Pneumococcal Septicemia after COVID-19 in Teenager: Clinical Case

There is a hypothesis among the scientists that COVID-19 and SARS-CoV-2 have negative effect on the immune system. Many reports on multisystem inflammatory syndrome (MIS) development after COVID-19 are published worldwide. Whereas, the impact of this infection on prevalence and course of any other infectious diseases has not been studied. The aim of the study is to draw attention to the estimation of other infectious diseases course after COVID-19 by analyzing the clinical case of pneumococcal septicemia in 17 years old boy who has undergone COVID-19. We obtained data about COVID-19 during the analysis of the child's medical history. The course of infectious process was similar to the course of multisystem inflammatory syndrome in children (MIS-C), compared by clinical manifestations. The results of ELISA blood assay have proved it: we revealed IgG to SARS-CoV-2. However, PCR test revealed S. pneumoniae DNA in blood, so MIS-C diagnosis was excluded. The role of SARS-CoV-2 in other diseases in postcovid period can not be excluded, and it requires further studies. Development of screening and rehabilitation principles is recommended despite the severity of COVID-19 in children.

Characteristics of routine coagulationtesting and thromboelastometry in polytrauma patients at admission

Background: polytrauma is one of the emergency surgeries with a high mortality rate. One of the leading causes of death is coagulopathy that is not detected early and treated promptly. Thromboelastometry (ROTEM) is a whole blood assay that evaluates the viscoelastic properties during clot formation and clot lysis. This method can detect coagulopathy rapidly and accurately, thereby improving the management of bleeding after trauma. Objectives: describing and evaluating the correlation between ROTEM parameters and routine coagulation tests in polytrauma patients at admission. Method: 110 patients admitted to the Emergency Department, Viet Duc University Hospital from May 2021 to July 2021 were diagnosed with polytrauma. All patients underwent routine coagulation testing and ROTEM parameters at admission. Result: the average age of patients is 41.4±14.7 years old, men accounts for 77.3%, average ISS score is 24.5±6.3. The proportion of the polytrauma patients with coagulopathy by routine coagulation testing was 50.9%. A significant correlation was found between routine coagulation parameters and ROTEM: between APTT with CFT-INTEM (r=0.65; p<0.01); between PT and CFT-EXTEM (r=0.64; p<0.01); between platelet count and MCF-INTEM (r=0.56; p=0.00) and MCF-EXTEM (r=0.57; p=0.00); between fibrinogen level and MCF-INTEM (r=0.71; p=0.00), MCF-EXTEM (r=0.71; p=0.00), and MCF-FIBTEM (r=0.91; p=0.00). Conclusion: proportion of the polytrauma patients with coagulopathy by routine coagulation testing was 50.9%. A significant correlation was found between routine coagulation parameters and ROTEM

A Novel Liquid Biopsy (NETest) identifies Paragangliomas and Pheochromocytomas with High Accuracy

Pheochromocytomas and paragangliomas (PHEOs/PGLs) represent diagnostically challenging and complex neuroendocrine tumors (NETs). Current biomarker tests for PHEOs/PGLs are technically complex or limited. We assessed the diagnostic utility of a NET-specific 51-marker gene blood assay (NETest) in patients with PHEOs/PGLs (n=81), including 10 pediatric patients, and age-/gender-matched controls (n=142) using a prospective case:control (1:2) analysis. mRNA was measured (qPCR) and results scaled 0-100 (ULN<20). Receiver Operating Curve (ROC) and non-parametric (Mann-Whitney) were used for analyses (2-tailed). All data is presented as mean±SEM. NETest accuracy for PHEO/PGL diagnosis was 100%. PHEO/PGL scores were 70±3 versus 8.5±1 in controls (p<0.0001) and ROC analysis was 0.99±0.004 (p<0.0001). Diagnostic metrics were 94% accurate, 100% sensitive, and 92% specific. Imaging correlation with 68Ga-PET-SSA was 100%. NETest levels in PHEOs (n=26) were significantly (p<0.0001) elevated (83±4) versus 66±4 in PGLs (n=40) and mixed PHEOs/PGLs (n=5: 37±3). Adrenal-derived tumors (n=30) exhibited higher scores (76±5) than extra-adrenal-derived tumors (66±4, p<0.05). Cluster 2 tumors exhibited significantly (p=0.034) elevated NETest levels (n=4: 92±2) versus Cluster 1 tumors (n=35: 69±4). Regulatory pathway analysis identified elevated RAS-RAF, metastatic, pluripotential, neural and secretory gene cluster levels (p<0.05) in PHEOs compared to PGLs. Cluster 2 PPGLs exhibited elevated (p=0.046) levels of growth-factor signaling genes compared to cluster 1. The PHEOs/PGLs in the pediatric cohort (n=10) were all NETest-positive (81±8) and exhibited a gene expression profile spectrum analogous to adults. Circulating NET transcript analysis identifies PHEOs/PGLs with 100% efficacy and is likely to have clinical utility in the diagnosis and management of PHEO/PGL patients.

