Biology subjects

Matos, T.

Publications and source records attributed to Matos, T..

2 recordsLinked to original sources

bioRxiv2024

Real-time monitoring of attenuated cytomegalovirus using Raman spectroscopy allows non-destructive characterization during flow

Real-time monitoring of viral particles can have a crucial impact on vaccine manufacturing and can alleviate public health by supporting continuous supply. Spectroscopic methods such as Raman spectroscopy can provide rapid and non-invasive measurements. Here, we have developed a Raman spectroscopy-based tool to monitor the quality and quantity of viral particles in a continuous flow set-up. The attenuated human cytomegalovirus (CMV) is characterized across a wide range of concentrations (4.50 x 109 to 2.90 x 1011 particles/mL) and flow rates (100 {micro}m/s to 1000 {micro}m/s) within a square quartz capillary. This process analytical technology (PAT) tool enables the detection of viral particles even at high flow rates such as 1000 {micro}m/s. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and dynamic light scattering (DLS) demonstrated that the samples maintain their integrity even after laser exposure, reiterating the non-invasive nature of Raman spectroscopy. To the best of our knowledge, this is the first report on characterizing CMV particles using Raman spectroscopy. We have also demonstrated the limit of detection (LoD) (2.36 x 1010 particles/mL) for CMV particles in continuous flow (via the Raman spectroscopy method) by addressing the effect of flow rate, concentration, and integrity of samples. This technology could enhance our understanding of the quality control in bio-manufacturing processes required in vaccine production.

bioengineering↗

bioRxiv2021

A tripartite structure, the complex nuclear receptor element (cNRE), is a cis-regulatory module of viral origin required for atrial chamber preferential gene expression

Optimal cardiac function requires appropriate contractile proteins in each heart chamber. Atria require slow myosins to act as variable reservoirs, while ventricles demand fast myosin for swift pumping functions. Hence, myosin is under chamber-biased cis-regulatory control to achieve this functional distribution. Failure in proper regulation of myosin genes can lead to severe congenital heart dysfunction. The precise regulatory input leading to cardiac chamber-biased expression remains uncharted. To address this, we computationally and molecularly dissected the quail Slow Myosin Heavy Chain III (SMyHC III) promoter that drives specific gene expression to the atria to uncover the regulatory information leading to chamber expression and understand their evolutionary origins. We show that SMyHC III gene states are autonomously orchestrated by a complex nuclear receptor cis-regulatory element (cNRE), a 32- bp sequence with hexanucleotide binding repeats. Using in vivo transgenic assays in zebrafish and mouse models, we demonstrate that preferential atrial expression is achieved by the combinatorial regulatory input composed of atrial activation motifs and ventricular repression motifs. Through comparative genomics, we provide evidence that the cNRE emerged from an endogenous viral element, most likely through infection of an ancestral host germline. Our study reveals an evolutionary pathway to cardiac chamber-specific expression.

developmental biology↗