Facile Synthesis of Needle-like Copper Sulfide Structures as an Effective Electrochemical Sensor Material for Neurotransmitter Detection in Biological Samples.

Electrochemical sensors, due to their excellent and unique features, are of high interest nowadays for the detection and monitoring of several biological compounds. In such a case, serotonin (SRN), an important neurotransmitter, was herein studied for its detection in biological fluids since its presence is more crucial to be monitored and detected in clinical and medical applications. Several study strategies have been used to determine the chemical and physical properties. The crystalline size of the constructed copper sulfide (Cu2S) material was measured to be 25.92 nm. The Cu2S was fabricated over the working surface and further analyzed for several sensor parameters to be optimized. The charge transfer resistance of the copper sulfide-modified glassy carbon electrode (Cu2S/GCE) was determined to be about 277.0 Ω. With the linear range from about 0.029 µM to 607.6 µM for SRN, the limit of detection (LOD) was calculated as 3.2 nM, with a good sensitivity of 13.23 µA µM-1 cm2. The sensor experienced excellent repeatability, reproducibility, and long-term stability. The fabricated electrode was selective with the presence of different interfering compounds. The real sample analysis, as determined with the regular addition method with human serum and urine samples, revealed a good recovery percentage. Thus, the employed fabricated electrode material will be highly effective in sensing other analytes of choice.

Altered expression of the L-arginine/nitric oxide pathway in ovarian cancer: metabolic biomarkers and biological implications.

MOTIVATION: Ovarian cancer (OC) is a highly lethal gynecological malignancy. Extensive research has shown that OC cells undergo significant metabolic alterations during tumorigenesis. In this study, we aim to leverage these metabolic changes as potential biomarkers for assessing ovarian cancer. METHODS: A functional module-based approach was utilized to identify key gene expression pathways that distinguish different stages of ovarian cancer (OC) within a tissue biopsy cohort. This cohort consisted of control samples (n = 79), stage I/II samples (n = 280), and stage III/IV samples (n = 1016). To further explore these altered molecular pathways, minimal spanning tree (MST) analysis was applied, leading to the formulation of metabolic biomarker hypotheses for OC liquid biopsy. To validate, a multiple reaction monitoring (MRM) based quantitative LCMS/MS method was developed. This method allowed for the precise quantification of targeted metabolite biomarkers using an OC blood cohort comprising control samples (n = 464), benign samples (n = 3), and OC samples (n = 13). RESULTS: Eleven functional modules were identified as significant differentiators (false discovery rate, FDR < 0.05) between normal and early-stage, or early-stage and late-stage ovarian cancer (OC) tumor tissues. MST analysis revealed that the metabolic L-arginine/nitric oxide (L-ARG/NO) pathway was reprogrammed, and the modules related to "DNA replication" and "DNA repair and recombination" served as anchor modules connecting the other nine modules. Based on this analysis, symmetric dimethylarginine (SDMA) and arginine were proposed as potential liquid biopsy biomarkers for OC assessment. Our quantitative LCMS/MS analysis on our OC blood cohort provided direct evidence supporting the use of the SDMA-to-arginine ratio as a liquid biopsy panel to distinguish between normal and OC samples, with an area under the ROC curve (AUC) of 98.3%. CONCLUSION: Our comprehensive analysis of tissue genomics and blood quantitative LC/MSMS metabolic data shed light on the metabolic reprogramming underlying OC pathophysiology. These findings offer new insights into the potential diagnostic utility of the SDMA-to-arginine ratio for OC assessment. Further validation studies using adequately powered OC cohorts are warranted to fully establish the clinical effectiveness of this diagnostic test.

Two dimensional architectures of graphitic carbon nitride with the substitution of heteroatoms for bifunctional electrochemical detection of nilutamide.

