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ZnO nanoparticles (NPs) were prepared by green synthesis using plant leaf extraction of Ochradenus baccatus and characterized by XRD, FESEM, HRTEM, and Raman spectroscopy techniques. Since elevated CO levels have been associated with inflammatory conditions, cardiovascular diseases, and respiratory disorders and the methane gas primarily produced by gut microbiota and linked to gastrointestinal disorders and other abnormal methane levels in breath samples, the nanoparticles were applied for gas sensor fabrication. Thus, the gas sensors fabricated using ZnO nanoparticles were investigated for CH4, H2, CO, and NO2 gases. The gas sensing was performed for the fabricated sensors at various operating temperatures and gas concentrations. Interestingly, leaf-extracted green synthesized ZnO NPs were more sensitive to CH4, CO, and NO2 gases than to H2. The results of sensing studies revealed that the nanoparticles exhibit a selectivity toward gas depending on the gas type. The sensor response was also studied against the humidity. These findings bridge between the laboratory and industry sectors for future gas sensors development, which can be used for exhaled breath analysis and serve as potential diagnostic tools for highly sensitive contagious diseases. [ABSTRACT FROM AUTHOR]

In recent times, the global landscape of disease detection and monitoring has been profoundly influenced by the convergence of nanotechnology and biosensing techniques. Biosensors have enormous potential to monitor human health, with flexible or wearable variants, through monitoring of biomarkers in clinical and biological behaviors and applications related to health and disease, with increasing biorecognition, sensitivity, selectivity, and accuracy. The emergence of nanomaterial-based biosensors has ushered in a new era of rapid and sensitive diagnostic tools, offering unparalleled capabilities in the realm of disease identification. Even after the declaration of the end of the COVID-19 pandemic, the demand for efficient and accessible diagnostic methodologies has grown exponentially. In response, the integration of nanomaterial biosensors into breathalyzer devices has gained considerable attention as a promising avenue for low-cost, non-invasive, and early detection of COVID-19. This review delves into the forefront of scientific advancements, exploring the potential of emerging nanomaterial biosensors within breathalyzers to revolutionize the landscape of COVID-19 detection, providing a comprehensive overview of their principles, applications, and implications. [ABSTRACT FROM AUTHOR]

Background: Selective cancer cell recognition is the most challenging objective in the targeted delivery of anti-cancer agents. Extruded specific cancer cell membrane coated nanoparticles, exploiting the potential of homotypic binding along with certain protein-receptor interactions, have recently proven to be the method of choice for targeted delivery of anti-cancer drugs. Prediction of the selective targeting efficiency of the cancer cell membrane encapsulated nanoparticles (CCMEN) is the most critical aspect in selecting this strategy as a method of delivery. Materials and methods: A probabilistic model based on binding scores and differential expression levels of Glioblastoma cancer cells (GCC) membrane proteins (factors and receptors) was implemented on python 3.9.1. Conditional binding efficiency (CBE) was derived for each combination of protein involved in the interactions. Selective propensities and Odds ratios in favour of cancer cells interactions were determined for all the possible combination of surface proteins for 'k' degree of interaction. The model was experimentally validated by two types of Test cultures. Results: Several Glioblastoma cell surface antigens were identified from literature and databases. Those were screened based on the relevance, availability of expression levels and crystal structure in public databases. High priority eleven surface antigens were selected for probabilistic modelling. A new term, Breakeven point (BEP) was defined as a characteristic of the typical cancer cell membrane encapsulated delivery agents. The model predictions lie within ±7% of the experimentally observed values for both experimental test culture types. Conclusion: The implemented probabilistic model efficiently predicted the directional preference of the exposed nanoparticle coated with cancer cell membrane (in this case GCC membrane). This model, however, is developed and validated for glioblastoma, can be easily tailored for any type of cancer involving CCMEN as delivery agents for potential cancer immunotherapy. This probabilistic model would help in the development of future cancer immunotherapeutic with greater specificity. [ABSTRACT FROM AUTHOR]

