Gel electrophoretic analysis of differently shaped interacting and non-interacting bioconjugated nanoparticles

The use of a simple gel electrophoretic method to study mixtures of differently shaped biofunctionalized nanoparticles (NP's) that undergo bioaffinity interactions is demonstrated. Both gold nanorods (NR's) and quasi-spherical nanoparticles (qNS's) were functionalized with an interacting antigen and antibody pairing (alpha-1 antitrypsin (AAT) protein and antiAAT) or non-interacting antibody controls (antiBNP). Gel-based measurements were accompanied with transmission electron microscopy (TEM) and UV-vis spectroscopy analysis before and after separation. Initial measurements of NR and qNS bioconjugates suspended individually were applied to optimize the gel separation conditions and it was demonstrated that higher particle uniformities could be obtained relative to the initial stock solutions. A series of NR and qNS mixtures prepared at various stoichiometric ratios were then compared for both interacting (antiAAT–AAT) and non-interacting (antiAAT–antiBNP) particle conjugates. Both gel images and extinction measurements were utilized to demonstrate reduced NP concentrations transported along the gel due to bioaffinity-induced NP assembly. This confirmed that gel electrophoresis can be extended to identifying particle aggregation associated with protein bioaffinity interactions as well as being an established tool for separating particles based on size, shape and surface chemistry

Femtomolar detection of Tau proteins in undiluted plasma using surface plasmon resonance

The ability to directly detect Tau protein and other neurodegenerative biomarkers in human plasma at clinically relevant concentrations continues to be a significant hurdle for the establishment of diagnostic tests for Alzheimer’s disease (AD). In this article, we introduce a new DNA aptamer/antibody sandwich assay pairing and apply it for the detection of human Tau 381 in undiluted plasma at concentrations as low as 10 fM. This was achieved on a multichannel surface plasmon resonance (SPR) platform with the challenge of working in plasma overcome through the development of a tailored mixed monolayer surface chemistry. In addition, a robust methodology was developed involving various same chip control measurements on reference channels to which the detection signal was normalized. Comparative measurements in plasma between SPR and enzyme-linked immunosorbent assay (ELISA) measurements were also performed to highlight both the 1000-fold performance enhancement of SPR and the ability to measure both spiked and native concentrations that are not achievable with ELISA

Tandem femto- and nanomolar analysis of two protein biomarkers in plasma on a single mixed antibody monolayer surface using surface plasmon resonance

The multiplexed detection of protein biomarkers in plasma present over a range of clinically relevant concentrations continues to be difficult for surface-based bioaffinity detection platforms such as surface plasmon resonance (SPR). As well as nonspecific adsorption, challenges include quantitative comparison between targets whose concentrations differ by orders of magnitude, regenerating SPR chips after plasma exposure, and the two- or four-channel limitation of many commercial SPR instruments limiting sample throughput. In this article, we explore an approach where two protein biomarkers alpha-1 antitrypsin (AAT) and Tau 381 are detected in tandem within a single SPR channel at micromolar and femtomolar concentrations, respectively. This was achieved by creating a mixed antibody (antiAAT and antiTau) monolayer on the chip surface. After the adsorption of AAT and/or Tau, further specificity was obtained via the adsorption of a DNA aptamer specific to each target. The detection range for each target was controlled via the relative surface density ratio of each antibody type as well as each aptamer concentration. Calibration measurements were performed in both buffer and spiked plasma with the detection of native concentrations of ∼39 fM (Tau) and ∼65 μM (AAT) in a human plasma sample. Finally, tandem measurements of both targets within the same SPR signal channel were demonstrated at these very different concentrations

The Microbiome in Systemic Sclerosis: Pathophysiology and Therapeutic Potential

Systemic sclerosis (SSc), also known as scleroderma, is an autoimmune disease with unknown etiology characterized by multi-organ fibrosis. Despite substantial investigation on SSc-related cellular and molecular mechanisms, effective therapies are still lacking. The skin, lungs, and gut are the most affected organs in SSc, which act as physical barriers and constantly communicate with colonized microbiota. Recent reports have documented a unique microbiome signature, which may be the pathogenic trigger or driver of SSc. Since gut microbiota influences the efficacy and toxicity of oral drugs, evaluating drug–microbiota interactions has become an area of interest in disease treatment. The existing evidence highlights the potential of the microbial challenge as a novel therapeutic option in SSc. In this review, we have summarized the current knowledge about molecular mechanisms of SSc and highlighted the underlying role of the microbiome in SSc pathogenesis. We have also discussed the latest therapeutic interventions using microbiomes in SSc, including drug–microbiota interactions and animal disease models. This review aims to elucidate the pathophysiological connection and therapeutic potential of the microbiome in SSc. Insights into the microbiome will significantly improve our understanding of etiopathogenesis and developing therapeutics for SSc

Evaluation of Efficient and Noble-Metal-Free NiTiO3 Nanofibers Sensitized with Porous gC3N4 Sheets for Photocatalytic Applications

