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Aside from evaporation of water from buffer solutions, precipitation of buffer salts can also easily occur when a buffer solution comes into contact with a pure organic solvent. [...]one should avoid- at all costs-a situation where a LC system component containing an aqueous buffer is flushed immediately with a pure organic solvent. Solvents and Buffers Figure 1 shows a picture of a buffer bottle I observed in an LC laboratory about a year ago. Many of the aqueous buffer solutions used in LC (and even high-performance liquid chromatography [HPLC] grade water, if given enough time and exposure to aboratory dust) are environments quite favorable to microbes, particularly those in the middle of the pH range.
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Colorimetric RT-LAMP Assay is a diagnostic method that has attracted much attention because of its rapidity, simplicity, and accuracy compared to other disease diagnosis methods. Despite having many advantages, the RT-LAMP Colorimetric Assay has disadvantages, especially for kits that use phenol red as an indicator. The disadvantage derives from the input RNA/DNA samples containing high buffer levels, which causes no color change and false-negative results. This study aimed to develop and optimize the colorimetric RT-LAMP method on high-buffered SARS-CoV-2 RNA samples. We found that a temperature of 69°C for 50 minutes with the addition of post-treatment in the form of heating at 80°C for 10 minutes is an optimal condition for high-buffered SARS-CoV-2 samples. The condition proved effective in changing the result's color from red (negative) to yellow (positive). We also classified the analysis results based on the correlation between the Cycle threshold (Ct) value of SARS-CoV-2 viruses and the Optical Density (OD) value, which was quantified using a spectrophotometer at 415 nm (with a correlation value of-0.9084), where yellow color indicated Ct below 20, amber color indicated Ct between 20 and 30, orange color indicated Ct between 30 and 35, and red indicated Ct more than 35 (negative). In conclusion, this study successfully detects the SARS-CoV-2 virus in high-buffered samples using Phenol Red Colorimetric RT-LAMP Assay, with a sensitivity of 85% for Ct Cutoff 40. © 2023, Bogor Agricultural University. All rights reserved.
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Background: Stem cell transplantation is a potentially life-saving intervention for patients with blood cancer. Research suggests that there are existing disparities in access, care and treatment experiences, with patients identifying with a minority ethnic heritage reporting greater disadvantage compared to patients from white British, Irish or Northern European backgrounds. Although we know that the COVID-19 pandemic has been challenging for patients, less is known about the experiences of patients from specific ethnic communities. Method(s): In this research, we undertook 1-1, semi-structured interviews with eight patients within the stem cell transplant pathway who identify with a minority ethnic background. Interviews took place online, between May and November 2021. The interview questions explored views on aspects of the pandemic, including access and experience of care delivery, intervention, mental health and financial impact. Interviews were transcribed verbatim and analysed using reflexive thematic analysis. Result(s): The majority of patient participants were female (75%), and identified with a Black African or Caribbean heritage (75%). Four themes were identified from the data: (1) 'Lack of suitable donors' described the challenges of finding a stem cell donor and the importance of raising awareness of stem cell donation specifically within minority ethnic communities;(2) 'Experiences of care' explored patients' experiences of the healthcare system from pre-diagnosis to post-treatment, including how the COVID-19 pandemic had made some aspects of care easier but acted to disrupt others;(3) 'Intense and unpredictable process' described the nature of living with blood cancer and how this has impacted upon patients' lives including the extent to which they had been able to access and make use of advice to help keep them safe during the pandemic;and (4) 'Coping mechanisms' identified the factors which helped patients' to understand and adjust to living with their diagnosis and the treatment process. Conclusion(s): Patients in the stem cell transplant pathway often experience intense treatment regimens, debilitating symptoms and long hospital stays, which has a significant impact on their lives. Minority ethnic patients describe facing additional challenges in relation to health inequalities both within and outside of the pandemic. Seeking support from family, peers or community organisations can help buffer the negative impacts of living with blood cancer and multiple disadvantages, but such support was more difficult to access during the pandemic. This had psychological consequences for patients who are already within an intense emotional journey. Pandemic recovery plans should address mental health support as a priority.
