A point-of-care lateral flow assay for neutralising antibodies against SARS-CoV-2

Background: As vaccines against SARS-CoV-2 are now being rolled out, a better understanding of immunity to the virus, whether from infection, or passive or active immunisation, and the durability of this protection is required. This will benefit from the ability to measure antibody-based protection to SARS-CoV-2, ideally with rapid turnaround and without the need for laboratory-based testing. Methods: We have developed a lateral flow POC test that can measure levels of RBD-ACE2 neutralising antibody (NAb) from whole blood, with a result that can be determined by eye or quantitatively on a small instrument. We compared our lateral flow test with the gold-standard microneutralisation assay, using samples from convalescent and vaccinated donors, as well as immunised macaques. Findings: We show a high correlation between our lateral flow test with conventional neutralisation and that this test is applicable with animal samples. We also show that this assay is readily adaptable to test for protection to newly emerging SARS-CoV-2 variants, including the beta variant which revealed a marked reduction in NAb activity. Lastly, using a cohort of vaccinated humans, we demonstrate that our whole-blood test correlates closely with microneutralisation assay data (specificity 100% and sensitivity 96% at a microneutralisation cutoff of 1:40) and that fingerprick whole blood samples are sufficient for this test. Interpretation: Taken together, the COVID-19 NAb-testTM device described here provides a rapid readout of NAb based protection to SARS-CoV-2 at the point of care

HIV integration and the establishment of latency in CCL19-treated resting CD4(+) T cells require activation of NF-κB

BACKGROUND: Eradication of HIV cannot be achieved with combination antiretroviral therapy (cART) because of the persistence of long-lived latently infected resting memory CD4(+) T cells. We previously reported that HIV latency could be established in resting CD4(+) T cells in the presence of the chemokine CCL19. To define how CCL19 facilitated the establishment of latent HIV infection, the role of chemokine receptor signalling was explored. RESULTS: In resting CD4(+) T cells, CCL19 induced phosphorylation of RAC-alpha serine/threonine-protein kinase (Akt), nuclear factor kappa B (NF-κB), extracellular-signal-regulated kinase (ERK) and p38. Inhibition of the phosphoinositol-3-kinase (PI3K) and Ras/Raf/Mitogen-activated protein kinase/ERK kinase (MEK)/ERK signalling pathways inhibited HIV integration, without significant reduction in HIV nuclear entry (measured by Alu-LTR and 2-LTR circle qPCR respectively). Inhibiting activation of MEK1/ERK1/2, c-Jun N-terminal kinase (JNK), activating protein-1 (AP-1) and NF-κB, but not p38, also inhibited HIV integration. We also show that HIV integrases interact with Pin1 in CCL19-treated CD4(+) T cells and inhibition of JNK markedly reduced this interaction, suggesting that CCL19 treatment provided sufficient signals to protect HIV integrase from degradation via the proteasome pathway. Infection of CCL19-treated resting CD4(+) T cells with mutant strains of HIV, lacking NF-κB binding sites in the HIV long terminal repeat (LTR) compared to infection with wild type virus, led to a significant reduction in integration by up to 40-fold (range 1-115.4, p = 0.03). This was in contrast to only a modest reduction of 5-fold (range 1.7-11, p > 0.05) in fully activated CD4(+) T cells infected with the same mutants. Finally, we demonstrated significant differences in integration sites following HIV infection of unactivated, CCL19-treated, and fully activated CD4(+) T cells. CONCLUSIONS: HIV integration in CCL19-treated resting CD4(+) T cells depends on NF-κB signalling and increases the stability of HIV integrase, which allow subsequent integration and establishment of latency. These findings have implications for strategies needed to prevent the establishment, and potentially reverse, latent infection

