Cardiovascular biomarker troponin I biosensor: Aptamer-gold-antibody hybrid on a metal oxide surface.

Myocardial infarction (MI) is highly related to cardiac arrest leading to death and organ damage. Radiological techniques and electrocardiography have been used as preliminary tests to diagnose MI; however, these techniques are not sensitive enough for early-stage detection. A blood biomarker-based diagnosis is an immediate solution, and due to the high correlation of troponin with MI, it has been considered to be a gold-standard biomarker. In the present research, the cardiac biomarker troponin I (cTnI) was detected on an interdigitated electrode sensor with various surface interfaces. To detect cTnI, a capture aptamer-conjugated gold nanoparticle probe and detection antibody probe were utilized and compared through an alternating sandwich pattern. The surface metal oxide morphology of the developed sensor was proven by microscopic assessments. The limit of detection with the aptamer-gold-cTnI-antibody sandwich pattern was 100 aM, while it was 1 fM with antibody-gold-cTnI-aptamer, representing 10-fold differences. Further, the high performance of the sensor was confirmed by selective cTnI determination in serum, exhibiting superior nonfouling. These methods of determination provide options for generating novel assays for diagnosing MI.

Room Temperature Ionic Liquids-Based Electrochemical Sensors: An Overview on Paracetamol Detection.

Paracetamol (PAR) is an effective antipyretic and analgesic drug utilized worldwide, safer at therapeutic levels but over-dosing and the chronic usage of PAR results in accumulation of toxic metabolites, which leads to kidney and liver damages. Hence, a simple, rapid, cost-effective, and sensitive analytical technique is needed for the accurate determination of PAR in pharmaceutical and biological samples. Though numerous techniques have been reported for PAR detection, electrochemical methods are being receiving more interest due to their advantages. Moreover, in the past few decades, room temperature ionic liquids (RTILs) have been utilized in electrochemical sensors due to their attractive properties. In this present review, authors gathered research findings available for the determination of PAR using RTIL-based electrochemical sensors and discussed. The advantages and limitations in these systems as well as the future research directions are summarized.

Continuous electrical lysis of cancer cells in a microfluidic device with passivated interdigitated electrodes.

Cell lysis is a critical step in genomics for the extraction of cellular components of downstream assays. Electrical lysis (EL) offers key advantages in terms of speed and non-interference. Here, we report a simple, chemical-free, and automated technique based on a microfluidic device with passivated interdigitated electrodes with DC fields for continuous EL of cancer cells. We show that the critical problems in EL, bubble formation and electrode erosion that occur at high electric fields, can be circumvented by passivating the electrodes with a thin layer (∼18 µm) of polydimethylsiloxane. We present a numerical model for the prediction of the transmembrane potential (TMP) at different coating thicknesses and voltages to verify the critical TMP criterion for EL. Our simulations showed that the passivation layer results in a uniform electric field in the electrode region and offers a TMP in the range of 5-7 V at an applied voltage of 800 V, which is well above the critical TMP (∼1 V) required for EL. Experiments revealed that lysis efficiency increases with an increase in the electric field (E) and residence time (tr): a minimum E ∼ 105 V/m and tr ∼ 1.0 s are required for efficient lysis. EL of cancer cells is demonstrated and characterized using immunochemical staining and compared with chemical lysis. The lysis efficiency is found to be ∼98% at E = 4 × 105 V/m and tr = 0.72 s. The efficient recovery of genomic DNA via EL is demonstrated using agarose gel electrophoresis, proving the suitability of our method for integration with downstream on-chip assays.

Controlled drug release for inflammatory bowel disease (IBD).

Controlled drug release in formulation is an important area of research. Formulations using crospovidone as super-disintegrants to achieve immediate release once it reaches the ileo- cecal region is relevant. The Eudragit L30D pH dependent polymer that allows drug release after a lag time of 4-5 hrs to achieve desired drug release from the drug delivery system is critical. Hence, pre-formulation to study drug-polymer interaction is essential. The linear correlation between the predicted and actual values for all the batches of optimization is shown with high correlation coefficient (r-value). Therefore, the designed formulation is promising for ileo-cecal targeted pulsatile drug delivery system in the management of Crohn's disease.

Graphitic carbon nitride/graphene nanoflakes hybrid system for electrochemical sensing of DNA bases in meat samples.