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Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infected more than 600 million people across 219 countries during the past three years. SARS-CoV-2 consists of a positive-strand RNA genome that encodes structural and nonstructural proteins and shares a 79% sequence homology with severe acute respiratory syndrome coronavirus 1 (SARS-CoV-1). Nonstructural proteins are necessary for viral replication and suppression of the host cell immune response. Nonstructural protein 1 (nsp1), a small protein conserved among most beta-coronaviruses, inhibits host messenger RNA (mRNA) translation by binding to ribosomal mRNA channels. Nsp1 also triggers degradation of host mRNA while viral RNA remains intact. We have previously shown that nsp1 localizes within stress granules (SGs), non-membranous vesicles of stalled mRNA that form in response to viral infection. We also found that upon induction of stress, SGs disperses within 60- 120 minutes in the presence of nsp1. Since SGs are known to store and protect translationally stalled mRNAs that are target of nsp1, we sought to analyze the level of mRNAs accumulation in SGs in the presence of nsp1. The goal of this project is to identify the impact of nsp1 on stress granule formation during SARS-CoV infection. We used human embryonic kidney cells (HEK293) and transfected them with DNA expressing SARSCoV- 1 nsp1 or a control plasmid. Cells were then incubated at 37degreeC under 5% CO2 concentration for 16 hours. Following incubation, cells were subjected to 30 min of oxidative stress using sodium arenite. Cells were collected and lysed using lysis buffer, then centrifuge at 18 000xg to collect SG pellets used for RNA isolation. Isolated mRNAs were quantified using quantitative RT-PCR. We specifically targeted mRNAs that tend to show a preferential accumulation in SGs without any viral infection. When nsp1 was expressed, we found majority of mRNAs have shown a 2-fold decrease in accumulation in SGs. These results suggest there is a direct effect of nsp1 in dispersing of RNA from SGs. We are currently investigating the effect of viral leader sequence in their accumulation in SGs in the presence of nsp1. This project was supported by the DRP award from SC INBRE (NIGMS, P20GM103499).Copyright © 2023 The American Society for Biochemistry and Molecular Biology, Inc.
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Background: People with cystic fibrosis (CF) are more likely to have anxiety and depression symptoms than the general population, with psychological distress being associated with negative health outcomes. The Cincinnati Children's Hospital Medical Center CF Center has been screening people with CF aged 12 and older since 2016 for anxiety and depressive symptoms. Little is known about longitudinal mental health trends for youth with CF, especially during the COVID-19 pandemic. Method(s): Chart review was conducted for individuals aged 12 and older seen for routine care at our center with at least one General Anxiety Disorder (GAD-7) or Patient Health Questionnaire (PHQ-9) screening result between January 2016 and December 2021. Data included demographic characteristics;dates and scores of GAD-7 and PHQ-9;mental health encounters 12 months before each screening date;and clinical variables of disease severity, including percentage predicted forced expiratory volume in 1 second (FEV1pp), body mass index, CF-related diabetes (CFRD), antibiotics in the 28 days prior, and exacerbations in the 12 months prior. Descriptive statistics were used to summarize demographic variables, logistic regression and linear mixed modeling were used to identify predictive relationships, and t-testswere used to compare impact of COVID with that of prior years. Result(s): The sample included 150 individuals with at least one screen across the 6 years. An average of 83 people completed at least one GAD-7 or PHQ-9 in each year. Across the 6-year time period, the percentage of individuals with low scores increased, and the percentage of people with moderate to severe scores was stable (Figure 1). Approximately 35% of individuals were rescreened at least once in a given year because of a previously high symptom score. For thosewho screened in the moderate to severe range (>=10) on initial screens per year, an average of 32% (GAD-7) and 37% (PHQ-9) had a lower score (<10) on their second screen per year. Individuals who scored 10 or higher on initial GAD-7 or PHQ-9 screens in any year were statistically more likely to have a CFRD diagnosis ( p = 0.02, GAD-7;p = 0.02, PHQ-9) and more psychology or psychiatry visits 12 months before the screening date ( p < 0.01, GAD-7;p < 0.01, PHQ-9) than those with minimal scores. In addition, PHQ-9 scores of 10 or greater were significantly associated with lower FEV1pp than low scores. Adherence to screening protocols consistently increased over time. Of all eligible individuals, 56% completed a GAD-7 and 55% a PHQ-9 in 2016, increasing to 92% and 94%, respectively, by 2021, despite the impact of the COVID-19 pandemic on CF care visit frequency. GAD-7 scores were not significantly different before COVID and during COVID ( p = 0.06);PHQ-9 scores were higher before than during COVID ( p = 0.02) despite similar numbers of screens conducted per year. (Figure Presented)Figure 1. Percentage of initial Patient Health Questionnaire (PHQ-9) and General Anxiety Disorder (GAD-7) scores per year from 2016 to 2021 of people with cystic fibrosis seen at Cincinnati Children's Hospital Medical Center Conclusion(s): These longitudinal trends in mental health symptom scores over time are reassuring,with increasing frequency of lowscores and stable moderate to severe scores. This may be because of greater awareness of mental health symptoms, more interventions through care teams, or improved access to resources. Similarly, although general population data suggest worsening of anxiety and depressive symptoms during the COVID- 19 pandemic, we hypothesize thatwewere able to buffer the impact of the pandemic on mental health in our center by screening and responding to screens. These results highlight the importance of consistent monitoring and support for mental health symptoms in people with CFCopyright © 2022, European Cystic Fibrosis Society. All rights reserved