The exploration of graphitic carbon nitride (g-C3N4), a two-dimensional (2D) metal-free polymer semiconducting material, is largely discussed due to its large specific surface area, high electrical conductivity, thermal stability, and adaptable electronic structure. The adaption of sulfur (S) and phosphorous (P) atoms into the layers of g-C3N4 increases the electrochemical performance of detecting nilutamide (NT). The aggregation severity can be decreased by integrating S/P into g-C3N4, thereby improving surface area and electrical conductance. The g-C3N4, S/gC3N4, P/g-C3N4, and S/P/g-C3N4 were studied with X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), Field emission scanning electron microscopy (FESEM), Transmission electron microscopy (TEM), Fourier transform infrared (FTIR), Ultraviolet visible spectroscopy (UV), Thermogravimetric analysis (TGA), and Brunauer-Emmett-Teller (BET). The well-assigned S/P/g-C3N4 exhibited a good crystalline structure with more active sites for improved electron transfer toward NT detection. Both differential pulse voltammetry (DPV) and amperometry (IT) was studied for NT detection. The electrochemical studies were done with a linear range of 0.019-1.17 µM to 5.36-1891.98 µM in DPV and 0.01 µM-158.3 µM in IT technique. The attained limit of detection in DPV analysis was 3.2 nM and with IT analysis 2.4 nM. The nanocomposite S/P/g-C3N4 shows good selectivity towards NT. The fabricated electrode showed excellent repeatability, reproducibility, and stability, with a significant recovery range in real sample analysis.

Single center blind testing of a US multi-center validated diagnostic algorithm for Kawasaki disease in Taiwan.

Background: Kawasaki disease (KD) is the leading cause of acquired heart disease in children. The major challenge in KD diagnosis is that it shares clinical signs with other childhood febrile control (FC) subjects. We sought to determine if our algorithmic approach applied to a Taiwan cohort. Methods: A single center (Chang Gung Memorial Hospital in Taiwan) cohort of patients suspected with acute KD were prospectively enrolled by local KD specialists for KD analysis. Our previously single-center developed computer-based two-step algorithm was further tested by a five-center validation in US. This first blinded multi-center trial validated our approach, with sufficient sensitivity and positive predictive value, to identify most patients with KD diagnosed at centers across the US. This study involved 418 KDs and 259 FCs from the Chang Gung Memorial Hospital in Taiwan. Findings: Our diagnostic algorithm retained sensitivity (379 of 418; 90.7%), specificity (223 of 259; 86.1%), PPV (379 of 409; 92.7%), and NPV (223 of 247; 90.3%) comparable to previous US 2016 single center and US 2020 fiver center results. Only 4.7% (15 of 418) of KD and 2.3% (6 of 259) of FC patients were identified as indeterminate. The algorithm identified 18 of 50 (36%) KD patients who presented 2 or 3 principal criteria. Of 418 KD patients, 157 were infants younger than one year and 89.2% (140 of 157) were classified correctly. Of the 44 patients with KD who had coronary artery abnormalities, our diagnostic algorithm correctly identified 43 (97.7%) including all patients with dilated coronary artery but one who found to resolve in 8 weeks. Interpretation: This work demonstrates the applicability of our algorithmic approach and diagnostic portability in Taiwan.

Strategic orchestration of MoSe2 microspheres on ß-cd functionalized rGO: A sustainable electrocatalyst for detection of rifampicin in real samples.

The ill effects of prolonged use of rifamycin antibiotics such as rifampicin accentuates its need for detection in the environment as well as in biological fluids. Antibiotics in water and soil are long-lasting, bio-accumulative, and hazardous to aquatic species as well as human health. To address this issue, a sensing platform has been developed using Molybdenum diselenide (MoSe2) embedded on reduced graphene oxide (rGO) functionalized with ß-cyclodextrin (ß-cd) polymer. The formation of hybrid composite was validated with X-ray diffraction analysis (XRD), Raman spectroscopy, fourier transform infrared spectroscopy (FT-IR), X-ray photoelectron spectroscopy (XPS), and field emission scanning electron microscopy (FE-SEM) with EDX analysis. The formation of microspheres were observed with hexagonal crystal system and P63/mmc space group. Furthermore, the composite was employed to fabricate an efficient electrochemical sensor for detecting the widely used antibiotic, rifampicin (RIF). The results reveal excellent activity of the sensor with a limit of detection (LOD) of 28 nM in a linear working range from 0.019 to 374.5 µM. The sensor also exhibited a high sensitivity of 11.64 µA µM-1 cm-2. Additionally, the sensor showed appreciable recovery range when monitored in real-samples such as human serum and urine, and industrial water, and fish samples.

Self-assembled nanorods with reduced graphene oxide as efficient nano-catalyst for dual modality sensing of hazardous phenolic compound.

