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Increasing evidence suggests that the presence and spatial localization and distribution pattern of tumor infiltrating lymphocytes (TILs) is associate with response to immunotherapies. Recent studies have identified TGF β activity and signaling as a determinant of T cell exclusion in the tumor microenvironment and poor response to PD-1/PD-L1 blockade. Here we coupled the artificial intelligence (AI)-powered digital image analysis and gene expression profiling as an integrative approach to quantify distribution of TILs and characterize the associated TGF β pathway activity. Analysis of T cell spatial distribution in the solid tumor biopsies revealed substantial differences in the distribution patterns. The digital image analysis approach achieves 74% concordance with the pathologist assessment for tumor-immune phenotypes. The transcriptomic profiling suggests that the TIL score was negatively correlated with TGF β pathway activation, together with elevated TGF β signaling activity observed in excluded and desert tumor phenotypes. The present results demonstrate that the automated digital pathology algorithm for quantitative analysis of CD8 immunohistochemistry image can successfully assign the tumor into one of three infiltration phenotypes: immune desert, immune excluded or immune inflamed. The association between "cold" tumor-immune phenotypes and TGF β signature further demonstrates their potential as predictive biomarkers to identify appropriate patients that may benefit from TGF β blockade. An artificial intelligence-powered digital image analysis approach has been established to characterize and classify the cancer immune phenotypes, which might be utilized as predictive biomarkers for TGF β blockade therapy. [Display omitted] [ABSTRACT FROM AUTHOR]

The aim of this study is to develop a nucleic acid sensor based on piezoelectric crystal microbalance system (QCM) for following hybridization. Piezoelectric quartz crystal surfaces were first treated in a glow–discharge apparatus with ethylene diamine (EDA) plasma at 15 W (discharge power), 2.5 min (incubation time) and 35 ml/min (monomer flow rate) to create amino groups on the crystal surfaces. The thickness of the EDA–plasma film formed was about 43±24 A˚. Then, the amino groups on the crystal surfaces were converted to aldehyde groups by reacting the amino groups with glutaraldehyde (GA) at different conditions. A GA concentration of 2.5% and an incubation time of 2 h were selected as optimal values at this step, corresponding to a GA surface concentration of about 270 ng/cm2. A double strand Oligonucleotides, having one extra base on 5′-end of one of the complementary strands, were immobilized through the amino groups of this base onto the GA-modified crystals. Optimal immobilization conditions were as follows: oligonucleotide concentration: 1 μg/ml; time: 3 h; pH: 9.2 carbonate buffer; ionic strength: 0.1; and temperature: 20 °C. The QCM sensor carrying the covalently bound strand was used in the hybridization experiments, which showed that equilibrium is achieved in about 5 min, and the frequency shift measured is related to the concentration of the target strand to be measured within the medium. [Copyright &y& Elsevier]

The aim of this study is to investigate complexation of metallothionein (MT) with cadmium and zinc ions. An oligopeptide (i.e. Lys-Cys-Thr-Cys-Cys-Ala), a fragment of MT was covalently immobilized onto piezoelectric crystals, which were first treated with ethylene diamine plasma in a glow-discharge apparatus, and then were chemically reacted with glutaraldehyde. Complexation of the immobilized MT with Zn(II) and Cd(II) ions in aqueous media was followed by recording the changes of the frequency shifts of the piezoelectric quartz crystals. The amount of Cd(II) ions interacted with the immobilized MT molecules was the highest at pH 7.4, and decreased with an increase in the pH of the medium, in parallel to the decrease in the amount of immobilized MT. The number of Zn(II) ions interacted with the immobilized MT molecules was higher than the number of Cd(II) ions when the adsorption was from solutions containing a single-metal ion with the same ion concentrations. In consecutive adsorption studies, we observed that the type of metal ions used in the first interaction is important. These experiments showed also that there is an exchange between the metal ions, and competition provokes adsorption of both ions due to synergistic–antagonistic effects. [Copyright &y& Elsevier]