Experiences in using spatial skills testing instruments with younger audiences

As graphics educators, we routinely test the 3-D spatial skills of our university-aged students using standardized instruments such as the Purdue Spatial Visualization Test: Rotations (PSVT:R), the Mental Cutting Test (MCT), the Mental Rotation Test (MRT), and the Differential Aptitude Test: Space Relations (DAT:SR). These tests were developed and validated primarily with older subjects and have not been used significantly with younger audiences. At Michigan Technological University, we have been offering a course aimed at improving the 3-D spatial skills of our first-year engineering students since 1993, but with proper training at the pre-college level, the need for our course should be diminished. Spatial skills are a part of the national standards for K-12 math education in the US; however, most teachers have been hesitant to include spatial skills training in their pre-college mathematics courses due to a lack of suitable materials for that audience. Further, with the current emphasis on testing in the US, teachers will need to be able to show that spatial skills interventions improve test scores. As part of a research study funded by the National Science Foundation in the US focused on improving the 3-D spatial skills of middle school and high school students, who typically range from 13-years old to 17-years old, the standardized testing instruments were modified and tested with these audiences. This paper describes the types of modifications made to the testing instruments and details the results obtained from using them to assess 3-D spatial skills among younger students

Sharp-tailed grouse lek attendance and fidelity in upper Michigan

To assess and improve existing monitoring protocols for sharp-tailed grouse (Tympanuchus phasianellus) in the eastern Upper Peninsula of Michigan, we used data from 58 radio-collared grouse (46 M, 12 F) monitored within 3 openland landscape types: a xeric, conifer-dominated site, a wetland-dominated site, and a site dominated by low-intensity agriculture. We used lek counts and radio telemetry to determine lek attendance rates, factors affecting lek attendance rates, lek fidelity, and inter-sexual variation in these parameters. Our analysis indicated lek attendance varied with respect to sex of bird, day of year, time after sunrise, and wind speed. Peak male lek attendance rates exceeded those of females by up to 40%, and peak lekking activity for both males and females occurred during the second and third weeks of April. Male lekking activity occurred earlier and was sustained longer than that of females. Lekking activity was negatively related to time of day and wind speed. We observed strong lek fidelity as radio-collared birds attended a primary lek 94% of the time, indicating a low probability of multiple counting of individual birds. We also proposed a method to adjust lek count data for the probability that birds are on a lek during lek counts. Our proposed method can be used by researchers and managers to improve estimates of the number of birds attending a lek by reducing the negative bias associated with observed counts

Remote sensing of biotic effects: Zebra mussels (Dreissena polymorpha) influence on water clarity in Saginaw Bay, Lake Huron

In this study, Advanced Very High Resolution Radiometer (AVHRR) remote sensing reflectance (Rrs), imagery from 1987-1993 is used to study changes in water clarity before and after zebra mussels (Dreissena polymorpha) were discovered in Saginaw Bay, Lake Huron. Spatial and temporal trends in the data indicate distinct and persistent increases in water clarity in the inner bay after the first large recruitment of zebra mussels in the fall of 1991. The pre-Dreissena imagery show that turbidity in the inner bay was influenced by the Saginaw River discharge in spring, biological production (plankton) in summer, and wind-driven resuspension in fall, with highest turbidity in spring and fall. Spatial patterns in the post-Dreissena images were more similar regardless of season, with low reflectances in the shallow regions of the inner bay where zebra mussel densities were highest. A regression model based on point data from 24 sampling stations over the 7-yr period indicates that reflectances varied significantly by site and zebra mussel densities, as well as seasonally. Trends in observed and predicted values of reflectances followed similar patterns at each station - highest values were found during 1991 and lowest during 1992 at all stations, with slightly higher Rrs in 1993 compared to 1992. Whereas AVHRR Rrs highlight the value of historical imagery for reconstructing seasonal and interannual turbidity patterns in near-shore waters, a new generation of operational ocean color satellites, such as SeaWiFS (Sea-viewing Wide Field-of-view Sensor) and the newly launched MODIS (moderate resolution imaging spectroradiometer), now provide for routine monitoring of important biological and physical processes from space

Leadership behavior in relation to dominance and reproductive status in gray wolves, Canis lupus

