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Today, we heavily rely on technology and increasingly utilize it to monitor our own health. The identification of sensitive, accurate biosensors that are capable of real-time cortisol analysis is one important potential feature for these technologies to aid us in the maintenance of our physical and mental wellbeing. Detection and quantification of cortisol, a well-known stress biomarker present in sweat, offers a noninvasive and potentially real-time method for monitoring anxiety. Molecularly imprinted polymers are attractive candidates for cortisol recognition elements in such devices as they can selectively rebind a targeted template molecule. However, mechanisms of imprinting and subsequent rebinding depend on the choice and composition of the prepolymerization mixture where the molecular interactions between the template, functional monomer, cross-linker, and solvent molecules are not fully understood. Here, we report the synthesis and evaluation of a molecularly imprinted polymer selective for cortisol detection. Molecular dynamics simulations were used to investigate the interactions between all components in the prepolymerization mixture of the as-synthesized molecularly imprinted polymer. Varying the component ratio of the prepolymerization mixture indicates that the number of cross-linker molecules relative to the template impacts the quality of imprinting. It was determined that a component ratio of 1:6:30 of cortisol, methacrylic acid, and ethylene glycol dimethacrylate, respectively, yields the optimal theoretical complexation of cortisol for the polymeric systems investigated. Experimental synthesis and rebinding results demonstrate an imprinting factor of up to 6.45. The trends in cortisol affinity predicted by molecular dynamics simulations of the prepolymerization mixture were also corroborated through experimental analysis of those modeled molecularly imprinted compositions, demonstrating the predictive capabilities of these simulations.

COVID-19 patients elicit strong responses to the nucleocapsid (N) protein of SARS-CoV-2 but binding antibodies are also detected in prepandemic individuals, indicating potential crossreactivity with common cold human coronaviruses (HCoV) and questioning its utility in seroprevalence studies. We investigated the immunogenicity of the full-length and shorter fragments of the SARS-CoV-2 N protein, and the crossreactivity of antibodies with HCoV. We identified a C-terminus region in SARS-CoV2 N of minimal sequence homology with HCoV that was more specific for SARS-CoV-2 and highly immunogenic. IgGs to the full-length SARS-CoV-2 N also recognized N229E N, and IgGs to HKU1 N recognized SARS-CoV-2 N. Crossreactivity with SARS-CoV-2 was stronger for alpha- rather than beta-HCoV despite having less sequence identity, revealing the importance of conformational recognition. Higher preexisting IgG to OC43 N correlated with lower IgG to SARS-CoV-2 N in rRT-PCR negative individuals, reflecting less exposure and indicating a potential protective association. Antibodies to SARS-CoV-2 N were higher in patients with more severe and longer duration of symptoms and in females. IgGs remained stable for at least 3 months, while IgAs and IgMs declined faster. In conclusion, N protein is a primary target of SARS-CoV-2-specific and HCoV crossreactive antibodies, both of which may affect the acquisition of immunity to COVID-19.

Although some evidence showed the activation of complement systems in COVID-19 patients, proinflammatory status and lectin pathway remain unclear. Thus, the present study aimed to demonstrate the role of MBL and ficolin-3 in the complement system activation and compared to pandemic Influenza A virus H1N1 subtype infection (H1N1pdm09) and control patients. A total of 27 lungs formalin-fixed paraffin-embedded samples (10 from H1N1 group, 6 from the COVID-19 group, and 11 from the control group) were analyzed by immunohistochemistry using anti-IL-6, TNF-alfa, CD163, MBL e FCN3 antibodies. Genotyping of target polymorphisms in the MBL2 gene was performed by real-time PCR. Proinflammatory cytokines such as IL-6 and TNF-alpha presented higher tissue expression in the COVID-19 group compared to H1N1 and control groups. The same results were observed for ICAM-1 tissue expression. Increased expression of the FCN3 was observed in the COVID-19 group and H1N1 group compared to the control group. The MBL tissue expression was higher in the COVID-19 group compared to H1N1 and control groups. The genotypes AA for rs180040 (G/A), GG for rs1800451 (G/A) and CC for rs5030737 (T/C) showed a higher prevalence in the COVID-19 group. The intense activation of the lectin pathway, with particular emphasis on the MBL pathway, together with endothelial dysfunction and a massive proinflammatory cytokines production, possibly lead to a worse outcome in patients infected with SARS-Cov-2. Moreover, 3 SNPs of our study presented genotypes that might be correlated with high MBL tissue expression in the COVID-19 pulmonary samples.

Highlights • We summarized the most important errors which may happen during nucleic acid tests for detection of SARS-CoV-2. • We explored the errors in three different analytic phases. • We showed that most important sources of error occurred during pre-analytic phase., The COVID-19 pandemic remains a significant problem involving health systems worldwide. Accurate and early detection of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection is critical for minimizing spread and initiating treatment. Among test methods, real-time reverse-transcriptase polymerase-chain-reaction (rRT-PCR) is considered the gold standard. Although this test has high specificity and relatively high sensitivity, the occurrence of falsely negative results in symptomatic patients and/or having a positive CT scan remains a challenge. Sources of error can be pre-analytical (sampling, storage and processing), analytical (RNA extraction, cDNA synthesis and amplification) and post-analytical (interpretation and analysis and test reporting). These potential sources of error and efforts to mitigate are reviewed in this article with an emphasis on the analytical phase.

In the present study, a facile, eco-friendly, and controlled synthesis of gold nanoparticles (Au NPs) using Prunus nepalensis fruit extract is reported. The biogenically synthesized Au NPs possess ultra-active intrinsic peroxidase-like activity for the oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB) in the presence of H2O2. Chemical analysis of the fruit extract demonstrated the presence of various bioactive molecules such as amino acids (l-alanine and aspartic acids), organic acids (benzoic acid and citric acid), sugars (arabinose and glucose), phenolic acid, and bioflavonoids (niacin and myo-inositol), which likely attributed to the formation of stable biogenic Au NPs with excellent peroxidase-mimicking activity. In comparison with the natural horseradish peroxidase (HRP) enzyme, the biogenic Au NPs displayed a 9.64 times higher activity with regard to the reaction velocity at 6% (v/v) H2O2, presenting a higher affinity toward the TMB substrate. The Michaelis-Menten constant (KM) values for the biogenic Au NPs and HRP were found to be 6.9 × 10-2 and 7.9 × 10-2 mM, respectively, at the same concentration of 100 pM. To investigate its applicability for biosensing, a monoclonal antibody specific for Mycobacterium bovis (QUBMA-Bov) was directly conjugated to the surface of the biogenic Au NPs. The obtained results indicate that the biogenic Au NPs-QUBMA-Bov conjugates are capable of detecting M. bovis based on a colorimetric immunosensing method within a lower range of 100 to 102 cfu mL-1 with limits of detection of 53 and 71 cfu mL-1 in an artificial buffer solution and in a soft cheese spiked sample, respectively. This strategy demonstrates decent specificity in comparison with those of other bacterial and mycobacterial species. Considering these findings together, this study indicates the potential for the development of a cost-effective biosensing platform with high sensitivity and specificity for the detection of M. bovis using antibody-conjugated Au nanozymes.

