Use of Graphene and Its Derivatives for the Detection of Dengue Virus.

Every year, the dengue virus and its principal mosquito vector, Aedes sp., have caused massive outbreaks, primarily in equatorial countries. The pre-existing techniques available for dengue detection are expensive and require trained personnel. Graphene and its derivatives have remarkable properties of electrical and thermal conductivity, and are flexible, light, and biocompatible, making them ideal platforms for biosensor development. The incorporation of these materials, along with appropriate nanomaterials, improves the quality of detection methods. Graphene can help overcome the difficulties associated with conventional techniques. In this review, we have given comprehensive details on current graphene-based diagnostics for dengue virus detection. We have also discussed state-of-the-art biosensing technologies and evaluated the advantages and disadvantages of the same.

Detection of a novel glycodelin biomarker using electrochemical immunosensor for endometriosis.

Endometriosis is one of the important issues in women worldwide, which decreases the quality of women's lives in their reproductive age. The diagnosis of endometriosis is carried out by the invasive procedure, which is expensive and painful. In the last few decades, researchers have given more attention to constructing a suitable biomarker-based biosensor for semi/non-invasive diagnosis of endometriosis. As a result, glycodelin (GLY) was found as a promising biomarker because of its selectivity and sensitivity. To the best of our knowledge, it was the first study that reported the detection of GLY biomarker using an electrochemical immunosensor. Briefly, a label-free electrochemical immunosensing platform was constructed through in-situ surface modification of cysteamine layer and immobilisation of antibody (anti-GLY) with help of glutaraldehyde. The interaction between antigen and antibody was measured using square wave voltammetry (SWV). The SWV signal could decrease proportionally with the increasing GLY concentration ranging from 1 to 1000 ng mL-1 (R2 = 0.9981) and a detection limit (LOD) of 0.43 ng mL-1. Moreover, an immunosensor could exhibit high sensitivity, selectivity, long-term stability, reproducibility and regeneration. Accuracy of the immunosensor was compared with enzyme-linked immunosorbent assay (ELISA), and satisfying results were obtained. The detection of GLY biomarker may be a new possibility for endometriosis diagnosis.

Polyaniline Based Electrochemical Sensor for the Detection of Dengue Virus Infection.

BACKGROUND: Dengue burden is increasing day-by-day globally. A rapid, sensitive, cost-effective early diagnosis kit is the need of the hour. In this study, a label-free electrochemical immunosensor was proposed for dengue virus detection. A modified Polyaniline (PANI) coated Glassy Carbon (GC) electrode, immobilized with DENV NS1 antibody was used to detect the circulating DENV NS1 antigen in both spiked and infected sample. METHODS: Cloning, purification and expression of DENV NS1 protein in Escherichia coli (E. coli) was performed and sensor design, PANI modification on GC electrode surface by electrochemical polymerization and immobilization of NS1 antibody on the modified electrode surface was done and finally the analytical performance of the electrochemical immunosensor was done using Cyclic Voltammetry (CV) and Electrochemical Impedance Spectroscopy (EIS). RESULTS: CV and EIS were used to study and quantitate the circulating DENV antigen. The calibration curve showed wide linearity, good sensitivity (Slope=13.8% IpR/ml.ng -1) and distribution of data with a correlation coefficient (R) of 0.997. A lower Limit of Detection (LOD) was found to be 0.33 ng.ml -1 which encourages the applicability of the sensor. CONCLUSION: Thus, a PANI based new electrochemical immunosensor has been developed which has the potential to be further modified for the development of cost effective, point of care dengue diagnostic kit.

Cytokines, Angiogenesis, and Extracellular Matrix Degradation are Augmented by Oxidative Stress in Endometriosis.