Development of an Improved Whole Blood Assay for Diagnosis of Latent and Active Tuberculosis Cases

Background: Latent TB infection (LTBI) is an infection where the presence of disease causing organism M. tuberculosis is there without any sign and symptoms of the disease hence mostly remains undiagnosed, though Tuberculin skin test (TST) and Interferon Gamma Release Assay (IGRA) were used to diagnose the LTBI. They have their limitations, TST gives major cross-reactivity with BCG vaccine and gives inaccurate results in individuals who have taken BCG and IGRA are very costly and variable sensitivity is repeated in various populations hence the modifications are needed in the IGRA for proper diagnosis of LTBI. Objectives: In the proposed study we aimed to develop an improved whole blood assay towards a diagnosis of latent and active TB infection as an alternative to the Quantiferon QFT assay Methodology: Synthetic antigenic peptides against latency specific antigens will be designed and synthesized. Designed peptides will be screened for LTBI specific cytokine by in-vitro experiments. Development & production of Whole assay using selected peptides evaluation of developed assay through ELISA in clinical samples. Expected Results: Latent specific peptides will be identified and peptide-based whole blood assay for detection and diagnosis of tuberculosis will be developed as an indigenous alternative for the existing QFT assay. Conclusion: An improved whole blood assay will be developed for screening of LTBI in the Indian population.

Proof of Concept of a Binary Blood Assay for Predicting Radiosensitivity

Radiation therapy (RT), either alone or in combination with surgery and/or chemotherapy is a keystone of cancers treatment. Early toxicity is common, sometimes leading to discontinuation of treatment. Recent studies stressed the role of the phosphorylated ATM (pATM) protein in RT-toxicity genesis and its ability in predicting individual radiosensitivity (IRS) in fibroblasts. Here we assessed the reliability of the pATM quantification in lymphocytes to predict IRS. A first retrospective study was performed on 150 blood lymphocytes of patients with several cancer types. Patients were divided into 2 groups, according to the grade of experienced toxicity. The global quantity of pATM molecules was assessed by ELISA on lymphocytes to determine the best threshold value. Then, the binary assay was assessed on a validation cohort of 36 patients with head and neck cancers. The quantity of pATM molecules in each sample of the training cohort was found in agreement with the observed Common Terminology Criteria for Adverse Events (CTCAE) grades with an AUC = 0.71 alone and of 0.77 combined to chemotherapy information. In the validation cohort, the same test was conducted with the following performances: sensitivity = 0.84, specificity = 0.54, AUC = 0.70 and 0.72 combined to chemotherapy. This study provides the basis of an easy to perform assay for clinical use.