Objectives: To use a case review approach for investigating the types of cognitive error identifiable following a complicated patient admission with a multisystem disorder in an acute care setting where diagnosis was difficult and delayed. Methods: A case notes review was undertaken to explore the cognitive factors associated with diagnostic error in the case of an 18-year-old male presenting acutely unwell with myalgia, anorexia and vomiting. Each clinical interaction was analysed and identified cognitive factors were categorised using a framework developed by Graber et al. Results: Cognitive factors resulting in diagnostic errors most frequently occurred within the first five days of hospital admission. The most common were premature closure; failure to order or follow up an appropriate test; over-reliance on someone else's findings or opinion; over-estimating or underestimating usefulness or salience of a finding, and; ineffective, incomplete or faulty history and physical examination. Cognitive factors were particularly frequent around transitions of care and patient transfers from one clinical area to another. The presence of senior staff did not necessarily mitigate against diagnostic error from cognitive factors demonstrated by junior staff or diagnostic errors made out-of-hours. Conclusions: Cognitive factors are a significant cause of diagnostic error within the first five days after admission, especially around transitions of care between different clinical settings and providers. Medical education interventions need to ensure clinical reasoning training supports individuals and teams to develop effective strategies for mitigating cognitive factors when faced with uncertainty over complex patients presenting with non-specific symptoms in order to reduce diagnostic error.

Urinary 5-hydroxyindoleacetic acid (5-HIAA), vanillylmandelic (VMA), homovanillic acid (HVA), catecholamines and metanephrines are produced in excess by catecholamine-producing tumors. These biogenic amines are unstable at low or high pH and require hydrochloric acid (HCl) to prevent their degradation. However, HCl addition may result in very low pH causing degradation or deconjugation of several metabolites. This study evaluated the buffering properties of sodium citrate to stabilize all biogenic amines. The metabolite concentrations were measured by LC-MS/MS or by a coulometric assay in 22 urine samples collected native and with HCl or sodium citrate. We studied the effect of pH, time (48 h, four weeks) and storage temperature at 22 °C, 4 °C, and −20 °C. We found that catecholamines degradation was prevented by HCl and citrate and that 5-HIAA was degraded in 5 out of 22 samples collected with HCl. All biogenic amines were efficiently stabilized by citrate for four weeks at 22 °C, except epinephrine (48 h at 4 °C, or four weeks at −20 °C). Sodium citrate did not cause quantification or analytical artefacts concerns. In conclusion, sodium citrate is a non-hazardous alternative to HCl for patients to send unfrozen urine samples to the laboratory which may safely store the sample for four weeks.

BACKGROUND: Although the SARS-CoV-2 virus is transmitted mainly through the respiratory tract, possible transmission by transfusion from asymptomatic carriers should be explored. As yet there are no reports of transfusion transmission of COVID-19. Haemovigilance findings within a three-month surveillance period during the new coronavirus pandemic are presented. MATERIALS AND METHODS: Due to great demand and shortage, blood sessions in outpatient facilities were organized during the high prevalence period of COVID-19, alongside a national plan to monitor the evolving public health situation by random molecular screening of high-risk groups of the population. Haemovigilance protocols were implemented as well as surveillance for any COVID-19 case reported post-transfusion. A 14-day quarantine and follow-up molecular and antibody testing of any COVID-19 positive case was obligatory. RESULTS: Post-donation, post-transfusion information and molecular testing of swab samples collected from three asymptomatic donors at risk for COVID-19, revealed the case of an immunosupressed patient who had been transfused with whole blood derived platelets from a donor subsequently diagnosed with COVID-19. The recipient exhibited no symptoms of the disease. Molecular and antibody testing results were negative. CONCLUSION: Haemovigilance provided information supporting the absence of transfusion transmission of COVID-19, thus strengthening the hypothesis that, even if it cannot yet be definitively ruled out, COVID-19 is not transmitted through blood transfusion. As of early June 2020, a perfect test does not exist, therefore haemovigilance along with the implementation of strict proactive measures is crucial to identify eluding asymptomatic individuals and ensure blood safety during the pandemic.

Viscosity is a pivotal factor for indicating the dysfunction of the mitochondria. To date, most of the fluorescent probes developed for mitochondrial viscosity have been designed using BODIPY, hemicyanine, or pyridine-based molecular rotors as part of the core structure. Our aim with this research was to extend the range of suitable fluorophores available for the construction of such fluorescent molecular rotors for evaluating the viscosity of mitocondria. Herein, we have developed a green fluorescent protein (GFP)-chromophore-based fluorescent probe (MIT-V) for the detection of mitochondrial viscosity in live cells. MIT-V exhibited a high sensitivity toward viscosity (from 7.9 cP to 438.4 cP). The "off-on"sensing mechanism of MIT-V was ascribed to the restricted rotation of single bonds and excited-state C= C double bonds of MIT-V. Cell studies indicated that MIT-V targets the mitochondria and that it was able to monitor real-time changes in the viscosity of live HeLa cell mitochondria. Therefore, we propose that MIT-V can be used as an effective chemosensor for the real-time imaging of mitochondrial viscosity in live cells. Our results clearly demonstrate the utility of such GFP-chromophore-based derivatives for the development of viscosity-sensitive systems.

Background The association between SARS-CoV-2 commercial serological assays and virus neutralization test (VNT) has been poorly explored in mild patients with COVID-19. Methods 439 serum specimens were longitudinally collected from 76 healthcare workers with RT-PCR-confirmed COVID-19. The clinical sensitivity (determined weekly) of 9 commercial serological assays were evaluated. Clinical specificity was assessed using 69 pre-pandemic sera. Correlation, agreement, and concordance with the VNT were also assessed on a subset of 170 samples. Area under the ROC curve (AUC) was estimated at 2 neutralizing antibody titers. Results The Wantai Total Ab assay targeting the receptor binding domain (RBD) within the S protein presented the best sensitivity at different times during the course of disease. The clinical specificity was greater than 95% for all tests except for the Euroimmun IgA assay. The overall agreement with the presence of neutralizing antibodies ranged from 62.2% (95%CI; 56.0–68.1) for bioMérieux IgM to 91.2% (87.0–94.2) for Siemens. The lowest negative percent agreement (NPA) was found with the Wantai Total Ab assay (NPA 33% (21.1–48.3)). The NPA for other total Ab or IgG assays targeting the S or the RBD was 80.7% (66.7–89.7), 90.3% (78.1–96.1), and 96.8% (86.8–99.3) for Siemens, bioMérieux IgG, and DiaSorin, respectively. None of the commercial assays have sufficient performance to detect a neutralizing titer of 80 (AUC Conclusions Although some assays show a better agreement with VNT than others, the present findings emphasize that commercialized serological tests, including those targeting the RBD, cannot substitute a VNT for the assessment of functional antibody response.

Background Reverse transcription-quantitative PCR on nasopharyngeal swabs is currently the reference COVID-19 diagnosis method but exhibits imperfect sensitivity. Methods We developed a multiplex reverse transcription-digital droplet PCR (RT-ddPCR) assay, targeting 6 SARS-CoV-2 genomic regions, and evaluated it on nasopharyngeal swabs and saliva samples collected from 130 COVID-19 positive or negative ambulatory individuals, who presented symptoms suggestive of mild or moderate SARS-CoV2 infection. Results For the nasopharyngeal swab samples, the results obtained using the 6-plex RT-ddPCR and RT-qPCR assays were all concordant. The 6-plex RT-ddPCR assay was more sensitive than RT-qPCR (85% versus 62%) on saliva samples from patients with positive nasopharyngeal swabs. Conclusion Multiplex RT-ddPCR represents an alternative and complementary tool for the diagnosis of COVID-19, in particular to control RT-qPCR ambiguous results. It can also be applied to saliva for repetitive sampling and testing individuals for whom nasopharyngeal swabbing is not possible.

