Advanced Optogenetic-Based Biosensing and Related Biomaterials

The ability to stimulate mammalian cells with light, brought along by optogenetic control, has significantly broadened our understanding of electrically excitable tissues. Backed by advanced (bio)materials, it has recently paved the way towards novel biosensing concepts supporting bio-analytics applications transversal to the main biomedical stream. The advancements concerning enabling biomaterials and related novel biosensing concepts involving optogenetics are reviewed with particular focus on the use of engineered cells for cell-based sensing platforms and the available toolbox (from mere actuators and reporters to novel multifunctional opto-chemogenetic tools) for optogenetic-enabled real-time cellular diagnostics and biosensor development. The key advantages of these modified cell-based biosensors concern both significantly faster (minutes instead of hours) and higher sensitivity detection of low concentrations of bioactive/toxic analytes (below the threshold concentrations in classical cellular sensors) as well as improved standardization as warranted by unified analytic platforms. These novel multimodal functional electro-optical label-free assays are reviewed among the key elements for optogenetic-based biosensing standardization. This focused review is a potential guide for materials researchers interested in biosensing based on light-responsive biomaterials and related analytic tools.

Development of a Tri-Functional Nanoprobe for Background-Free SERS Detection of Sialic Acid on the Cell Surface

Sialic acid (SA) on the surface of cells is indispensable in numerous physiological and pathological processes, and sensitive and reproducible detection of SA is crucial for diagnosis and therapy in many diseases. Here, we developed a tri-functional nanoprobe as a sensitive and straightforward surface-enhanced Raman spectroscopy (SERS) nanoprobe for sialoglycan detection on cell surfaces. The reporter was designed to provide three key functionalities that make it ideal for SA detection. First, we employed two recognition groups, phenylboronic acid and an ammonium group, that enhance SA recognition and capture efficiency. Second, we used cyano as the Raman reporter because it emits in the cellular Raman silent region. Finally, thiol acted as an anchoring agent to conjugate the reporter to silver nanocubes to provide SERS enhancement. Our molecular nanoprobe design demonstrated the ability to detect SA on the cell surface with high sensitivity and spatial resolution, opening up new routes to cellular diagnostics.

THE DIFFERENT SCREENING METHODS FOR THE CERVICAL LESION DIAGNOSTIC AND THE ECONOMIC AND SOCIAL IMPLICATIONS OF SCREENING AMONG FEMALE POPULATION

"The study aims to compare different screening methods that are currently being used to confirm a cervical intraepithelial lesion (CIN) underlying the inherent advantages of the varied examination procedures. In this study we are looking at the quality contrast of the different paraclinical examinations relative to the cost, invasiveness and cultural acceptance of such procedures. Every year, more than 100,000 women in EU countries are diagnosed with cervix uteri cancers (CCU). This type of cancer can be prevented if precancerous cells are detected and treated. HPV is found in over 90% of cervix uteri cancers. In Europe, more than half of the countries have implemented screening programs for cervical cancer, and most European countries now have national HPV vaccination programs, however target populations vary depending on the epidemiological evidence and the budgetary level of each countries health system. In this study we evaluated and compared the current and modern techniques used for cervix cellular diagnostics. Also, this study helps improve the understanding on the economic and social implications of screening among female population and the impact it has on healthcare system relieving. Romania recorded an incidence of 22.6 cases of cervix uteri cancers / 100.000 women (age standardized rate), and a mortality of 9.6 / 100.000, ranking second highest in the EU zone."

The Artificial Intelligence in Digital Pathology and Digital Radiology: Where Are We?

Thanks to the incredible changes promoted by Information and Communication Technology (ICT) conveyed today by electronic-health (eHealth) and mobile-health (mHealth), many new applications of both organ and cellular diagnostics are now possible [...]

Nanobodies Right in the Middle: Intrabodies as Toolbox to Visualize and Modulate Antigens in the Living Cell

In biomedical research, there is an ongoing demand for new technologies to elucidate disease mechanisms and develop novel therapeutics. This requires comprehensive understanding of cellular processes and their pathophysiology based on reliable information on abundance, localization, post-translational modifications and dynamic interactions of cellular components. Traceable intracellular binding molecules provide new opportunities for real-time cellular diagnostics. Most prominently, intrabodies derived from antibody fragments of heavy-chain only antibodies of camelids (nanobodies) have emerged as highly versatile and attractive probes to study and manipulate antigens within the context of living cells. In this review, we provide an overview on the selection, delivery and usage of intrabodies to visualize and monitor cellular antigens in living cells and organisms. Additionally, we summarize recent advances in the development of intrabodies as cellular biosensors and their application to manipulate disease-related cellular processes. Finally, we highlight switchable intrabodies, which open entirely new possibilities for real-time cell-based diagnostics including live-cell imaging, target validation and generation of precisely controllable binding reagents for future therapeutic applications.