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Raman spectroscopy probes the vibrational modes of a molecule. In recent years, surface-enhanced Raman spectra (SERS) of oligonucleotides on gold or silver nanoparticles have yielded significantly stronger signals. Raman spectra of DNA are high throughput, quantitative, and label-free and show distinct features created by vibrational modes such as ring deformation, backbone bending, and hydrogen bond stretching. Here we are using gold nanoparticles to probe various structural changes in a short helical DNA designed to mimic SL1 of coronavirus and SL of U6 snRNA. Phosphate buffers containing 1 M KCl or 10 mM magnesium chloride were utilized at two different pH (5.5 and 7). Differences in peak intensity are being observed between canonically paired helical DNA and DNA of similar composition with modifications containing non-canonical A*C base pair. We are comparing ion binding, pH-related, and temperature-variable conditions to observe changes in DNA structures.Copyright © 2023 The American Society for Biochemistry and Molecular Biology, Inc.
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The field-deployable point-of-care diagnostic test for rapid detection of SARS-COV-2 is needed for implementation of the control measures. In this direction, recently developed CRISPR technology combined with isothermal recombinase polymerase amplification assay is a versatile highly sensitive detection platform for rapid diagnosis of infectious diseases. Here we report the development of RT-RPA-CRISPR based LFA assay for detection of SARS-CoV-2 targeting conserved RdRp and E genes. Various sets of primers and gRNAs were designed targeting conserved regions of the RdRp and E genes of different lineages of SARS-CoV-2 viruses. The isothermal RT-RPA based amplification reactions were standardized using invitro transcribed RNAs of the target regions. The optimum amplification was observed at 42degreeC for 30 min as confirmed by visualization of the amplicons in agarose gel. Subsequently, CRISPRCAS12 reaction was implemented for specific detection of amplicons. Different sets of gRNAs targeting RdRp and E genes were designed and synthesized by in-vitro transcription. The CRISP/CAS12-gRNA complex and single stranded fluorescence probe were added to the RT-RPA amplicons for cleavage of fluorescence probe in positive reaction. Subsequently, the cleaved probes were detected in precoated LFA strips. Upon probe cleavage reaction, the product was mixed with buffer and loaded into LFA strips. In positive reaction, test line showed strong band in test line and light band in control line. The standardized RT-RPA-CRISPR-LFA assay was tested for detection of SARS-CoV-2 using previously isolated RNAs from clinical cases of human SARS-CoV-2 infections. The developed assay successfully detected the positive cases. In conclusion, the developed assay could serve as versatile POC platform for rapid detection of SARS-CoV-2 nucleic acids in human as well as animals.
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Purpose of Study: Antibiotic resistance remains one of the largest healthcare and public health challenges. Several studies have documented that the spread of antibiotic resistant bacteria in nosocomial settings has been exacerbated worldwide due to increased rates of hospitalization and intubation in the wake of the COVID-19 pandemic. One way to address antibiotic resistance is to identify novel compounds that inhibit essential microbial processes. Two-component regulatory systems are important mediators of signal transduction that allow bacteria to communicate with and respond to changes in their environment. The WalRK system is a two-component system that is conserved and essential for viability in many Gram-positive human pathogens. We hypothesize that a ligand that specifically binds with the DNA-interaction surface of the WalR protein can lead to cell death and can serve as a lead compound for future drug development efforts. Methods Used: We describe the development process of an assay to identify WalR binding compounds. In silico molecular dynamics docking approaches were utilized to identify potential WalR binding compounds from virtual compound libraries. To assess their WalR-binding capacity in vitro, overexpression strains for several WalR recombinant constructs were engineered and protein constructs were purified to homogenicity. Isothermal titration calorimetry (ITC) is a technique that measures heat release or absorption when two molecules interact. A MicroCal PEAQ ITC instrument was utilized to develop a WalR-binding assay. Summary of Results: WalR is a two-domain protein featuring a regulatory and a DNA-binding domain. Two constructs, a truncated DNA-binding domain and a full-length protein construct proved soluble, and pure quantities necessary to conduct ITC measurements could be successfully obtained (12 mg full-length protein and 23 mg truncated protein). These proteins were amenable to ITC experiments. We found that experiments were best run with at least a two-fold increase of ligand concentration to protein concentration supplied in identical buffer conditions over nineteen injections. We are currently assessing the binding affinities of our in silico hit compounds. Conclusion(s): Our results show that ITC enables the detailed, rapid, and reproducible characterization of the binding relationship between the DNA-binding domain of the WalR protein and any potential ligands. The protocol discussed herein will enable further drug discovery studies on the WalR response regulator protein to identify and characterize inhibitors, providing insight towards the development of novel antimicrobial compound.