Highlights • Non-severe COVID-19 patients have abnormal laboratory investigations. • Patients with prolonged pretreatment APTT have a longer isolation length. • Patients with elevated eosinophils after treatment have a shorter isolation length. • A nomogram could help to predict isolation probability at 11-, 16- and 21-day., Background Majority coronavirus disease 2019 (COVID-19) patients are classified as mild and moderate (non-severe) diseases. We aim to develop a model to predict isolation length for non-severe patients. Methods Among 188 non-severe patients, 96 patients were enrolled as training cohort to identify factors associated with isolation length via Cox regression model and develop a nomogram. Other 92 patients formed as validation cohort to validate nomogram. Concordance index (C-index), area under the curve (AUC) and calibration curves were used to evaluated nomogram. Results Increasing absolute eosinophil count (AEC) after admission was correlated with shorter isolation length (P = 0.02). Baseline activated partial thromboplastin time (APTT) > 30 s was correlated with longer isolation length (P = 0.03). A nomogram to predict isolation probability at 11-, 16- and 21-day was developed and validated. The C-indices of training and validation cohort were 0.604 and 0.682 respectively. Both cohorts showed a good discriminative ability (AUC, 11-day: 0.646 vs 0.730; 16-day: 0.663 vs 0.750; 21-day: 0.711 vs 0.783; respectively) and calibration power. Conclusions Baseline APTT and dynamic change of AEC were two significant factors associated with isolation length of non-severe patients. Nomogram could predict isolation probability for each patient to estimate appropriate quarantine length.

Highlights • COVID-19 biosafety action plans can maintain stat laboratory quality. • Test volume initially declined, and then increased to surpass pre-pandemic levels. • Turnaround time was maintained, and pre-analytical errors were stable., Objective It is unclear if implementation of biosafety action plans in response to the COVID-19 pandemic has affected laboratory quality metrics. Methods This retrospective study used quality data, including turnaround time (TAT) and number/type of unacceptable specimens from a stat laboratory supporting an outpatient medical clinic serving predominantly elderly cancer patients. Four months of data from the height of the COVID-19 pandemic (March-June 2020) were compared to the same months in 2019. Results March-May 2020 test volumes were decreased compared to 2019. June 2020 test volume was slightly increased compared to 2019. TATs in 2020 were similar/ slightly improved compared to the same months in 2019, due to shortened collect to receive and receive to verify TATs. The number and types of unacceptable specimens were similar in 2020 and 2019. Conclusions Despite the challenges to the system caused by the pandemic, laboratory quality metrics were maintained.

Sequencing the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) genome is a crucial task for controlling the ongoing coronavirus disease (COVID-19) pandemic. However, elucidating the pathological mechanisms of SARS-CoV-2 in humans has been challenging. A comprehensive analysis of the molecular characteristics of SARS-CoV-2 and molecular changes in COVID-19 patients may have practical significance in developing assays for the detection of SARS-CoV-2 and formulating clinical treatment strategies against COVID-19. The omics approach for studying biochemical mechanisms can be used to elucidate the molecular characteristics and pathophysiology of SARS-CoV-2. The omics-scale research on COVID-19 has been carried out rapidly, bringing hope for developing a robust diagnostic assay, discovering reliable biomarkers to assess disease progression, and developing therapeutic drugs and vaccines. In this review, we summarize, from an omics perspective, the strategies for the detection of SARS-CoV-2 antigens and antibodies against the virus, the metabolomic and proteomic changes in COVID-19 patients, and the progress of research on anti-SARS-CoV-2 drugs with their potential clinical applications.

The COVID-19 pandemic has taken a major toll on the economy and funding for public education. For that reason, the pandemic has a worrisome effect on the sustainability of university/college based Medical Laboratory Sciences MLS training programs. Stakeholders of university-based MLS programs include university administrators, students, clinical affiliates and faculty. Each group has specific goals and challenges that affect the sustainability of the program. This report details strategies that can be used to satisfy the goals specific to key stakeholders that lead to sustainability. These strategies apply in pandemic times and in the back-to-normal future.

Objective Early detection of patients with COVID-19 who will need mechanical invasive ventilation (MIV) may aid in delivering proper care and optimizing the use of limited resources. Methods In this single-center retrospective observational study, we aimed to identify simple laboratory parameters that in combination with ferritin (a surrogate marker of severe inflammation) may help predict early (first 48 hours) MIV. A total of 160 patients with COVID-19 in whom serum ferritin, absolute lymphocyte count (ALC), platelet count, C-reactive protein (CRP), and lactate dehydrogenase (LDH) had been analyzed at admission were included. Results We found that ferritin, LDH, ALC, and CRP predicted with 88% accuracy the probability of early MIV. Results indicated that LDH showed the greater area under the curve (AUC), with a value of 89.1%. Using the AUC, we established cutoff values for clinical application. Finally, we developed a classification tree based on LDH for its clinical use. Conclusion Ferritin, LDH, ALC, and CRP predict with 88% accuracy the probability of early MIV.

Bacterial infection is a universal threat to public health, which not only causes many serious diseases but also exacerbates the condition of the patients of cancer, pandemic diseases, e.g., COVID-...

An outbreak of COVID-19, caused by infection with SARS-CoV-2 in Wuhan, China in December 2019, spread throughout the country and around the world, quickly. The primary detection technique for SARS-CoV-2, the reverse-transcription polymerase chain reaction (RT-PCR)–based approach, requires expensive reagents and equipment and skilled personnel. In addition, for SARS-CoV-2 detection, specimens are usually shipped to a designated laboratory for testing, which may extend the diagnosis and treatment time of patients with COVID-19. The latest research shows that clustered regularly interspaced short palindromic repeats (CRISPR)–based approaches can quickly provide visual, rapid, ultrasensitive, and specific detection of SARS-CoV-2 at isothermal conditions. Therefore, CRISPR-based approaches are expected to be developed as attractive alternatives to conventional RT-PCR methods for the efficient and accurate detection of SARS-CoV-2. Recent advances in the field of CRISPR-based biosensing technologies for SARS-CoV-2 detection and insights into their potential use in many applications are reviewed in this article.

Highlights • Digital PCR is a sensitive new technique for SARS-CoV-2 detection in clinical applications. • Digital PCR would substantially reduce the rates of false-negative COVID-19 test results, in particular those pertaining to asymptomatic carriers. • Real-time reverse-transcriptase (RT)-PCR is a fast and convenient method for viral nucleic acid detection, but it’s results can be compromised by numerous factors and can yield false-negative results., The global coronavirus disease 2019 (COVID-19) pandemic has posed great challenges in people’s daily lives. Highly sensitive laboratory techniques played a critical role in clinical COVID-19 diagnosis and management. In this study the feasibility of using a new digital PCR-based detection assay for clinical COVID-19 diagnosis was investigated by comparing its performance with that of RT-PCR. Clinical patient samples and samples obtained from potentially contaminated environments were analyzed. The study included 10 patients with confirmed COVID-19 diagnoses, 32 validated samples of various types derived from different clinical timepoints and sites, and 148 environmentally derived samples. SARS-CoV-2 nucleic acids were more readily detected in respiratory tract samples (35.0%). In analyses of environmentally derived samples, the positivity rate of air samples was higher than that of surface samples, probably due to differences in virus concentrations. Digital PCR detected SARS–CoV–2 in several samples that had previously been deemed negative, including 3 patient-derived samples and 5 environmentally derived samples. In this study digital PCR exhibited higher sensitivity than conventional RT-PCR, suggesting that it may be a useful new method for clinical SARS-CoV-2 detection. Improvement of SARS-CoV-2 detection would substantially reduce the rates of false-negative COVID-19 test results, in particular those pertaining to asymptomatic carriers.