This research presents a simple, fast and simultaneous electrochemical quantitative determination of nucleobases, for example guanine (G), adenine (A), and thymine (T) in a beef and chicken livers samples to measure the quality of food products based on hybrids of graphitic carbon nitride/Graphene nanoflakes (g-C3N4/GNF) modified electrode. Graphitic carbon nitride (g-C3N4) made of graphite-like covalent link connects nitrogen, nitride, and carbon atoms in the structural design with improved the electrical properties and low band gap semiconductor. The g-C3N4/GNF nanocomposite was synthesized by the hydrothermal treatment to form a porous g-C3N4 interconnected three dimensional (3D) network of g-C3N4 and GNF. The 3D g-C3N4/GNF/GCE was utilized for the detection of nucleic acid bases with a well resolved oxidation peak for the individual analyte. The electrocatalytic current was established to be a linear range from 0.3 × 10-7 to 6.6 × 10-6, 0.3 × 10-7 to 7.3 × 10-6, and 5.3 × 10-6 to 63.3 × 10-4 M for G, A, and T with a detection limit of 4.7, 3.5 and 55 nM, respectively. The diffusion co-efficient and the kinetic parameters were derived from the chronoamperometry technique. The proposed sensing strategy has been effectively used for the application in real sample analysis and observed that the electrode free from the surface fouling.

D-glucosamine chitosan base molecule-assisted synthesis of different shape and sized silver nanoparticles by a single pot method: A greener approach for sensor and microbial applications.

Silver nanoparticle was synthesized using D-glucosamine chitosan base as green reducing agent at elevated temperature in alkaline pH ranges. The excess of D-glucosamine chitosan base was used as it is both stabilizing and reducing agent at different pHs, regulates the shape and size of the silver nanoparticles. The progressive growth of silver nanoparticles was monitored by UV-Visible spectral studies. A sharp peak at 420 nm indicates the formation of spherical silver nanoparticles. The size and shape of silver nanoparticles were observed from Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM) methods. The anisotropically grown nanoparticles were used as probe for Surface Enhanced Raman Studies (SERS) using ATP (4-aminothiophenol) as a model system. The catalytic behavior of silver nanoparticles was exploited for 4-nitrophenol reduction and observed that the reduction reaction follows pseudo first order kinetics with a rate constant 0.65 min. The antibacterial activity of silver nanoparticles was also tested for both gram-positive and -negative microorganisms, in which higher zone of inhibition was observed for gram negative microorganism.

How to Design Donor-Acceptor Based Heterocyclic Conjugated Polymers for Applications from Organic Electronics to Sensors.

Over the past few years, significant progress has been made in the design of organic semi-conducting conjugated polymers that readily transport holes or electrons and can result in light emission. The conjugated backbone consist mainly of electron-donating (donor) and electron-withdrawing (acceptor) units as alternating groups in a conjugated oligomer or polymer that can be regulated by physical properties such as π conjugation length, monomer alteration, inter/intramolecular interactions and energy levels. Certainly, it is notable today that the highest occupied molecular orbital level of the producing material is localized predominantly on the electron-donating moiety and lowest unoccupied molecular orbital level on the electron-accepting moiety. Conjugated oligomers or polymers are used in many detecting fields due to their exceptional ability to sense toxic chemicals, metal ions and biomolecules. The conjugated polymers have unique delocalized π-electronic "molecular wires" that can expand the fluorescence intensity considerably. The fluorescence intensity of polymers can be quenched by particular quenching molecules. In this review, the fluorescence intensity, detecting of multiple metal ions, solubility, photochemical stability and optoelectronic properties of these conjugated polymers, and how they can be regulated by different functional groups, are discussed in detail.

Non-inertial lift induced migration for label-free sorting of cells in a co-flowing aqueous two-phase system.

Label free sorting of circulating tumor cells (CTCs) often remains a challenge due to their rarity in peripheral blood and identical morphology to white blood cells. We present a novel label-free passive microfluidic technique for isolation of cancer cells (EpCAM+ and CD45-) from peripheral blood mononuclear cells (PBMCs) (CD45+ and EpCAM-) in aqueous two-phase system (ATPS). Our technique involves non-inertial lift induced lateral cell migration across liquid-liquid interface that is employed for sorting cells of different size and stiffness. The interplay between lift force and interfacial tension (IFT) force governs cell migration phenomena. We estimate the order of magnitude of the lift force and find it to be higher than the IFT for cancer cells above a critical strain rate parameter ([small gamma, Greek, dot above]/h). The effect of spreading parameter and viscoelastic force was found to have negligible effect on lateral migration of cells. We demonstrated isolation of two different types of cancer cells, namely, MCF-7 and MDA-MB-231 from PBMCs and quantify our sorting results by tagging the cells with EpCAM and CD45 and using fluorescence imaging. With 102-104 cancer cells in 105-107 PBMCs, we achieved a processing rate of >25 000 cells per min at a sorting efficiency of ∼99%. Moreover, we demonstrated that cancer cells isolated from PBMCs using the proposed technique remain viable and can be cultured for downstream analysis.