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Background: The potential aerosol spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV- 2) has been suggested. While indoor air sampling for SARS-CoV- 2 has demonstrated detectable viral RNA and has been related to virus transmission, the contribution of outdoor air to the spread of the viral infection is not yet known. We aimed at developing a methodology to detect the virus in outdoor air. Method(s): T he s ampling w as p erformed u sing a C HEMVOL v olumetric impactor (Butraco) equipped with 2 stages (PM > 10 & 2.5 > PM > 10um). Filters were collected and preserved at -80 degreeC. Total RNA extraction was performed directly from the collected filters with the Phenol-Chloroform method using TRItidy GTM reagent according to the manufacturer's instructions. For total RNA purification samples were purified with the commercial kit E.Z.N.A. Total RNA Kit I. Real-Time Reverse Transcription PCR was executed to detect the N gene from the Sarbecovirus family and RdRp gene from SARS-CoV- 2 using the ViroReal Kit SARS-CoV- 2 Multiplex. A protein-rich fraction was obtained with ammonium bicarbonate buffer extraction followed by lyophilization. SARS-CoV- 2 spike protein was assessed by specific immunological detection (SARS-CoV- 2 Antigen Test Kit). Result(s): RT-PCR for N gene results, identifying Sarbecovirus family, were positive and Cq > 33, in the samples from the last week of December 2020 and the first and second weeks of January 2021, in both PM > 10 and 2.5 > PM > 10. The RdRp gene was undetectable, probably due to low virus concentration. The protein samples from the same days tested positive for the specific antigen spike protein. All results combined confirm the detection of SARS-CoV- 2 in outdoor air. Conclusion(s): Airborne SARS-CoV- 2 was detected in ambient air. These results will contribute to an early detection of SARS-Cov- 2 in ambient air, thus eventually providing the base for early alert systems allowing the implementation of preventive measures to control outbreaks.
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Motivation: The low solubility, weak acid drug, niclosamide is a host cell modulator with broad-spectrum anti-viral cell-activity against many viruses, including stopping the SARS-CoV-2 virus from infecting cells in cell culture. As a result, a simple universal nasal spray preventative was proposed and investigated in earlier work regarding the dissolution of niclosamide into simple buffers. However, starting with pharmaceutical grade, niclosamide represents a new 505(b)(2) application. The motivation for this second paper in the series was therefore to explore if and to what extent niclosamide could be extracted from commercially available and regulatory-approved niclosamide oral tablets that could serve as a preventative nasal spray and an early treatment oral/throat spray, with possibly more expeditious testing and regulatory approval. Experimental: Measurements of supernatant niclosamide concentrations were made by calibrated UV-Vis for the dissolution of niclosamide from commercially available Yomesan crushed into a powder for dissolution into Tris Buffer (TB) solutions. Parameters tested were as follows: time (0-2 days), concentration (300 µM to -1 mM), pH (7.41 to 9.35), and anhydrous/hydrated state. Optical microscopy was used to view the morphologies of the initial crushed powder, and the dissolving and equilibrating undissolved excess particles to detect morphologic changes that might occur. Results: Concentration dependence: Niclosamide was readily extracted from powdered Yomesan at pH 9.34 TB at starting Yomesan niclosamide equivalents concentrations of 300 µM, 600 µM, and 1 mM. Peak dissolved niclosamide supernatant concentrations of 264 µM, 216 µM, and 172 µM were achieved in 1 h, 1 h, and 3 h respectively. These peaks though were followed by a reduction in supernatant concentration to an average of 112.3 µM ± 28.4 µM after overnight stir on day 2. pH dependence: For nominal pHs of 7.41, 8.35, 8.85, and 9.35, peak niclosamide concentrations were 4 µM, 22.4 µM, 96.2 µM, and 215.8 µM, respectively. Similarly, the day 2 values all reduced to 3 µM, 12.9 µM, 35.1 µM, and 112.3 µM. A heat-treatment to 200 °C dehydrated the niclosamide and showed a high 3 h concentration (262 µM) and the least day-2 reduction (to 229 µM). This indicated that the presence, or formation during exposure to buffer, of lower solubility polymorphs was responsible for the reductions in total solubilities. These morphologic changes were confirmed by optical microscopy that showed initially featureless particulate-aggregates of niclosamide could grow multiple