Highlights • A synthetic positive control template system for COVID-19 diagnosis was developed. • The low detection limits of SPT were 10−12 to >10−15 M for different gene targets. • One picogram of patient RNA allowed convincing detection with new positive controls., Backgrounds The coronavirus disease 2019 (COVID-19) pandemic is still ongoing. Real-time reverse transcription polymerase chain reaction (real-time RT-PCR) is regarded as a gold-standard method for the diagnosis of COVID-19. However, unexpected contamination of synthesized positive control samples included in COVID-19 test kits have increased the inconclusiveness of disease interpretation. Therefore, it is important to establish new methods for the preparation of reliable positive controls that are not affected by contamination for the accurate for diagnosis of COVID-19, but it still remains a challenge. Methods A new approach for producing synthetic positive controls using synthetic positive template (SPT) oligonucleotides was designed. SPT oligonucleotides contain probe binding and virus-irrelevant regions were used as templates for real-time PCR to evaluate the expression level of SARS-CoV-2 genes (RdRP, E, and N). The limit of detection (LOD) for individual SARS-CoV-2 genes by Ct values with different concentrations of SPT templates and genomic RNAs from SARS-CoV-2 infected samples was determined. Results LODs with SPT templates were >10−15 (atto) M for RdRP, 10−12 (femto) to 10−13 (100 atto) M for E gene, and 10−12 to 10−14 (10 atto) M for N gene, respectively. Real-time RT-PCR assay using serially diluted genomic RNAs prepared from SARS-CoV-2 virus infected cultures showed that picogram quantities of RNAs is resulted in the LOD. The sensitivity of RdRP and E genes based on Ct values was less than that of N gene with this platform. Conclusion This method significantly reduces the risk of false-positive reactions resulting from contamination in the synthesis procedures of positive control materials. Therefore, this approach could be integrated into the currrently available COVID-19 test kits and will provide a general method for preparing positive controls in the diagnosis of emerging RNA virus infections.

Background Coronavirus disease 2019 (COVID-19) caused by a novel betacoronavirus named severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has attracted top health concerns worldwide within a few months after its appearance. Since viruses are highly dependent on the host small RNAs (microRNAs) for their replication and propagation, in this study, top miRNAs targeting SARS-CoV-2 genome and top miRNAs targeting differentially expressed genes (DEGs) in lungs of patients infected with SARS-CoV-2, were predicted. Methods All human mature miRNA sequences were acquired from miRBase database. MiRanda tool was used to predict the potential human miRNA binding sites on the SARS-CoV-2 genome. EdgeR identified differentially expressed genes (DEGs) in response to SARS-CoV-2 infection from GEO147507 data. Gene Set Enrichment Analysis (GSEA) and DEGs annotation analysis were performed using ToppGene and Metascape tools. Results 160 miRNAs with a perfect matching in the seed region were identified. Among them, there was 15 miRNAs with more than three binding sites and 12 miRNAs with a free energy binding of −29 kCal/Mol. MiR-29 family had the most binding sites (11 sites) on the SARS-CoV-2 genome. MiR-21 occupied four binding sites and was among the top miRNAs that targeted up-regulated DEGs. In addition to miR-21, miR-16, let-7b, let-7e, and miR-146a were the top miRNAs targeting DEGs. Conclusion Collectively, more experimental studies especially miRNA-based studies are needed to explore detailed molecular mechanisms of SARS-CoV-2 infection. Moreover, the role of DEGs including STAT1, CCND1, CXCL-10, and MAPKAPK2 in SARS-CoV-2 should be investigated to identify the similarities and differences between SARS-CoV-2 and other respiratory viruses., Highlights • miR-29 family had 11 binding site on the SARS-COV-2 genome. • miR-29a/b, 21, 761, 3130, 3167 and miR-3175 bound to the spike coding sequence. • miR-16, 146a, 21, 615 and let-7b/e targeted SARS-CoV-2 induced DEGs. • miR-146a, 203a, 24, 615 and miR-16 targeted DEGs involved in viral prosesess

Graphical abstract, Highlights • Evaluating the single-, dual-, triple-target detection kits based on RT-qPCR. • The number of targets in a detection kit is not a key factor affecting sensitivity. • The amount of sample loading may influence the performance of a detection kit. • Providing a guide when developing a nucleic acid detection kit for SARS-CoV-2. • Digital PCR can be used to review clinically suspected samples., Background The coronavirus disease 2019 (COVID-19) has become a pandemic. Reverse transcription quantitative PCR (RT-qPCR) has played a vital role in the diagnosis of COVID-19, but the rates of false negatives is not ideal in dealing with this highly infectious virus. It is thus necessary to systematically evaluate the clinical performance of the single-, dual-, triple-target detection kits to guide the clinical diagnosis of this disease. Methods A series of reference materials calibrated by droplet digital PCR (ddPCR) and 57 clinical samples were used to evaluate the clinical performance of six single-, dual-, triple-target SARS-CoV-2 nucleic acid detection kits based on RT-qPCR. Results The dual-target kits, kit B and kit C had the highest and the lowest detection sensitivity, which was 125 copies/mL and 4000 copies/mL, respectively. Among the 57 clinical samples from patients with COVID-19, 47 were tested positive by the kit B, while 35, 29, 28, 30, and 29 were found positive by the kits A, C, D, E, and F, respectively. The number of targets in a detection kit is not a key factor affecting sensitivity, while the amount of sample loading may influence the performance of a detection kit. Conclusions This study provides a guide when choosing or developing a nucleic acid detection kit for the diagnosis of COVID-19. Also, the absolute-quantification feature and high-sensitivity performance of ddPCR, suggesting that it can be used to review clinically suspected samples.

Endemic species of coronavirus (HCoV-OC43, HCoV-229E, HCoV-NL63, and HCoV-HKU1) are frequent causes of upper respiratory tract infections. Three highly pathogenic coronaviruses have been associated with outbreaks and epidemics and have challenged clinical microbiology laboratories to quickly develop assays for diagnosis. Their initial characterization was achieved by molecular methods. With the great advance in metagenomic whole-genome sequencing directly from clinical specimens, diagnosis of novel coronaviruses could be quickly implemented into the workflow of managing cases of pneumonia of unknown cause, which will markedly affect the time of the initial characterization and accelerate the initiation of outbreak control measures.

Background Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has infected over 21 million people worldwide since August 16, 2020. Compared to PCR and serology tests, SARS-CoV-2 antigen assays are underdeveloped, despite their potential to identify active infection and monitor disease progression. Methods We used Single Molecule Array (Simoa) assays to quantitatively detect SARS-CoV-2 spike, S1 subunit, and nucleocapsid antigens in the plasma of patients with coronavirus disease (COVID-19). We studied plasma from 64 patients who were COVID-19 positive, 17 who were COVID-19 negative, and 34 prepandemic patients. Combined with Simoa anti-SARS-CoV-2 serological assays, we quantified changes in 31 SARS-CoV-2 biomarkers in 272 longitudinal plasma samples obtained for 39 patients with COVID-19. Data were analyzed by hierarchical clustering and were compared to longitudinal RT-PCR test results and clinical outcomes. Results SARS-CoV-2 S1 and N antigens were detectable in 41 out of 64 COVID-19 positive patients. In these patients, full antigen clearance in plasma was observed a mean ± 95% CI of 5 ± 1 days after seroconversion and nasopharyngeal RT-PCR tests reported positive results for 15 ± 5 days after viral-antigen clearance. Correlation between patients with high concentrations of S1 antigen and ICU admission (77%) and time to intubation (within 1 day) was statistically significant. Conclusions The reported SARS-CoV-2 Simoa antigen assay is the first to detect viral antigens in the plasma of patients who were COVID-19 positive to date. These data show that SARS-CoV-2 viral antigens in the blood are associated with disease progression, such as respiratory failure, in COVID-19 cases with severe disease.

