Molecular Diagnostics on Smartphone

During the COVID-19 pandemic, point-of-care genetic testing (POCT) devices were used for on-time and on-site detection of the virus, which helped to prevent and control the spread of the pandemic. Smartphones, which are widely used electronic devices with many functions, have the potential to be used as a molecular diagnostic platform for universal healthcare monitoring. Several integrated diagnostics platforms for the real-time and end-point detection of COVID-19 were developed using the functions of smartphones, such as the operating system, power, sound, camera, data storage, and display. These platforms use the 5V output power of smartphones, which can be amplified to power a micro-capillary electrophoresis system or a thin-film heater, and the CMOS camera of smartphones can capture the color change during a colorimetric loop-mediated isothermal amplification test and detect fluorescence signals. Smartphones can also be used with self-written web-based apps to enable automatic and remote pathogen analysis on POCT platforms. Our lab developed a handheld micro-capillary electrophoresis device for end-point detection of SARS-CoV-2, as well as an integrated smartphone-based genetic analyzer for the qualitative and quantitative colorimetric detection of foodborne pathogens with the help of a custom mobile app. © 2023 SPIE.

The evaluation of practical learning instruction management and student satisfaction with the "photocolorimetric methodology platform for measuring egg yolk color" during the COVID-19 pandemic situation

Online teaching management has been widely used during the COVID-19 pandemic situation and it has a direct impact on practical teaching management because students do not have access to equipment, chemicals, and tools. This study's purpose is to evaluate practical learning instruction management and student satisfaction with "photocolorimetric methodology platform for measuring egg yolk color" during the COVID-19 pandemic. This study compared the student satisfaction and effectiveness of a learning instruction platform for measuring egg yolk color using a laboratory machine and an online teaching management platform using photocolorimetric methodology. The results of this experiment revealed that the two platforms evaluated yolk colors L*, a*, and b* similarly (P > 0.05). Furthermore, the students were satisfied with the learning instruction with the photocolorimetric methodology platform for measuring egg yolk color at 4.76 points or the most level.

Pterin interactions with gold clusters: A theoretical study

Gold (Au) nanoclusters (NCs) are novel materials with low cytotoxicity and high chemical stability. These properties are in high demand during the bioimaging. Moreover, the optical properties of gold clusters allow to use them as colorimetric and luminescent bionanosensors. Pterins are low molecular weight organic compounds, which are used in medicine as biomarkers of phenylketonuria, vitiligo, inflammation and immune system activation, cancer, COVID-19, etc. We have investigated the possibility of gold nanosensors usage to detect pterin (Ptr). Ptr-Aunq structures (n = 1–6;q = 0–2) Gibbs energy of complexation (Eb) have been obtained using density functional theory. The highest Eb was determined for the complexes of Au62+ and Au32+ in acidic and alkaline aqueous solution, respectively. The detection of pterin with gold clusters seems to be prospective using both colorimetric and fluorescent detection because of the intense S0→S1 transition in the absorption spectrum of the Au5+ complex. Raman detection of pterin should be performed at alkaline pH because of the dramatic changes in the spectrum of Ptr−1 upon the addition of Au clusters. We believe that these tunable changes of the pterin spectra due to Au clusters and nanoparticles attachment could be exploited in further studies on nanosensor design. © 2023

Multipurpose advanced split T7 promoter-based transcription amplification for ultrasensitive molecular diagnostics

An accurate, convenient, and rapid diagnostic platform, which can be applied in facility-limited or point-of-care (POC) settings, is essential to help prevent the spread of infectious diseases and enable the most effective treatment to be selected. In this study, we describe the development of a new isothermal molecular diagnostic system named multipurpose advanced split T7 promoter-based transcription amplification (MASTER) for the rapid and ultrasensitive detection of various pathogens containing single-stranded RNA and double-stranded DNA. MASTER produces a large number of RNA amplicons in the presence of target pathogens, which generate fluorescence or colorimetric signals based on light-up RNA aptamers or lateral flow assays. Implementing MASTER at 37 °C for<1 h achieved the detection of a single copy per reaction without cross-reactivity. Moreover, the testing of 40 clinical samples revealed that MASTER exhibited excellent accuracy with 100% sensitivity and specificity for SARS-CoV-2 diagnosis. Furthermore, a one-pot MASTER system capable of accelerating practical applications was demonstrated, indicating that the MASTER system is a promising platform for the effective surveillance of various pathogens. © 2023 Elsevier B.V.