The modern development in the agricultural production has huge influential factors being highly beneficial and also includes some health hazards. Under the class of chlorophenols, 2,4,6-trichlorophenol is a widely used chemical which remains as a major pollutant in the environment. The detection of 2,4,6-trichlorophenol was initiated as a controlling measure to decrease the seriousness prevailing in the ecosystem. The electrochemical and UV-vis absorption sensing platform are simple and low-cost detection techniques with precise and sensitive analysis. Cadmium tin oxide integrated with the reduced graphene oxide was employed as a nanohybrid for the construction of the working electrode. The structural and morphological analysis confirmed the high degree of crystallinity of the nanocomposite with nanorod formation. The high surface area, with high charge carrier mobility, and increased electrical conductivity of the material boosted the 2,4,6-trichlorophenol detection. The active surface area was calculated to be 0.068 cm-1, 0.089 cm-1, 0.118 cm-1 and 0.146 cm-1 for all the modified electrodes. The resistance of the electrodes was about 91.4 Ω, 72.9 Ω, 48.8 Ω and 41.6 Ω. The linear range of 2,4,6-trichlorophenol was 0.019 µM-0.299 µM and 1.299 µM-1678.97 µM in electrochemical sensing and 10.99 µM-24.84 µM in UV detection. The obtained limit of detection with the formulation 3σ/SD was about 3.05 nM and 80 nM with sensitivity about 14.01 µA µM-1 cm-2. The real sample detection in environmental real samples showed good recovery results. The specific selectivity, good repeatability, reproducibility and stability analysis proves the good sensing parameters. Thus, the fabricated electrode is highly sufficient of sensing 2,4,6-trichlorophenol. These excellent features of the material can be applied for several other applications which will provide good performances.

Investigation of T site variation in spinel aluminates TAl2O4 (T= Mg, Zn & Cu), and formation of electrocatalyst CuAl2O4/carbon for efficient sensing application.

Spinel structured aluminates TAl2O4 (T = Mg, Zn, and Cu) were synthesized by a facile hydrothermal method. The resultant enhancement in the electrochemical behavior was achieved due to the covalent synergism among the elements coexisting together. Structural and morphological characterizations were performed by X-ray diffraction, Fourier transform infrared spectroscopy, Raman spectroscopy, and field emission scanning electron microscopy. MgAl2O4, ZnAl2O4 and CuAl2O4 has displayed same space group Fd3m of Laue class lattice type of the cubic structure as they were synthesized at same temperature (600 °C). CuAl2O4 spinel structure displayed a nanoneedle like structure along with the small sized cylindrical particles alongside to which CuAl2O4 spinel is combined with activated carbon (CuAl/C) and was applied to develop a facile sensor for the electrochemical detection of Acetaminophen (ACAP) using cyclic voltammetry (CV) and differential pulse voltammetry (DPV), which exhibited maximum conductivity, and a substantial electroactive surface area. Finally, the defect-rich composite, CuAl/C, showed excellent sensor performance towards DPV with 21.5 nM limit of detection (LOD) in a wide linear working range of 0.199 µM-165.88 µM ACAP concentration, with a high sensitivity of 19.1221 µA µM-1cm2. Additionally, the sensor showed excellent recovery results in real-time analysis for environmental aquatic samples like industrial wastewater and Tuna Fish.

Multi-Omics Signatures Link to Ticagrelor Effects on Vascular Function in Patients With Acute Coronary Syndrome.

