State-of-the-Art Glycomics Technologies in Glycobiotechnology.

Glycosylation affects the properties of biologics; thus regulatory bodies classified it as critical quality attribute and force biopharma industry to capture and control it throughout all phases, from R&D till end of product lifetime. The shift from originators to biosimilars further increases importance and extent of glycoanalysis, which thus increases the need for technology platforms enabling reliable high-throughput and in-depth glycan analysis. In this chapter, we will first summarize on established glycoanalytical methods based on liquid chromatography focusing on hydrophilic interaction chromatography, capillary electrophoresis focusing on multiplexed capillary gel electrophoresis, and mass spectrometry focusing on matrix-assisted laser desorption; we will then highlight two emerging technologies based on porous graphitized carbon liquid chromatography and on ion-mobility mass spectrometry as both are highly promising tools to deliver an additional level of information for in-depth glycan analysis; additionally we elaborate on the advantages and challenges of different glycoanalytical technologies and their complementarity; finally, we briefly review applications thereof to biopharmaceutical products.

Baicalein: A metabolite with promising antineoplastic activity.

Cancer, being a multifactorial disease has diverse presentation in different subgroups which is mainly attributed to heterogenous presentation of tumor cells. This cancer cell heterogeneity is the major reason for variable response to standard chemotherapeutic regimes owing to which high relapse rate and multi-drug resistance has increasingly been reported over the past decade. Interestingly, the research on natural compounds in combination with standard therapies have reported with interesting and promising results from the pre-clinical trials and few of which have also been tested in other phases of clinical trials.

Acute stress induces the rapid and transient induction of caspase-1, gasdermin D and release of constitutive IL-1β protein in dorsal hippocampus.

The proinflammatory cytokine interleukin (IL)-1β plays a pivotal role in the behavioral manifestations (i.e., sickness) of the stress response.

Near-field Terahertz Sensing of HeLa Cells and Based on Monolithic Integrated Metamaterials with a Spintronic Terahertz Emitter.

Label-free biosensors operating within the terahertz (THz) spectra have helped to unlock a myriad of potential THz applications, ranging from biomaterial detection to point-of-care diagnostics. However, the THz wave diffraction limit and the lack of emitter-integrated THz biosensors hinder the proliferation of high-resolution near-field label-free THz biosensing. Here, a monolithic THz emission biosensor (TEB) is achieved for the first time by integrating asymmetric double-split ring resonator metamaterials with a ferromagnetic heterojunction spintronic THz emitter.

Garcinol Exhibits Anti-Neoplastic Effects by Targeting Diverse Oncogenic Factors in Tumor Cells.

Garcinol, a polyisoprenylated benzophenone, is the medicinal component obtained from fruits and leaves of () and has traditionally been extensively used for its antioxidant and anti-inflammatory properties. In addition, it has been also been experimentally illustrated to elicit anti-cancer properties. Several in vitro and in vivo studies have illustrated the potential therapeutic efficiency of garcinol in management of different malignancies.

In vivo intrathecal IL-1β quantification in rats: Monitoring the molecular signals of neuropathic pain.

Background: Neuropathic pain, or pain after nerve injury, is a disorder with a significant reliance on the signalling of cytokines such as IL-1β. However, quantifying the cytokine release repeatedly over time in vivo is technically challenging.

Aim: To evaluate if changes in IL-1β are correlated with the presentation of mechanical allodynia over time, by repeatedly quantifying intrathecal IL-1β concentrations following chronic constriction injury of the sciatic nerve in rats.

Capillary-assisted microfluidic biosensing platform captures single cell secretion dynamics in nanoliter compartments.

Cancer cells continuously secrete inflammatory biomolecules which play significant roles in disease progression and tumor metastasis toward secondary sites. Despite recent efforts to capture cancer cells' intercellular secretion heterogeneity using microfluidics, the challenges in operation of these systems as well as the complexity of designing a biosensing assay for long-term and real-time measurement of single cell secretions have become grand research barriers. Here, we present a new capillary-based microfluidic biosensing approach to easily and reliably capture ~500 single cells inside isolated dead-end nanoliter compartments using simple pipette injection, and quantify their individual secretion dynamics at the single cell resolution over a long period of culture (~16 h).

Biomedical Applications of Integrating Sphere: A Review.

Integrating sphere remains a valuable tool for biomedical optics research. Specifically, it has been used to determine the optical properties (i.e.