A real-time monitoring platform of colorimetric LAMP for developing rapid visual detection kits of SARS-CoV-2

Visual detection of nucleic acids is important to diagnose the serious acute infectious diseases such as coronavirus disease 2019 (COVID-19). During this pandemic, reliable visual detection kits have been in high demand for screening and prevention of the virus. While developing these visual detection kits, a real-time monitoring platform is usually applied to study the amplification and detection processes of nucleic acids and optimize the detecting conditions. Herein, we developed a real-time monitoring platform of colorimetric loop-mediated isothermal amplification (LAMP) to investigate the amplification and detection processes of nucleic acids. Using this platform, we could obtain the real-time amplification curves, and optimize the reaction temperature, color change, and detection time. Based on the optimized conditions, a visual detection kit for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was successfully developed with a sensitivity of 102 copies µL−1 in 12 min. This real-time monitoring platform has advantages of simple construction, steady performance, high sensitivity, and outstanding anti-pollution capability, and could replace the traditional colorimetric methods by photographing and reading values. This platform would accelerate the development of visual detection kits for colorimetric LAMP, help to explore the amplification and transcription of nucleic acids, and provide support for the prevention of emerging biological threats. © 2023

Loop-Mediated Isothermal Amplification on Paper Microfluidic Chips for Highly Sensitive and Specific Zika Virus Detection Using Smartphone.

Zika virus (ZIKV) infection may cause serious birth defects and is a critical concern for women of child-bearing age in affected regions. A simple, portable, and easy-to-use ZIKV detection method would enable point-of-care testing, which may aid in prevention of the spread of the virus. Herein, we describe a reverse transcription isothermal loop-mediated amplification (RT-LAMP) method that detects the presence of ZIKV RNA in complex samples (e.g., blood, urine, and tap water). Phenol red is the colorimetric indicator of successful amplification. Color changes based on the amplified RT-LAMP product from the presence of viral target are monitored using a smartphone camera under ambient light conditions. A single viral RNA molecule per µL can be detected in as quickly as 15 min using this method with 100% sensitivity and 100% specificity in blood and tap water, while 100% sensitivity and 67% specificity in urine. This platform can also be used to identify other viruses including SARS-CoV-2 and improve the current state of field-based diagnostics.

Association of Interleukin-6 (IL-6) and High-Density Lipoprotein Cholesterol (HDL-C) with Disease Severity of Coronavirus Disease 2019 (COVID-19)

Introduction: IL-6 is the key molecule of cytokine storm in COVID-19. Dyslipidemia is a common complication in patients with Coronavirus disease 2019 (COVID-19), but the association of dyslipidemia with the severity of COVID-19 is still unclear. In this study, we aimed to investigate the biochemical alterations of High-Density Lipoprotein Cholesterol (HDL-C), and Interleukin-6 (IL-6) in COVID-19 patients and their relationships with the disease severity. Material(s) and Method(s): We conducted a retrospective single-institutional study of 99 consecutive confirmed cases of COVID-19. Serum IL-6 and HDL-C concentrations, demographic and clinical profile were collected during hospital stay. Duration of study was from September 2020 to August 2021. Descriptive statistics were applied to summarize the demographic data. Results are reported as mean with standard deviation. Receiver operating characteristic curve (ROC) analysis was used to compare biochemical markers. Result(s): Serum HDL-C levels had a significant positive correlation with SpO2 with correlation coefficient r = 0.589. Serum IL-6 had a negative correlation with SpO2 with correlation coefficient r =-0.632. The AUC for IL6 and HDL-C in predicting COVID severity is 0.982 and 0.985 respectively. Conclusion(s): HDL-C is decreased and IL-6 is increased with the disease severity.Copyright © 2023, Dr Yashwant Research Labs Pvt Ltd. All rights reserved.

