Graphene-Conductive Ink Coated Laser Engraved Kapton Electrochemical Biosensor for the Detection of Dopamine and Immune Sensing

Novel and flexible disposable laser-engraved Kapton (LEK) electrodes modified with graphene conductive inks have been developed for dopamine and Interleukin-6 (IL-6) detection. The LEK sensors exhibit high reproducibility (RSD=0.76%, N=5) and stability (RSD=4.39%, N=15)after multiple bendings, and thus make the sensors ideal for wearable and stretchable bioelectronics applications. We have developed graphene conductive ink- PEDOT:PSS (G-PEDOT:PSS) and polyaniline (G-PANI) based electrode coatings for working electrode modification to improve the sensitivity (41.41 times higher for screen printed electrode with G-PEDPT:PSS ink modification),and limit of detection (LOD) . We have further compared the performance of the fabricated electrodes with commercially available screen-printed electrodes (SPE), screen-printed electrodes modified with G-PEDOT:PSS (SPE/G-PEDOT:PSS) and G-PANI (SPE/G-PANI). SPE/G-PANI has a lower LOD of 0.632 μM, compared to SPE/G-PEDOT:PSS (0.867 μM) and SPE/G-PANI(1.974 μM). The lowest LOD of the LEK/G-PANI sensor (0.4084 μM, S/N=3) suggests that it can be a great alternative to measure dopamine levels in a physiological medium. Additionally, LEK/G-PANI electrode has excellent LOD (2.6234 pg/mL) to detect IL-6

Hydrogel and Graphene Embedded Piezoresistive Microcantilever Sensor for Solvent and Gas Flow Detection

Piezoresistive microcantilever sensor is widely used in sensing applications including liquid and gas flow detection. Microcantilevers can function as an embedded system if they are coated with polymers or nanomaterials to improve sensing performance. In this paper, we investigated the performance of piezoresistive microcantilevers (PMC) with and without additional coating. We studied the sensitivity of the PMC sensor after coating it with a three-dimensional porous hydrogel and piezoresistive graphene oxide layer. Hydrogel embedded piezoresistive microcantilever (EPM) showed better results than PMC during solvent sensing application. The resistance change for hydrogel embedded PMC was higher compared to bare PMC by 430% (3.2% to 17%) while detecting isopropyl alcohol (IPA), by approximately 1.5 orders of magnitude (0.19% to 5.7%) while detecting the presence of deionized water. Graphene Oxide coated PMC showed a wider detection range by 30 milliliter/min and 24% better sensitivity than bare PMC during the gas detection experiment. Additionally, we compared the experiment result with COMSOL simulation to develop a model for our embedded PMC sensing. Simulation shows significantly higher deflection of the EPM compared to the bare PMC (66.67% higher while detecting IPA, consistent with the trend observed during the experiment). The facile drop casting-based embedded microcantilever fabrication technique can lead to improved performance in different sensing applications. Our future work will focus on detecting biomolecules by using our constructed embedded systems

Graphene-conductive polymer-based electrochemical sensor for dopamine detection

The central nervous system\u27s (CNS) dopaminergic system dysfunction has been linked to neurological illnesses like schizophrenia and Parkinson\u27s disease. As a result, sensitive and selective detection of dopamine is critical for the early diagnosis of illnesses associated with aberrant dopamine levels. In this research, we have investigated the performance of electrochemical screen-printed sensors for different concentrations of dopamine detection using graphene-based conductive PEDOT: PSS(G-PEDOT: PSS) and Polyaniline(GPANI) inks on the working electrode and compared the sensitivity. SEM characterization technique has been performed to visualize the microstructures of the proposed inks. We have investigated cyclic voltammetry (CV) electrochemical techniques with ferri/ferrocyanide redox couple to assess the efficiency of the designed electrodes in detecting dopamine. GPANI ink has shown to have better LOD and stability to detect dopamine with screen-printed electrodes. Further, we have also studied electrochemical analysis for the selective detection of dopamine without the interference of Ascorbic Acid (AA)

Non-Destructive Infrared Thermographic Curing Analysis of Polymer Composites

Infrared (IR) thermography is a non-contact method of measuring temperature that analyzes the infrared radiation emitted by an object. Properties of polymer composites are heavily influenced by the filler material, filler size, and filler dispersion, and thus thermographic analysis can be a useful tool to determine the curing and filler dispersion. In this study, we investigated the curing mechanisms of polymer composites at the microscale by capturing real-time temperature using an IR Thermal Camera. Silicone polymers with fillers of Graphene, Graphite powder, Graphite flake, and Molybdenum disulfide (MoS 2) were subsequently poured into a customized 3D printed mold for thermography. The nanocomposites were microscopically heated with a Nichrome resistance wire, and real-time surface temperatures were measured using different Softwares. This infrared thermal camera divides the target area into 640x480 pixels, allowing measurement and analysis of the sample with a resolution of 65 micrometers. Depending on the filler material, the temperature rises to a certain maximum point before curing, and once curing is complete, polymer composites exhibit a rapid temperature change indicating a transition from viscous fluid to solid. MoS 2 , Polydimethylsiloxane (PDMS) without filler, and PDMS with larger filler are ranked in order of maximum constant temperature. PDMS (without filler) cures in 500s, while PDMS-Graphene and PDMS Graphite Powder cure in about 800s. The curing time for PDMS Graphite flake is slightly longer (950s), while MoS 2 is around 520s. Therefore, this technique can indicate the influence of fillers on the curing of composites at the microscale, which is difficult to achieve by conventional methods such as differential scanning calorimetry. This nondestructive, low-cost, fast infrared thermography can be used to analyze the properties of polymer composites with different fillers and dispersion qualities in a variety of applications including precision additive manufacturing and quality control of curable composite inks

