Novel Concept of Micro Patterned Micro Titer Plates Fabricated via UV-NIL for Automated Neuronal Cell Assay Read-Out

The UV-nanoimprint lithography(UV-NIL) fabrication of a novel network of micron-sized channels, forming an open channel microfluidic system is described. Details about the complete manufacturing process, from mastering to fabrication in small batches and in high throughput with up to 1200 micro titer plates per hour is presented. Deep insight into the evaluation of a suitable UV-curable material, mr-UVCur26SF is given, presenting cytotoxic evaluation, cell compatibility tests and finally a neuronal assay. The results indicate how the given pattern, in combination with the resist, paves the way to faster, cheaper, and more reliable drug screening.This project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement NO. 646260. The research was also partially supported by NextGenMicrofluidics project under HORIZON2020 with grant agreement no 862092

Roll-to-Roll pilot line for large-scale manufacturing of microfluidic devices

Roll-to-roll (R2R) technologies with roller-based nanoimprinting methods enable manufacturing of highly cost-effective and large-scale sheets of flexible polymer film with precise structures on a micro- and nanoscale 1. Areas that can benefit strongly from such large scale technologies are microfluidics, biosensors, and lab-on-chip products for point of care diagnostics, drug discovery and food control. Here, R2R fabrication could greatly reduce production costs and increase manufacturing capacity with respect to currently used products. A pilot line with this technology is investigated in the European Horizon 2020 project R2R Biofluidics and its capabilities are tested on two Demonstrators: - Demonstrator 1: In-vitro diagnostic chip with imprinted microfluidic channels based on optical chemiluminescence measurement by photodetectors. - Demonstrator 2: Neuronal cell culture plate with imprinted cavities and channels for controlled culturing and fluorescence imaging of neurons, for high throughput drug screening. Please click Additional Files below to see the full abstract

Activity of bisnaphthalimidopropyl derivatives against trypanosoma brucei.

Current treatments for African trypanosomiasis are either toxic, costly, difficult to administer, or prone to elicit resistance. This study evaluated the activity of bisnaphthalimidopropyl (BNIP) derivatives against Trypanosoma brucei. BNIPDiaminobutane (BNIPDabut), the most active of these compounds, showed in vitro inhibition in the single-unit nanomolar range, similar to the activity in the reference drug pentamidine, and presented low toxicity and adequate metabolic stability. Additionally, using a murine model of acute infection and live imaging, a significant decrease in parasite load in BNIPDabut-treated mice was observed. However, cure was not achieved. BNIPDabut constitutes a new scaffold for antitrypanosomal drugs that deserves further consideration

Multicolor fluorescent biosensors for multiplexed detection of GPCR receptor activation

We describe a new live-cell assay that uses fluorescent biosensors co-expressed with several GPCRs for measuring the second messenger concentration changes that occurs after GPCR activation. The molecular structure of the Biosensors comprises: a membrane localization peptide, a second messenger transduction protein binding peptide, a reticulum retention signal and a fluorescent peptide. The biosensors are normally localized in the Plasma Membrane but an increase in the second messenger concentration leads to a change in the structural folding of the Biosensor that promotes its vesicularization. The second messenger transduction protein binding peptide could be replaced depending on the second messenger, resulting three different versions of the biosensors: cAMP; Calcium and DAG Nomad Biosensor. In addition, we also present here the development of multiplexed assays using the different versions of the Biosensor enabling the analysis both different intracellular signaling and receptor internalization, simultaneously.<br

Myocardial fibrosis : biomedical research from bench to bedside

Myocardial fibrosis refers to a variety of quantitative and qualitative changes in the interstitial myocardial collagen network that occur in response to cardiac ischaemic insults, systemic diseases, drugs, or any other harmful stimulus affecting the circulatory system or the heart itself. Myocardial fibrosis alters the architecture of the myocardium, facilitating the development of cardiac dysfunction, also inducing arrhythmias, influencing the clinical course and outcome of heart failure patients. Focusing on myocardial fibrosis may potentially improve patient care through the targeted diagnosis and treatment of emerging fibrotic pathways. The European Commission funded the FIBROTARGETS consortium as a multinational academic and industrial consortium with the primary aim of performing a systematic and collaborative search of targets of myocardial fibrosis, and then translating these mechanisms into individualized diagnostic tools and specific therapeutic pharmacological options for heart failure. This review focuses on those methodological and technological aspects considered and developed by the consortium to facilitate the transfer of the new mechanistic knowledge on myocardial fibrosis into potential biomedical applications.(VLID)483915

Antifibrotic effect of novel neutrophil gelatinase-associated lipocalin inhibitors in cardiac and renal disease models

International audienceNeutrophil gelatinase-associated lipocalin (NGAL) is involved in cardiovascular and renal diseases. Gene inactivation of NGAL blunts the pathophysiological consequences of cardiovascular and renal damage. We aimed to design chemical NGAL inhibitors and investigate its effects in experimental models of myocardial infarction (MI) and chronic kidney disease induced by 5/6 nephrectomy (CKD) on respectively 8-12 weeks old C57Bl6/j and FVB/N male mice. Among the 32 NGAL inhibitors tested, GPZ614741 and GPZ058225 fully blocked NGAL-induced inflammatory and profibrotic markers in human cardiac fibroblasts and primary mouse kidney fibroblasts. The administration of GPZ614741 (100 mg/kg/day) for three months, was able to improve cardiac function in MI mice and reduced myocardial fibrosis and inflammation. The administration of GPZ614741 (100 mg/kg/day) for two months resulting to no renal function improvement but prevented the increase in blood pressure, renal tubulointerstitial fibrosis and profibrotic marker expression in CKD mice. In conclusion, we have identified new compounds with potent inhibitory activity on NGAL-profibrotic and pro-inflammatory effects. GPZ614741 prevented interstitial fibrosis and dysfunction associated with MI, as well as tubulointerstitial fibrosis in a CKD model. These inhibitors could be used for other diseases that involve NGAL, such as cancer or metabolic diseases, creating new therapeutic options