Comparative DNA Analysis of Solid Tumors by Flow Cytometric and Image Analyses of Touch Imprints and Flow Cell Suspensions

Comparative DNA analysis by flow cytometric (FCM) and image analyses (IA) has shown a high concordance rate. When present, discordance has been attributed to the presence of aneuploid cell populations detected only by IA, yet missed by FCM. This phenomenon has been explained by loss of aneuploid cells during FCM cell processing, differences in sampling area, or misinterpretation of the DNA histograms. To determine which factors are responsible for the discordance between IA and FCM, 82 fresh solid tumors from various sites were examined. Flow cytometric analysis was performed on cell suspensions isolated from the tumors, whereas IA was performed on touch imprints (IAT) and on cytosmears of the same cell suspension used for FCM (IAF). Comparison between IAT and IAF (IAT/IAF) assessed cell processing and sampling area differences, whereas IAF/FCM comparison assessed differences in apparatus and methodology as possible contributing factors to discordance. Furthermore, DNA histograms of IAT, IAF, and FCM were analyzed in the discordant cases to determine whether the discordance was due primarily to different cell populations detected (true discordance) or due to differences in histogram interpretation of the same cell populations (false discordance). IAT/IAF and IAF/FCM concordance rates (90% and 88%) were not significantly different from that of IAT/FCM (87%). False discordance accounted for most of the discordant cases in IAT/FCM comparison (six cases, 67%), whereas true discordance was seen in three cases. In all three truly discordant cases, the DNAaneuploid cell populations detected only by IAT yet missed by FCM were also detected by IAF. This study demonstrates that discordance between IA and FCM is probably not due to cell loss during FCM cell processing or sampling area differences, but may be due to differences in assessing DNA ploidy in the interpretation of IA histograms and/or dilution of aneuploid cells by normal diploid cells in FCM

Remote Sensing of Ploidy Level in Quaking Aspen (Populus Tremuloides Michx.)

Ploidy level in plants may influence ecological functioning, demography and response to climate change. However, measuring ploidy level typically requires intensive cell or molecular methods. We map ploidy level variation in quaking aspen, a dominant North American tree species that can be diploid or triploid and that grows in spatially extensive clones. We identify the predictors and spatial scale of ploidy level variation using a combination of genetic and ground‐based and airborne remote sensing methods. We show that ground‐based leaf spectra and airborne canopy spectra can both classify aspen by ploidy level with a precision‐recall harmonic mean of 0.75–0.95 and Cohen\u27s kappa of c. 0.6–0.9. Ground‐based bark spectra cannot classify ploidy level better than chance. We also found that diploids are more common on higher elevation and steeper sites in a network of forest plots in Colorado, and that ploidy level distribution varies at subkilometer spatial scales. Synthesis. Our proof‐of‐concept study shows that remote sensing of ploidy level could become feasible in this tree species. Mapping ploidy level across landscapes could provide insights into the genetic basis of species\u27 responses to climate change

Simultaneous free-volume modeling of the self-diffusion coefficient and dynamic viscosity at high pressure

International audienceA free-volume model of the dynamic viscosity and the self-diffusion coefficients was discussed. The temperature-pressure variations of the dynamic viscosity and the self-diffusion coefficients of small molecules were predicted. The compounds, carbon tetrachloride, cyclohexane, benzene, chlorotrifluoromethane, tetramethylsilane and methylcyclohexane were used for the investigation. The relation between microstructure, free volume and different complex thermophysical properties were emphasized by the model

Ventilatory muscle strength, diaphragm thickness and pulmonary function in world-class powerlifters.

Resistance training activates the ventilatory muscles providing a stimulus similar to ventilatory muscle training. We examined the effects of elite powerlifting training upon ventilatory muscle strength, pulmonary function and diaphragm thickness in world-class powerlifters (POWER) and a control group (CON) with no history of endurance or resistance training, matched for age, height and body mass

Control of intestinal stem cell function and proliferation by mitochondrial pyruvate metabolism.