The role of reactive nitrogen species as biomarkers of disease severity in COVID-19

Reactive nitrogen species (RNS) such as nitric oxide (NO) can be produced by local pulmonary cells and inflammatory cells, and they play a role in the pathogenesis of chronic pulmonary diseases and pulmonary infections. NO is an unstable compound turning to a more stable nitrite and nitrate rapidly. The objective of our study was to investigate the association between plasma nitrate+nitrite, nitrate, and nitrite levels and the severity of coronavirus disease (COVID)-19. Blood plasma samples of mild, moderate, and severe COVID-19 patients were collected from 2 different hospitals. Plasma of healthy subjects, who had never COVID-19 was used as controls (n=20 for each group). Samples were isolated by centrifugation following ultrafiltration prior to the commercial nitrate/nitrite colorimetric assay kit. Plasma nitrate+nitrite and nitrate levels, respectively, were significantly increased in severe patients (medians=34.4muM and 33.4muM), as compared to mild groups (medians=22.3muM and 20.6muM;p<0.05). In contrast, nitrite levels were significantly lower in severe patients (median= 0.8muM) than mild patients (median= 1.6muM;p<0.0001). Patients with severe disease were older (64.9 years) than the mild patients (50.6 years;p<0.05). In severe patients, age was positively correlated with nitrate+nitrite and nitrate levels, respectively (r=0.502, p=0.024;r=0.489, p=0.029). Our findings suggest that RNS may play a role in the pathogenesis of COVID-19 and that it may be considered as a marker of severity.

Is The Co-administration of Metformin and Clomiphene Superior to Induce Ovulation in Infertile Patients with Poly Cystic Ovary Syndrome and Confirmed Insulin-resistance: A Double Blind Randomized Clinical Trial

Objective: This study aimed to compare the effects of clomiphene citrate (CC) combined with metformin or placebo on infertile patients with poly cystic ovary syndrome (PCOS) and insulin resistance (IR). Material(s) and Method(s): We included 151 infertile women with PCOS and IR in a university hospital from November 2015 to April 2022 in this prospective, double-blind, randomized, placebo-controlled trial. Patients were randomized into two groups;group A: received CC plus metformin (n = 76) and group B: received CC plus placebo (n = 75). The ovulation rate was the main outcome measure. Clinical pregnancy, ongoing pregnancy, live birth and abortion rates were secondary outcome measures. Result(s): There was no remarkable difference in ovulation rate in two groups. Moreover, no significant changes were observed in clinical pregnancy, ongoing pregnancy, live birth and abortion rates between two groups. A larger proportion of women in group A suffered from side effects of metformin (9.3% versus 1.4%;p=0.064), although this was not significant. Conclusion(s): In IR infertile women with PCOS, metformin pre-treatment did not increase the ovulation, clinical pregnancy and live birth rates in patients on clomiphene citrate.Copyright © 2023 Tehran University of Medical Sciences.

Malachite Green-Based Detection of SARS-CoV-2 by One-Step Reverse Transcription Loop-Mediated Isothermal Amplification

The pandemic of severe acute respiratory syndrome 2 (SARS-CoV-2) revealed the necessity of diagnosis of the infected people to prevent the prevalence infection cycle. Many commercial pathogen diagnosis methods are based on the detection of genomic materials. Isothermal amplification methods such as loop-mediated-isothermal amplification (LAMP) are the method of choice in these cases. Reverse transcription steps are efficiently coupled to LAMP for the detection of pathogens with genomic RNAs such as SARS-CoV-2. Many detection systems for LAMP include fluorescent readout systems. Although such systems result in desirable limits of detection, the need for special instrumentation is the main dispute of such systems to become real point of care assays. In contrast, colorimetric detection methods would reduce costs and improve the applicability of the system. In this study one-step reverse transcription-LAMP reaction was established that enables visual detection of the SARS-CoV-2 genome. Nasopharyngeal RNA samples were first validated by reverse transcription quantitative polymerase chain reaction and then subjected to RT-LAMP. To lower the cost associated with the readout system equipment, malachite green (MG) was used. The color change of MG to blue allowed visual detection of the virus. Firstly, experiments were set up as two-step RT-LAMP reaction to identify the best primer sets. In addition, MG concentration was optimized with the significant colorimetric signal for the positive samples. Next, a one-step colorimetric method was developed for the detection of SARS-CoV-2 based on MG color shift in 2 h.

