The current practice for diagnosis of COVID-19, based on SARS-CoV-2 PCR testing of pharyngeal or respiratory specimens in a symptomatic patient at high epidemiologic risk, likely underestimates the ...true prevalence of infection. Serologic methods can more accurately estimate the disease burden by detecting infections missed by the limited testing performed to date. Here, we describe the validation of a coronavirus antigen microarray containing immunologically significant antigens from SARS-CoV-2, in addition to SARS-CoV, MERS-CoV, common human coronavirus strains, and other common respiratory viruses. A comparison of antibody profiles detected on the array from control sera collected prior to the SARS-CoV-2 pandemic versus convalescent blood specimens from virologically confirmed COVID-19 cases demonstrates near complete discrimination of these two groups, with improved performance from use of antigen combinations that include both spike protein and nucleoprotein. This array can be used as a diagnostic tool, as an epidemiologic tool to more accurately estimate the disease burden of COVID-19, and as a research tool to correlate antibody responses with clinical outcomes.
Under physiological conditions, most Ca2+-ATPase (SERCA) molecules bind ATP before binding the Ca2+ transported. SERCA has a high affinity for ATP even in the absence of Ca2+, and ATP accelerates ...Ca2+ binding at pH values lower than 7, where SERCA is in the E2 state with low-affinity Ca2+-binding sites. Here we describe the crystal structure of SERCA2a, the isoform predominant in cardiac muscle, in the E2·ATP state at 3.0-Å resolution. In the crystal structure, the arrangement of the cytoplasmic domains is distinctly different from that in canonical E2. The A-domain now takes an E1 position, and the N-domain occupies exactly the same position as that in the E1·ATP·2Ca2+ state relative to the P-domain. As a result, ATP is properly delivered to the phosphorylation site. Yet phosphoryl transfer never takes place without the filling of the two transmembrane Ca2+-binding sites. The present crystal structure explains what ATP binding itself does to SERCA and how nonproductive phosphorylation is prevented in E2.
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Adolescent athletes' values regarding health behaviors, including their attitudes toward doping, are largely derived from those of their parents. Therefore, clarifying the factors that affect ...parents' intentions regarding their children's medicine intake and nutrition can help elucidate the process of forming values of healthy behaviors in young athletes.
Between March 8 and March 9, 2021, an online questionnaire survey was conducted via an Internet research company; data from 2,000 residents in Japan were collected. Participants were male and female residents aged 30-59 years with children in elementary or high school and belonging to sports clubs. The survey items included respondent's and child's basic information, respondent's health literacy, and level of sports in which the respondent and child were (or are) engaged. Respondents were also asked if they would like their children to receive prescription drugs, over-the-counter drugs, herbal medicines, vaccines, supplements, or energy drinks. Logistic regression analysis was performed to analyze the relationship between respondents' basic information and health literacy and their intention to receive prescription and over-the-counter drugs, herbal medicines, vaccines, supplements, and energy drinks.
Higher parental health literacy was associated with higher children's willingness to receive prescription drugs (odds ratio OR = 1.025, 95% confidence interval CI: 1.016-1.035), over-the-counter drugs (OR = 1.012, 95% CI: 1.003-1.021), prescription herbal medicines (OR = 1.021, 95% CI: 1.021-1.030), over-the-counter herbal medicines (OR = 1.012, 95% CI: 1.003-1.021), and vaccines (OR = 1.025, 95% CI: 1.016-1.035). Conversely, the children's intention to receive energy drinks (OR = 0.990, 95% CI: 0.980-1.000) decreased significantly. As the child's athletic level increased, parents' willingness for their children to receive oral prescription medicines decreased (OR = 0.886, 95% CI: 0.791-0.992) and that to receive supplements (OR = 1.492, 95% CI: 1.330-1.673) and energy drinks significantly increased (OR = 1.480, 95% CI: 1.307-1.676).
Health literacy of adolescent athletes' parents is associated with their children's willingness to receive medicines. Healthcare providers should counsel parents of adolescent athletes to allow their children to receive necessary drug treatments and prevent doping violations caused by supplement intake.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
To detect the presence of antibodies in blood against SARS-CoV-2 in a highly sensitive and specific manner, here we describe a robust, inexpensive ($200), 3D-printable portable imaging platform ...(TinyArray imager) that can be deployed immediately in areas with minimal infrastructure to read coronavirus antigen microarrays (CoVAMs) that contain a panel of antigens from SARS-CoV-2, SARS-1, MERS, and other respiratory viruses. Application includes basic laboratories and makeshift field clinics where a few drops of blood from a finger prick could be rapidly tested in parallel for the presence of antibodies to SARS-CoV-2 with a test turnaround time of only 2-4 h. To evaluate our imaging device, we probed and imaged coronavirus microarrays with COVID-19-positive and negative sera and achieved a performance on par with a commercial microarray reader 100× more expensive than our imaging device. This work will enable large scale serosurveillance, which can play an important role in the months and years to come to implement efficient containment and mitigation measures, as well as help develop therapeutics and vaccines to treat and prevent the spread of COVID-19.