needle-shaped crystals and form needle masses, especially in the presence of Tris-buffered sodium chloride, where new red needles were rapidly made. Scale up: A scaled-up 1 L solution of niclosamide was made achieving 165 µM supernatant niclosamide in 3 h by dissolution of just one fifth (100 mg niclosamide) of a Yomesan tablet. Conclusion: These comprehensive results provide a guide as to how to utilize commercially available and approved tablets of niclosamide to generate aqueous niclosamide solutions from a simple dissolution protocol. As shown here, just one 4-tablet pack of Yomesan could readily make 165 L of a 20 µM niclosamide solution giving 16,500 10 mL bottles. One million bottles, from just 60 packs of Yomesan, would provide 100 million single spray doses for distribution to mitigate a host of respiratory infections as a universal preventative-nasal and early treatment oral/throat sprays throughout the world. Graphical Abstract: pH dependence of niclosamide extraction from crushed Yomesan tablet material into Tris buffer (yellow-green in vial) and Tris-buffered saline solution (orange-red in vial). Initial anhydrous dissolution concentration is reduced by overnight stirring to likely monohydrate niclosamide; and is even lower if in TBSS forming new niclosamide sodium needle crystals grown from the original particles. Supplementary Information: The online version contains supplementary material available at 10.1186/s41120-023-00072-x.
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Objective: Many Chinese teenagers are experiencing high mental stress levels due to epidemic-related restrictions and closures. Mental stress can induce numerous associated symptoms, and physical exercise is considered to buffer mental stress. However, it remains unclear whether health motivation regulates the relationships among mental stress, physical exercise, and stress symptoms. This study examined whether mental stress events during the epidemic can predict stress symptoms, whether physical exercise can buffer mental stress, and whether the mental stress buffer effect is enhanced when health motivation regarding physical exercise is high. Methods: In total, 2,420 junior high school students (1,190 boys and 1,230 girls; 826 seventh-grade students, 913 eighth-grade students, and 681 ninth-grade students) from nine provinces nationwide were selected to investigate mental stress events, symptoms, health motivation, and physical exercise in adolescents. The hypothesis was tested with a multiple regression analysis. Results: A positive relationship between adolescent mental stress events and stress symptoms was observed, and an interactive relationship was found among health motivation, physical exercise, and mental stress factors. Specifically, the mental stress-buffering effect of physical exercise was significant only when health motivation was high. Conclusion: In the post-epidemic period, the influence of mental stress events on stress symptoms in adolescents was found to be buffered by physical exercise only in terms of high health motivation. This result highlighted the role of health motivation in the buffering effect of physical exercise on mental stress during an epidemic.
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COVID-19 , Motivation , Male , Female , Humans , Adolescent , COVID-19/epidemiology , Exercise/physiology , Students , Stress, PsychologicalABSTRACT
The COVID-19 pandemic, unleashed at the end of 2019, seriously affected not only the health of thousands of Peruvians but also the economy and production systems. This is reflected in the reduction of the GDP, which during that year suffered a reduction of 11.12%. Regarding the chocolate industry, consumption, production, and importation were growing in previous years, but had a significant decrease during 2020. Due to government sanitary restrictions, many Peruvian businesses were forced to keep their inventory on hand almost all year. Despite the fact that some experts affirm that it is convenient to maintain a greater number of inventories when there is dynamism in the labor market, it is very risky to keep such levels of stocks permanently. In the present investigation, an inventory management improvement model is proposed using the DDMRP and PA techniques, with which the optimal quantities to be produced and kept in inventories will be determined at the different levels of the supply chain. In this way, the impact generated by the Inventory Maintenance Cost (IMC) will be reduced, and the Inventory Turnover Ratio (ITR) will be increased to 3.2. The model will be validated by simulation to verify that the levels are optimal for the system. © 2022 ACM.
