from Year 1 to Year 4, 233 and 228 participants had at least one episode of hospitalized VCD in the vaccinated (n = 22 603) and placebo (n = 11 301) groups, respectively (RR = 0.511, 95% CI 0.42-0.62). Among these, 48 and 47 cases, respectively, were classified as clinically severe. In children aged ≥9 years, 88 and 136 participants had at least one episode of hospitalized VCD in the vaccinated (n = 17 629) and placebo (n = 8821) groups, respectively (RR = 0.324; 95% CI 0.24-0.43). In vaccinated participants aged <9 years, particularly in those aged 2-5 years, there were more hospitalized VCD cases compared with the control participants in Year 3 but not in Year 4. The overall RR in those aged <9 years for Year 1 to Year 4 was 0.786 (95% CI 0.60-1.03), with a higher protective effect in the 6-8 year olds than in the 2-5 year olds. CONCLUSIONS: The overall benefit-risk remained positive in those aged ≥9 years up to year 4, although the protective effect was lower in years 3 and 4 than in years 1 and 2.

Anti-biofouling implantable catheter using thin-film magnetic microactuators.

Here we report on the development of polyimide-based flexible magnetic actuators for actively combating biofouling that occurs in many chronically implanted devices. The thin-film flexible devices are microfabricated and integrated into a single-pore silicone catheter to demonstrate a proof-of-concept for a self-clearing smart catheter. The static and dynamic mechanical responses of the thin-film magnetic microdevices were quantitatively measured and compared to theoretical values. The mechanical fatigue properties of these polyimide-based microdevices were also characterized up to 300 million cycles. Finally, the biofouling removal capabilities of magnetically powered microdevices were demonstrated using bovine serum albumin and bioconjugated microbeads. Our results indicate that these thin-film microdevices are capable of significantly reducing the amount of biofouling. At the same time, we demonstrated that these microdevices are mechanically robust enough to withstand a large number of actuation cycles during its chronic implantation.

Three Decades of Dengue Surveillance in Five Highly Endemic South East Asian Countries.

We described and quantified epidemiologic trends in dengue disease burden in 5 Asian countries (Indonesia, Thailand, Malaysia, Philippines, and Vietnam) and identified and estimated outbreaks impact over the last 3 decades. Dengue surveillance data from 1980 to 2010 were retrieved from DengueNet and from World Health Organization sources. Trends in incidence, mortality, and case fatality rate (CFR) were systematically analyzed using annual average percent change (AAPC), and the contribution of epidemic years identified over the observation period was quantified. Over the 30-year period, incidence increased in all countries (AAPC 1980-2010: 6.7% in Thailand, 10.4% in Vietnam, 12.0% in Indonesia, 18.1% in Malaysia, 24.4% in Philippines). Mortality also increased in Indonesia, Malaysia, and Philippines (AAPC: 6.8%, 7.0%, and 29.2%, respectively), but slightly decreased in Thailand and Vietnam (AAPC: -1.3% and -2.5%), and CFR decreased in all countries (AAPC: -4.2% to -8.3%). Epidemic years, despite representing less than a third of the observation period, contributed from 1 to 3 times more cases versus nonepidemic years. Implementation of more sensitive surveillance methods over the study period may have contributed to a reporting or ascertainment bias in some countries. Nonetheless, these data support the urgent need for novel, integrated, or otherwise effective dengue prevention and control tools and approaches.

PIK3CA mutations can initiate pancreatic tumorigenesis and are targetable with PI3K inhibitors.

Aberrations in the phosphoinositide 3-kinase (PI3K) signaling pathway have a key role in the pathogenesis of numerous cancers by altering cell growth, metabolism, proliferation and apoptosis. Interest in targeting the PI3K signaling cascade continues, as new agents are being clinically evaluated. PIK3CA mutations result in a constitutively active PI3K and are present in a subset of pancreatic cancers. Here we examine mutant PIK3CA-mediated pancreatic tumorigenesis and the response of PIK3CA mutant pancreatic cancers to dual PI3K/mammalian target of rapamycin (mTOR) inhibition. Two murine models were generated expressing a constitutively active PI3K within the pancreas. An increase in acinar-to-ductal metaplasia and pancreatic intraepithelial neoplasms (PanINs) was identified. In one model these lesions were detected as early as 10 days of age. Invasive pancreatic ductal adenocarcinoma developed in these mice as early as 20 days of age. These cancers were highly sensitive to treatment with dual PI3K/mTOR inhibition. In the second model, PanINs and invasive cancer develop with a greater latency owing to a lesser degree of PI3K pathway activation in this murine model. In addition to PI3K pathway activation, increased ERK1/2 signaling is common in human pancreatic cancers. Phosphorylation of ERK1/2 was also investigated in these models. Phosphorylation of ERK1/2 is demonstrated in the pre-neoplastic lesions and invasive cancers. This activation of ERK1/2 is diminished with dual PI3K/mTOR inhibition. In summary, PIK3CA mutations can initiate pancreatic tumorigenesis and these cancers are particularly sensitive to dual PI3K/mTOR inhibition. Future studies of PI3K pathway inhibitors for patients with PIK3CA mutant pancreatic cancers are warranted.