The TinyArray imager, a robust inexpensive portable imaging device, can detect antibodies against SARS-CoV-2 based on coronavirus antigen microarrays.
A coronavirus antigen microarray (COVAM) was constructed containing 11 SARS-CoV-2, 5 SARS-1, 5 MERS, and 12 seasonal coronavirus recombinant proteins. The array is designed to measure immunoglobulin ...isotype and subtype levels in serum or plasma samples against each of the individual antigens printed on the array. We probed the COVAM with COVID-19 convalescent plasma (CCP) collected from 99 donors who recovered from a PCR+ confirmed SARS-CoV-2 infection. The results were analyzed using two computational approaches, a generalized linear model (glm) and random forest (RF) prediction model, to classify individual specimens as either Reactive or non-reactive against the SARS-CoV-2 antigens. A training set of 88 pre-COVID-19 specimens (PreCoV) collected in August 2019 and102 positive specimens from SARS-CoV-2 PCR+ confirmed COVID-19 cases was used for these analyses. Results compared with an FDA emergency use authorized (EUA) SARS-CoV2 S1-based total Ig chemiluminescence immunoassay (Ortho Clinical Diagnostics VITROS Anti-SARS-CoV-2 Total, CoV2T) and with a SARS-CoV-2 S1-S2 spike-based pseudovirus micro neutralization assay (SARS-CoV-2 reporter viral particle neutralization titration (RVPNT) showed high concordance between the three assays. Three CCP specimens that were negative by the VITROS CoV2T immunoassay were also negative by both COVAM and the RVPNT assay. Concordance between VITROS CoV2T and COVAM was 96%, VITROS CoV2T and RVPNT 93%, and RVPNT and COVAM 91%. The discordances were all weakly reactive samples near the cutoff threshold of the VITROS CoV2T immunoassay. The multiplex COVAM allows CCP to be grouped according to antibody reactivity patterns against 11 SARS-CoV-2 antigens. Unsupervised K-means analysis, via the gap statistics, as well as hierarchical clustering analysis revealed three main clusters with distinct reactivity intensities and patterns. These patterns were not recapitulated by adjusting the VITROS CoV2T or RVPNT assay thresholds. Plasma classified by COVAM reactivity patterns offers potential to improve CCP therapeutic efficacy CoV2T alone. The use of a SARS-CoV-2 antigen array can qualify CCP for administration as a treatment for acute COVID-19, and interrogate vaccine immunogenicity and performance in preclinical, clinical studies, and routine vaccination to identify antibody responses predictive of protection from infection and disease.
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Collaborations between hospital and community pharmacists play a key role in ensuring consistent continuation of pharmacotherapy within official medical care plans designed to promote community-based ...healthcare. A previous study conducted by the authors clarified the constructs of collaboration between the hospital and community pharmacists from the hospital pharmacist's perspective. In this study, a similar questionnaire was used to survey a group of pharmacies with 424 outlets nationwide, and 244 responses were collected. Factor analysis from the community pharmacists' perspective extracted five latent factors comprising 18 variables, and structural equation modeling yielded a model with a high goodness-of-fit. In the latter model, variables representing "Organizational climate" and "Basic policy on collaboration" formed the foundation of the collaboration between hospital and community pharmacists, while variables representing "Understanding of healthcare policy", "Resources for collaboration", and "Community support systems" represented concepts that flexibly compensated for fluctuations in medical policy. These trends were similar to those of the constructs previously indicated by hospital pharmacists. We performed multiple regression analysis and structural equation modeling to confirm the impact of the inclusion of "Need for collaboration" as a dependent variable on the proposed constructs of hospital-community pharmacist collaboration using different analytical methods. Our results indicated that "Organizational climate", "Basic policy on collaboration", and "Community support systems" affected the "Need for collaboration". Our findings indicate that future studies are needed to confirm and clarify the causal relationships demonstrated by the constructs of hospital-community pharmacist collaboration seen within the current study.