Most differentiated cells convert glucose to pyruvate in the cytosol through glycolysis, followed by pyruvate oxidation in the mitochondria. These processes are linked by the mitochondrial pyruvate carrier (MPC), which is required for efficient mitochondrial pyruvate uptake. In contrast, proliferative cells, including many cancer and stem cells, perform glycolysis robustly but limit fractional mitochondrial pyruvate oxidation. We sought to understand the role this transition from glycolysis to pyruvate oxidation plays in stem cell maintenance and differentiation. Loss of the MPC in Lgr5-EGFP-positive stem cells, or treatment of intestinal organoids with an MPC inhibitor, increases proliferation and expands the stem cell compartment. Similarly, genetic deletion of the MPC in Drosophila intestinal stem cells also increases proliferation, whereas MPC overexpression suppresses stem cell proliferation. These data demonstrate that limiting mitochondrial pyruvate metabolism is necessary and sufficient to maintain the proliferation of intestinal stem cells

Interleukin-17A Mediates Acquired Immunity to Pneumococcal Colonization

Although anticapsular antibodies confer serotype-specific immunity to pneumococci, children increase their ability to clear colonization before these antibodies appear, suggesting involvement of other mechanisms. We previously reported that intranasal immunization of mice with pneumococci confers CD4+ T cell–dependent, antibody- and serotype-independent protection against colonization. Here we show that this immunity, rather than preventing initiation of carriage, accelerates clearance over several days, accompanied by neutrophilic infiltration of the nasopharyngeal mucosa. Adoptive transfer of immune CD4+ T cells was sufficient to confer immunity to naïve RAG1−/− mice. A critical role of interleukin (IL)-17A was demonstrated: mice lacking interferon-γ or IL-4 were protected, but not mice lacking IL-17A receptor or mice with neutrophil depletion. In vitro expression of IL-17A in response to pneumococci was assayed: lymphoid tissue from vaccinated mice expressed significantly more IL-17A than controls, and IL-17A expression from peripheral blood samples from immunized mice predicted protection in vivo. IL-17A was elicited by pneumococcal stimulation of tonsillar cells of children or adult blood but not cord blood. IL-17A increased pneumococcal killing by human neutrophils both in the absence and in the presence of antibodies and complement. We conclude that IL-17A mediates pneumococcal immunity in mice and probably in humans; its elicitation in vitro could help in the development of candidate pneumococcal vaccines

Reconsidering the Tribal-State Compact Process

This essay evaluates the tribal‐state compact process, as one of several alternative, nonadversarial processes, warranting attention. It argues that, because of its binding character and relatively low cost (in contrast to litigation), and because it is based in the idea of tribes and states exhibiting mutual respect, the compact process is an advanced version of negotiation and bargaining that tribes and states should consider where appropriate

How Well Does Societal Mobility Restriction Help Control the COVID-19 Pandemic? Evidence from Real-Time Evaluation

One of the most widely implemented policy response to the novel coronavirus (SARS-CoV-2) pandemic has been the imposition of restrictions on mobility (1). These restrictions have included both incentives, encouraging working from home, supported by a wide range of online activities such as meetings, lessons, and shopping, and sanctions, such as stay at home orders, restrictions on travel, and closure of shops, offices, and public transport (2-5). The measures constitute a major component of efforts to control the COVID-19 pandemic. Compared to previous epidemic responses, they are unprecedented in both scale and scope (6). The rationale underpinning these public health measures is that restricting normal activities decreases the number, duration, and proximity of interpersonal contacts and thus the potential for viral transmission. Transmission simulations using complex mathematical modelling have built on past experience such as the 1918 influenza epidemic (7), as well as assumptions about the contemporary scale and nature of contact in populations (8). However, the initial models were not always founded on empirical evidence from behavioral scientists on the feasibility or sustainability of mass social and behavior change in contemporary society. While reductions in interpersonal contact and increases in physical distancing are known to decrease respiratory infection spread (9), the paucity of recent examples of large-scale restrictions on mobility has limited the scope for research on their impact on transmission. Where restrictions have been imposed, as with Ebola, they have involved diseases with a different mode of transmission. Nonetheless, the rapidity of progression of this pandemic has forced many governments into trialing various approaches to containment with limited evidence of effectiveness (10). More conventional public health prevention measures (such as quarantine of contacts, isolation of infected individuals and contact tracing) and control measures in health systems (such as patient flow segregation, negative pressure ventilation, and use of personal protective equipment) (11-14), have been applied widely to control the epidemic in many countries as part of a portfolio of policy responses. However, mobility restriction as a new large-scale mass behavioral and social prescription has incurred considerable costs (15, 16). Estimates suggest global GDP growth has fallen by as much as 10% (17), at least in part due to mobility restriction policies. Although views differ, not least because of the lack of information of what would happen if the disease was unchecked and the emerging evidence of persisting disability in survivors, some have argued that this is greater than would be accounted for by the economic impact of direct illness and deaths from COVID-19 (18, 19). To inform decisions on large scale restrictions of mobility, there is an urgent need to assess their effectiveness in limiting pandemic spread. To this end, we examined the association of mobility with COVID-19 incidence in Organization of Economic Cooperation and Development (OECD) countries and equivalent economies such as Singapore and Taiwan

River histories : A thematic review

Peer reviewe