No specific treatment for COVID-19 infection is available up till now, and there is a great urge for effective treatment to reduce morbidity and mortality during this pandemic. We aimed to evaluate the effect of combining chloroquine/hydroxychloroquine (CQ/HCQ) and zinc in the treatment of COVID-19 patients. This was a randomized clinical trial conducted at three major University hospitals in Egypt. One hundred ninety-one patients with a confirmed diagnosis of COVID-19 infection were randomized into two groups: group I (96) patients received both HCQ and zinc, and group II (95) received HCQ only. The primary endpoints were the recovery within 28 days, the need for mechanical ventilation, and death. The two groups were matched for age and gender. They had no significant difference regarding any of the baseline laboratory parameters or clinical severity grading. Clinical recovery after 28 days was achieved by 79.2% in the zinc group and 77.9% in zinc-free treatment group, without any significant difference (p = 0.969). The need for mechanical ventilation and the overall mortality rates did not show any significant difference between the 2 groups either (p = 0.537 and 0.986, respectively). The age of the patient and the need for mechanical ventilation were the only risk factors associated with the patients’ mortality by the univariate regression analysis (p = 0.001 and

Introduction Early diagnosis and identification of potential critical cases for timely treatment are crucial for COVID‐19 patients. The aim of this study was to analyze the diagnostic and prognostic implications of WBC and cell population data (CPD) abnormalities related to COVID‐19 at disease onset. Methods Baseline WBC counts and CPD data were analyzed in one hundred COVID‐19 patients presenting to emergency department and subsequently discharged (n = 49), admitted (n = 51) or deceased (n = 22), and in 47 healthy subjects. Results Lymphopenia and eosinopenia were observed in all COVID‐19 patients, with more intensity in the admitted and deceased groups, that also presented increased WBC and neutrophil counts. On CPD analysis, COVID‐19 was associated with increased volume of neutrophils, lymphocytes, and monocytes, whereas conductivity was decreased for neutrophils and increased for lymphocytes. The ROC curve analysis showed good performance for lymphocyte counts in predicting COVID‐19 diagnosis (AUC = 0.858), for neutrophil counts in predicting admission for COVID‐19 (AUC = 0.744) and for monocytes volume in predicting COVID‐19 diagnosis (AUC = 0.837). Conclusion WBC counts and CPD parameters at disease onset in COVID‐19 patients can improve diagnostic characterization and aid in the discrimination between severe and nonsevere presentations.

The long evolutionary battle between humans and pathogens has played an important role in shaping the current network of host-pathogen interactions. Each organ brings new challenges from the perspective of a pathogen to establish a suitable niche for survival while subverting the protective mechanisms of the host. Lungs, the organ for oxygen exchange, have been an easy target for pathogens due to its accessibility. The organ has evolved diverse capabilities to provide the flexibility required for an organism’s health and at the same time maintain protective functionality to prevent and resolve assault by pathogens. The pathogenic invasions are strongly challenged by healthy lung architecture which includes the presence and activity of the epithelium, mucous, antimicrobial proteins, surfactants, and immune cells. Competitively, the pathogens in the form of viruses, bacteria, and fungi have evolved an arsenal of strategies that can over-ride the host’s protective mechanisms. While bacteria such as Mycobacterium tuberculosis (M. tuberculosis) can survive in dormant form for years before getting active in humans, novel pathogens can wreak havoc as they pose a high risk of morbidity and mortality in a very short duration of time. Recently, a coronavirus strain SARS-CoV-2 has caused a pandemic which provides us an opportunity to look at the host manipulative strategies used by respiratory pathogens. Their ability to hide, modify, evade, and exploit cell’s processes are key to their survival. While pathogens like M. tuberculosis have been infecting humans for thousands of years, SARS-CoV-2 has been the cause of the recent pandemic. Molecular understanding of the strategies used by these pathogens could greatly serve in design of predictive, preventive, personalized medicine (PPPM). In this article, we have emphasized on the clinically relevant evasive strategies of the pathogens in the lungs with emphasis on M. tuberculosis and SARS-CoV-2. The molecular basis of these evasive strategies illuminated through advances in genomics, cell, and structural biology can assist in the mapping of vulnerable molecular networks which can be exploited translationally. These evolutionary approaches can further assist in generating screening and therapeutic options for susceptible populations and could be a promising approach for the prediction, prevention of disease, and the development of personalized medicines. Further, tailoring the clinical data of COVID-19 patients with their physiological responses in light of known host-respiratory pathogen interactions can provide opportunities to improve patient profiling and stratification according to identified therapeutic targets.

Highlights • RT-qPCR and ddPCR were used for SARS-CoV-2 nucleic acids detection. • ddPCR shows higher sensitivity and lower limit of detection than RT-qPCR. • ddPCR successfully detected the dynamic changes in viral load while RT-qPCR failed to detect it. • Low-viral-load samples were not uncommon in clinical SARS-CoV-2 nucleic acids testing., Background Qualitative and quantitative detection of nucleic acids of SARS-CoV-2, the pathogen that causes coronavirus disease 2019 (COVID-19), plays a significant role in COVID-19 diagnosis, surveillance, prevention, and control. Methods A total of 117 samples from 30 patients with confirmed COVID-19 and 61 patients without COVID-19 were collected. Reverse transcriptase quantitative PCR (RT-qPCR) and droplet digital PCR (ddPCR) were used for qualitative and quantitative analyses of these samples to evaluate the diagnostic performance and applicability of the two methods. Results The positive detection rates of RT-qPCR and ddPCR were 93.3% and 100%, respectively. Among the 117 samples, 6 samples were tested single-gene positive by RT-qPCR but positive by ddPCR, and 3 samples were tested negative by RT-qPCR but positive by ddPCR. The viral load of samples with inconsistent results were relatively low (3.1–20.5 copies/test). There were 17 samples (37%) with a viral load below 20 copies/test among the 46 positive samples, and only 9 of them were successfully detected by RT-qPCR. A severe patient was dynamically monitored. All 6 samples from this patient were tested negative by RT-qPCR, but 4 samples were tested positive by ddPCR with a low viral load. Conclusion Qualitative analysis of COVID-19 samples can meet the needs of clinical screening and diagnosis, while quantitative analysis provides more information to the research community. Although both ddPCR and RT-qPCR can provide qualitative and quantitative results, ddPCR showed higher sensitivity and lower limit of detection than RT-qPCR, and it does not rely on the standard curve to quantify viral load. Therefore, ddPCR offers greater advantages than RT-qPCR.

Resume Pour faire face a l’emergence du Sars-CoV-2 responsable d’une pandemie mondiale, de tres nombreux tests diagnostiques ont ete developpes et mis sur le marche dans un delai tres court. La RT-PCR sur prelevement rhino-pharynge est la methode de reference pour le diagnostic et le depistage de l’infection a Sars-CoV-2 mais les tests developpes presentent de grande variabilite en termes de sensibilite et de delai de rendu des resultats. Les tests antigeniques presentent en general une sensibilite plus faible mais presentent l’avantage d’une mise en œuvre plus simple et plus rapide. Devant des tableaux evoca-teurs de Covid-19 avec un resultat de RT-PCR negatif, une serologie peut etre recommandee avec le dosage des IgM et des IgG. La serologie est egalement un outil pertinent pour les etudes epidemiologiques. Neanmoins il est important de rappeler que le taux d’anticorps anti-Sars-Cov-2 decroit avec le temps et peut ainsi impacter les resultats d’etudes sero-epidemiologiques. Malgre la necessite de repondre rapidement a un besoin diagnostique urgent, il demeure indispensable de valider les methodes choisies sur un panel d’echantillons bien caracterise, les performances de certains tests etant parfois insatisfaisantes pour assurer un diagnostic fiable.

