Plasma cell-free DNA promise monitoring and tissue injury assessment of COVID-19.

Coronavirus 2019 (COVID-19) is a complex disease that affects billions of people worldwide. Currently, effective etiological treatment of COVID-19 is still lacking; COVID-19 also causes damages to various organs that affects therapeutics and mortality of the patients. Surveillance of the treatment responses and organ injury assessment of COVID-19 patients are of high clinical value. In this study, we investigated the characteristic fragmentation patterns and explored the potential in tissue injury assessment of plasma cell-free DNA in COVID-19 patients. Through recruitment of 37 COVID-19 patients, 32 controls and analysis of 208 blood samples upon diagnosis and during treatment, we report gross abnormalities in cfDNA of COVID-19 patients, including elevated GC content, altered molecule size and end motif patterns. More importantly, such cfDNA fragmentation characteristics reflect patient-specific physiological changes during treatment. Further analysis on cfDNA tissue-of-origin tracing reveals frequent tissue injuries in COVID-19 patients, which is supported by clinical diagnoses. Hence, our work demonstrates and extends the translational merit of cfDNA fragmentation pattern as valuable analyte for effective treatment monitoring, as well as tissue injury assessment in COVID-19.

Signal Quality Assessment Model for Wearable EEG Sensor on Prediction of Mental Stress.

Electroencephalogram (EEG) plays an important role in E-healthcare systems, especially in the mental healthcare area, where constant and unobtrusive monitoring is desirable. In the context of OPTIMI project, a novel, low cost, and light weight wearable EEG sensor has been designed and produced. In order to improve the performance and reliability of EEG sensors in real-life settings, we propose a method to evaluate the quality of EEG signals, based on which users can easily adjust the connection between electrodes and their skin. Our method helps to filter invalid EEG data from personal trials in both domestic and office settings. We then apply an algorithm based on Discrete Wavelet Transformation (DWT) and Adaptive Noise Cancellation (ANC) which has been designed to remove ocular artifacts (OA) from the EEG signal. DWT is applied to obtain a reconstructed OA signal as a reference while ANC, based on recursive least squares, is used to remove the OA from the original EEG data. The newly produced sensors were tested and deployed within the OPTIMI framework for chronic stress detection. EEG nonlinear dynamics features and frontal asymmetry of theta, alpha, and beta bands have been selected as biological indicators for chronic stress, showing relative greater right anterior EEG data activity in stressful individuals. Evaluation results demonstrate that our EEG sensor and data processing algorithms have successfully addressed the requirements and challenges of a portable system for patient monitoring, as envisioned by the EU OPTIMI project.

An alternative approach to synthesize cDNA bypassing traditional reverse transcription.

cDNAs of certain target genes are difficult to obtain by traditional reverse transcription. Herein we describe a novel method to synthesize cDNA based upon the use of the class IIS restriction enzymes. Briefly, the exons of a certain gene are separately PCR-amplified, each using the primers containing a recognition sequence of a certain class IIS restriction enzyme. All the fragments are restricted using the enzyme(s), resulting in the cohesive end of each exon that is complementary to the one in its adjacent exon. Then the fragments can be assembled together in their naturally occurring order. We successfully applied this method to acquire the coding sequence of Hoxa7 gene. This approach is simple, highly efficient, less error prone and cost-effective, and can also be used to fuse different PCR-fragments from distinct genes to create a chimeric gene or to perform site-directed mutagenesis.

Cytokine production following experimental implantation of xenogenic dermal collagen and polypropylene grafts in mice.

AIM: We earlier showed that xenogenic Pelvicol (Bard, Olen, Belgium) implants induce a lesser inflammatory response than Prolene (Johnson and Johnson, Dilbeek, Belgium). The purpose of this study was to determine cytokine profiles in the host immune responses to Pelvicol in a mouse model. The hypothesis was that Pelvicol would induce a "T-helper2" (Th2) rather than T-helper1 (Th1) type of inflammatory response. METHODS: Mice were implanted subcutaneously with Pelvicol or Prolene and the graft sites were harvested at 3 to 28 days. Histopathology was done and cytokine levels were determined by immunohistochemistry and RT-PCR. Flow cytometry was used to identify which cell population contributed to the observed cytokine production profiles. RESULTS: Pelvicol induced a decreased inflammation and displayed an increase in IL-10 and TGF-beta, but reduce of TNF-alpha and IFN-gamma, indicating a Th2 type dominated response as examined by immunohistochemistry and RT-PCR. Flow cytometry showed that the monocytes/maceophages were the main cell population responsible for production of these cytokines. Monocytes/maceophages from Pelvicol explants showed upregulated expression of IL-10 while Prolene explants expressed TNF-alpha. CONCLUSION: Pelvicol induced a Th2 type cytokine-dominated immune response after subcutaneous implantation in mice.