Gold-silver alloy hollow nanoshells-based lateral flow immunoassay for colorimetric, photothermal, and SERS tri-mode detection of SARS-CoV-2 neutralizing antibody.

Although many approaches have been developed for the quick assessment of SARS-CoV-2 infection, few of them are devoted to the detection of the neutralizing antibody, which is essential for assessing the effectiveness of vaccines. Herein, we developed a tri-mode lateral flow immunoassay (LFIA) platform based on gold-silver alloy hollow nanoshells (Au-Ag HNSs) for the sensitive and accurate quantification of neutralizing antibodies. By tuning the shell-to-core ratio, the surface plasmon resonance (SPR) absorption band of the Au-Ag HNSs is located within the near infrared (NIR) region, endowing them with an excellent photothermal effect under the irradiation of optical maser at 808 nm. Further, the Raman reporter molecule 4-mercaptobenzoic acid (MBA) was immobilized on the gold-silver alloy nanoshell to obtain an enhanced SERS signal. Thus, these Au-Ag HNSs could provide colorimetric, photothermal and SERS signals, with which, tri-mode strips for SARS-CoV-2 neutralizing antibody detection were constructed by competitive immunoassay. Since these three kinds of signals could complement one another, a more accurate detection was achieved. The tri-mode LFIA achieved a quantitative detection with detection limit of 20 ng/mL. Moreover, it also successfully detected the serum samples from 98 vaccinated volunteers with 79 positive results, exhibiting great application value in neutralizing antibody detection.

A triple-target reverse transcription loop-mediated isothermal amplification (RT-LAMP) for rapid and accurate detection of SARS-CoV-2 virus.

The spreading of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) across the world has impacted people's health and lives worldwide in recent years. Rapid and accurate diagnosis is crucial for curbing the pandemic of coronavirus disease 2019 (COVID-19). Reverse transcription loop-mediated isothermal amplification (RT-LAMP) has great potential for SARS-CoV-2 detection but fails to completely replace conventional PCR due to the high false-positive rate (FPR). We proposed a triple-target RT-LAMP method for dual-signal, sensitive, and simultaneous detection of conserved genes of SARS-CoV-2. Multiple LAMP primer sets were designed for N, E, and M genes and their amplification efficacy were screened. Then, using artificial plasmids and RNA, the optimal primer set for each gene was examined on specificity, sensitivity, and detection range. The RT-LAMP initiated by these primer sets exhibited better specificity and sensitivity than that of RT-qPCR, and the triple-target RT-LAMP could determine different variants of SARS-CoV-2. By testing 78 artificial RNA samples, the total FPR of triple-target RT-LAMP was eliminated compared with that of mono-target RT-LAMP. The triple-target RT-LAMP method precisely identified throat swab specimens through colorimetry and fluorescent signals within 60 min, and the limit of detection (LOD) was as low as 187 copies/reaction. In the future, the triple-target RT-LAMP can be applied to in-field and on-site diagnosis of symptomatic and asymptomatic virus carriers.

Increase of nutritional security in Sub-Saharan Africa through the production of dried products from underutilized crops