NAD Repletion Rescues Female Fertility during Reproductive Aging.

Reproductive aging in female mammals is an irreversible process associated with declining oocyte quality, which is the rate-limiting factor to fertility. Here, we show that this loss of oocyte quality with age accompanies declining levels of the prominent metabolic cofactor nicotinamide adenine dinucleotide (NAD). Treatment with the NAD metabolic precursor nicotinamide mononucleotide (NMN) rejuvenates oocyte quality in aged animals, leading to restoration in fertility, and this can be recapitulated by transgenic overexpression of the NAD-dependent deacylase SIRT2, though deletion of this enzyme does not impair oocyte quality.

A Method for in Vivo Quantification Of Cytokine IL-1β In The Rat Intrathecal Space.

IL-1β is a potent pro-inflammatory cytokine critical to multiple pathologies in the central nervous system (CNS). Quantification of IL-1β in vivo is challenging due to pM range of IL-1β released in the spinal cord and also the terminal nature of cerebrospinal fluid (CSF) sampling in rodents. Herein we developed a robust in vivo device on stainless steel suitable for detection of IL-1β in the spinal cord of rats.

Non-destructive, label free identification of cell cycle phase in cancer cells by multispectral microscopy of autofluorescence.

Background: Cell cycle analysis is important for cancer research. However, available methodologies have drawbacks including limited categorisation and reliance on fixation, staining or transformation. Multispectral analysis of endogenous cell autofluorescence has been shown to be sensitive to changes in cell status and could be applied to the discrimination of cell cycle without these steps.

On-chip structure-switching aptamer-modified magnetic nanobeads for the continuous monitoring of interferon-gamma ex vivo.

Cytokines are cell signaling molecules that indicate the health status of the body. In this study, we developed a microfluidic device integrated with structure-switching aptamers capable of continuously tracking the concentration of the cytokine interferon gamma (IFN-γ) in cell culture medium and blood serum. First, a ferrocene (Fc)-labeled structure-switching signaling aptamer with a hairpin structure targeting IFN-γ was immobilized on magnetic nanobeads by the strongest noncovalent interactions between streptavidin and biotin.

A Nanoparticle-Based Affinity Sensor that Identifies and Selects Highly Cytokine-Secreting Cells.

We developed a universal method termed OnCELISA to detect cytokine secretion from individual cells by applying a capture technology on the cell membrane. OnCELISA uses fluorescent magnetic nanoparticles as assay reporters that enable detection on a single-cell level in microscopy and flow cytometry and fluorimetry in cell ensembles. This system is flexible and can be modified to detect different cytokines from a broad range of cytokine-secreting cells.

Polymer brush based fluorescent immunosensor for direct monitoring of interleukin-1β in rat blood.

A sandwich immunosensor was successfully developed for monitoring of interleukin-1β (IL-1β) in rat whole blood. The substrate stainless steel (SS) was first coated with a polydopamine layer and subsequently grafted with poly(ethylene glycol) methacrylate brushes, onto which a sandwich immunosensor was modified for detection of IL-1β. The device has been successfully applied for monitoring of IL-1β with a limit of detection of 4.

Microfluidic Actuation via 3D-Printed Molds toward Multiplex Biosensing of Cell Apoptosis.

Multiplexed analysis of biochemical analytes such as proteins, enzymes, and immune products using a microfluidic device has the potential to cut assay time, reduce sample volume, realize high-throughput, and decrease experimental error without compromising sensitivity. Despite these huge benefits, the need for expensive specialized equipment and the complex photolithography fabrication process for the multiplexed devices have, to date, prevented widespread adoption of microfluidic systems. Here, we present a simple method to fabricate a new microfluidic-based multiplexed biosensing device by taking advantage of 3D-printing.

Advances in biosensors for the detection of ochratoxin A: Bio-receptors, nanomaterials, and their applications.

Ochratoxin A (OTA) is a class of mycotoxin mainly produced by the genera Aspergillus and Penicillium. OTA can cause various forms of kidney, liver and brain diseases in both humans and animals although trace amount of OTA is normally present in food. Therefore, development of fast and sensitive detection technique is essential for accurate diagnosis of OTA.

CRISPR/Cas Multiplexed Biosensing: A Challenge or an Insurmountable Obstacle?