Highlights • We summarized the existing laboratory testing methods for SARS-CoV-2, including nucleic acid tests, antibody tests and genomic sequencing. • We analyzed the different clinical specimens used for SARS-CoV-2 testing to provide a reference for the clinical selection of appropriate specimens. • We compared the advantages and disadvantages of different laboratory testing methods for COVID-19, and provided an outlook of the improvement and development of detection technology. • The specificity and sensitivity of Ig subclass antibody detection were also compared., The outbreak of Coronavirus Disease-2019 (COVID-19) caused by Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) has threatened health worldwide. As of the end of 2020, there were nearly 10 million confirmed cases and nearly 5 million deaths associated with COVID-19. Rapid and early laboratory diagnosis of COVID-19 is the main focus of treatment and control. Molecular tests are the basis for confirmation of COVID-19, but serological tests for SARS-CoV-2 are widely available and play an increasingly important role in understanding the epidemiology of the virus and in identifying populations at higher risk for infection. Point-of-care tests have the advantage of rapid, accurate, portable, low cost and non-specific device requirements, which provide great help for disease diagnosis and detection. This review will discuss the performance of different laboratory diagnostic tests and platforms, as well as suitable clinical samples for testing, and related biosafety protection. This review shall guide for the diagnosis of COVID-19 caused by SARS-CoV-2.

A significant amount of clinical and research interest in thrombosis is focused on large vessels (e.g., stroke, myocardial infarction, deep venous thrombosis, etc.); however, thrombosis is often present in the microcirculation in a variety of significant human diseases, such as disseminated intravascular coagulation, thrombotic microangiopathy, sickle cell disease, and others. Further, microvascular thrombosis has recently been demonstrated in patients with COVID-19, and has been proposed to mediate the pathogenesis of organ injury in this disease. In many of these conditions, microvascular thrombosis is accompanied by inflammation, an association referred to as thromboinflammation. In this review, we discuss endogenous regulatory mechanisms that prevent thrombosis in the microcirculation, experimental approaches to induce microvascular thrombi, and clinical conditions associated with microvascular thrombosis. A greater understanding of the links between inflammation and thrombosis in the microcirculation is anticipated to provide optimal therapeutic targets for patients with diseases accompanied by microvascular thrombosis.

Highlights • The role of serological testing is crucial in the clinical management of COVID-19. • We discuss the utility of anti–SARS–CoV–2 serology testing in a clinical setting. • Utilizing anti–SARS–CoV2 detection in the advanced stage of infection is recommended., Serologic tests are one of the available diagnostic tools in COVID–19. Growing literature highlights their role in the clinical management of the disease. Unfortunately, due to the limited availability of commercial tests and the lack of reliable trials establishing the sensitivity and specificity of the diagnostic method, the clinical application of the test needs to be precisely determined. In this paper, we discuss the utility of anti–SARS–CoV–2 serology testing in a clinical setting and propose diagnostic algorithms that include serological tests in patients with confirmed or suspected COVID–19.

Highlights • Severe acute respiratory syndrome CoV-2 can interacts with cardiovascular system. • COVID-19 potentiates existing cardiovascular diseases. • Risk factor for cardiovascular disease are common in COVID-19. • A number of patients with COVID-19 manifest an increase in cardiac biomarkers release. • Cardiovascular complications were independently reported with COVID-19 infections., The coronavirus disease COVID-19 is a public health emergency caused by a novel coronavirus named severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). SARS-CoV-2 infection uses the angiotensin-converting enzyme 2 (ACE2) receptor, and typically spreads through the respiratory tract. Invading viruses can elicit an exaggerated host immune response, frequently leading to a cytokine storm that may be fueling some COVID-19 death. This response contributes to multi-organ dysfunction. Accumulating data points to an increased cardiovascular disease morbidity, and mortality in COVID-19 patients. This brief review explores potential available evidence regarding the association between COVID-19, and cardiovascular complications.

Background Rapid, reliable, and widespread testing is required to curtail the ongoing COVID-19 pandemic. Current gold-standard nucleic acid tests are hampered by supply shortages in critical reagents including nasal swabs, RNA extraction kits, personal protective equipment, instrumentation, and labor. Methods To overcome these challenges, we developed a rapid colorimetric assay using reverse-transcription loop-mediated isothermal amplification (RT-LAMP) optimized on human saliva samples without an RNA purification step. We describe the optimization of saliva pretreatment protocols to enable analytically sensitive viral detection by RT-LAMP. We optimized the RT-LAMP reaction conditions and implemented high-throughput unbiased methods for assay interpretation. We tested whether saliva pretreatment could also enable viral detection by conventional reverse-transcription quantitative polymerase chain reaction (RT-qPCR). Finally, we validated these assays on clinical samples. Results The optimized saliva pretreatment protocol enabled analytically sensitive extraction-free detection of SARS-CoV-2 from saliva by colorimetric RT-LAMP or RT-qPCR. In simulated samples, the optimized RT-LAMP assay had a limit of detection of 59 (95% confidence interval: 44–104) particle copies per reaction. We highlighted the flexibility of LAMP assay implementation using 3 readouts: naked-eye colorimetry, spectrophotometry, and real-time fluorescence. In a set of 30 clinical saliva samples, colorimetric RT-LAMP and RT-qPCR assays performed directly on pretreated saliva samples without RNA extraction had accuracies greater than 90%. Conclusions Rapid and extraction-free detection of SARS-CoV-2 from saliva by colorimetric RT-LAMP is a simple, sensitive, and cost-effective approach with broad potential to expand diagnostic testing for the virus causing COVID-19.

Highlights • Elevated LDH and CRP serum concentrations are associated to respiratory failure in CoVID-19 patients., Objective The dramatic worldwide CoVID-19 infection requires the identification of a reliable and inexpensive tool to quickly discriminate patients with a more unfavorable outcome. Methods We performed routine laboratory tests suitable to identify tissue damage and inflammatory status in 123 consecutive CoVID-19 patients admitted to the Emergency Department of the hospital of Piacenza (Emilia-Romagna, Northern Italy). The results were correlated with patients’ respiratory function evaluated by the partial pressure of arterial oxygen to fraction of inspired oxygen ratio (PaO2/FiO2). Results: The most common laboratory abnormalities were lymphocytopenia and elevated values of C-reactive protein (CRP) and lactate dehydrogenase (LDH). Aspartate aminotransferase (AST), alanine aminotransferase (ALT) and creatine kinase (CK) were also increased. The respiratory performance (PaO2/FiO2) showed a strong inverse correlation with LDH (r= 0.62, r2 0.38, p value< 0.0001) and CRP (r= 0.55, r2 0.31, p value< 0.0001). PaO2/FiO2 values also showed a significant inverse correlation with age (r= -0.37, p< 0.0001), AST (r= -0.31, p