BACKGROUND: Long-term antiplatelet agents including the potent P2Y12 antagonist ticagrelor are indicated in patients with a previous history of acute coronary syndrome. We sought to compare the effect of ticagrelor with that of aspirin monotherapy on vascular endothelial function in patients with prior acute coronary syndrome. METHODS: This was a prospective, single center, parallel group, investigator-blinded randomized controlled trial. We randomized 200 patients on long-term aspirin monotherapy with prior acute coronary syndrome in a 1:1 fashion to receive ticagrelor 60 mg BD (n=100) or aspirin 100 mg OD (n=100). The primary end point was change from baseline in brachial artery flow-mediated dilation at 12 weeks. Secondary end points were changes to platelet activation marker (CD41_62p) and endothelial progenitor cell (CD34/133) count measured by flow cytometry, plasma level of adenosine, IL-6 (interleukin-6) and EGF (epidermal growth factor), and multi-omics profiling at 12 weeks. RESULTS: After 12 weeks, brachial flow-mediated dilation was significantly increased in the ticagrelor group compared with the aspirin group (ticagrelor: 3.48±3.48% versus aspirin: -1.26±2.85%, treatment effect 4.73 [95% CI, 3.85-5.62], P<0.001). Nevertheless ticagrelor treatment for 12 weeks had no significant effect on platelet activation markers, circulating endothelial progenitor cell count or plasma level of adenosine, IL-6, and EGF (all P>0.05). Multi-omics pathway assessment revealed that changes in the metabolism and biosynthesis of amino acids (cysteine and methionine metabolism; phenylalanine, tyrosine, and tryptophan biosynthesis) and phospholipids (glycerophosphoethanolamines and glycerophosphoserines) were associated with improved brachial artery flow-mediated dilation in the ticagrelor group. CONCLUSIONS: In patients with prior acute coronary syndrome, ticagrelor 60 mg BD monotherapy significantly improved brachial flow-mediated dilation compared with aspirin monotherapy and was associated with significant changes in metabolomic and lipidomic signatures. REGISTRATION: URL: https://www. CLINICALTRIALS: gov; Unique identifier: NCT03881943.

Preparation of substrates for microarray protein chips with different ending functional groups.

Protein microarray chips are composed of three components, these are pre-treatment substrates, surface chemical modification, and immobilizing protein on substrate surfaces. In this study, self-assembly monolayers are used for surface chemical modification. Using this method, silanization on a glass and silicon chip is achieved, forming the terminal group substrates. Modification of the substrate surface to provide COOH and NH2 terminal functional groups provides a mechanism to proteins to immobilize on the substrate surface. To observe immobilized proteins on the substrate surface, they are first labeled with Cy5 fluorescent dye before analysis using a GenePix 4000B Microarray Scanner. The scanner induces fluorescence in the labelling dye and the resulting light is analyzed to provide information concerning both the quantity of immobilized protein, and the orientation of attachment. The antigen of the HSV-1 virus, a common human virus, was used in this study, performing an antigen-antibody analysis to determine the efficacy of the method under test for clinical diagnosis.

Vanadium selenide decorated reduced graphene oxide nanocomposite: A co-active catalyst for the detection of 2,4,6 - Trichlorophenol.

Transition metal chalcogenides (TMCs) have great potential in diverse electrochemical technologies owing to their unique characteristics. In the present work, we portray the design and synthesis of Vanadium selenide (V2Se9)/reduced graphene oxide (rGO) forming a two-dimensional (2D) hybrid nanocomposite via a simple hydrothermal method. The successfully synthesized nanocomposite underwent in-depth surface and morphological characterizations by XRD, Raman spectroscopy, XPS, TEM, STEM and its potential as an electro catalyst was investigated by using glassy carbon electrode (GCE) for the detection of 2,4,6-trichlorophenol (TCP). The structural features favored a high charge transfer ratio, high surface area as well as excellent conductivity and catalytic activity. The V2Se9/rGO/GCE modified electrode showed a low charge transfer resistance (Rct) of 54.057 Ω cm2, a decent detection limit (LOD) of 35.07 nM and a very high sensitivity of 22 µA µM-1 cm-2 in a working range of 0.001 µM-1150 µM. This is due to the active proton interaction, surface enhancement, and positive synergistic effect between rGO and V2Se9. The proposed sensor has good detection potential in agricultural soil, river water, fish, and beverage samples like wine and apple juice. The obtained results from our investigation would elucidate the application of the catalyst in electrochemical sensors.

Facile Synthesis of Ag/AgVO3/N-rGO Hybrid Nanocomposites for Electrochemical Detection of Levofloxacin for Complex Biological Samples Using Screen-Printed Carbon Paste Electrodes.