Currently, an estimated 20% of the population in Sub-Saharan Africa is food insecure with the incidence of hunger and malnutrition still rising. This trend is amplified by the socio-economic consequences of the COVID-19 pandemic. In contrast, more than a third of the harvestable perishable produce is lost due to a lack of preservation or failure to utilize preservation as is the case for underutilized crops (UCs). Moreover, some of the preservation techniques utilized are poor, leading to the deterioration of food quality, especially the micronutrients. In this study, we thus exemplarily investigated the impact of different drying settings on the quality of two highly nutritious UCs, namely cocoyam and orange-flesh sweet potato (OFSP) (40, 60, and 80 °C for cocoyam and 40, 50, 60, and 70 °C for OFSP) to deduce the optimum quality retention and further develop a theoretical design of processing units and processing guidelines for decentralized food processing. Drying cocoyam at 80 °C and OFSP at 60 °C, respectively resulted in a relatively shorter drying time (135 and 210 min), a lower total color difference (2.29 and 11.49-13.92), greater retentions for total phenolics (0.43 mg GAE/100 gDM and 155.0-186.5 mg GAE/100 gDM), total flavonoid (128 mg catechin/100 gDM and 79.5-81.7 mg catechin/100 gDM) and total antioxidant activity (80.85% RSA and 322.58-334.67 mg AAE/100 gDM), respectively for cocoyam and OFSP. The β-carotene, ascorbic acid and vitamin A activity per 100 gDM of the OFSP flours ranged between 6.91- 9.53 mg, 25.90 − 35.72 mg, and 0.53 − 0.73 mg RAE, respectively. © 2022 The Author(s). Published with license by Taylor and Francis Group, LLC.

Development of a colorimetric assay for quantification of favipiravir in human serum using ferrihydrite

We developed a colorimetric analytical method for favipiravir (FPV), a promising treatment for COVID-19. FPV forms yellow complexes with ferrihydrite (Fh) by a ligand substitution reaction with the iron (III) hydroxyl surface groups in Fh. Fh-coated microbeads were packed into a capillary tube with an inner diameter of 1 mm. FPV-spiked serum after pretreatment was drawn into the capillary packed with Fh-coated microbeads. The intensities of the yellow-color complexes on the microbeads were transformed into red-green-blue (RGB) pixels using Image-J software 1.8, and the difference between green and blue was used as the analytical signal. The signal increased with increasing FPV concentration. The proposed method showed good linearity (R2 = 0.9907) over a concentration range of 25–200 μg/mL. The RSD (n = 3) and the LOD (3σ) values were estimated to be 2.8–15.4% and 16.91 μg/mL, respectively. The proposed method enables us to quantify FPV rapidly and easily without the need for expensive equipment. © 2022 Elsevier B.V.

Dual-mode visual detection strategies of viable pathogens for point-of-care testing

Here, we introduce a power-free foldable poly(methyl methacrylate) (PMMA) microdevice fully integrating DNA extraction, amplification, and visual detection, realized in novel dual modes – colorimetric and aggregate formation – using 4-Aminoantipyrine (4-AP) for monitoring pathogens. The microdevice contains two parts: reaction and detection zones. A sealing film was utilized to connect the two zones and make the device foldable. The FTA card was deposited in the reaction zone for DNA extraction, followed by loop-mediated isothermal amplification (LAMP) at 65 °C for 45 min. When the detection zone is folded toward the reaction zone, paper discs modified with 4-AP placed in the detection zone are delivered to the reaction zone. Specifically, in the presence of LAMP amplicons, 4-AP is oxidized into antipyrine red or generates aggregates by interacting with copper sulfate, forming copper hybrid nanostructure (Cu-hNs). In the absence of LAMP amplicons, 4-AP is not oxidized and maintains yellow color or fails to form aggregates. Furthermore, we introduced the ethidium homodimer-1 (EthD-1) to identify viable bacteria. EthD-1 penetrated the compromised membranes of nonviable cells and prevented further DNA amplification by intercalating with the DNA. In this way, only samples containing viable cells displayed color change or formed aggregates upon reaction with 4-AP. Using this method, SARS-CoV-2 RNA and Enterococcus faecium were identified by naked eye, with the limit of detection of 103 copies/μL and 102 CFU/mL, respectively, within 60 min. The introduced microdevice can be used for rapidly monitoring viable pathogens and controlling outbreaks of infectious disease in resource-limited settings. © 2022 Elsevier B.V.

Colorimetric and Raman dual-mode lateral flow immunoassay detection of SARS-CoV-2 N protein antibody based on Ag nanoparticles with ultrathin Au shell assembled onto Fe3O4 nanoparticles.