COVID-19 infection is associated with elevated oxidative stress, systemic hyper-inflammatory responses, endothelial dysfunction, and red blood cell membrane deformability. Nigella sativa extract is widely used in alternative and complementary medicine systems in a large population, due to its highly therapeutic, economic, natural, and safe nature. The aim of this study was to evaluate the effect of N. sativa extract on oxidative stress, hemolysis, proteolysis, and glycation through in vitro studies, as well as to find out its anti-viral potential against severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) using in silico studies. N. sativa seed extract (at 600 µg/mL) displayed 67.33% scavenging activity in the 2,2-diphenyl-1-picrylhydrazyl (DPPH) test, and 70.28% hydrogen peroxide reducing activity. N. sativa exhibited anti-proteolytic activity by decreasing heat-induced denaturation of bovine serum albumin (BSA) and egg albumin by 63.14% and 57.95%, respectively, and exhibited anti-proteinase potential of 66.28% at 600 μg/mL. In addition, heat-induced hemolysis and hypersalinity-induced hemolysis were inhibited by 57.86% and 61.7%, respectively, by the N. sativa seeds. N. sativa also inhibited browning intensity by 56.38%, and percent aggregation index by 51.38%, amyloid structure by 48.28%, and AGE-specific fluorescence by 52.18%, thereby protecting the native structure of BSA from glycation. The binding interactions between bioactive molecules of N. sativa seed with SARS-CoV-2 spike glycoprotein were proven by using in silico molecular docking tools. The functional amino acids involved in the interactions are Asp467, Thr108, Thr114, Ile468, Asn234, Gln155, Glu465, Arg466, Gly232, and Ile233, indicating the inhibiting property of N. sativa on SARS-CoV-2. Finally, we may infer that phytoconstituents of N. sativa seeds have the potential to protect against the spike protein of SARS-CoV-2. Studies on N. sativa seeds might act as a path to develop a potent alternative therapy against viral infections, especially COVID-19 infection, in the future. However, the limitations linked with the use of natural products are also needed to be considered in this regard. [ABSTRACT FROM AUTHOR]

Lymphoma is a chronic inflammatory disease in which the immune system is highly affected. Increased oxidative stress is one of the common conditions of cancer and affects macromolecules. Histone modifications affect the chromatin structure and functions. In this study, histone H1 (His-H1) protein was modified by reactive oxygen species (ROS), and structural and chemical changes were studied. Hodgkin lymphoma (HL) and non-Hodgkin lymphoma (NHL) patients were selected, and oxidative stress markers, inflammatory cytokines, and serum autoantibodies were analyzed using biochemical and immunological assays. Furthermore, the formation of antigen-antibody immune complexes was assessed by the Langmuir plot. ROS-modified His-H1 (ROS-His-H1) showed substantial structural perturbation in protein (UV-hyperchromicity and increased intrinsic fluorescence) compared to the native His-H1 protein. A possible explanation for the changes is suggested by the exposure of the aromatic chromophore to the solvent. In-depth structural analysis by circular dichroism (CD) exhibited major changes in α-helix (−21.43%) and turns (+33%), reflecting changes in the secondary structure of histone H1 protein after ROS exposure. ELISA and competitive ELISA findings revealed high recognitions of serum autoantibodies to ROS-His-H1 from NHL, followed by HL subjects. Healthy controls showed negligible binding. Non-modified His-H1 did not show any binding with serum samples from either cohort. High apparent association constants (ACCs) were calculated for ROS-His-H1 using purified IgGs from NHL (1.46 × 10–6 M) compared to HL (1.33 × 10–6 M) patients. Non-modified His-H1 exhibited a hundred times less ACC for NHL (2.38 × 10–8 M) and HL (2.46 × 10–8 M) patients. Thus, ROS modifications of histone H1 cause structural changes and expose cryptic neo-epitopes on the protein against which autoantibodies were generated. These perturbations might affect the histone DNA interaction dynamics and potentially be correlated with gene dysregulation. These subtle molecular changes with an immune imbalance might further aggravate the disease. [ABSTRACT FROM AUTHOR]

Advanced stage cancers are aggressive and difficult to treat with mono-therapeutics, substantially decreasing patient survival rates. Hence, there is an urgent need to develop unique therapeutic approaches to treat cancer with superior potency and efficacy. This study investigates a new approach to develop a potent combinational therapy to treat advanced stage leukemia. Biologically active α-amino amide analogs (RS)-N-(2-(cyclohexylamino)-2-oxo-1-phenylethyl)-N-phenylpropiolamide (α-AAA-A) and (RS)-N-(2-(cyclohexylamino)-2-oxo-1-phenylethyl)-N-phenylbut2-enamide (α-AAA-B) were synthesized using linear Ugi multicomponent reaction. Cytotoxicities and IC50 values of α-AAA-A and α-AAA-B against leukemia cancer cell lines (HL-60 and K562) were analyzed though MTT assay. Cytotoxic assay analyzed percent killing of leukemia cell lines due to the effect of γδ T cells alone or in combination with α-AAA-A or α-AAA-B. Synthesized biologically active molecule α-AAA-A exhibited increased cytotoxicity of HL-60 (54%) and K562 (44%) compared with α-AAA-B (44% and 36% respectively). Similarly, α-AAA-A showed low IC50 values for HL-60 (1.61 ± 0.11 μM) and K562 (3.01 ± 0.14 μM) compared to α-AAA-B (3.12 ± 0.15 μM and 6.21 ± 0.17 μM respectively). Additive effect of amide analogs and γδ T cells showed significantly high leukemia cancer cell killing as compared to γδ T cells alone. A unique combinational therapy with γδ T cells and biologically active anti-cancer molecules (α-AAA-A/B), concomitantly may be a promising cancer therapy. [ABSTRACT FROM AUTHOR]