Highlights • Blood group was the risk factors for COVID-19. • Blood group was related to the clinical characteristics of patients. • Patients with blood group A had an increased risk for infection with SARS-CoV-2, whereas blood group O was associated with a decreased risk., Objective To explore ABO blood group distribution and clinical characteristics in patients with COVID-19. Methods The clinical data of 187 patients with COVID-19 seen between January 20, 2020 and March 5, 2020 at the First Hospital of Changsha were retrospectively analyzed. The differences in the ABO blood group distribution between COVID-19 patients and the control group (1991 cases) were analyzed. The relationship between blood type and clinical characteristics was analyzed. Results Of the 187 patients with COVID-19, 69 had type A (36.90%), 63 had type B (33.69%), 41 had type O (21.92%), and 14 had type AB blood (7.49%). The proportion of patients with type A blood in the COVID-19 group was significantly higher than that in the control group (36.90% vs. 27.47%, P = 0.006), while the proportion of patients with type O blood in the COVID-19 group was significantly lower than that in the control group (21.92% vs. 30.19%, P = 0.018). The risk of COVID-19 was higher for individuals with blood group A than for those with blood group O (OR = 1.849, 95% CI = 1.228–2.768, P = 0.003). The risk of COVID-19 was higher for patients with blood group A than for those with a blood group other than A (OR = 1.544, 95% CI = 1.122–2.104, P = 0.006). Patients with blood group O had a lower risk of COVID-19 than non-O blood group patients (OR = 0.649, 95% CI = 0.457–0.927, P = 0.018). The ABO blood group distribution was related to COVID-19 status. Conclusions Patients with blood group A had an increased risk for infection with SARS-CoV-2, whereas blood group O was associated with a decreased risk, indicating that certain ABO blood groups were correlated with SARS-CoV-2 susceptibility. Blood type was related to some clinical characteristics of patients with COVID-19.

Les infections asymptomatiques ne sont pas rares ; les signes cliniques associent le plus souvent des signes généraux et des signes respiratoires ; ils sont polymorphes et non spécifiques. La tomodensitométrie thoracique est un outil important d’aide au diagnostic et de triage pour les patients dont l’état justifie une hospitalisation. Les deux complications principales de la maladie sont d’une part la détresse respiratoire par aggravation de la pneumonie, qui survient le plus souvent après sept à dix jours d’évolution dans le cadre d’un orage cytokinique, et la maladie thromboembolique d’autre part. Le taux de létalité est de l’ordre de 2 %. La prise en charge en milieu hospitalier comporte l’oxygénation le plus souvent et toujours la prévention de la maladie thromboembolique. Les surinfections bactériennes sont rares. Le remdésivir permettrait de raccourcir la durée des signes cliniques chez des patients hospitalisés et sous oxygénothérapie. La dexaméthasone permettrait de réduire la létalité chez les patients dont l’état justifie une ventilation mécanique. Le développement de nombreux candidats vaccins suscite beaucoup d’espoir pour accélérer la sortie de pandémie.

Highlights • Fever was associated with poor outcomes in patients with COVID-19. • Leukocytosis was associated with severe disease. • Leukopenia was associated with a better prognosis. • CRP was associated with poor prognosis. • Leuocytosis and CRP on arrival may predict severe COVID-19., Background Since December 2019, coronavirus 2019 (COVID-19) has spread worldwide. Identifying poor prognostic factors is helpful for risk stratification. In this meta-analysis, we investigated the association between severe COVID-19 and a change in white blood cell (WBC) count, an elevation of C-reactive protein (CRP), and fever. Moreover, we aimed to evaluate the diagnostic accuracy of leukocytosis and an elevation of CRP. Methods We performed a systematic search of PubMed, EMBASE, Scopus, and the Cochrane Library through April 20th, 2020. The odds ratios (ORs) and 95% confidence intervals (CIs) were calculated. A sensitivity analysis was conducted according to the study size (>200 or 55 or

Highlights • The detection rate of hock-a-loogie saliva was higher than nasopharyngeal swabs. • Hock-a-loogie saliva sample had a higher viral load than throat swab. • Hock-a-loogie saliva sampling for SARS-CoV-2 detection is reliable., To clarify the effect of different respiratory sample types on SARS-CoV-2 detection, we collected throat swabs, nasal swabs and hock-a-loogie saliva or sputum, and compared their detection rates and viral loads. The detection rates of sputum (95.65%, 22/23) and hock-a-loogie saliva (88.09%, 37/42) were significantly higher than those in throat swabs (41.54%, 27/65) and nasal swabs (72.31%, 47/65) (P

BACKGROUND: Ensuring steady stream of safe blood is the ultimate goal of blood transfusion practice. The current COVID-19 pandemic has affected almost every part of life and economy. Consequently, this study sets off to assess the effect of the pandemic on blood supply and blood transfusion in the University of Calabar Teaching Hospital. METHODS: Data from the Donor Clinic and Blood Group Serology Unit of the University of Calabar Teaching Hospital were retrospectively extracted to evaluate supply and use of blood before and during COVID-19 pandemic. RESULT: A total of 1638 donors were recorded within the study period. Age range 15-29 and 30-44 years constituted majority of the subjects (58.9% and 33.4%, respectively). The donor pool were male-dominated. Commercial donors (61.7%) and family replacement donors (30.6%) constituted majority of the donor pool. Most of the donor pool were students (37.1%), public servants (22.8%) and artisans (18.6%). A concomitant decrease of 26.1% and 18.9% were recorded in blood donation and request during the COVID-19 pandemic. CONCLUSION: Blood supply was not significantly affected in our study center as both requests and donations decreased. Consideration for improving family replacement donation was advised.

Highlights • Evaluation of EDITM ELISAs for detection of SARS-CoV-2 IgM and IgG in human plasma. • Antibodies were measured in 64 SARS-CoV-2 RT-PCR confirmed COVID-19 patients. • with serial blood samples collected at different time points from symptom onset, • in 200 healthy blood donors, and in 256 intensive care unit patients. • High “true” vs. low “false” positivity rates for the SARS-CoV-2 IgM and IgG ELISAs., Background Besides SARS-CoV-2 RT-PCR testing, serological testing is emerging as additional option in COVID-19 diagnostics. Aim of this study was to evaluate novel immunoassays for detection of SARS-CoV-2 antibodies in human plasma. Methods Using EDITM Novel Coronavirus COVID-19 Enzyme Linked Immunosorbent Assays (ELISAs), we measured SARS-CoV-2 IgM and IgG antibodies in 64 SARS-CoV-2 RT-PCR confirmed COVID-19 patients with serial blood samples (n=104) collected at different time points from symptom onset. Blood samples from 200 healthy blood donors and 256 intensive care unit (ICU) patients collected before the COVID-19 outbreak were also used. Results The positivity rates in the COVID-19 patients were 5.9% for IgM and 2.9% for IgG ≤5 days after symptom onset; Between day 5 and day 10 the positivity rates were 37.1% for IgM and 37.1% for IgG and rose to 76.4% for IgM and 82.4% for IgG after >10-15 days. After 15-22 days the “true” positivity rates were 94.4% for IgM and 100% for IgG. The “false” positivity rates were 0.5% for IgM and 1.0% for IgG in the healthy blood donors, 1.6% for IgM and 1.2% for IgG in ICU patients. Conclusions This study shows high “true” vs. low “false” positivity rates for the EDITM SARS-CoV-2 IgM and IgG ELISAs.