Exciton kinetics, quantum efficiency, and efficiency droop of monolayer MoS2 light-emitting devices.

We have investigated the quantum efficiency of monolayer MoS2 light-emitting devices through detailed temperature and power-dependent photoluminescence studies and rate equation analysis. The internal quantum efficiency can reach 45 and 8.3% at 83 and 300 K, respectively. However, efficiency droop is clearly measured with increasing carrier injection due to the unusually large Auger recombination coefficient, which is found to be ∼10(-24) cm(6)/s at room temperature, nearly 6 orders of magnitude higher than that of conventional bulk semiconductors. The significantly elevated Auger recombination in the emerging two-dimensional (2D) semiconductors is primarily an indirect process and is attributed to the abrupt bounding surfaces and the enhanced correlation, mediated by magnified Coulomb interactions, between electrons and holes confined in a 2D structure.

Role of nitrification in the biodegradation of selected artificial sweetening agents in biological wastewater treatment process.

The biodegradation of the six artificial sweetening agents including acesulfame (ACE), aspartame (ASP), cyclamate (CYC), neohesperidindihydrochalcone (NHDC), saccharin (SAC), and sucralose (SUC) by nitrifying activated sludge was first examined. Experimental results showed that ASP and NHDC were the most easily degradable compounds even in the control tests. CYC and SAC were efficiently biodegraded by the nitrifying activated sludge, whereas ACE and SUC were poorly removed. However, the biodegradation efficiencies of the ASs were increased with the increase in initial ammonium concentrations in the bioreactors. The association between nitrification and co-metabolic degradation was investigated and a linear relationship between nitrification rate and co-metabolic biodegradation rate was observed for the target artificial sweeteners (ASs). The contribution of heterotrophic microorganisms and autotrophic ammonia oxidizers in biodegradation of the ASs was elucidated, of which autotrophic ammonia oxidizers played an important role in the biodegradation of the ASs, particularly with regards to ACE and SUC.

Photoemission and absorption spectroscopy of carbon nanotube interfacial interaction.

Element-specific techniques including near edge X-ray absorption fine structure, extended X-ray absorption fine structure and X-ray photoemission spectroscopy for the characterization of the carbon nanotube interfacial interactions are reviewed. These techniques involve soft and hard X-rays from the laboratory-based and synchrotron radiation facilities. The results provided information of how the nano-particles of catalysts are involved in the initial stage of nanotube growth, the nanotube chemical properties after purification, functionalization, doping and composite formation.

Structural-chemical evolution within exfoliated clays.

The exfoliated (delaminated) structures of lamellar clays offer potential as precursors for the formation of various nanostructured materials. In this article, Lucentite and Laponite phyllosilicate clays, which both have empirical formulas of Na(0.33)[Mg(2.67)Li(0.33)Si4O10(OH)2] but differ in nanodimensions, have been exfoliated. Experiments were carried out for mixtures containing approximately 1 wt % phyllosilicate in a 5% aqueous solution of poly(acrylic acid) at different temperatures. X-ray diffraction and photoemission spectroscopy measurements for the solid products recovered after stirring the mixtures at 20 degrees C showed that the fully extended chains of poly(acrylic acid) were intercalated within the interlayer spaces between the silicate plates of the clays. At 85 degrees C, however, the clays were exfoliated and/or partially exfoliated. Photoemission spectroscopy also indicated that the exfoliated structures primarily consisted of silica nanoplates. 29Si nuclear magnetic resonance and oxygen K-edge near-edge X-ray absorption fine structure indicated that the surfaces of the plates were terminated by high concentrations of the silanol (-SiOH) groups, which created structural branches during intercalation. A model was developed in which intercalation and the removal of ions from the clays after the poly(acrylic acid) interactions reduced the electrostatic van der Waals forces between the plates. It was also shown that the formation of branches created a steric effect that inhibited the stacking of the plates. Together these resulted in exfoliation.

Dispersion of organically modified clays within n-alcohols.

A method for formation of polymer-clay nanocomposites involves dispersion of the nanometer silicate layers of clays into a solvent, followed by dispersion into polymers. The dispersion of layered silicates within solvents affects the structure and properties of the nanocomposites. We report the dispersion of organically modified clays, used for formation of nanocomposites with organic polymers, within a range of alcohol solvents. Experiments involved stirring a mixture containing approximately 1 wt% of alkylammonium-modified clays in n-alcohols with general molecular structure RnOH, where n represents the number of carbons of alkyl chains, varying from 2 to 8. The clays precipitated from the dispersion when RnOH solvents with n<5 were used, however, they formed gels for solvents with n5. The increased dispersion was related to the decrease of polarity and hydrogen bonding force within solvents. X-ray diffraction for the dispersed clays indicated that the interlayer spaces (1.8 nm), formed by regular stacking of the silicate layers, expanded to a maximum of 3.0 nm after treatment with RnOH with n5. The interlayer expansion was due to the intercalation of n-alcohol molecules within the interlayer spaces. It is suggested that the alkyl chains of n-alcohols remain parallel to the silicate surface in the intercalate. Preliminary experiments on the influence of these alcohol solvents on the intercalation of polyol (polyether) are also reported.