The last few months of 2019 witnessed the emergence, rise and rapid spread of a novel coronavirus known as severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), causing an acute respiratory disease called coronavirus disease 2019 or Covid-19. Severe pathological manifestations of the disease in the infected population with comorbidities are linked to acute respiratory distress syndrome (ARDS), associated with an exaggerated synthesis and expression of cytokines, leading to a systemic inflammatory response also known as a cytokine storm (CS). Elderly patients (> 60 years of age) showed more deaths in Covid-19 infection. Age-related immune imbalance increases patient susceptibility to CS. In acute Covid-19 infection, it is difficult to minimize or control the overproduction of cytokines; hence, limited medical treatments are effective. This review aims to provide an overview of the current knowledge of involvement of cytokines in SARS-CoV-2 infection, susceptibility factors for the accompanying cytokine storm in severe Covid-19 cases and possible treatment strategies. [ABSTRACT FROM AUTHOR]

Aim: Elevated levels of 16α‐hydroxyestrone (16α‐OHE1) have been described in endometrial cancer (EC) and estrogen receptors (ER) expressed in endometrial tissue, but research on their combined role is lacking. We aimed to investigate the affinity and binding specificity of EC antibodies against the 16α‐OHE1‐ERα aggregate in the serum of EC patients. Specificities of EC antibodies were also evaluated according to various clinical characteristics found in these cancer patients. Methods: The binding specificity and affinity of EC antibodies against 16α‐OHE1‐ERα in the serum of 120 EC patients were evaluated by direct binding and competition ELISA and quantitative precipitation titration. Binding of EC antibodies was also determined according to various clinical characteristics in EC patients through competition ELISA. Results: Antibodies from EC patients demonstrated high recognition of 16α‐OHE1‐ERα compared to ERα (P < 0.05) or 16α‐OHE1 (P < 0.001). The relative affinity of EC IgG was 1.49 × 10−7 M, 1.34 × 10−6 M and 1.13 × 10−6 M for 16α‐OHE1‐ERα, ERα and 16α‐OHE1, respectively. Several factors, such as obesity, postmenopausal status, use of hormonal therapy, ER and progesterone receptor (PR) status, low 2‐OHE1/16α‐OHE1 ratio, chemotherapy and hypertension, augment the production of antibodies against 16α‐OHE1‐ERα in EC patients. Conclusion: 16α‐OHE1‐ERα is a high‐affinity antigen for EC antibodies in the serum of EC patients and might function as a biomarker for this disease. Furthermore, several factors enhanced the production of antibodies against 16α‐OHE1‐ERα in the sera of these EC patients. [ABSTRACT FROM AUTHOR]

Abstract Mentally depressed breast cancer (MDBC) patients expressed estrogen receptor (ER) and 16α‑hydroxyestrone (16α‑OHE 1) is directly responsible for causing breast cancer (BC). This study aimed to identify whether depression in breast cancer patients enhanced the production of autoantibodies against 16α‑OHE 1 -ER adduct in breast cancer patients. The antibodies in the serum of 65 breast cancer patients (including 35 MDBC) and 40 control subjects were screened by direct binding, inhibition enzyme-linked immunosorbent assay (ELISA), and quantitative precipitin titration. Competition ELISA was also utilized for the estimation of 16α‑OHE 1 in the serum of 30 cancer patients. Autoantibodies from MDBC showed strong recognition to 16α‑OHE 1 -ER in comparison to overall breast cancer patients (p < 0.05) and control subjects (p < 0.001). Although breast cancer sera showed high binding to 16α‑OHE 1 -ER in comparison to ER (p < 0.05) or 16α‑OHE 1 (p < 0.001), the relative affinities of autoantibodies for 16α‑OHE 1 -ER were found to be 1.38 × 10−7 and 1.23 × 10−7 for breast cancer and MDBC patients respectively. No significant difference, either in the level of 16α‑OHE 1 or 2‑hydroxyestrone/16α‑OHE 1 ratio, was observed in the serum of cancer patients compared with controls, although inflammatory cytokines (IL-6, TNF-α) were significantly high in these patients. Depression in breast cancer patients augments the production of autoantibodies against 16α‑OHE 1 -ER through the generation of inflammatory conditions. Depression in these patients increased the release of pro-inflammatory cytokines that generate more autoantibodies and show strong binding with 16α‑OHE 1 -ER. Highlights • Depression increases autoantibodies production in breast cancer through inflammatory conditions. • Depression increases the release of inflammatory cytokine in breast cancer. • 16α‑Hydroxyestrone and estrogen receptor generate neo-epitopes, well recognized by breast cancer patient's IgGs. [ABSTRACT FROM AUTHOR]