BACKGROUND: The effect of the interplay among inflammation, angiogenesis, extracellular matrix (ECM) degradation, and oxidative stress (OS) on the pathogenesis of endometriosis remains unclear. Previously, we demonstrated the role of OS in endometriosis. Here, we performed a comprehensive investigation of several molecules involved in inflammation, angiogenesis, and ECM degradation in women with endometriosis to study their interplay with OS. METHODS: Blood samples were collected from women with endometriosis (N=80), as well as from women with tubal factor infertility as controls (N=80). Interleukin (IL)-1ß, tumor necrosis factor-alpha, interferon-gamma, transforming growth factor-beta, IL-4, -10, -2, -6, -8, vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP)-2, -9, tissue inhibitor of metalloproteinases (TIMP)-1, -2, and cyclooxygenase (COX)-2 levels in serum samples were measured using an ELISA. Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) in peripheral blood mononuclear cells was measured using flow cytometry. RESULTS: Cytokines, VEGF, MMPs, and COX-2 were significantly higher and TIMPs were significantly lower in patients with endometriosis. Multivariate statistical analysis indicated that IL-10 was the most significant variable capable of discriminating endometriosis samples from controls. CONCLUSIONS: Deregulation of NF-κB activation by OS affects the expression of various cytokines in endometriosis. Elevated cytokine levels further up-regulate IL-10, which subsequently activates the MMPs, leading to excessive ECM degradation and angiogenesis. Moreover, IL-10 emerged as the most important molecule involved in the pathogenesis of endometriosis. Measurement of these molecules may help in better management of the patients with endometriosis.

Author Correction: Metabolomics reveals perturbations in endometrium and serum of minimal and mild endometriosis.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Biomedical Application of Doxorubicin Coated Hydroxyapatite-Poly(lactide-co-glycolide) Nanocomposite for Controlling Osteosarcoma Therapeutics.

Nano drug delivery systems are widely used in cancer treatment nowadays. It is used to accomplish a remarkable drug therapeutic index to increase the efficacy of nanocomposites against cancer cells without affecting the other cells. Ceramic nanoparticles are well-known to carry chemotherapeutic drugs to the infected sites. Interest in them is aroused by their potential for application as promising biomaterials, especially in various orthopaedic applications. In the current study, Hydroxyapatite (HAp) was prepared by a simple in situ precipitation method and coated with a potent anticancer drug doxorubicin (DOX) using poly(lactide-co-glycolide) (PLGA) polymer. The interfacial strength of the composite is enhanced by the use of polymer in the nanocomposite preparation. An interaction between HAp particle and PLGA matrix has been noticed, which leads to improve the physicochemical properties of the prepared composites. Such a novel nanocomposite is further physicochemically characterized using Fourier Transform Infrared Spectroscopy (FTIR), X-ray Diffraction (XRD), Transmission Electron Spectroscopy (TEM) and Particle Size Distribution (PSD). In addition, the biocompatibility and the anticancer activity of the nanocomposite were evaluated by a colorimetric assay (MTT assay). The synthesized DOX-HAp-PLGA nanocomposite shows a significant cytotoxicity towards osteosarcoma cells, which may be potentially used as an anticancer agent against osteosarcoma diseases.

Label-Free Electrochemical Immunosensor Based on One-Step Electrochemical Deposition of AuNP-RGO Nanocomposites for Detection of Endometriosis Marker CA 125.

Endometriosis is the third most prominent gynecological disorder. Cancer antigen 125 (CA 125) is the primary serum marker used for late-stage endometriosis diagnosis and management. Herein, we developed a label-free immunosensor for electrochemical detection of CA 125 for endometriosis blood serum samples. The sensor was fabricated by one-step electrochemical deposition of highly conductive gold nanoparticles (AuNPs) and reduced graphene oxide (RGO) nanocomposite, via one-step electrochemical deposition. This method involved in situ reduction of HAuCl3 and graphene oxide and increased electrocatalytic performance. Different analytical techniques confirmed the morphology and structure of the AuNP/RGO nanocomposite. In addition, the antibody (Ab) was immobilized on the modified electrode surface through the self-assembly monolayer. The square wave voltammetry method has been utilized to measure the interaction of Ab and antigen (Ag). The as-fabricated sensor demonstrates a dynamic linear range of 0.0001 → 300 U mL-1 and lower limit of detection is 0.000042 U mL-1 toward CA125 detection. The developed sensor provides acceptable stability, high selectivity, and reproducibility. The proposed immunosensor has been applied to the CA 125 detection in endometriosis patient blood samples, and the results confirm the reliability of the as-fabricated sensor that is further associated with the standard ELISA analysis. The AuNP/RGO-based sensor can be used as an excellent tool for future prospective clinical diagnostics applications.