Highlights • We sorted out the values of laboratory indexes in the collected literature and calculated out the final statistical values. • We compared the laboratory indexes of severe and non-severe patients infected with COVID-19, which facilitates early diagnosis and treatment of severe and critically ill patients. • We also described the combined analysis of different types of laboratory indexes, which played an important role in predicting the development of patients' condition., Background The pandemic coronavirus disease 2019 (COVID-19) has threaten the global health. The characteristics of laboratory findings of coronavirus are of great significance for clinical diagnosis and treatment. We found indicators that may most effectively predict a non-severe COVID-19 patient develop into a severe patient. Methods We conducted a meta-analysis to compare the laboratory findings of severe patients with non-severe patients with COVID-19 from searched articles. Results Through the analysis of laboratory examination information of patients with COVID-19 from 35 articles (5912 patients), we demonstrated that severe cases possessed higher levels of leukocyte (1.20-fold), neutrophil (1.33-fold), CRP (3.04-fold), PCT (2.00-fold), ESR (1.44-fold), AST (1.40-fold), ALT (1.34-fold), LDH (1.54-fold), CK (1.44-fold), CK-MB (1.39-fold), total bilirubin(1.14-fold), urea (1.28-fold), creatine (1.09-fold), PT (1.03-fold) and D-dimer (2.74-fold), as well as lower levels of lymphocytes (1.44-fold), eosinophil (2.00-fold), monocyte (1.08-fold), Hemoglobin (1.53-fold), PLT (1.15-fold), albumin (1.15-fold), and APTT (1.02-fold). Lymphocyte subsets and series of inflammatory cytokines were also different in severe cases with the non-severe ones, including lower levels of CD4 T cells (2.10-fold) and CD8 T cells (2.00-fold), higher levels of IL-1β (1.02-fold), IL-6 (1.93-fold) and IL-10 (1.55-fold). Conclusions Some certain laboratory inspections could predict the progress of the COVID-19 changes, especially lymphocytes, CRP, PCT, ALT, AST, LDH, D-dimer, CD4 T cells and IL6, which provide valuable signals for preventing the deterioration of the disease.

Highlights • Dysregulated and excessive cytokine storm is essential for critically ill COVID-19 development. • Mechanisms of SARS-CoV-2-infected included ACE2 receptor-mediated inflammatory response, Cell Pyroptosis, Delayed IFN-α and -β response, and Anti-S-IgG-mediated inflammatory response. • Infected lung epithelial cells and macrophages mainly release cytokines involved in innate immunity and adaptive immunity. • Endothelial cells damage and microvascular thrombosis played an important role in pathophysiological changes of COVID-19 involved respiratory failure, hypotension, liver and kidney injury. • Individualized targeted cytokine drug therapy might an effective measure to alleviate the violent inflammation besides Remdesivir, Chloroquine, Lopinavir and Ritonavir, Favipiravir etc., A novel coronavirus disease 2019 (COVID-19) triggered by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is currently spreading globally, causing severe pneumonia and acute lung injury in many patients. Even worse, severe respiratory may develop into acute respiratory distress syndrome and multiple organ dysfunction syndrome in COVID-19. The cytokine storm caused by immune over-activation due to virus infection may be an important cause of death in the late period of progress, but the pathogenesis of cytokine storm is still unclear. This article reviews the mechanisms of SARS-CoV-2-induced cytokine storm in detail based on the current discovered researches, and put forward some valuable medication ideas for the targeted cytokines drug researches and treatment. The goal of this work will be helpful for reducing excessive immune response.

Copyright © 2022 Morais, Medeiros, Carvalho, Morello, Teixeira, Maciel, Nhantumbo, Balau, Rosa, Nogueira, Rodrigues, Carvalho, Antunes and Arez. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms., Mechanisms of malaria parasite interaction with its host red blood cell may provide potential targets for new antimalarial approaches. Pyruvate kinase deficiency has been associated with resistance to malaria in both experimental models and population studies. Two of the major pyruvate kinase deficient-cell disorders are the decrease in ATP and the increase in 2,3-biphosphoglycerate (2,3-BPG) concentration. High levels of this metabolite, only present in mammalian red blood cell, has an inhibitory effect on glycolysis and we hypothesized that its accumulation may also be harmful to the parasite and be involved in the mechanism of protection provided by that enzymopathy. We examined the effect of a synthetic form, 2,3-DPG, on the Plasmodium falciparum intraerythrocytic developmental cycle in vitro. Results showed an impairment of parasite growth with a direct effect on parasite maturation as significant lower progeny emerged from parasites that were submitted to 2,3-DPG. Further, adding the compound to the culture medium did not result in any effect on the host cell, but instead the metabolic profile of an infected cell became closer to that of a non-infected cell., This research was funded by Fundação para a Ciência e Tecnologia (https://www.fct.pt/index.phtml.en), projects PTDC_BIA-CEL_28456_2017 and GHTM - UID/04413/2020 (https://ghtm.ihmt.unl.pt/).

Despite the good biocompatibility and antibacterial activity of zinc sulfide nanoparticles (ZnS NPs), whether they possess antiviral activity is still unclear. Here, GSH-modified ZnS NPs (GSH-ZnS NPs) were synthesized and their significant antiviral activity was demonstrated using the

Recent evidence suggests that proinflammatory cytokines, such as tumor necrosis factor α (TNF-α), play a pivotal role in the development of inflammatory-related pathologies (covid-19, depressive disorders, sepsis, cancer, etc.,). More importantly, the development of TNF-α biosensors applied to biological fluids (e.g. sweat) could offer non-invasive solutions for the continuous monitoring of these disorders, in particular, polydimethylsiloxane (PDMS)-based biosensors. We have therefore investigated the biofunctionalization of PDMS surfaces using a silanization reaction with 3-aminopropyltriethoxysilane, for the development of a human TNF-α biosensor. The silanization conditions for 50 μm PDMS surfaces were extensively studied by using water contact angle measurements, electron dispersive X-ray and Fourier transform infrared spectroscopies, and fluorescamine detection. Evaluation of the wettability of the silanized surfaces and the Si/C ratio pointed out to the optimal silanization conditions supporting the formation of a stable and reproducible aminosilane layer, necessary for further bioconjugation. An ELISA-type immunoassay was then successfully performed for the detection and quantification of human TNF-α through fluorescent microscopy, reaching a limit of detection of 0.55 μg/mL (31.6 nM). Finally, this study reports for the first time a promising method for the development of PDMS-based biosensors for the detection of TNF-α, using a quick, stable, and simple biofunctionalization process.

Scientific reports 12, 19452 (2022). doi:10.1038/s41598-022-23702-y, Published by Macmillan Publishers Limited, part of Springer Nature, [London]

INTRODUCTION: The novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a pandemic. Many studies have shown that several laboratory parameters are related to disease severity and mortality in SARS-CoV-2 cases. This meta-analysis aimed to determine the relationship of a prognostic factor, D-dimer, with disease severity, need for intensive care unit (ICU) care, and mortality in SARS-CoV-2 patients. METHODS: A systematic search for all observational studies and trials involving adult patients with SARS-CoV-2 that had any data related to D-dimer on admission was conducted using PubMed, Science Direct, Scopus, ProQuest, and MedRxiv databases. We performed random-effects inverse-variance weighting analysis using mean difference (MD) of D-dimer values for outcomes such as disease severity, mortality, and need for ICU care. RESULTS: A total of 29 studies (4,328 patients) were included in this meta-analysis, which revealed a higher mean of D-dimer levels on admission in severe patients than in nonsevere patients (MD = 0.95, [95% confidence interval (CI): 0.61-1.28], P

Objective Reorganization of the emergency department (ED) during the COVID-19 pandemic implied closure of the ED-dedicated laboratory and manual transport of all specimens to the dislocated central laboratory. The impact of such reorganization on laboratory turnaround time (TAT) was examined. Methods The TAT from blood sampling to specimen reception (TAT1), from specimen reception to test reporting (TAT2), and from sampling to test reporting (TAT3) were compared between the pandemic peak month in 2020 and the same month in 2019. We evaluated whether TAT2 fulfills the recommended 60-minute criteria. Results A statistically significant difference was observed for all comparisons (P Conclusion An ED reorganization compromised the usual laboratory services for patients in the ED, with manual specimen delivery being the main cause for TAT prolongation.