Biology subjects

Garcia-Cordero, J.

Publications and source records attributed to Garcia-Cordero, J..

2 recordsLinked to original sources

bioRxiv2025

Integrating microfluidic automation into thermoplastic devices for analysis of small volumes of blood

Microfluidic devices hold considerable promise for miniaturizing and automating laboratory scale workflows involving sample preparation and biomarker analysis. By-and-large microfluidic automation has been implemented using polydimethyl siloxane (PDMS) - a material that presents challenges for scalable manufacturing. In this paper, we fabricated an automated microfluidic device composed of thermoplastic material, poly (methyl methacrylate) (PMMA), and demonstrated a sophisticated workflow consisting of plasma extraction, sample aliquoting and sample-reagent mixing steps performed in the device. Automation was accomplished using multiple on-board microvalves consisting of a pressure sensitive adhesive (PSA) membrane sandwiched between PMMA flow and control layers. In addition to novel microvalves, our device included features designed to address challenges arising from the gas impermeable nature of PMMA. These features included a bubble trap and air micro-tanks to supply O2 for enzymatic reactions. We rigorously characterized the performance of the microfluidic device and demonstrated that the quality of plasma extracted in the device is comparable to a standard plasma isolation method involving centrifugation. We also showed close agreement between glucose levels measured in the device and by standard assay. As a final demonstration of utility, we loaded 5 {micro}L of blood, isolated plasma and detected three analytes simultaneously - glucose, lactate and H2O2. In addition to minimizing the volume of blood and eliminating standard laboratory equipment (e.g., centrifuge), our device enabled a rapid response (<10 min). The use of thermoplastic materials may pave the way for inexpensive, mass-produced automated microfluidic devices for rapid detection of blood biomarkers at the point of care.

bioengineering↗

bioRxiv2021

ADAM17 inhibition prevents neutrophilia and lung injury in a mouse model of Covid-19

Severe coronavirus disease 2019 (Covid-19) is characterized by lung injury, cytokine storm and increased neutrophil-to-lymphocyte ratio (NLR). Current therapies focus on reducing viral replication and inflammatory responses, but no specific treatment exists to prevent the development of severe Covid-19 in infected individuals. Angiotensin-converting enzyme-2 ACE-2) is the receptor for SARS-CoV-2, the virus causing Covid-19, but it is also critical for maintaining the correct functionality of lung epithelium and endothelium. Coronaviruses induce activation of a disintegrin and metalloprotease 17 (ADAM17) and shedding of ACE-2 from the cell surface resulting in exacerbated inflammatory responses. Thus, we hypothesized that ADAM17 inhibition ameliorates Covid-19-related lung inflammation. We employed a pre-clinical mouse model using intra-tracheal instillation of a combination of polyinosinic:polycytidylic acid (poly-I:C) and the receptor-binding domain of the SARS-CoV-2 spike protein (RBD-S) to mimic lung damage associated with Covid-19. Histological analysis of inflamed mice confirmed the expected signs of lung injury including edema, fibrosis, vascular congestion and leukocyte infiltration. Moreover, inflamed mice also showed an increased NLR as observed in critically ill Covid-19 patients. Administration of the ADAM17 inhibitors apratastat and TMI-1 significantly improved lung histology and prevented leukocyte infiltration. Reduced leukocyte recruitment could be explained by reduced production of pro-inflammatory cytokines and lower levels of the endothelial adhesion molecules ICAM-1 and VCAM-1. Additionally, the NLR was significantly reduced by ADAM17 inhibition. Thus, we propose inhibition of ADAM17 as a novel promising treatment strategy in SARS-CoV-2-infected individuals to prevent the progression towards severe Covid-19.

pathology↗