Milk and dairy products are one of the chief sources of nutrition for human beings particularly for infant and children. Aflatoxin M1 (AFM1) a hydroxylated metabolite of aflatoxin B1 found in milk and milk products causes serious health issues for human beings. The objective of this study was to evaluate the effect of storage on the level of aflatoxin M1 in milk and other dairy products sold at retail stores of Tripoli Province, Libya. Selected samples (Skimmed and cream milk, infant milk formula, butter, cheese, Cheddar, spread and slice) were evaluated by using specialized RIDASCREEN AFM1 competitive enzyme linked immune sorbent assay (ELISA) technique. Our investigation revealed that, the concentration of AFM1 increased with the duration of storage. Furthermore, we found that the newly manufactured samples had very low concentration of AFM1 and within the permitted range. Moreover, AFM1 concentration in skimmed and cream milk having 6 month shelf life had 5.00 ngkg-1 and 5.03 ngkg-1 respectively. Furthermore, both the expired skimmed and cream milk had AFM1 concentrations 121.8 ngkg-1 and 108.18 ngkg-1, respectively. In addition to that, we found that the levels of AFM1 in different dairy products varies with different shelf lives (12 and 1 month), such as cheddar (5.0 and 72.79 ngkg-1), Spread (5.30 and 60.03 ngkg-1), Slice (5.50 and 61.18 ngkg-1). Additionally, infant milk formula with shelf life of 24 months and expired samples had AFM1 less than 5.00 ngkg-1 and 60.8 ngkg-1, respectively. Based on our investigation, we found that the presence AFM1 in milk and milk products at high concentration may cause serious illness to consumers' health and the consequent economic losses. [ABSTRACT FROM AUTHOR]

Background: Depression and smoking contribute to the prognosis of autoimmune rheumatoid arthritis (RA). Advanced glycation end products (AGEs) are also detected in RA patients. This study correlates RA in patients with various levels of depression and a history of smoking through the detection of antibodies against AGEs of proteins. Methods: Sixty RA subjects were selected and divided into 4 groups based on their levels of depression and smoking habits. The division was as follows: group I consisted of RA patients classified as depressed (RA-D); group II consisted of RA patients with a history of smoking (RA-S); group III consisted of RA patients suffering from depression who were also smokers (RA-DS); and group IV consisted of patients with RA alone (RA-A), i.e., not depressed and non-smokers. In vitro human serum albumin (HSA) was modified by glucose, and the modifications were studied by biochemical and biophysical techniques. Glycated (G)-HSA was used as an antigen, and autoantibodies against G-HSA (G-HSA-Abs) were screened in serum samples of different groups of RA subjects. Oxidative stress levels in all patients and healthy individuals were analyzed by protein-bound carbonyl content estimations. Results: Significant biochemical and biophysical changes were detected in G-HSA when compared to native (N)-HSA. All patients and control subjects were screened for circulating G-HSA-Abs and N-HSA-Abs. From the cohort of different samples, serum autoantibodies from RA-DS showed a high recognition of G-HSA-Abs titres compared to RA-D or RA-S. RA-A exhibited the least binding of circulating G-HSA-Abs of all the groups. The oxidative stress marker, the carbonyl content also exhibited highest levels in RA-DS, followed by RA-D and RA-S. Band shift assay showed the highest titres of immunoglobulin G in the serum samples of RA-DS. Conclusions: Smoking and concomitant depression in RA subjects may lead to enhanced oxidative stress levels responsible for the gradual formation and/or exposing of cryptic epitopes on HSA that induce the production of G-HSA-Abs. Hence, we postulate that by reducing depression and corresponding oxidative stress, it may be possible to control or limit the severity of the precipitation of RA disease activity and improve prognosis. [ABSTRACT FROM AUTHOR]

The inherent resistance of cancer stem cells (CSCs) to existing therapies has largely hampered the development of effective treatments for advanced malignancy. To help develop novel immunotherapy approaches that efficiently target CSCs, an experimental model allowing reliable distinction of CSCs and non-CSCs was set up to study their interaction with non-MHC-restricted γδ T cells and antigen-specific CD8+ T cells. Stable lines with characteristics of breast CSC-like cells were generated from ras-transformed human mammary epithelial (HMLER) cells as confirmed by their CD44hi CD24lo GD2+ phenotype, their mesenchymal morphology in culture and their capacity to form mammospheres under non-adherent conditions, as well as their potent tumorigenicity, self-renewal and differentiation in xenografted mice. The resistance of CSC-like cells to γδ T cells could be overcome by inhibition of farnesyl pyrophosphate synthase (FPPS) through pretreatment with zoledronate or with FPPS-targeting short hairpin RNA. γδ T cells induced upregulation of MHC class I and CD54/ICAM-1 on CSC-like cells and thereby increased the susceptibility to antigen-specific killing by CD8+ T cells. Alternatively, γδ T-cell responses could be specifically directed against CSC-like cells using the humanised anti-GD2 monoclonal antibody hu14.18K322A. Our findings identify a powerful synergism between MHC-restricted and non-MHC-restricted T cells in the eradication of cancer cells including breast CSCs. Our research suggests that novel immunotherapies may benefit from a two-pronged approach combining γδ T-cell and CD8+ T-cell targeting strategies that triggers effective innate-like and tumour-specific adaptive responses. [ABSTRACT FROM AUTHOR]