Serological antibody tests are useful complements of nuclei acid detection for SARS-CoV-2 diagnosis, which can significantly improve diagnostic accuracy. However, antibody detection in serum or plasma remains challenging to do with high sensitivity. In this study, Ag nanoparticles with ultra-thin Au shells embedded with 4-mercaptobenzoic acid (MBA) (AgMBA@Au) were manufactured and then assembled onto Fe3O4 surface by electrostatic interaction to construct the Fe3O4-AgMBA@Au nanoparticles (NPs) with magnetic-Raman-colorimetric properties. Based on the composite nanoparticles, a colorimetric and Raman dual-mode lateral flow immunoassay (LFIA) for ultrasensitive identification of SARS-CoV-2 nucleocapsid (N) protein antibody was constructed. The magnetic nanoparticles (Fe3O4 NPs) were acted as the core and coated a layer of AgMBA@Au particles on the surface by electrostatic interaction to prepare Fe3O4-AgMBA@Au NPs, which can amplify the SERS signal due to multiple AgMBA@Au particles concentrated on a single magnetic nanoparticle. Moreover, the Fe3O4-AgMBA@Au NPs facilitated pre-purifying sample using magnetic separation, and complex matrix interference would be greatly decreased in the detection. The Fe3O4-AgMBA@Au NPs modified with N protein recognized and bound with N protein antibodies, which were trapped on the T-line, forming color band for observing detection. Under optimal conditions, the N protein antibodies could be qualitatively detected in colorimetric mode with the visual limit of 10-8 mg/mL and quantitatively detected by SERS signals between 10-6 and 10-10 mg /mL with 0.08 pg/mL detection limit. The coefficients variations (CV) of intra-assay was 8.0%, whereas of inter-assay was 11.7%, confirming of good reproducibility. Finally, this approach was able to discriminate between positive, negative, and weakly positive samples when detecting 107 clinical serum samples. The process enables highly sensitive quantitative assays that are valuable for evaluating disease processes and guiding treatment. Colorimetric and Raman dual-mode LFIA detection of SARS-CoV-2 N protein antibody based on Fe3O4-AgMBA@Au nanoparticles.

PART 1: ULTRAVIOLET-C (UVC) EXPOSURE AND ITS EFFECT ON COLOR CHANGE FOR POLYSTYRENE

Due to the recent and ongoing pandemic - COVID-19 - there was an urgency to determine a method to delay the continuously rapid development of the new virus. As a result, Ultraviolet-C (UVC) light, also known as Ultraviolet Germicidal Irradiation (UVGI), has been in higher demand because of its known ability to disinfect quickly and effectively. However, because of its short wavelength/higher energy, either 222nm or 254nm, material degradation is usually much more accelerated than Ultraviolet-A (UVA) or Ultraviolet-B (UVB). At this moment, this study only observed color change when exposing polystyrene to UVC light, and it is believed that this is one of the first studies, if not the first, conducted with this material. Polystyrene was selected because of its availability, abundance of relevant research (ie. UVA/UVB exposure results), and its use in weathering standards. Additionally, since there are no standards specifically about UVC exposure, this preliminary research may provide some direction. © 2022 Society of Plastics Engineers. All rights reserved.