Abstract
We analyzed data from two ongoing COVID-19 longitudinal serological surveys in Orange County, CA., between April 2020 and March 2021. A total of 8476 finger stick blood specimens were ...collected before and after a vaccination campaign. IgG levels were determined using a multiplex antigen microarray containing antigens from SARS-CoV-2, SARS, MERS, Common CoV, and Influenza. Twenty-six percent of specimens from unvaccinated Orange County residents in December 2020 were SARS-CoV-2 seropositive; out of 852 seropositive individuals 77 had symptoms and 9 sought medical care. The antibody response was predominantly against nucleocapsid (NP), full length, and S2 domain of spike. Anti-receptor binding domain (RBD) reactivity was low and not cross-reactive against SARS S1 or SARS RBD. A vaccination campaign at the University of California Irvine Medical Center (UCIMC) started on December, 2020 and 6724 healthcare workers were vaccinated within 3 weeks. Seroprevalence increased from 13% pre-vaccination to 79% post-vaccination in January, 93% in February, and 99% in March. mRNA vaccination induced higher antibody levels than natural exposure, especially against the RBD domain and cross-reactivity against SARS RBD and S1 was observed. Nucleocapsid protein antibodies can be used to distinguish vaccinees to classify pre-exposure to SARS-CoV-2 Previously infected individuals developed higher antibody titers to the vaccine than non pre-exposed individuals. Hospitalized patients in intensive care with severe disease reach significantly higher antibody levels than mild cases, but lower antibody levels compared to the vaccine. These results indicate that mRNA vaccination rapidly induces a much stronger and broader antibody response than SARS-CoV-2 infection.
This cross-sectional study explored the association between medication non-adherence and its factors in patients with non-communicable diseases (NCDs) using an online structured questionnaire emailed ...to 30,000 people (aged over 20 years who lived in Japan at the time of the survey). The questions concerned respondents' characteristics, medication non-adherence, health beliefs, lifestyles, and trouble taking medication. Factors related to non-adherence were analyzed among patients with lifestyle-related NCDs categorized into two age groups: 20-59, and >60 years. Unintentional (
< 0.001) and intentional (
< 0.001) non-adherence were more common among patients aged 20-59 than in older adults. NCD patients aged 20-59 experienced significantly more trouble taking medication than older adults. Multiple regression analysis showed that for patients aged 20-59 with NCDs, unintentional non-adherence was significantly and positively associated with current smoking habits (β = 0.280,
< 0.001), while intentional non-adherence was significantly and positively associated with alcohol consumption (β = 0.147,
= 0.020) and current smoking habits (β = 0.172,
= 0.007). In patients aged 20-59, unhealthy eating habits (β = -0.136,
= 0.034) and lack of exercise (β = -0.151,
= 0.020) were negatively associated with intentional non-adherence. In conclusion, factors affecting medication non-adherence in patients with lifestyle-related diseases are related to health awareness, lifestyle, and medication barriers.
The effects of adjuvants for increasing the immunogenicity of influenza vaccines are well known. However, the effect of adjuvants on increasing the breadth of cross-reactivity is less well ...understood. In this study we have performed a systematic screen of different toll-like receptor (TLR) agonists, with and without a squalene-in-water emulsion on the immunogenicity of a recombinant trimerized hemagglutinin (HA) vaccine in mice after single-dose administration. Antibody (Ab) cross-reactivity for other variants within and outside the immunizing subtype (homosubtypic and heterosubtypic cross-reactivity, respectively) was assessed using a protein microarray approach. Most adjuvants induced broad IgG profiles, although the response to a combination of CpG, MPLA and AddaVax (termed 'IVAX-1') appeared more quickly and reached a greater magnitude than the other formulations tested. Antigen-specific plasma cell labeling experiments show the components of IVAX-1 are synergistic. This adjuvant preferentially stimulates CD4 T cells to produce Th1>Th2 type (IgG2c>IgG1) antibodies and cytokine responses. Moreover, IVAX-1 induces identical homo- and heterosubtypic IgG and IgA cross-reactivity profiles when administered intranasally. Consistent with these observations, a single-cell transcriptomics analysis demonstrated significant increases in expression of IgG1, IgG2b and IgG2c genes of B cells in H5/IVAX-1 immunized mice relative to naïve mice, as well as significant increases in expression of the IFNγ gene of both CD4 and CD8 T cells. These data support the use of adjuvants for enhancing the breath and durability of antibody responses of influenza virus vaccines.
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