Cell-based immunotherapy strategies target tumors directly (via cytolytic effector cells) or aim at mobilizing endogenous anti-tumor immunity. The latter approach includes dendritic cells (DC) most frequently in the form of in vitro cultured peripheral blood monocytesderived DC. Human blood γδT cells are selective for a single class of non-peptide agonists ("phosphoantigens") and develop into potent antigen-presenting cells (APC), termed gdTAPC within 1-3 days of in vitro culture. Availability of large numbers of γδT-APC would be advantageous for use as a novel cellular vaccine. We here report optimal gdT cell expansion (>107 cells/ml blood) when peripheral blood mononuclear cells (PBMC) from healthy individuals and melanoma patients were stimulated with zoledronate and then cultured for 14 days in the presence of IL-2 and IL-15, yielding gdT cell cultures of variable purity (77±21 and 56±26%, respectively). They resembled effector memory αβT (TEM) cells and retained full functionality as assessed by in vitro tumor cell killing as well as secretion of pro-inflammatory cytokines (IFNg,TNFα) and cell proliferation in response to stimulation with phosphoantigens. Importantly, day 14 γδT cells expressed numerous APC-related cell surface markers and, in agreement, displayed potent in vitro APC functions. Day 14 γδT cells from PBMC of patients with cancer were equally effective as their counterparts derived from blood of healthy individuals and triggered potent CD8C αβT cell responses following processing and cross-presentation of simple (influenza M1) and complex (tuberculin purified protein derivative) protein antigens. Of note, and in clear contrast to peripheral blood γδT cells, the ability of day 14 γδT cells to trigger antigen-specific abT cell responses did not depend on re-stimulation.We conclude that day 14 gdT cell cultures provide a convenient source of autologous APC for use in immunotherapy of patients with various cancers. [ABSTRACT FROM AUTHOR]

Estrogen metabolites have been implicated in rheumatoid arthritis (RA) and cancer, although the mechanism remains unestablished. Some estrogen metabolites, which are used for the assessment of cancer risk, play an important role in RA. The pathways by which malignancies associated with RA remain elusive. Possible mechanism involves enzymatic or nonenzymatic oxidation of estrogen into catecholestrogen metabolites through semiquinone and quinone redox cycle to produce free radicals that can cause DNA modifications. Modifications of DNA alter its immunogenicity and trigger various immune responses leading to elevated levels of cancer and RA antibodies. However, the role of different estrogen metabolites as a mediator of immune response cannot be ruled out in various immune-related diseases. [ABSTRACT FROM AUTHOR]

Background: Autoantibodies against glutamate decarboxylase-65 (GAD65Abs) are thought to be a major immunological tool involved in pathogenic autoimmunity development in various diseases. GAD65Abs are a sensitive and specific marker for type 1 diabetes (T1D). These autoantibodies can also be found in 6-10% of patients classified with type 2 diabetes (T2D), as well as in 1-2% of the healthy population. The latter individuals are at low risk of developing T1D because the prevalence rate of GAD65Abs is only about 0.3%. It has, therefore, been suggested that the antibody binding to GAD65 in these three different GAD65Ab-positive phenotypes differ with respect to epitope specificity. The specificity of reactive oxygen species modified GAD65 (ROS-GAD65) is already well established in the T1D. However, its association in secondary complications of T1D has not yet been ascertained. Hence this study focuses on identification of autoantibodies against ROS-GAD65 (ROS-GAD65Abs) and quantitative assays in T1D associated complications. Results: From the cohort of samples, serum autoantibodies from T1D retinopathic and nephropathic patients showed high recognition of ROS-GAD65 as compared to native GAD65 (N-GAD65). Uncomplicated T1D subjects also exhibited reactivity towards ROS-GAD65. However, this was found to be less as compared to the binding recorded from complicated subjects. These results were further proven by competitive ELISA estimations. The apparent association constants (AAC) indicate greater affinity of IgG from retinopathic T1D patients (1.90 × 10-6 M) followed by nephropathic (1.81 × 10-6 M) and uncomplicated (3.11 × 10-7 M) T1D patients for ROS-GAD65 compared to N-GAD65. Conclusion: Increased oxidative stress and blood glucose levels with extended duration of disease in complicated T1D could be responsible for the gradual formation and/or exposing cryptic epitopes on GAD65 that induce increased production of ROS-GAD65Abs. Hence regulation of ROS-GAD65Abs could offer novel tools for analysing and possibly treating T1D complications. [ABSTRACT FROM AUTHOR]