Detection of SARS-CoV-2 by RT-LAMP assay in Human COVID-19 patients

Background: The novel COVID-19 outbreak has infected human population all around the world. Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) diagnosis in a rapid manner remains challenging for health care professionals. Currently, RT-qPCR technique is extensively practiced in SARS-CoV-2 diagnosis and is considered as gold standard. The constraints of RT-qPCR, high cost and need for trained technician, longer detection time, highlighted the need for alternate healthcare diagnostic approaches. They follow the WHO assured standard and offer the health-care sector optimism. One of them is the Loop Mediated isothermal amplification system (LAMP). There is no need for costly equipment like thermal cycler since LAMP assay is performed at a fixed temperature. It can also be implemented as a point of care testing device. RT-LAMP is one of the extensively used isothermal amplification system in pathogen diagnostics.Aims: The current study aims to validate and standardize RT-LAMP assay for rapid diagnosis of SARS-CoV-2 in both lab and field conditions. The reactions can be carried out using a heating vessel including the use of a water bath and end-point detection by colorimetry. A rising middle ground of tiny, more portable technology, that provides most of the capability at less cost and time.Methods and Results: 20 Samples were taken from COVID-19 positive patients. RNA extraction from COVID-19 samples was followed up by one-step reverse transcription and loop-mediated isothermal amplification (LAMP). LAMP primers were designed to amplify the conserved regions of SARS-COV-2 specific genes. The target regions for primer design were selected after genome-wide sequence alignment of SARS-CoV-2 strains isolated in various regions of the world i.e., Europe, Africa, Asia, and North America. RT-LAMP assays were performed at the specific incubation temperature (60°C) for 50 minutes. Assay was optimized as per consumable compatibility, COVID template integrity, primer concentration, template concentration, primer ratio, testing time etc. Sensitivity and specificity of the assay was elucidated. Finally, different end-point analysis i.e., Agarose Gel Electrophoresis and Colorimetry have been used to interpret the results.Conclusion: RT-LAMP assay has shown to be a quick and accurate diagnostic method that can be put to use for SARS-CoV-2 detection in laboratories and Point-of- Care settings. © 2022 IEEE.

CINEMATOGRAPHIC COLORIMETRY APPLIED TO NEW TELEVISION JOURNALISTIC FORMATS: THE CASE OF THE SPANISH PROGRAM LO DE ÉVOLE

With the irruption of multi-screens that the Netflix era has brought about, a good part of the old visual formulas, especially television and reporter ones, have become outdated. This is especially observable in television formats strongly linked to reporter journalism, such as Spaniards around the world, Street travelers and others. In recent years, a trend of adaptation of cinematographic codes to television products has already been perceived, but the Covid-19 pandemic and the ban on theatrical exhibition in cinemas around the world has precipitated a change in the model of consumption. This article presents an investigation on a nuclear element: the cinematographic colorimetry used in journalistic television programs, specifically, one of the most watched journalistic programs on Spanish television: Lo de Évole. © 2022 BY-NC.

Coronavirus but not influenza virus infection induces the expression of both cellular and soluble urokinase plasminogen activator receptor

Background: Urokinase plasminogen activator receptor (uPAR/PLAUR) is a cell surface receptor on monocytes and macrophages (MO/MPs), which when cleaved, forms soluble uPAR (suPAR). Previous work has shown that uPAR is differentially induced in MO/MPs after coronavirus (CoV) infection and contributes to hyperfibrinolysis. In contrast to cellular uPAR's fibrinolytic function (implicated in CoV-induced coagulopathy), suPAR is a MP chemotactic factor implicated in perpetuating tissue injury after CoV infection. However, it is unclear if induced levels of cellular uPAR correlate with the elevated levels of suPAR after CoV infection and how suPAR expression is regulated after influenza virus (IAV) infection. Method(s): Published scRNA sequencing data (GSE149689) of PBMCs from patients (COVID-positive and IAV-positive) and healthy controls was analyzed. In vitro, we performed infections of immortalized murine MPs (RAW264.7) with the CoVs MHVA59 or MHV1, or the IAV (A/PR/8/34;H1N1) at the indicated MOI for 24 hours. uPAR and urokinase levels were measured by ELISA. Urokinase activity of conditioned media was measured using a commercially available colorimetric assay. Result(s): ScRNA analysis revealed differential expression of PLAUR in MOs from COVID and IAV-positive relative to healthy controls, with the highest PLAUR expression in COVID-positive samples (Fig. A-B). Infection of RAW264.7 cells with CoV and IAV induced cellular uPAR expression, but only CoV infection induced suPAR expression (Fig. C). In contrast to uPAR, the expression of both cellular and soluble urokinase levels was induced after infection with either CoVs or IAV (Fig. D). Finally, induction of urokinase activity of conditioned media after viral stimulation was observed after infection with either CoVs or IAV (Fig. E). Conclusion(s): CoV but not IAV infection induces the expression of both cellular and soluble uPAR in MO/MPs. Both CoV and IAV induce the expression of cellular and soluble urokinase, and urokinase activity. Understanding this virus specific effect on uPAR/suPAR in innate immune cells may reveal the pathogenesis of virus-associated coagulopathy and tissue injury.