Performing multiplex detection is still an elusive goal for molecular diagnostics. CRISPR/Cas-based biosensing has demonstrated potential for multiplex detection. Instead of being an insurmountable obstacle, CRISPR/Cas multiplexed biosensing is a realistic challenge with some recent successful applications.

Monitorable Mitochondria-Targeting DNAtrain for Image-Guided Synergistic Cancer Therapy.

It is highly desirable to realize real-time monitoring of the drug delivery/release process in cancer treatment. Herein, a monitorable mitochondria-specific DNAtrain (MitoDNAtrs) was developed for image-guided drug delivery and synergistic cancer therapy. In this system, mitochondria-targeting Cy5.

Signal amplification strategies for paper-based analytical devices.

Paper-based analytical devices (PADs) are very popular for point-of-care diagnostics, which provide a fast, cost-effective and possible multiplexed detection of a spectrum of molecules. They have been matching forward proudly to contribute to the modern analytical science and life science. Accompanying with their advantages and huge potentials, low detection sensitivity is continuing to challenge the application of PADs from bench to bedside.

CRISPR/Cas Systems towards Next-Generation Biosensing.

Beyond its remarkable genome editing ability, the CRISPR/Cas9 effector has also been utilized in biosensing applications. The recent discovery of the collateral RNA cleavage activity of the Cas13a effector has sparked even greater interest in developing novel biosensing technologies for nucleic acid detection and promised significant advances in CRISPR diagnostics. Now, along with the discovery of Cas12 collateral cleavage activities on single-stranded DNA (ssDNA), several CRISPR/Cas systems have been established for detecting various targets, including bacteria, viruses, cancer mutations, and others.

Turn-On Fluorescence Aptasensor on Magnetic Nanobeads for Aflatoxin M1 Detection Based on an Exonuclease III-Assisted Signal Amplification Strategy.

In order to satisfy the need for sensitive detection of Aflatoxin M1 (AFM1), we constructed a simple and signal-on fluorescence aptasensor based on an autocatalytic Exonuclease III (Exo III)-assisted signal amplification strategy. In this sensor, the DNA hybridization on magnetic nanobeads could be triggered by the target AFM1, resulting in the release of a single-stranded DNA to induce an Exo III-assisted signal amplification, in which numerous G-quadruplex structures would be produced and then associated with the fluorescent dye to generate significantly amplified fluorescence signals resulting in the increased sensitivity. Under the optimized conditions, this aptasensor was able to detect AFM1 with a practical detection limit of 9.

Simultaneous voltammetric determination of cadmium(II), lead(II), mercury(II), zinc(II), and copper(II) using a glassy carbon electrode modified with magnetite (FeO) nanoparticles and fluorinated multiwalled carbon nanotubes.

A method is described for the simultaneous voltammetric determination of the heavy metal ions cadmium(II), lead(II), mercury(II), zinc(II), and copper(II) using a glassy carbon electrode (GCE) modified with magnetite (FeO) nanoparticles and fluorinated multiwalled carbon nanotubes (FeO/F-MWCNTs). The FeO/F-MWCNT composite was synthesized by a hydrothermal method and characterized by X-ray photoelectron spectroscopy, X-ray diffraction, transmission electron microscopy, scanning electron microscopy, elemental mapping, electrochemical impedance spectroscopy, and square wave stripping voltammetry. Under the optimum conditions, the electrode displays excellent response to the ions.

Application of CdTe/CdS/ZnS quantum dot in immunoassay for aflatoxin B1 and molecular modeling of antibody recognition.

In order to develop a sensitive immunoassay for Aflatoxin B1 (AFB1) monitoring, a hybridoma secreting anti-AFB1 monoclonal antibody with high binding affinity was screened. A new type of CdTe/CdS/ZnS quantum dot was synthesized and conjugated with an artificial antigen for use as a fluorescent probe in a simple one-step fluorescence immunoassay (FLISA). The developed FLISA allowed a sensitive determination of AFB1 in cereal samples in a wide linear range, from 0.

Graphene Oxide Based Recyclable in Vivo Device for Amperometric Monitoring of Interferon-γ in Inflammatory Mice.

Cytokine sensing is challenging due to their typically low abundances in physiological conditions. Nanomaterial fabricated interfaces demonstrated unique advantages in ultrasensitive sensing. Here, we demonstrate an amperometric sensing device based on graphene oxide (GO) and structure-switching aptamers for long-term detection of cytokines in a living organism.