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In light of the COVID-19 pandemic, getting infected through the built environment is being studied. The measures that should be taken to reduce infection through the built environment are essential;not only for COVID-19, but this idea is present at all times of widespread diseases. The purpose of this research is to systematically review the relationship between the built environment and the spread of infection to create a potential guideline to reduce the transmission rate. Articles and studies on the relationship between infectious disease and the built environment were reviewed. Articles matching the selection criteria were identified. Most articles described peer reviews, consensus statements, and reports. The articles have provided data that can be used as guidance for reducing the transmission of infection within the built environment. It was found that evidence has been created such as ventilation, buffer spaces, flooring, and surfaces that can reduce the infection of COVID-19.Copyright © 2022 Mohan R, et al.
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To enhance biosafety and reliability in SARS-CoV-2 molecular diagnosis, virus lysis/transport buffers should inactivate the virus and preserve viral RNA under various conditions. Herein, we evaluated the SARS-CoV-2-inactivating activity of guanidine hydrochloride (GuHCl)- and surfactant (hexadecyltrimethylammonium chloride (Hexa-DTMC))-based buffer, Prep Buffer A, (Precision System Science Co., Ltd., Matsudo, Japan) and its efficacy in maintaining the stability of viral RNA at different temperatures using the traditional real-time one-step RT-PCR and geneLEAD VIII sample-to-result platform. Although Prep Buffer A successfully inactivated SARS-CoV-2 in solutions with high and low organic substance loading, there was considerable viral genome degradation at 35 °C compared with that at 4 °C. The individual roles of GuHCl and Hexa-DTMC in virus inactivation and virus genome stability at 35 °C were clarified. Hexa-DTMC alone (0.384%), but not 1.5 M GuHCl alone, exhibited considerable virucidal activity, suggesting that it was essential for potently inactivating SARS-CoV-2 using Prep Buffer A. GuHCl and Hexa-DTMC individually reduced the viral copy numbers to the same degree as Prep Buffer A. Although both components inhibited RNase activity, Hexa-DTMC, but not GuHCl, directly destroyed naked viral RNA. Our findings suggest that samples collected in Prep Buffer A should be stored at 4 °C when RT-PCR will not be performed for several days.
Subject(s)
COVID-19 , Surface-Active Agents , Humans , Cetrimonium , Chlorides , Genome, Viral , Guanidine/pharmacology , Lipoproteins , Reproducibility of Results , RNA, Viral/genetics , Saliva , SARS-CoV-2/genetics , Surface-Active Agents/pharmacology , Virus Activation , Biological TransportABSTRACT
To harness the antimicrobial properties of a crude methanolic extract of Henna (Lawsonia inermis) leaf as a potential alternative sanitiser, there is the need to test its performance in different solutions. In this work, the effects of distilled water (dH20), Acetate-HCL (AH) Buffer (pH 4.6), Phosphate Buffer Saline (PBS) (pH 7.2) and Tris-HCL (TBH) Buffer (pH 8.6) on the antibacterial and antiviral activity of the extract were assessed. Through standard phytochemical screening and HPLC-MS (LCMS STANDARD 7.M), it was found that the extract consisted of about 30 different compounds including flavonoids. The extent of the antimicrobial activity of the extract in solutions was in the increasing order of AH > dH2O >>>> TBH > PBS. Under the same conditions, reduced antibacterial activity and complete cessation of the antiviral activity of the extract in TBH and PBS was observed. However, in AH and dH20, within 1-5 min, 1 mg ml-1, 0.125 mg ml-1 and 0.0625 mg ml-1 of the extract caused complete inactivation of E.coli (reductions of 8.2 log CFU ml-1), B. subtilis (reductions of 8.2 log CFU ml-1) and MS2 (reductions of 9.7 log PFU ml-1) respectively. The fluorescence microscopy images of the live/dead staining of the inactivated bacterial samples validated the extent of the inactivation. The broad spectrum and high antimicrobial activity of the extract, coupled with the plant not a staple food, has long history of safe use by humans as a medicine and cosmetic, cheaply available in abundance in many regions of the world, thus making the extract a potential candidate as an alternative sanitiser in the time of COVID-19 Pandemic and beyond.