High surface graphene nanoflakes as sensitive sensing platform for simultaneous electrochemical detection of metronidazole and chloramphenicol.

We demonstrate a mediator-free electrochemical sensitive sensing for detection of metronidazole (MNZ) and chloramphenicol (CAP) in real samples. A few-layered defect-free graphene nanoflakes (GNFs) were synthesized by surfactant-assisted exfoliation method, which was characterized by several methods like XRD, Raman spectroscopy, FE-SEM and TEM images. Cyclic voltammetry and electrochemical impedance spectroscopy were studied to understand the electron transfer behavior of GNFs modified glassy carbon electrode (GCE). Electrochemical behavior of MNZ and CAP was investigated at GNFs/GCE in PBS and observed peak potentials at -0.32 V and - 0.51 V vs. Ag/AgCl. A wider reduction potential window of MNZ and CAP were observed by differential pulse voltammetry which was found to be 280 mV at GNFs/GCE. Amperometry method was employed for detection of the unknown concentration of analytes under hydrodynamic condition. A linear calibration plot was obtained by plotting the concentrations of MNZ and CAP in ranging from 0.5 to 5.5 nM with R2 of 0.9983 and 0.9980, respectively. The detection limit of MNZ and CAP were found to be 0.15 nM and 0.38 nM, this system have shown good stability, excellent reproducibility and free interference from GNFs/GCE. The present method was successfully applied for the determination of MNZ and CAP in drugs and urine samples.

Angular photogrammetric analysis of the soft-tissue facial profile of Indian adults.

INTRODUCTION:: Soft-tissue analysis has become an important component of orthodontic diagnosis and treatment planning. Photographic evaluation of an orthodontic patient is a very close representation of the appearance of the person. The previously established norms for soft-tissue analysis will vary for different ethnic groups. Thus, there is a need to develop soft-tissue facial profile norms pertaining to Indian ethnic groups. AIM AND OBJECTIVES:: The aim of this study is to establish the angular photogrammetric standards of soft-tissue facial profile for Indian males and females and also to compare sexual dimorphism present between them. MATERIALS AND METHODS: The lateral profile photographs of 300 random participants (150 males and 150 females) between ages 18 and 25 years were taken and analyzed using FACAD tracing software. Inclusion criteria were angles Class I molar occlusion with acceptable crowding and proclination, normal growth and development with well-aligned dental arches, and full complements of permanent teeth irrespective of third molar status. This study was conducted in Indian population, and samples were taken from various cities across India. Descriptive statistical analysis was carried out, and sexual dimorphism was evaluated by Student's t-test between males and females. RESULTS: The results of the present study showed statistically significant (P < 0.05) gender difference in 5 parameters out of 12 parameters in Indian population. CONCLUSION: In the present study, soft-tissue facial measurements were established by means of photogrammetric analysis to facilitate orthodontists to carry out more quantitative evaluation and make disciplined decisions. The mean values obtained can be used for comparison with records of participants with the same characteristics by following this photogrammetric technique.

Solid-State Synthesis of POPD@AgNPs Nanocomposites for Electrochemical Sensors.