Multifunctional Graphene–Polymer Nanocomposite Sensors Formed by One-Step In Situ Shear Exfoliation of Graphite

Graphene nanocomposites are a promising class of advanced materials for sensing applications; yet, their commercialization is hindered due to impurity incorporation during fabrication and high costs. The aim of this work is to prepare graphene–polysulfone (G−PSU) and graphene–polyvinylidene fluoride (G−PVDF) nanocomposites that perform as multifunctional sensors and are formed using a one-step, in situ exfoliation process whereby graphite is exfoliated into graphene nanoflakes (GNFs) directly within the polymer. This low-cost method creates a nanocomposite while avoiding impurity exposure since the raw materials used in the in situ shear exfoliation process are graphite and polymers. The morphology, structure, thermal properties, and flexural properties were determined for G−PSU and G−PVDF nanocomposites, as well as the electromechanical sensor capability during cyclic flexural loading, temperature sensor testing while heating and cooling, and electrochemical sensor capability to detect dopamine while sensing data wirelessly. G−PSU and G−PVDF nanocomposites show superior mechanical characteristics (gauge factor around 27 and significantly enhanced modulus), thermal characteristics (stability up to 500 °C and 170 °C for G−PSU and G−PVDF, respectively), electrical characteristics (0.1 S/m and 1 S/m conductivity for G−PSU and G−PVDF, respectively), and distinguished resonant peaks for wireless sensing (~212 MHz and ~429 MHz). These uniquely formed G−PMC nanocomposites are promising candidates as strain sensors for structural health monitoring, as temperature sensors for use in automobiles and aerospace applications, and as electrochemical sensors for health care and disease diagnostics

Hepatitis B Virus-Encoded HBsAg Contributes to Hepatocarcinogenesis by Inducing the Oncogenic Long Noncoding RNA LINC00665 through the NF-κB Pathway

Clinical and in vivo studies have demonstrated a role for hepatitis B virus (HBV)-encoded HBsAg (hepatitis B surface antigen) in HBV-related hepatocellular carcinoma (HCC); however, the underlying mechanisms remain largely unknown. Here, we investigated the role of HBsAg in regulating long noncoding RNAs (lncRNAs) involved in HCC progression. Our analysis of microarray data sets identified LINC00665 as an HBsAg-regulated lncRNA. Furthermore, LINC00665 is upregulated in liver samples from HBV-infected patients as well as in HCC, specifically in HBV-related HCC liver samples. These findings were supported by our in vitro data demonstrating that HBsAg, as well as HBV, positively regulates LINC00665 in multiple HBV cell culture models. Next, we evaluated the oncogenic potential of LINC00665 by its overexpression and CRISPR interference (CRISPRi)-based knockdown in various cell-based assays. LINC00665 promoted cell proliferation, migration, and colony formation but inhibited cell apoptosis in vitro. We then identified the underlying mechanism of HBsAg-mediated regulation of LINC00665. We used immunofluorescence assays to show that HBsAg enhanced the nuclear translocation of NF-κB factors in HepG2 cells, confirming that HBsAg activates NF-κB. Inhibition of NF-κB signaling nullified HBsAg-mediated LINC00665 upregulation, suggesting that HBsAg acts through NF-κB to regulate LINC00665. Furthermore, the LINC00665 promoter contains NF-κB binding sites, and their disruption abrogated HBsAg-induced LINC00665 upregulation. Finally, HBsAg facilitated the enrichment of the NF-κB factors NF-κB1, RelA, and c-Rel in the LINC00665 promoter. Taken together, this work shows that HBsAg can drive hepatocarcinogenesis by upregulating oncogenic LINC000665 through the NF-κB pathway, thereby identifying a novel mechanism in HBV-related HCC. IMPORTANCE Hepatitis B virus (HBV) is a major risk factor for hepatocellular carcinoma (HCC). Numerous reports indicate an oncogenic role for HBV-encoded HBsAg; however, the underlying mechanisms are not well understood. Here, we studied the role of HBsAg in regulating lncRNAs involved in hepatocarcinogenesis. We demonstrate that HBsAg, as well as HBV, positively regulates oncogenic lncRNA LINC00665. The clinical significance of this lncRNA is highlighted by our observation that LINC00665 is upregulated in liver samples during HBV infection and HBV-related HCC. Furthermore, we show LINC00665 can drive hepatocarcinogenesis by promoting cell proliferation, colony formation, and cell migration and inhibiting apoptosis. Taken together, this work identified LINC00665 as a novel gene through which HBsAg can drive hepatocarcinogenesis. Finally, we show that HBsAg enhances LINC00665 levels in hepatocytes by activating the NF-κB pathway, thereby identifying a novel mechanism by which HBV may contribute to HCC.ISSN:2165-049