Background: Glycated proteins present new immunological epitopes on their surface against which autoantibodies are generated that have a possible role in immunopathogenesis in diabetic complications. Methods: In the present study, in vitro glycation- and reactive oxygen species (ROS)-modified human serum albumin (HSA) has been studied by different spectroscopic techniques (UV and fluorescence) and thermal denaturation profiles. The binding characteristics of circulating autoantibodies in diabetic patients and diabetic patients with secondary complications against native HSA (N-HSA) and ROS-modified glycated HSA (RG-HSA) were assessed by direct and competition enzyme-linked immunosorbent assay (ELISA). In another approach, antibodies against RG-HSA (RG-HSA-Abs) induced in experimental animals were used as an immunochemical probe for the detection of gluco-oxidative lesions in blood proteins of patients (n = 8) with diabetic retinopathy. Results: Modified RG-HSA showed marked structural changes. High recognition of RG-HSA was shown by diabetic serum autoantibodies. Diabetic patients with retinopathy, nephropathy and atherosclerosis showed significantly (p < 0.001) stronger binding to RG-HSA over N-HSA. Normal human sera exhibited negligible binding with either antigen. Competitive inhibition ELISA results show significantly high binding of RG-HSA-Abs to albumin, immunoglobulin G and red blood cell membrane isolated from diabetic retinopathic patients. Conclusion: In conclusion, these results suggest that hyperglycemia together with ROS may contribute to the immunopathogenesis of diabetes-associated complications. Copyright © 2010 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Glutamic acid decarboxylase-65 (GAD65) is an immunological marker of type 1 autoimmune diabetes. High titre of autoantibodies against GAD65 (GAD65Abs) have also been detected in some other autoimmune diseases. In search of a potential immunological marker of type 1 diabetes, in vitro GAD65 was modified by hydroxyl radical followed by the study of structural and conformational perturbed protein by different spectroscopic techniques (UV, fluorescence and CD) and thermal denaturation profile. Binding studies of circulating autoantibodies from diabetic groups (type 1 and type 2) with native and reactive oxygen species (ROS) modified GAD65, exhibited high recognition of type 1 diabetic serum autoantibodies with modified antigen (p < 0.001) over unmodified GAD65. Binding specificity of isolated IgG of patients (n = 17) from each diabetic group and control group (n = 10) was checked by inhibition enzyme linked immunosorbent assay (ELISA) and quantitative precipitin titration assay. Relative affinity of ROS-GAD65Abs for modified and native GAD65 was in the order of 1.56 × 10- 6 and 2.72 × 10- 7 M, as calculated by Langmuir plot. In coherence, ROS oxidation of GAD65 causes conformational perturbation, generating highly immunogenic unique neoepitopes that may be one of the factors in antigen-driven induction of type 1 diabetes autoantibodies that can serve as a potential marker in early diagnosis/prognosis of the disease. [ABSTRACT FROM AUTHOR]

• Depression linked to higher antibodies production against estogenized insulin in T1D. • Depression enhance the release of IL-1β and IL-17 in T1D. • Depression generates inflammatory conditions in T1D. Depression has been commonly associated with type 1 diabetes (T1D) and insulin covalently modified with catecholestrogens (CEs) was found in serum of these T1D patients. This study aimed to know whether depression link to higher antibodies against estrogenized insulin in T1D. ELISA (direct binding and competition) and quantitative precipitin titration were used to detect antibodies and their affinities against estrogenized insulin in the serum of 66 depressed T1D (DT1D) patients (out of 110 T1D) and 41 control subjects. Antibodies from DT1D patients showed high binding specificity to estrogenized insulin (2-hydroestradiol-insulin; 2-OHE 2 -Ins) in comparison to overall T1D patients (p < 0.05) or control subjects (p < 0.001). However, T1D sera demonstrate high recognition to 2-OHE 2 -Ins as compared to Ins (p < 0.05) or 2-OHE 2 (p < 0.001). The affinity of antibodies from DT1D and T1D patients was 1.32 × 10-7 M and 1.43 × 10-7 M, respectively. Depression linked to higher antibodies production against estrogenized insulin in T1D. Furthermore, depression in T1D generates inflammatory conditions that further increased antibodies production in T1D patients. [ABSTRACT FROM AUTHOR]