Metabolomics reveals perturbations in endometrium and serum of minimal and mild endometriosis.

Endometriosis is a common benign gynecological disease, characterized by growth and proliferation of endometrial glands and stroma outside the uterus. With studies showing metabolic changes in various biofluids of endometriosis women, we have set upon to investigate whether endometrial tissue show differences in their metabolic profiles. 1H NMR analysis was performed on eutopic endometrial tissue of women with endometriosis and controls. Analysis was performed on spectral data and on relative concentrations of metabolites obtained from spectra using multivariate and univariate data analysis. Analysis shows that various energy, ketogenic and glucogenic metabolites have significant altered concentrations in various stages of endometriosis. In addition, altered tissue metabolites in minimal and mild stages of endometriosis were explored in serum of these patients to assess their role in disease diagnosis. For Stage I diagnosis alanine was found to have 90% sensitivity (true positives) and 58% specificity (true negatives). For Stage II diagnosis alanine, leucine, lysine, proline and phenylalanine showed significant altered levels in serum. While sensitivity of these serum metabolites varied between 69.2-100% the specificity values ranged between 58.3-91.7%. Further, a regression model generated with this panel of serum markers showed an improved sensitivity and specificity of 100% and 83%, respectively for Stage II diagnosis.

Preparation of albumin based nanoparticles for delivery of fisetin and evaluation of its cytotoxic activity.

Fisetin is a well known flavonoid that shows several properties such as antioxidant, antiviral and anticancer activities. Its use in the pharmaceutical field is limited due to its poor aqueous solubility which results in poor bioavailability and poor permeability. The aim of our present study is to prepare fisetin loaded human serum albumin nanoparticles to improve its bioavailability. The nanoparticles were prepared by a desolvation method and characterized by spectroscopic and microscopic techniques. The particles were smooth and spherical in nature with an average size of 220 ± 8 nm. The encapsulation efficiency was found to be 84%. The in vitro release profile showed a biphasic pattern and the release rate increases with increase in ionic strength of solution. We have also confirmed the antioxidant activity of the prepared nanoparticles by a DPPH (2,2-diphenyl-1-picrylhydrazyl) assay. Further its anticancer activity was evaluated using MCF-7 breast cancer cell lines. Our findings suggest that fisetin loaded HSA nanoparticles could be used to transfer fisetin to target areas under specific conditions and thus may find use as a delivery vehicle for the flavonoid.

HOXA-11 mediated dysregulation of matrix remodeling during implantation window in women with endometriosis.

PURPOSE: To investigate the Homeobox genes HOXA-10 and HOXA-11 mediated endometrial molecular defects during implantation window in endometriosis-associated infertility cases. METHODS: Endometrial biopsies were obtained during implantation window from 31 infertile women with endometriosis (age < 35 years) and 26 age and BMI-matched infertile women without endometriosis were included in the study for comparison purposes. Endometrial expression of HOXA-10 and HOXA-11 genes, MMP-2, -9, α(v)ß(3) integrin, leukemia inhibitory factor and surface characteristics including average roughness and topology were assessed. RESULTS: A significantly lower expression of HOXA-10 and HOXA-11 were observed in endometriotic women compared to non-endometriotic controls. Further, a significantly higher endometrial expression of MMP-2 and -9 were observed in women with endometriosis when compared with controls. Interestingly, endometrial surface were observed to be grossly affected in terms of average roughness and topology in women with endometriosis compared to controls. CONCLUSIONS: The findings suggest that aberrant expression of HOXA-10 and -11 genes adversely affects endometrial remodelling and expression of receptivity markers.

Proinflammatory cytokines induced altered expression of cyclooxygenase-2 gene results in unreceptive endometrium in women with idiopathic recurrent spontaneous miscarriage.