Graphene Nanocomposite Ink Coated Laser Transformed Flexible Electrodes for Selective Dopamine Detection and Immunosensing

Novel and flexible disposable laser-induced graphene (LIG) sensors modified with graphene conductive inks have been developed for dopamine and interleukin-6 (IL-6) detection. The LIG sensors exhibit high reproducibility (relative standard deviation, RSD = 0.76%, N = 5) and stability (RSD = 4.39%, N = 15) after multiple bendings, making the sensors ideal for wearable and stretchable bioelectronics applications. We have developed electrode coatings based on graphene conductive inks, poly(3,4-ethylenedioxythiophene):polystyrene sulfonate (G-PEDOT:PSS) and polyaniline (G-PANI), for working electrode modification to improve the sensitivity and limit of detection (LOD). The selectivity of LIG sensors modified with the G-PANI ink is 41.47 times higher than that of the screen-printed electrode with the G-PANI ink modification. We have compared our fabricated bare laser-engraved Kapton sensor (LIG) with the LIG sensors modified with G-PEDOT (LIG/G-PEDOT) and G-PANI (LIG/G-PANI) conductive inks. We have further compared the performance of the fabricated electrodes with commercially available screen-printed electrodes (SPEs) and screen-printed electrodes modified with G-PEDOT:PSS (SPE/G-PEDOT:PSS) and G-PANI (SPE/G-PANI). SPE/G-PANI has a lower LOD of 0.632 μM compared to SPE/G-PEDOT:PSS (0.867 μM) and SPE/G-PANI (1.974 μM). The lowest LOD of the LIG/G-PANI sensor (0.4084 μM, S/N = 3) suggests that it can be a great alternative to measure dopamine levels in a physiological medium. Additionally, the LIG/G-PANI electrode has excellent LOD (2.6234 pg/mL) to detect IL-6. Also, the sensor is successfully able to detect ascorbic acid (AA), dopamine (DA), and uric acid (UA) in their ternary mixture. The differential pulse voltammetry (DPV) result shows peak potential separation of 229, 294, and 523 mV for AA–DA, DA–UA, and UA–AA, respectively

Genome profiling of uropathogenic E. coli from strictly defined community-acquired UTI in paediatric patients: a multicentric study

Abstract Background Urinary tract infection (UTI) in children is a common bacterial infection. The emergence of extended-spectrum beta-lactamases (ESBLs) poses a major challenge against the treatment of uropathogens. We aimed to characterize the E. coli isolates recovered from children with UTI for their resistance profile and circulating sequence types (ST). Methods Children (> 1.5–18 years of age) from different community health centres of India with symptoms of UTI were enrolled. Isolates causing significant bacteriuria were identified by Matrix-Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (MALDI-TOF MS) and tested for antimicrobial susceptibility by the automated system, VITEK-2 (Biomeriux, Durhum, US). Nineteen E. coli isolates (15 ESBL positive and 4 ESBL negative) were sequenced in Oxford Nanopore platform followed by core-genome phylogeny, accessory genome cluster analysis, identification of sequence types, mobile genetic elements, genetic antimicrobial resistance markers. The correlation between detection of antimicrobial resistance genes with phenotypic resistance profiles was also investigated. Results Eleven percent of children had significant bacteriuria [male:female—1:1, > 50% were 11–18 years of age group]. E. coli was predominant (86%) followed by K. pneumoniae (11%). Susceptibility of E. coli was highest against fosfomycin (100%) followed by carbapenems (90.7%) and nitrofurantoin (88.8%). ST131 (15.8%) and ST167 (10.5%) found as high-risk clones with the presence of plasmid [IncFIB (63.1%), IncFIA (52.6%)], and composite transposon [Tn2680 (46.6%)] in many isolates. Few isolates coharboured multiple beta-lactamases including bla NDM-5 (33.3%), bla OXA-1 (53.3%), bla CTX-M-15 (60%) and bla TEM-4 (60%). Conclusions This study highlights horizontal transmission of resistance genes and plasmids in paediatric patients at community centers across the nation harbouring multidrug-resistant genes such as bla NDM-5 and bla CTX-M-15 associated with high-risk clones ST131 and ST167. The data is alarming and emphasizes the need for rapid identification of resistance markers to reduce the spread in community. To our knowledge, this is the first multicentric study targeting paediatric UTI patients from the community setting of India