Silver vanadate nanorods (ß-AgVO3) with silver nanoparticles (Ag-NPs) decorated on the surface of the rods were synthesized by using simple hydrothermal technique and later anchored onto nitrogen-doped reduced graphene oxide (N-rGO) to make a novel nanocomposite. Experimental analyses were carried out to identify the electronic configuration by X-ray diffraction analysis, Fourier transform infrared spectroscopy, and X-ray photoelectron spectroscopy analysis, which revealed monoclinic patterns of the C12/m1 space group with Wulff construction forming beta silver vanadate (ß-AgVO3) crystals with optical density and phase transformations. Ag nucleation showed consistent results with metallic formation and electronic changes occurring in [AgO5] and [AgO3] clusters. Transmission electron microscopy and field-emission scanning electron microscopy with elemental mapping and EDX analysis of the morphology reveals the nanorod structure for ß-AgVO3 with AgNPs on the surface and sheets for N-rGO. Additionally, a novel electrochemical sensor is constructed by using Ag/AgVO3/N-rGO on screen-printed carbon paste electrodes for the detection of antiviral drug levofloxacin (LEV) which is used as a primary antibiotic in controlling COVID-19. Using differential pulse voltammetry, LEV is determined with a low detection limit of 0.00792 nm for a linear range of 0.09-671 µM with an ultrahigh sensitivity of 152.19 µA µM-1 cm-2. Furthermore, modified electrode performance is tested by real-time monitoring using biological and river samples.

Rational design and preparation of copper vanadate anchored on sulfur doped reduced graphene oxide nanocomposite for electrochemical sensing of antiandrogen drug nilutamide using flexible electrodes.

Copper vanadate nanoparticles (Cu2V2O7) are synthesized by using a simple hydrothermal method and later anchored with sulfur-doped reduced graphene oxide (S-rGO) by using ultrasonication to form a hybrid nanocomposite. The synthesized composite underwent characterizations like X-ray diffraction analysis (XRD), Raman spectroscopy, Fourier transform infrared spectroscopy (FT-IR), Dynamic ray scattering-Ultra violet-visible spectroscopy (DRS-UV-visible) and X-ray photoelectron spectroscopically revealed the triclinic pattern of the P 1̅ space group of α-Cu2V2O7 and the reduced oxygen deficiency state of metal centers (Cu+ or V4+) resulting with oxides of mixed-valence oxidative states and forming of Cu-O bond. Morphological analysis was carried out by using transmission electron microscopy (TEM) and Field emission scanning electron microscopy (FE-SEM) with elemental mapping and EDX analysis. Furthermore, a novel electrochemical sensor is prepared by using the hybrid sCu2V2O7/S-rGO nanocomposite on to a disposable screen-printed carbon paste electrode (SPCE) for electrochemical sensing of antiandrogen drug nilutamide (NLT). This report reveals excellent activity in determining NLT with a low detection limit of 0.00459 nM for the linear range of 0.001-15 µM with high sensitivity of 26.2605 µA µM-1 cm-2. Further, electrode performance showed appreciable performance in real-time monitoring of biological samples like human blood serum, urine samples.

Synthesis and cytotoxic analysis of thiolated xylose derivatives decorated on gold nanoparticles.

The rapid development of metal nanoparticles capped by an organic monolayer offers the possibility to create a whole new variety of products with novel characteristic, functions and applications. Among these, nanoparticles covered with carbohydrates (glyconanoparticles) constitute a good bio-mimetic model of carbohydrate presentation at the cell surface and are currently centered on many glycobiological and biomedical applications. In this study, a series of novel D-xylose gold nanoparticles (AuNPs) with linkages of alkyl or polyethylene glycol have been synthesized via D-xylosethiols, forming self-assembled monolayers on gold nanoparticles. The nano-gold solution, two carbohydrate derivatives and modified nano-gold solution were tested for cytotoxicity to check the biocompatibility. The MTT assay on NIH 3T3 cell lines confirmed that all the test materials showed no toxicity with the more than 90 % of cell viability in both low concentration (1 µM) and high concentration (100 µM), compared with the control.

Kinetics of SARS-CoV-2 positivity of infected and recovered patients from a single center.