Insulin (Ins) covalently modified by catecholestrogens (CEs) was commonly found in diabetic patients who have developed insulin resistance. Estrogenization of insulin altered its molecular function and effect carbohydrates metabolisms in these patients. Insulin resistance is a common phenomenon in diabetes but the exact mechanism remains unknown. In this study, binding specificity and affinity of autoantibodies against estrogenized insulin (4-hydroxyestradiol-insulin; 4-OHE 2 -Ins) were assayed in the serum of type 1 diabetes (T1D) patients in order to explain the phenomena behind insulin resistance. Specificity and affinity of autoantibod i es from the sera of 66 T1D patients and 41 controls were analyzed by direct binding, competition ELISA and quantitative precipitin titration. Insulin was also estimated in the serum of T1D patients by ELISA. Estrogenized insulin (4-OHE 2 -Ins) exhibited high affinity and specificity to T1D autoantibodies in comparison to Ins (p <.05) or 4-OHE 2 (p <.001). Estrogenization of insulin alters its interaction with the insulin receptor (IR). The affinity constant of 4-OHE 2 -Ins with the T1D autoantibodies was found to be 1.41 × 10−7 M. Estrogenization of insulin by catecholestrogen makes these molecules highly antigenic and produced high - affinity autoantibodies in T1D patients. As a result, patients develop insulin resistance and presented this molecule as a potential biomarker for T1D. • Estrogenized insulin (4-OHE 2 -Ins) exhibited high affinity and specificity for T1D autoantibodies. • Estrogenization of insulin alters the normal functions of the insulin in T1D patients • Estrogenization of insulin in the blood produced autoantibodies; as a result insulin resistance is developed in T1D patients [ABSTRACT FROM AUTHOR]

Depression is the common and early symptoms associated with early onset of SLE, 16α-hydroxyestrone (16α-OHE 1) levels were found to be significantly higher in serum and urine in patients with SLE. This study was carried out in order to know whether depression and its related parameters in the SLE patients enhanced the production of autoantibodies against 16α-OHE 1 -albumin (A) complexes. The autoantibodies in the serum of 100 SLE [including 65 depressed SLE (DSLE)] patients and 37 control subjects were detected by using direct binding, inhibition ELISA and quantitative precipitin titration. Autoantibodies from DSLE patients (and also the patients who were taken anti-depressant and with neurological symptoms) showed high binding to 16α-OHE 1 -A in contrast to SLE (p < 0.05) and control subjects (p < 0.001). Although, SLE sera showed high recognition to 16α-OHE 1 -A in comparison to A (p < 0.05) or 16α-OHE 1 (p < 0.001). The affinity of autoantibodies for 16α-OHE 1 -A was found to be high for DSLE (1.16 × 10−7 M) and SLE (1.24 × 10−7 M) patients as detected by Langmuir plot. The concentration of 16α-OHE 1 (p < 0.05) and inflammatory cytokines (IL-6, p < 0.05 and IL-17, p < 0.001) in the serum of SLE patients was found to be significantly higher than controls. Depression and its related parameters in SLE enhanced the production of autoantibodies against 16α-OHE 1 -A through the generation of inflammatory conditions. Depression in SLE patients increased the release of pro-inflammatory cytokine (IL-6 and IL-17) that in turn generating more autoantibodies and showed strong recognition to 16α-OHE 1 -A. • Depression and its related parameters increase the production of SLE autoantibodies through inflammatory conditions. • They increase the release of inflammatory cytokine (IL-6 and IL-17) in SLE patients. • 16α-Hydroxyestrone and albumin generate unique epitopes, well recognized by SLE autoantibodies. [ABSTRACT FROM AUTHOR]

The early immune response to microbes is dominated by the recruitment of neutrophils whose primary function is to clear invading pathogens. However, there is emerging evidence that neutrophils play additional effector and regulatory roles. The present study demonstrates that human neutrophils assume Ag cross-presenting functions and suggests a plausible scenario for the local generation of APC-like neutrophils through the mobilization of unconventional T cells in response to microbial metabolites. Vγ9/Vδ2 T cells and mucosal-associated invariant T cells are abundant in blood, inflamed tissues, and mucosal barriers. In this study, both human cell types responded rapidly to neutrophils after phagocytosis of Gram-positive and Gram-negative bacteria producing the corresponding ligands, and in turn mediated the differentiation of neutrophils into APCs for both CD4+ and CD8+ T cells through secretion of GM-CSF, IFN-γ, and TNF-α. In patients with acute sepsis, circulating neutrophils displayed a similar APC-like phenotype and readily processed soluble proteins for cross-presentation of antigenic peptides to CD8+ T cells, at a time when peripheral Vγ9/Vδ2 T cells were highly activated. Our findings indicate that unconventional T cells represent key controllers of neutrophil-driven innate and adaptive responses to a broad range of pathogens. [ABSTRACT FROM AUTHOR]

Non-enzymatic glycation of biomolecules by reducing sugars led to several products, including the advanced glycation end products (AGEs), the accumulation of which has been linked to various life-threatening diseases. The binding of AGEs to their respective protein receptors for advanced glycation end products (RAGE) can initiate a cascade of reactions, which may alter physiological conditions. The present work investigates the potential of 4-thiazolidinones as RAGE inhibitors. We performed an extensive computational study to identify the structural requirements needed to act as RAGE inhibitors. To achieve this goal, 4-thiazolidinone-based compounds available in PubChem, ZINC15, ChEMBL, and ChEBI databases were screened against RAGE (PDB: 4LP5), leading to the identification of top five drug-like candidates with a high binding affinity to RAGE V-domain catalytic region. Drug likeness, absorption, distribution, metabolism, excretion, and toxicity (ADMET) of the top-scoring compounds have been studied and discussed. Global molecular descriptors, chemical reactivity, hardness, softness, etc., have been estimated. Finally, molecular dynamics (MD) simulations at 100 ns were carried out to check the stability and other properties. Overall, we believe that the identified compounds can potentially attenuate RAGE-AGE interactions.Communicated by Ramaswamy H. Sarma.