In the present work, Poly(o-phenylenediamine) (POPD) stabilized silver nanoparticles (POPD@AgNPs) nanocomposites was synthesized by solid state oxidative polymerization method using o-phenylenediamine dihydrochloride (oPD-HCl) as monomer and silver nitrate (AgNO3) used as metal precursor as well as oxidizing agent no other external oxidizing agent was used. POPD@AgNPs nanocomposites were characterized by various instrumental techniques to confirm their size, shape and its composition. The electrocatalytic activity of POPD and POPD@AgNPs modified electrode was investigated over the oxidation of hydrazine (N2H4) and reduction of hydrogen peroxide (H2O2) using Cyclic Voltammetry (CV), Differential Pulse Voltammetry (DPV) and Chronoamperometry techniques. POPD and POPD@AgNPs were characterized using HR-TEM, FE-SEM, XRD, UV-Visible, FT-IR, Micro Raman spectroscopy and those results were confirmed their chemical purity, particle size, shape and its elemental compositions. Moreover, the DPV and chronoamperometry reveals that POPD@AgNPs is a good sensor for the electrochemical gas detection of N2H4 and H2O2 because it has good stability, easy-operation, excellent reproducibility, high sensitivity and good limit of detection when compared to with pure POPD. This system shows good stability, excellent sensitivity, response and the detection limit was obtained for the detection of N2H4 and H2O2 in trace level gases, which was lower than some of the modified electrodes.

Sunlight assisted synthesis of silver nanoparticles in zeolite matrix and study of its application on electrochemical detection of dopamine and uric acid in urine samples.

Sunlight assisted reduction of silver ions were accomplished for the synthesis of silver nanoparticles incorporated within the mesoporous silicate framework of zeolite Y. The zeolite-Y and AgNP/Zeo-Y were characterized by field emission scanning electron microscopy, transmission electron microscopy, N2 adsorption-desorption BET isotherm and X-ray diffraction techniques. The incorporation of silver nanoparticles within the porous framework was further confirmed by cyclic voltammetry and electrochemical impedance spectroscopy. An enhanced electrocatalytic oxidation of biologically important molecules like dopamine and uric acid using AgNP/Zeo-Y modified glassy carbon electrode has been developed. A simultaneous oxidation of DA and UA peaks were obtained at +0.31V and +0.43V (vs. Ag/AgCl) using AgNP/Zeo-Y/GCE under the optimum experimental condition. A well-resolved peak potential window (~120mV) for the oxidation of both DA and UA were observed at AgNP/Zeo-Y/GCE system. The calibration curves for DA and UA were obtained within the dynamic linear range of 0.02×10(-6) to 0.18×10(-6)M (R(2)=0.9899) and 0.05×10(-6) to 0.7×10(-6)M (R(2)=0.9996) and the detection limits were found to be 1.6×10(-8)M and 2.51×10(-8)M by using differential pulse voltammetry (DPV) method. The proposed method was successfully applied for the determination of both DA and UA in human urine samples with a related standard deviation was <3%, and n=5 using the standard addition method.

Enhanced amperometric detection of metronidazole in drug formulations and urine samples based on chitosan protected tetrasulfonated copper phthalocyanine thin-film modified glassy carbon electrode.

An enhanced electrocatalytic reduction of metronidazole antibiotic drug molecule using chitosan protected tetrasulfonated copper phthalocyanine (Chit/CuTsPc) thin-film modified glassy carbon electrode (GCE) has been developed. An irreversible reduction occurs at -0.47V (vs. Ag/AgCl) using Chit/CuTsPc modified GCE. A maximum peak current value is obtained at pH1 and the electrochemical reduction reaction is a diffusion controlled one. The detection limit is found to be 0.41nM from differential pulse voltammetry (DPV) method. This present investigation method is adopted for electrochemical detection of metronidazole in drug formulation and urine samples by using DPV method.

Amperometric detection of Sudan I in red chili powder samples using Ag nanoparticles decorated graphene oxide modified glassy carbon electrode.

A simple and sensitive electrochemical method was developed to determine the concentration of Sudan I in chili powder based on silver nanoparticles decorated graphene oxide modified glassy carbon electrode (AgNPs@GO/GCE). The voltammetry behaviour of Sudan I on modified GCE was investigated in phosphate buffer medium (PBS) with various pH ranges and the electron transfer properties were studied. It is found that the AgNPs@GO/GCE can catalyse the reduction of azo group, -N=N- followed by electrochemical oxidation of (-)OH group present in Sudan I dye molecule. Quantitative detection of Sudan I present in food products was carried out by amperometry method in which reduction potential was fixed at -0.77 V vs. Ag/AgCl. The amperometry method showed an excellent performance with a sensitivity of 6.83 µA mM(-1) and a detection limit of 11.4 × 10(-7)ML(-1). A linear calibration graph was constructed in the ranging 3.90 × 10(-6) to 3.19 × 10(-5)ML(-1). The method was successfully applied for the determination of Sudan I in red chili powder samples.