Recurrence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) positive detection in infected but recovered individuals has been reported. Patients who have recovered from coronavirus disease 2019 (COVID-19) could profoundly impact the health care system. We sought to define the kinetics and relevance of PCR-positive recurrence during recovery from acute COVID-19 to better understand risks for prolonged infectivity and reinfection. A series of 414 patients with confirmed SARS-Cov-2 infection, at The Second Affiliated Hospital of Southern University of Science and Technology in Shenzhen, China from January 11 to April 23, 2020. Statistical analyses were performed of the clinical, laboratory, radiologic image, medical treatment, and clinical course of admission/quarantine/readmission data, and a recurrence predictive algorithm was developed. 16.7% recovered patients with PCR positive recurring one to three times, despite being in strict quarantine. Younger patients with mild pulmonary respiratory syndrome had higher risk of PCR positivity recurrence. The recurrence prediction model had an area under the ROC curve of 0.786. This case series provides characteristics of patients with recurrent SARS-CoV-2 positivity. Use of a prediction algorithm may identify patients at high risk of recurrent SARS-CoV-2 positivity and help to establish protocols for health policy.

Influence of temperature variation on spinel-structure MgFe2O4 anchored on reduced graphene oxide for electrochemical detection of 4-cyanophenol.

The effect of annealing temperature variance on magnesium ferrites (MgFe2O4) later anchored on reduced graphene oxide (rGO) forming hybrid nanocomposite is demonstrated and its electrochemical performance investigated by using a screen-printed carbon paste electrode (SPCE) for detection of the environmental hazardous phenolic compound 4-cyanophenol (4-CY). The MgFe2O4 (MFO-600 °C) displayed an enhanced charge transfer ratio with high conductivity and electrocatalytic activity. To confirm the structural and morphological parameters of the rGO-MFO-2 hybrid micro/nanocomposite, X-ray diffraction (XRD), Raman spectroscopy, X-ray photoelectron microscopy (XPS), and field-emission electron microscopy (FE-EM) with EDX mapping have been utilized. The rGO/MFO-2/SPCE electrode displayed high catalytic performance in detecting 4-CY with good sensitivity of 6.836 µA µM-1 cm-2 in a working range 0.001 to 700 µM with a limit of detection of 0.0012 µM by using differential pulse voltammetry (DPV). This is achieved for the active interaction between rGO and MFO-2 active surface site areas resulting in good electrochemical activity and high electron transfer rate. Moreover, 4-CY detection has been performed in the presence of various interferents and through real-time analysis in samples like tap water, industrial river water, and fish which resulted in admirable recovery.Graphical abstract.

Nano assembly of NiFe spheres anchored on f-MWCNT for electrocatalytic reduction and sensing of nitrofurantoin in biological samples.

The current study reports a facile simple, low-cost electrochemical sensor in the detection of nitrofurantoin (NFT) by using NiFe/f-MWCNT hybrid composite as a promising electrocatalyst. NFT is an antibiotic drug that is extensively using in pharmaceuticals and also in animal food production which causes a severe threat for both human and animal environments. Extending the residues of NFT are left into rivers, soils, lakes, and groundwaters either found or discharged leading health issues. To this NiFe/f-MWCNT composite was synthesized using a hydrothermal mechanism and then ultrasonicated to form a hybrid composite for catalytic evaluation and electrochemical detection of NFT for the very first time. Furthermore, the physicochemical properties of NiFe nanospheres conjugated on f-MWCNT are scrutinized using various analytical and spectroscopical techniques. Resulting transmission electron microscopy (TEM) displays a chain like NiFe nanospheres anchored on f-MWCNT with a well-defined spherical shape, without any comprehensive agglomeration. The NiFe/f-MWCNT screen printed carbon paste electrode (SPCE) displayed an excellent electrocatalytic activity for NFT with a LOD of 0.03 µM and a sensitivity of 11.45 µA µM-1 cm-2. establishing a new selectivity and with the existence of co-interfering compounds. To enhance the practical abilities analysis were performed in Human serum and urine samples which resulted in satisfactory recoveries with high precision and linear accuracy illustrated in Scheme 1.

Analyzing polymeric matrix for fabrication of a biodegradable microneedle array to enhance transdermal delivery.

Traditional drug delivery systems, using invasive, transdermal, and oral routes, are limited by various factors, such as the digestive system environment, skin protection, and sensory nerve stimulation. To improve the drug delivery system, we fabricated a polysaccharide-based, dissolvable microneedle-based array, which combines the advantages of both invasive and transdermal delivery systems, and promises to be an innovative solution for minimally invasive drug delivery. In this study, we designed a reusable aluminum mold that greatly improved the efficiency and convenience of microneedle fabrication. Physical characterization of the polysaccharides, individual or mixed at different ratios, was performed to identify a suitable molecule to fabricate the dissolvable microneedle. We used a vacuum deposition-based micro-molding method at low temperature to fabricate the model. Using a series of checkpoints from material into product, a systematic feedback mechanism was built into the "all-in-one" fabrication step, which helped to improve production yields. The physical properties of the fabricated microneedle were assessed. The cytotoxicity analysis and animal testing of the microneedle demonstrated the safety and compatibility of the microneedle, and the successful penetration and effective release of a model protein.