One-dimensional nickel titanate nanofibers (NiTiO3 NFs) were synthesized and loaded with acetic acid-treated exfoliated and sintered sheets of graphitic carbon nitride (AAs-gC3N4) to fabricate a unique heterogeneous structure. This novel fabrication method for porous AAs-gC3N4 sheets using acetic acid-treated exfoliation followed by sintering provided gC3N4 with a surface area manifold larger than that of bulk gC3N4, with an abundance of catalytically active sites. Hybrid photocatalysts were synthesized through a two-step process. Firstly, NiTiO3 NFs (360 nm in diameter) were made by electrospinning, and these NiTiO3 NFs were sensitized with exfoliated gC3N4 sheets via a sonication process. Varying the weight ratio of NiTiO3 fibers to porous AAs-gC3N4 established that NiTiO3 NFs containing 40 wt% of porous AAs-gC3N4 exhibited optimal activity, i.e., removal of methylene blue and H2 evolution. After 60 min exposure to visible light irradiation, 97% of the methylene blue molecules were removed by the hybrid photocatalyst, compared with 82%, 72%, and 76% by pristine AAs-gC3N4, NiTiO3 NFs, and bulk gC3N4, respectively. The optimal structure also displayed excellent H2 evolution performance. The H2 evolution rate in the optimal sample (152 μmol g−1) was 2.2, 3.2 and 3-fold higher than that in pure AAs-gC3N4 (69 μmol g−1), NiTiO3 NFs (47 μmol g−1) and bulk gC3N4 (50 μmol g−1), respectively. This clearly shows that the holey AAs-gC3N4 nanosheets interacted synergistically with the NiTiO3 NFs. This extended the lifetime of photogenerated charge carriers and resulted in superior photocatalytic activity compared with pristine NiTiO3 NFs and bulk gC3N4. The higher Brunauer-Emmett-Teller surface area and the presence of many catalytically active sites also enhanced the photocatalytic performance of the hybrid sample. Moreover, through photoluminescence and photocurrent response analysis, a significant decrease in the recombination losses of the hybrid photocatalysts was also confirmed. Thus, this is a novel strategy to fabricate highly efficient photocatalysts with precisely tunable operating windows and enhanced charge separation

The Braking-Pressure and Driving-Direction Determination System (BDDS) Using Road Roughness and Passenger Conditions of Surrounding Vehicles

A fully autonomous vehicle must ensure not only fully autonomous driving but also the safety and comfort of its passengers. However, the self-driving technology that is currently completed focuses only on perfect driving and does not guarantee the safety and comfort of passengers. This paper proposes a braking-pressure and driving-direction determination system (BDDS), which computes the brake pressure and steering angle optimized for passenger safety by utilizing more diverse information than existing autonomous vehicles. The BDDS proposed in this paper consists of two modules. The road roughness classification module (RRCM) classifies the roughness of the road by using the pressure data applied to the suspension and the K-NN algorithm and computes the optimal brake pressure. The passenger recognition and sharing module (PRSM) identifies the current occupant status of the vehicle by using a body pressure sensor and CNN, shares the information with surrounding vehicles, and computes the optimal steering angle using passenger information and road information. As a result of the simulations described in this paper, the parameters of AI models were optimized. In addition, the RRCS was about 7% more accurate than the K-means clustering algorithm, and PRS was about 9% more accurate than the existing seat recognition system

Molecular Clustering and Velocity Increase in Converging-Diverging Nozzle in MD Simulation

A Molecular dynamics simulation in a converging-diverging nozzle was performed to study molecular collisions and their influence to average flow velocity according to a variety of vacuum levels. The static pressures and the dynamic pressure exerted by the molecule collision on the selected walls were compared to figure out the intensity variances of the directional flows. With pressure differences constant between the entrance and the exit of the nozzle, The numerical experiment was performed for molecular velocities and directional flows. The result shows that the velocities are increased at the nozzle exit as the vacuum level gets higher in that area because of less molecular collision

Direct Detection of α‑1 Antitrypsin in Serum Samples using Surface Plasmon Resonance with a New Aptamer–Antibody Sandwich Assay

The challenges associated with performing surface plasmon resonance (SPR) based measurements in serum and other biofluids have continued to limit the applicability of this valuable sensing technology for sensitive bioaffinity measurements of proteins in clinically relevant samples. In this paper, a new sandwich assay is introduced for the quantitative SPR analysis of α-1 antitrypsin (AAT), which is a recognized biomarker for Alzheimer’s disease. Detection was performed via the specific adsorption of AAT onto a gold chip surface modified with a DNA aptamer. The measurement dynamic range and also sensitivity in serum were improved with the subsequent surface binding of antiAAT. A methodology was established to measure the target protein in serum, albumin and immunoglobulin G (IgG) solutions with the results correlated with measurements in buffer only. A comparison between SPR and enzyme-linked immunosorbent assay (ELISA) measurements was also made. The detection of AAT in serum at clinically relevant concentrations was demonstrated with target concentrations as low as 10 fM readily achievable

Gold Nanostar Enhanced Surface Plasmon Resonance Detection of an Antibiotic at Attomolar Concentrations via an Aptamer-Antibody Sandwich Assay

A new sandwich assay for tetracycline (TC) involving a DNA aptamer and antibody pair is demonstrated in conjunction with gold nanostar (GNS) enhanced surface plasmon resonance (SPR) to achieve detection in the low attomolar range. GNS particles were covalently functionalized with the antibody probe (antiTC) and integrated into a surface sandwich assay in conjunction with a SPR gold chip modified with the TC-specific aptamer. After it was demonstrated that both affinity probes can bind simultaneously to TC, optimization of the assay was performed using either antiTC only or GNS-antiTC conjugates to interact with aptamer/TC complexes present on the chip surface. Target concentrations as low as 10 aM could be detected using GNS-antiTC’s, which was >10³ times greater in performance than when using antiTC only. In addition, good selectivity was achieved with respect to other tetracycline derivative antibiotics, such as oxytetracycline (OTC) and chlortetracycline (CTC), both which are structurally similar to TC. As a demonstration of trace antibiotic analysis in environmental samples, the GNS enhanced sandwich assay was applied to analyze TC added to aliquots of local river water and the results validated by comparing to conventional high-performance liquid chromatography (HPLC) analysis