Ex Vivo Antioxidant Capacities of Fruit and Vegetable Juices. Potential In Vivo Extrapolation

Background: In support of claims that their products have antioxidant properties, the food industry and dietary supplement manufacturers rely solely on the in vitro determination of the ORAC (oxygen radical antioxidant capacity) value, despite its acknowledged lack of any in vivo relevance. It thus appears necessary to use tests exploiting biological materials (blood, white blood cells) capable of producing physiological free radicals, in order to evaluate more adequately the antioxidant capacities of foods such as fruit and vegetable juices. Materials: Two approaches to assessing the antioxidant capacities of 21 commercial fruit and vegetable juices were compared: the ORAC assay and the “PMA–whole blood assay,” which uses whole blood stimulated by phorbol myristate acetate to produce the superoxide anion. We described in another paper the total polyphenol contents (TPCs) and individual phenolic compound contents of all the juices investigated here (Matute et al. Antioxidants 2020, 9, 1–18). Results: Ranking of the juices from highest to lowest antioxidant capacity differed considerably according to the test used, so there was no correlation (r = 0.33, p = 0.13) between the two assays when considering all juices. Although the results of the ORAC assay correlated positively with TPC (r = 0.50, p = 0.02), a much stronger correlation (r = 0.70, p = 0.004) emerged between TPC and % superoxide anion inhibition. In the PMA–whole blood assay, peonidin-3-O-glucoside, epigallocatechin gallate, catechin, and quercetin present in juices were found to inhibit superoxide anion production at concentrations below 1 µM, with a strong positive correlation. Conclusions: Associated with the determination of total and individual phenolic compounds contained in fruit and vegetable juices, the PMA–whole blood assay appears better than the ORAC assay for evaluating juice antioxidant capacity.

Stimulated Immune Response by TruCulture® Whole Blood Assay in Patients With European Lyme Neuroborreliosis: A Prospective Cohort Study

IntroductionBorrelia burgdorferi sensu lato complex (B. burgdorferi) can cause a variety of clinical manifestations including Lyme neuroborreliosis. Following the tick-borne transmission, B. burgdorferi initially evade immune responses, later symptomatic infection is associated with occurrence of specific antibody responses. We hypothesized that B. burgdorferi induce immune hyporesponsiveness or immune suppression and aimed to investigate patients with Lyme neuroborreliosis ability to respond to immune stimulation.MethodsAn observational cohort study investigating the stimulated immune response by standardized whole blood assay (TruCulture®) in adult patients with Lyme neuroborreliosis included at time of diagnosis from 01.09.2018-31.07.2020. Reference intervals were based on a 5-95% range of cytokine concentrations from healthy individuals (n = 32). Patients with Lyme neuroborreliosis and references were compared using Mann-Whitney U test. Heatmaps of cytokine responses were generated using the webtool Clustvis.ResultsIn total, 22 patients with Lyme neuroborreliosis (19 definite, 3 probable) were included. In the unstimulated samples, the concentrations of cytokines in patients with Lyme neuroborreliosis were comparable with references, except interferon (IFN)-α, interleukin (IL)-17A, IL-1β and IL-8, which were all significantly below the references. Patients with Lyme neuroborreliosis had similar concentrations of most cytokines in all stimulations compared with references. IFN-α, IFN-γ, IL-12 and IL-17A were lower than references in multiple stimulations.ConclusionIn this exploratory cohort study, we found lower or similar concentrations of circulating cytokines in blood from patients with Lyme neuroborreliosis at time of diagnosis compared with references. The stimulated cytokine release in blood from patients with Lyme neuroborreliosis was in general slightly lower than in the references. Specific patterns of low IL-12 and IFN-γ indicated low Th1-response and low concentrations of IL-17A did not support a strong Th17 response. Our results suggest that patients with Lyme neuroborreliosis elicit a slightly suppressed or impaired immune response for the investigated stimulations, however, whether the response normalizes remains unanswered.

A novel whole-blood stimulation assay to detect and quantify memory T-cells in COVID-19 patients

SARS-CoV-2 specific T-cells responses are essential for virus clearance. We present a novel and simple whole-blood assay allowing the detection of interferon-gamma-producing antiviral T-cells following peptide stimulation. We show that unlike neutralizing antibodies, antiviral memory T-cells persist at least 6 months in convalescent Covid-19 individuals.