Glucose biosensor based on glucose oxidase immobilized at gold nanoparticles decorated graphene-carbon nanotubes.

Biopolymer pectin stabilized gold nanoparticles were prepared at graphene and multiwalled carbon nanotubes (GR-MWNTs/AuNPs) and employed for the determination of glucose. The formation of GR-MWNTs/AuNPs was confirmed by scanning electron microscopy, X-ray diffraction, UV-vis and FTIR spectroscopy methods. Glucose oxidase (GOx) was successfully immobilized on GR-MWNTs/AuNPs film and direct electron transfer of GOx was investigated. GOx exhibits highly enhanced redox peaks with formal potential of -0.40 V (vs. Ag/AgCl). The amount of electroactive GOx and electron transfer rate constant were found to be 10.5 × 10(-10) mol cm(-2) and 3.36 s(-1), respectively, which were significantly larger than the previous reports. The fabricated amperometric glucose biosensor sensitively detects glucose and showed two linear ranges: (1) 10 µM - 2 mM with LOD of 4.1 µM, (2) 2 mM - 5.2 mM with LOD of 0.95 mM. The comparison of the biosensor performance with reported sensors reveals the significant improvement in overall sensor performance. Moreover, the biosensor exhibited appreciable stability, repeatability, reproducibility and practicality. The other advantages of the fabricated biosensor are simple and green fabrication approach, roughed and stable electrode surface, fast in sensing and highly reproducible.

Synthesis of zinc oxide nanoparticles on graphene-carbon nanotube hybrid for glucose biosensor applications.

Synthesis of zinc oxide nanoparticles incorporated graphene-carbon nanotubes hybrid (GR-CNT-ZnO) through a simple, one-pot method is demonstrated. The as-synthesized GR-CNT-ZnO composite is applied to fabricate an enzyme based glucose biosensor. The GOx immobilized on GR-CNT-ZnO composite exhibits well-defined redox peaks with a peak potential separation (ΔEp) of about 26 mV with enhanced peak currents, indicating a fast electron transfer at the modified electrode surface. The cyclic voltammetry measurements revealed that the modified film has high electrocatalytic ability towards glucose detection in the presence of oxygen. The proposed sensor has a wide linear detection range from 10 µM to 6.5 mM of glucose with a limit of detection (LOD) of 4.5 (±0.08) µM. In addition, the sensor possessed appreciable repeatability, reproducibility and remarkable stability for the sensitive determination of glucose. The practicality of this sensor has been demonstrated in human serum samples, with results being in good agreement with those determined using a standard photometric method.

An intermolecular disulfide bond is required for thermostability and thermoactivity of ß-glycosidase from Thermococcus kodakarensis KOD1.

Scientists are interested in understanding the molecular origin of protein thermostability and thermoactivity for possible biotechnological applications. The enzymes from extremophilic organisms have been of particular interest in the last two decades. ß-glycosidase, Tkßgly is a hyperthermophilic enzyme from Thermococcus kodakarensis KOD1. Tkßgly contains two conserved cysteine residues, C88 and C376. The protein tertiary structure obtained through homology modeling suggests that the C88 residue is located on the surface whereas C376 is inside the protein. To study the role of these cysteine residues, we substituted C88 and C376 with serine residues through site-directed mutagenesis. The wild-type and C376S protein existed in dimeric form and C88S in monomeric form, in an SDS-PAGE gel under non-reducing conditions. Optimal temperature experiments revealed that the wild-type was active at 100 °C whereas the C88S mutant exhibited optimal activity at 70 °C. The half-life of the enzyme at 70 °C was drastically reduced from 266 h to less than 1 h. Although C88 was not present in the active site region, the kcat/Km of C88S was reduced by 2-fold. Based on the structural model and biochemical properties, we propose that C88 is crucial in maintaining the thermostability and thermoactivity of the Tkßgly enzyme.