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Background At the Urbino hospital, all patients tested with antigen tests for SARS-CoV-2, if positive, are confirmed with a molecular test. Among the molecular tests in use in our laboratory, VitaPCRTM SARS-CoV-2 Assay is the fastest and easiest to use, as it takes about half an hour from preparation to the result and does not require a biosecurity laboratory, because performed from inactivated samples. Aim of the study Given the rapidity of VitaPCRTM SARS-CoV-2 Assay, it was decided to use it for the confirmation of samples found positive to LIAISON SARS-CoV-2 Ag test. This last test is performed by dedicated tubes with inactivating buffer based on detergent, therefore it does not interfere in the processes of extraction and amplification of nucleic acids. Materials and methods For evaluation, 48 samples tested positive with LIAISON SARS-CoV-2 Ag (Diasorin) test were analyzed, coming from various departments of the Urbino hospital, and confirmed by SimplexaTM COVID-19 Direct (Diasorin) molecular test. From the tubes of all the samples, 200 uL were taken and transferred to dedicated tubes for analysis with VitaPCRTM SARS-CoV-2 Assay (Menarini) to be analyzed, after eliminating 2.2 mL of buffer. Results VitaPCRTM SARS-CoV-2 Assay was able to detect SARSCoV- 2 RNA in all 48 tested antigenically positive samples. In particular, even in all positive samples with low values of TCID50/mL (<1000), 11 out of 48, results with Ct<28.8 were found, although not maintaining a direct correspondence between quantity of antigen and quantity of detected RNA. Conclusions The results obtained, despite requiring an increase in the number of cases, demonstrate the potential use of VitaPCRTM SARS-CoV-2 Assay for the confirmation of positive samples directly from the inactivated buffer used for LIAISON SARS-CoV-2 Ag assay. VitaPCRTM instrument, given the speed, simplicity of use and the possibility of use outside biosafety laboratories, it is an excellent diagnostic tool to support the confirmation procedures of samples tested positive for the SARSCoV- 2 N antigen.
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Background: Adverse life events such as life-threatening accidents, domestic and/or sexual violence, organic diseases (i.e., cancer), or COVID-19 can have a strong traumatic impact - generating reactions as intrusive thoughts, hyperarousal, and avoidance. Indeed, the traumatic impact of COVID-19 seems to lead individuals to experience anxiety and depression. However, the Anxiety-Buffer Hypothesis suggests that self-esteem could be considered a shield (buffer) against traumatic experiences and their outcomes (i.e., anxiety and depression). The present study has two objectives. First, to develop a measure of the impact of the traumatic event considering the aforementioned reactions. Second, to test the process - triggered by COVID19-related traumatic experience - in which self-esteem buffers the path that leads to anxiety and depression. Method: In Study 1 (N = 353), the Post-Traumatic Symptom Questionnaire (PTSQ) was developed and a deep investigation of its psychometric properties was conducted. In Study 2 (N = 445), a structural equation model with latent variables was performed to assess the buffering effect of self-esteem. Results: The PTSQ has excellent fit indices and psychometric properties. According to the ABH, results confirm the buffering effect of self-esteem in the relationships between traumatic symptoms and both anxiety and depression. Conclusion: On the one hand, the PTSQ is a solid and reliable instrument. On the other hand, that self-esteem is a protective factor against anxiety and depression related to a traumatic experience - such as COVID-19. Targeted psychological interventions should be implemented to minimize the psychological burden of the illness while promoting adaptation and positive aspects of oneself. Supplementary Information: The online version contains supplementary material available at 10.1007/s40653-022-00503-z.
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Ireland had one of the highest levels of COVID-19 in Europe and the numbers of cases and deaths were high especially in residential care settings. Since the onset of the pandemic, entire healthcare and research operations had to be reset, to prevent and manage COVID-19 in a Huntington's Disease (HD) residential care facility in Ireland. Lean Management principles were successfully used initially to manage the clinical caseload and eventually research during the pandemic. The strategy was to cut out the eight areas of 'wastes' as per the Lean Methodology. Initially these principles were applied in HD clinical care and eventually in research when it could be resumed during the pandemic. The first waste 'Waiting' was dealt with by creating of COVID-19 specific ward and buffer zone, 'Inventory' was managed by optimising the procurement and management of Personal protective equipment(PPE), the third waste 'defects' were minimized by continuous in-house training of personnel relevant to minimizing errors and optimizing care for COVID-19, the fourth waste 'transportation' was managed by transitioning the clinical and research visits to remote mode by deploying technology, the waste 'motion' was managed with segregated staff accommodation rosters and technology, 'Overproduction' was managed by optimizing clinical operations using emergency clinical governance initiatives, 'Overprocessing' was managed by eliminating repetitive, redundant, or less than valuable processes and finally 'untapped human potential' was tapped into by retraining and redeployment of staff in transdisciplinary roles. Research operations were completely optimised so that pandemic related restrictions had little effect on achieving research deliverables successfully.