We analyzed the leadership behavior of breeding and nonbreeding gray wolves (Canis lupus) in three packs during winter in 1997-1999. Scent-marking, frontal leadership (time and frequency in the lead while traveling), initiation of activity, and nonfrontal leadership were recorded during 499 h of ground-based observations in Yellowstone National Park. All observed scent-marking (N = 158) was done by breeding wolves, primarily dominant individuals. Dominant breeding pairs provided most leadership, consistent with a trend in social mammals for leadership to correlate with dominance. Dominant breeding wolves led traveling packs during 64% of recorded behavior bouts (N = 591) and 71% of observed travel time (N = 64 h). During travel, breeding males and females led packs approximately equally, which probably reflects high parental investment by both breeding male and female wolves. Newly initiated behaviors (N = 104) were prompted almost 3 times more often by dominant breeders (70%) than by nonbreeders (25%). Dominant breeding females initiated pack activities almost 4 times more often than subordinate breeding females (30 vs. 8 times). Although one subordinate breeding female led more often than individual nonbreeders in one pack in one season, more commonly this was not the case. In 12 cases breeding wolves exhibited nonfrontal leadership. Among subordinate wolves, leadership behavior was observed in subordinate breeding females and other individuals just prior to their dispersal from natal packs. Subordinate wolves were more often found leading packs that were large and contained many subordinate adults

Does a STEM researcher’s role orientation predict his or her ethical sensitivity to responsible conduct of research?

A role orientation inventory provides a record of the way in which an individual orients himself or herself with respect to a particular role that he or she plays within society. Role orientation inventories were originally developed to measure an individual’s professional role orientation,for instance, with regard to being a dentist, doctor, social worker, etc. A role orientation inventory can answer questions such as: Does a doctor see her role as primarily that of providing her patient with a service in exchange for payment? Does a dentist understand his role as fundamentally consisting of serving the best interests of his patients? Does a social worker see her role as an authority figure?A test of ethical sensitivity measures an individual’s ability to recognize and take account of the ethically relevant elements of ethically-charged situations, e.g., the rights and obligations of those involved, applicable principles and ethical guidelines, and the consequences of particular courses of action, etc. Also originally developed for the field of professional ethics, ethical sensitivity tests provide information about an individual’s ability to map out and size up a situation involving ethics prior to making an ethical judgment. In fact, one’s sensitivity to an ethical situation provides the material basis upon which one makes ethical judgments and chooses courses of action.As part of an NSF sponsored project, we have developed and validated a role orientation inventory for science, technology, engineering, and mathematics (STEM) researchers and a test of an individual’s ethical sensitivity to situations involving the responsible conduct of research (RCR). Our role orientation inventory instrument is an adaptation and extension of previous instruments that were originally designed with professionals in mind. We have created an instrument appropriate to the role of a STEM researcher. Our ethical sensitivity instrument embodies an innovative approach toward measuring ethical sensitivity that relies upon comparing an individual’s ethical sensitivity to situations involving RCR with his or her ethical sensitivity to common, everyday ethical situations.In order to prepare for the final study that will be the culmination of our NSF grant project and to further validate the instruments that we have designed, we have conducted a study designed to examine the relationships between a STEM researcher’s role orientation and his or her ethical sensitivity to RCR. As ethical sensitivity is considered to be the bedrock of ethical behavior,understanding the relationships between a STEM researcher’s role orientation and ethical sensitivity to RCR is an important step in understanding the psychology behind Stem-researchers’ behavior with respect to RCR. But it also illuminates aspects of ethical behavior that ethics educators should take into account and suggests possibilities of new approaches that ethics educators can take toward forming responsible researchers. For instance, if there is correlation between a STEM researcher’s role orientation and his or her ethical sensitivity to RCR, then it is important that ethics educators explore the possibility of developing pedagogical methods for influencing researcher’s role orientation in order to enhance their ethical sensitivity to RCR

MOESM4 of HIV integration and the establishment of latency in CCL19-treated resting CD4+ T cells require activation of NF-ÎşB

Additional file 4: Figure S4. Integration site selection and gene activation in chemokine treated cells. A, Gene expression was determined by Illumina bead array in unactivated, CCL19-treated or PHA-IL2 activated CD4+ T cells after 6 or 72 h. The ratio of expression of genes at the sites of integration was determined in each in vitro condition. B, Expression of individual genes at the site of HIV integration in CCL19-treated resting CD4+ T cells (x-axis) compared to unactivated (y-axis; upper panel) or PHA-IL2 activated CD4+ T cells (y-axis; lower panel). C, The distance of integration sites to specific genomic elements including LINE, H4K20me3 and H4R3me following HIV infection of unactivated, CCL19-treated and PHA-IL2 activated CD4+ T cells, or CD4+ T cells from HIV-infected patients on cART or randomly selected sites. Log distance is shown as box plots (median and quartiles) with violin plot of the kernel distribution. The means are shown as a red horizontal line