Ellagic acid encapsulated chitosan nanoparticles for drug delivery system in human oral cancer cell line (KB).

Ellagic acid (EA), a naturally occurring polyphenolic compound is well documented for its anticancer property in numerous pre-clinical models. The properties like poor water solubility and limited oral bio-availability of ellagic acid has hampered its clinical applications. The present study, reports the preparation of ellagic acid encapsulated chitosan nanoparticles (EA@CS-NP) by ionic gelation method as an effective drug delivery for oral cancer treatment. The synthesized ellagic acid nanoparticle is spherical shaped with an average particle size of 176nm. The drug-encapsulation and loading-efficiency of the nanoparticles were 94±1.03% and 33±2.15% respectively. The in vitro drug release profile in the PBS medium shows sustained release of EA from EA@CS-NP. Further, this study evaluates the therapeutic efficacy of EA@CS-NP in human oral cancer cell line (KB) using MTT and DNA fragmentation analysis. EA@CS-NP exhibit significant cytotoxicity in KB cells in a dose-dependent manner with a very low IC50 value compared to the free EA. The results of the present study strengthen our hypothesis and hope that this novel formulation could possibly overcome the current limitations of ellagic acid and can open a new avenue for oral cancer therapy.

Effect of early cyclosporine levels on kidney allograft rejection.

Acute rejection is the greatest risk factor for development of biopsy-proven chronic rejection and late kidney allograft loss. We previously noted that low cyclosporine (CsA) levels were a risk factor for early acute rejection in pediatric recipients (1). In our current study, we used logistic regression to identify risk factors for acute rejection in 726 adult kidney transplant recipients on triple therapy (prednisone, azathioprine, CsA). Variables considered for logistic regression analysis were donor organ source (cadaver vs. living), degree of HLA mismatch (1 to 6 vs. 0 antigen mismatch), transplant number (primary vs. retransplant), CsA levels (< 125 vs. > or = 125 ng/ml, < 150 ng/ml vs. > or = 150 ng/ml, and < 175 vs. > or = 175 ng/ml), and acute rejection episodes (0 vs. > or = 1). Of 726 recipients, 401 (55%) received cadaver kidneys; 325 (45%), living related. Overall, 572 (79%) had a primary transplant; 154 (21%), a retransplant. The vast majority of acute rejection episodes occurred within the first 2 months posttransplant; 68% of recipients had no acute rejection episodes by 2 months and 58% had none by 60 months posttransplant. Logistic regression analysis revealed that a cadaver donor kidney (vs. living) (p = 0.004), a 1 to 6 antigen mismatch (vs. 0 mismatch) (p = 0.001), and CsA levels < 150 ng/ml (vs. > or = 150 ng/ml) correlated with biopsy-proven acute rejection. The correlation for CsA levels < 150 ng/ml (vs. > or = 150 ng/ml) held true for levels at 1 wk (p < 0.05), 1 month (p = 0.0001), 2 months (p = 0.01), and 3 months (p = 0.02) posttransplant. Similar correlation was found for CsA levels < 125 ng/ml (vs. > or = 125 ng/ml) and < 175 ng/ml (vs. > or = 175 ng/ml). Comparative analyses were made (by Chi-square) of acute and chronic rejection rates when recipients were divided into 3 groups by CsA level (< 125 ng/ml, > or = 125 to < 150 ng/ml, and > or 150 ng/ml). At each time point (1 wk, 2 wk, 1 month, 2 months, 3 months), CsA levels < 125 ng/ml (vs. > or = 125 to < 150 ng/ml and > or = 150 ng/ml) were associated with the greatest increased risk of acute rejection--for both cadaver and living related recipients (all p < 0.05). CsA levels < 125 ng/ml at each time point (1 wk, 2 wk, 1 month, 2 months, 3 months) were also associated with a significantly increased risk of chronic rejection (all p < 0.001). The incidence of both acute and chronic rejection was reduced in the group with CsA levels > or = 125 to < 150 ng/ml and further reduced in the > or = 150 ng/ml group. Our data indicate that maintaining CsA levels > or = 150 ng/ml as part of triple therapy reduces the incidence of both acute and chronic rejection. Because chronic rejection is the leading cause of late allograft loss, maintaining adequate CsA levels should improve long-term graft survival. Based on this analysis, we have modified our own immunosuppressive regimens to achieve higher CsA levels earlier posttransplant.