ABSTRACT
Background: The rapid spread and recurrent infections of SARS-CoV-2 has led to increased use and availability of at-home antigen testing, but widespread testing of antibodies against spike and nucleocapsid to monitor vaccine-induced immunity and exposure to the virus is lacking. Most serological tests require a serum sample from a venous blood draw, increasing risk of exposure to COVID-19 and limiting availability and scalability of testing for many patients. This is especially the case for individuals who are immunocompromised, such as those with inflammatory bowel diseases (IBD), which are frequently on medications that might alter their immune response and impact vulnerability to SARS-CoV-2. Use of easily acquired and stably stored dried fingerstick blood serves a promising specimen source for at-home, remote testing for SARS-CoV-2 antibodies. Aim(s): Validate the use of fingerstick blood (dried and then eluted) versus serum as a specimen for the measurement of quantitative spike and nucleocapsid antibodies to SARS-CoV-2 in a diverse cohort of healthy and immunocompromised patients. Method(s): Patients were consented and enrolled into the Pediatric Gastrointestinal Tissue, Stool, Saliva and Blood Registry prior to having an endoscopic procedure. Five mL of blood was obtained by venipuncture and 10 muL of fingerstick blood was collected and dried on a Neoteryx Mitra device. Blood was eluted from the Neoteryx Mitra samples in 200 muL of dilution buffer (1% BSA, 0.05% Tween-20, 140 mM NaCl, 50 mM Tris (pH 8.0), 0.025% sodium azide) and placed on an orbital shaker at a speed of 500 RPM for 3 h. Paired serum and fingerstick blood eluate specimens were run on the quantitative Roche Elecsys SARS-CoV-2 spike antibody assay and the qualitative Roche Elecsys SARS-CoV-2 nucleocapsid antibody assay. Linear regression were performed on each assay with exclusion of values that were above the upper limit of detection of the assay. Result(s): We observed an excellent correlation in both SARS-CoV-2 antibody assays when comparing fingerstick blood eluates and serum. The linear regression for the nucleocapsid antibody assay had a slope of 15.5, intercept of 4.05, and R2 of 0.92, indicating that a Neoteryx value of 1.00 COI (cut-off index) equates to a serum value of 19.6 COI. The linear regression for the spike assay had a slope of 13.6, intercept of 953, and R2 of 0.95, indicating that a value of 1,000 U/mL from a fingerstick sample equates to a serum value of 14,544 U/mL. Conclusion(s): These data demonstrate that fingerstick blood collected on Neoteryx Mitra devices can be used as a specimen source in Roche Elecsys SARS-CoV-2 antibody assays to calculate the serum levels of spike and nucleocapsid antibodies. This can serve as a platform to remotely and reliably monitor the durability of antibody responses to natural infection with and immunization against SARS-CoV-2 in patients. Chart comparisons of nucleocapsid (top) and spike (bottom) protein antibody levels detected via remote fingerstick collection (Neoteryx, x-axis) and venous blood serum (y-axis).
ABSTRACT
The Vaccines for Children Program (VFC) is a federally funded program in the United States, providing vaccines to children who lack health insurance or who otherwise cannot afford the vaccination cost. The VFC program was created in 1993 and is required to be a new entitlement of each State's Medicaid plan. The program was officially implemented in October 1994 and served eligible children in all United States (US). Other countries, the United Nations (UN), and the World Health Organization (WHO), have similar programs. A critical aspect of these programs is the guidance surrounding the environmental monitoring of the materials. To best maintain the integrity of these products, specific storage parameters are required. It is necessary to store most vaccines at refrigeration or freezing temperatures. To best assure the efficacy of the vaccines, monitoring standards and equipment are specified. The technology and methodologies may be adequate for these programs' materials; these same methods are not for the COVID vaccine. [1] When reviewing the guidance recommendations worldwide, one may observe commonalities in the program. Each guidance calls for the use of digital data loggers (DDL), sampling rates of 15 to 30 minutes, daily check-in (during business hours), and the use of a temperature buffer, each without specificity. [2] The inadequacies of the VFC program monitoring methodologies fall far short when monitoring COVID vaccines. Herein considerations for the transport, storage, and distribution of the COVID vaccine cold chain will be discussed.