The inherent resistance of cancer stem cells (CSCs) to existing therapies has largely hampered the development of effective treatments for advanced malignancy. To help develop novel immunotherapy approaches that efficiently target CSCs, an experimental model allowing reliable distinction of CSCs and non-CSCs was set up to study their interaction with non-MHC-restricted γδ T cells and antigen-specific CD8 + T cells. Stable lines with characteristics of breast CSC-like cells were generated from ras-transformed human mammary epithelial (HMLER) cells as confirmed by their CD44 hi CD24 lo GD2 + phenotype, their mesenchymal morphology in culture and their capacity to form mammospheres under non-adherent conditions, as well as their potent tumorigenicity, self-renewal and differentiation in xenografted mice. The resistance of CSC-like cells to γδ T cells could be overcome by inhibition of farnesyl pyrophosphate synthase (FPPS) through pretreatment with zoledronate or with FPPS-targeting short hairpin RNA. γδ T cells induced upregulation of MHC class I and CD54/ICAM-1 on CSC-like cells and thereby increased the susceptibility to antigen-specific killing by CD8 + T cells. Alternatively, γδ T-cell responses could be specifically directed against CSC-like cells using the humanised anti-GD2 monoclonal antibody hu14.18K322A. Our findings identify a powerful synergism between MHC-restricted and non-MHC-restricted T cells in the eradication of cancer cells including breast CSCs. Our research suggests that novel immunotherapies may benefit from a two-pronged approach combining γδ T-cell and CD8 + T-cell targeting strategies that triggers effective innate-like and tumour-specific adaptive responses.

Peritonitis remains the major obstacle for the maintenance of long-term peritoneal dialysis and dysregulated host peritoneal immune responses may compromise local anti-infectious defense, leading to treatment failure. Whilst, tissue mononuclear phagocytes, comprising macrophages and dendritic cells, are central to a host response to pathogens and the development of adaptive immune responses, they are poorly characterized in the human peritoneum. Combining flow cytometry with global transcriptome analysis, the phenotypic features and lineage identity of the major CD14 + macrophage and CD1c + dendritic cell subsets in dialysis effluent were defined. Their functional specialization was reflected in cytokine generation, phagocytosis, and antigen processing/presentation. By analyzing acute bacterial peritonitis, stable (infection-free) and new-starter patients receiving peritoneal dialysis, we identified a skewed distribution of macrophage to dendritic cell subsets (increasing ratio) that associated with adverse peritonitis outcomes, history of multiple peritonitis episodes, and early catheter failure, respectively. Intriguingly, we also noted significant alterations of macrophage heterogeneity, indicative of different maturation and activation states that were associated with different peritoneal dialysis outcomes. Thus, our studies delineate peritoneal dendritic cells from macrophages within dialysate, and define cellular characteristics associated with peritoneal dialysis treatment failure. These are the first steps to unravelling the detrimental adaptive immune responses occurring as a consequence of peritonitis., (Copyright © 2016 International Society of Nephrology. Published by Elsevier Inc. All rights reserved.)

IMMUNOTHERAPY IS A FAST ADVANCING METHODOLOGY INVOLVING ONE OF TWO APPROACHES: (1) compounds targeting immune checkpoints and (2) cellular immunomodulators. The latter approach is still largely experimental and features in vitro generated, live immune effector cells, or antigen-presenting cells. γδ T cells are known for their efficient in vitro tumor killing activities. Consequently, many laboratories worldwide are currently testing the tumor killing function of γδ T cells in clinical trials. Reported benefits are modest; however, these studies have demonstrated that large γδ T-cell infusions were well tolerated. Here, we discuss the potential of using human γδ T cells not as effector cells but as a novel cellular vaccine for treatment of cancer patients. Antigen-presenting γδ T cells do not require to home to tumor tissues but, instead, need to interact with endogenous, tumor-specific αβ T cells in secondary lymphoid tissues. Newly mobilized effector αβ T cells are then thought to overcome the immune blockade by creating proinflammatory conditions fit for effector T-cell homing to and killing of tumor cells. Immunotherapy may include tumor antigen-loaded γδ T cells alone or in combination with immune checkpoint inhibitors.