OBJECTIVE: To investigate the expression pattern of proinflammatory, anti-inflammatory, and angiogenic cytokines and their effect on various mediators of endometrial receptivity in women with idiopathic recurrent spontaneous miscarriage (IRSM). DESIGN: A prospective study. SETTING: Tertiary care hospital and reproductive health research unit. PATIENT(S): Thirty-six women with IRSM (<35 years) and 30 fertile women as controls matched by age and body mass index undergoing sterilization. INTERVENTION(S): Endometrial biopsies in all women corresponding to the window of implantation. MAIN OUTCOME MEASURE(S): Assessment of endometrial expression of proinflammatory, anti-inflammatory, and angiogenic cytokines, mediators of matrix turnover and angiogenesis, markers of receptivity. RESULT(S): A statistical significantly higher level of proinflammatory cytokines, mediators of matrix turnover and angiogenesis, and a reduced expression of anti-inflammatory and angiogenic cytokines were observed in women with IRSM. Additionally, the markers of endometrial receptivity were poorly expressed in women with IRSM. CONCLUSION(S): Aberrant expression of proinflammatory, anti-inflammatory, and angiogenic cytokines during implantation window in women with IRSM is one of the key factors that adversely affect endometrial development, as evidenced by the inadequate expression of various endometrial receptivity markers.

Studies on the phenolic profiling, anti-oxidant and cytotoxic activity of Indian honey: in vitro evaluation.

Commercial honey types were screened for phenolic profile and anti-oxidant capacity. Phenolic profiling was done using high performance liquid chromatography, which was further corroborated with electro spray ionisation-mass spectroscopy. Dihydroxy benzoic acid, caffeic acid, ferulic acid and cinnamic acid were the major phenolic constituents found in the honey samples. The anti-oxidant content and free-radical scavenging effect of honey were established using various assays. Total anti-oxidant potential and free-radical scavenging ability varied among the honey varieties and exhibited significant correlation with their phenolic content. Further, honey samples with richly abundant phenolic content were found to limit oxidant-induced cell death more effectively. Cytotoxic studies of a selected sample on a breast cancer cell displayed growth inhibition, depending on the concentration used. Cell cycle analysis indicated increasing accumulation of cells at the sub-G(1) phase. These results summarise the phenolic profile and anti-oxidant and cytotoxic potential of Indian honey samples for the first time.

Cut-off value of reactive oxygen species for predicting semen quality and fertilization outcome.

Though the role of reactive oxygen species (ROS) in male infertility is widely investigated worldwide, there is a lack of adequate information on the cut-off value of ROS beyond which ART outcome may be adversely affected. The objective of the present study is to establish an upper level of ROS in semen samples which can be considered as a potential marker of good semen quality. Semen ejaculates were randomly collected from 338 male partners of infertile couples. The upper critical limit (UCL) of ROS was calculated from the control chart of normozoospermic patients and found to be 0.75x10(6) counted photons per minute (cpm)/10 million cells. This was further validated by estimating ROS in 17 proven fertile men and 38 patients undergoing intra cytoplasmic sperm injection (ICSI). As expected, all abnormal semen samples exhibited higher ROS as compared to nomozoospermic and proven fertile samples. All semen samples from proven fertile volunteers were found to be <0.075x10(6) cpm/10 million cells. On the basis of the calculated UCL, ICSI patients were divided into two groups: Group I (UCL). The semen parameters, fertilization rate and pregnancy outcome were found to be significantly affected in Group II. Significant difference in live birth-rates was observed between Group I (47.6%) and Group II (17.6%) while no live-birth was recorded for ROS level >0.1x10(6) cpm/10 million cells. It is concluded that the upper cut-off value of normal semen samples that correlates with good semen quality is, therefore, in the order of 0.075-0.1x10(6) cpm/10 million cells. In addition to the WHO [1999] semen analysis, this range is expected to assist andrologists and clinicians in predicting semen quality and fertilization outcome in patients with male factor infertility undergoing ICSI.