The aim of this study is to develop an immunoaffinity sensor based on piezoelectric crystals for human serum albumin (HSA) detection in aqueous media. Quartz crystals were treated with ethylene diamine (EDA) plasma in a glow-discharge apparatus in order to substitute amino groups on their surfaces. Then anti-HSA antibodies were immobilized via these amino groups by using glutaraldehyde (GA) as cross-linker. Immobilization of the antibody on the quartz crystal was examined for different pH, antibody concentration and treatment time. The optimum conditions for anti-HSA immobilization were evaluated by the measurements of the activity of the surface against HSA. The optimum values of pH, antibody concentration and treatment time were found 6.2, 0.15 mg/ml and 2 h, respectively. For detection of HSA into the solution, two methods were used. In the first (dip and dry) method, the frequency shifts were measured in air after the 1 h interaction of the anti-HSA immobilized crystals with HSA solution. In the other (direct) method, the frequency shifts were followed continuously for 60 min. while the probe was immersed in the HSA solution. An increase for the frequency shifts was observed with increasing of HSA concentration of 16–128 μg/ml. Both the immobilization and antibody-interaction conditions were found important on the extend of these specific interaction. The relations between the HSA concentrations and frequency shifts were exponential in both methods. [Copyright &y& Elsevier]

In the current study, we report on the design and development of a novel electrochemical immunosensor for the detection of cancer antigen 125 (CA125) oncomarker. Polyamidoamine/gold nanoparticles (PAMAM/AuNPs) were used to increase the conductivity and enhance the number of antibodies (Abs) immobilized on the electrode surface. Three-dimensional reduced graphene oxide-multiwall carbon nanotubes (3DrGO-MWCNTs) were used to modify the glassy carbon electrode to improve the electrode conductivity and specific surface area. Ab and toluidine blue attached to O -succinyl-chitosan-magnetic nanoparticles (Suc-CS@MNPs) as a tracer. The poor solubility of chitosan (CS) was improved by succinic anhydride using a novel modification method. Under optimum condition, the developed immunosensor exhibited a wide linear range (0.0005–75 U/mL) and an excellent limit of detection around 6 μU/mL. The reliability of the engineered immunosensor in detecting CA125 was verified by standard addition recovery method, which was further compared to enzyme-linked immunosorbent assay (ELISA). The proposed immunosensor exhibited excellent stability, high selectivity and sensitivity, and good reproducibility. Based on the great performance of the engineered immunosensor, it is proposed as a robust and reliable diagnostic tool for the detection of CA125 in the clinic. Image 1 • An immunosensor was developed for detecting CA125 with a wide linear range (0.0005–75 U/mL) and an excellent LOD (6 μU/mL). • PAMAM/AuNPs were synthesized through a one-step method using PAMAM as a stabilizing and reducing agent. • Chitosan water solubility in natural pH was increased using a novel modification method. • Succinyl-chitosan-coated magnetic nanoparticles were synthesized with good dispersity and high surface functional groups. [ABSTRACT FROM AUTHOR]

• Use of reduced graphene oxide as a photothermal label for lateral flow assay. • Reduced graphene oxide shows higher photothermal properties than graphene oxide. • Sensitivity improvement of a lateral flow assay by photothermal quantification. Lateral flow immunoassay (LFA) is a well-known point-of-care technology for the detection of various analytes. However, low sensitivity and lack of quantitative results are some of its critical drawbacks. Here we report a photothermal enhanced lateral flow sensor on the basis of the photothermal properties of reduced graphene oxide (rGO) for the detection of E-coli O157:H7 as a model pathogen. The calibration curve of the photothermal method exhibited a linear range from 5 × 10⁵ to 5 × 10⁷ CFU/ml with a correlation coefficient of R2 = 0.96 and a regression equation of y = 8.1x-43 for standard bacteria solutions in phosphate buffer. The limit of detection was ∼5 × 10⁵ CFU/ml for standard bacteria solutions, which was a 10-fold enhancement in sensitivity compared to the qualitative results. Specificity experiments showed that the photothermal method can only detect the target bacteria among 6 types of bacteria strains. It was confirmed that the developed technique could be a highly potential method for the rapid detection field because it can provide fast quantitative results with improved sensitivity. [ABSTRACT FROM AUTHOR]