Function of gamma delta (γδ) T cell in cancer with special emphasis on cervical cancer.

Cervical cancer is the second-most prevalent gynecologic cancer in India. It is typically detected in women between the ages of 35 and 44. Cervical cancer is mainly associated with the human papillomavirus (HPV). The report shows that 70 % of cervical cancer is caused by HPV 16 and 18. There are few therapeutic options and vaccines available for cervical cancer treatment and γδ T cell therapy is one of them. This therapy can kill various types of cancers, including cervical cancer. The major γδ T cell subset is the Vγ9Vδ2 T cell, mainly distributed in peripheral blood which recognize non-MHC peptide antigens and can eliminate MHC-downregulated cancer. Moreover, γδ T cells can express different types of receptors that bind to the molecules of stressed cells, often produced on cancerous cells but absent from healthy tissue. γδ T cells possess both direct and indirect cytotoxic capabilities against malignancies and show potential antitumoral responses. However, γδ T cells also encourage the progression of cancer. Cancer immunotherapy using γδ T cells will be a potential cancer treatment, as well as cervical cancer. This review focused on the γδ T cell and its function in cancer, with special emphasis on cervical cancer. It also focused on the ligand recognition site of γδ T cells, galectin-mediated therapy and pamidronate-treated therapy for cervical cancer. Instead of the great potential of γδ T cell for the eradication of cervical cancer, no comprehensive in-depth review is available to date, so there is a need to jot down the various roles and modes of action and different applications of γδ T cells for cancer research, which we believe will be a handy tool for the researchers and the readers.

Bio-nanocomposite based highly sensitive and label-free electrochemical immunosensor for endometriosis diagnostics application.

In this research, for the first time, a bio-nanocomposites based highly sensitive and label-free electrochemical immunosensor is reported with the aim of endometriosis diagnostics application. Multiwalled carbon nanotube and magnetite nanoparticle (MWCNT-Fe3O4) was dispersed in chitosan (CS) to fabricate a bio-nanocomposite to immobilize very monoclonal specific antibody (via cross-linking using glutaraldehyde) for selective electrochemical immuno-sensing of carbohydrate antigen 19-9 (CA19-9), a potential biomarker for endometriosis diagnostics. Well-characterized Anti-AbsCA19-9/CS-MWCNT-Fe3O4 immune-electrode fabricated on glassy carbon electrode (GCE) successfully detect CA 19-9 and exhibited a high sensitivity as (2.55 µA pg-1 cm-1), a detection limit of 0.163 pg mL-1, detection range from 1.0 pg mL-1 to 100 ng mL-1. Our fabricated electrochemical AbsCA19-9/CS-MWCNT-Fe3O4 immunosensor performed CA19-9 sensing in physiological range and at a very level which suggest it application for early-stage diagnostics, diseases monitoring, and optimization of therapy. To claim the clinical application, our sensor was tested using real samples and sensing performance was validated using enzyme-linked immune-sorbent assay (ELISA). The results of the studies projected AbsCA19-9/CS-MWCNT-Fe3O4 electrochemical CA19-9 immunosensor as a potential and affordable alternate of conventional techniques like ELISA. We believe that our fabricated sensor can be the plane of a disease's management program due to affordable, rapid, label-free, and sensitive detection of a targeted biomarker.

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.

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.

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.

Regulating, Measuring, and Modeling the Viscoelasticity of Bacterial Biofilms

Biofilms occur in a broad range of environments under heterogeneous physicochemical conditions, such as in bioremediation plants, on surfaces of biomedical implants, and in the lungs of cystic fibrosis patients. In these scenarios, biofilms are subjected to shear forces, but the mechanical integrity of these aggregates often prevents their disruption or dispersal. Biofilms' physical robustness is the result of the multiple biopolymers secreted by constituent microbial cells which are also responsible for numerous biological functions. A better understanding of the role of these biopolymers and their response to dynamic forces is therefore crucial for understanding the interplay between biofilm structure and function. In this paper, we review experimental techniques in rheology, which help quantify the viscoelasticity of biofilms, and modeling approaches from soft matter physics that can assist our understanding of the rheological properties. We describe how these methods could be combined with synthetic biology approaches to control and investigate the effects of secreted polymers on the physical properties of biofilms. We argue that without an integrated approach of the three disciplines, the links between genetics, composition, and interaction of matrix biopolymers and the viscoelastic properties of biofilms will be much harder to uncover.

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.

Upper control limit of reactive oxygen species in follicular fluid beyond which viable embryo formation is not favorable.

Though the role of reactive oxygen species (ROS) in female infertility has been a subject of rigorous research worldwide, there is inadequate information on the cut-off value of ROS in the oocyte microenvironment beyond which ART outcome may be adversely affected. An upper ROS level in follicular fluid (FF) samples of women undergoing IVF beyond which good quality embryo formation is unlikely, is established. ROS, lipid peroxidation and total antioxidant capacity were estimated. The upper cut-off ROS level beyond which viable embryo formation is not favorable was found to be approximately 107 cps/400 microl FF. This level, determined in women with tubal factor infertility, was further validated in women with endometriosis and PCOS and correlated with fertilization and pregnancy rate and embryo quality. Summarizing, a threshold level in FF has been established for the first time beyond which ROS may be considered toxic for viable embryo formation and pregnancy outcome.