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Designing effective antileukemic immunotherapy will require understanding mechanisms underlying tumor control or resistance. Here, we report a mechanism of escape from immunologic targeting in an acute myeloid leukemia (AML) patient, who relapsed 1 year after immunotherapy with engineered T cells expressing a human leukocyte antigen A*02 (HLA-A2)–restricted T cell receptor (TCR) specific for a Wilms’ tumor antigen 1 epitope, WT1126–134(TTCR-C4). Resistance occurred despite persistence of functional therapeutic T cells and continuous expression of WT1 and HLA-A2 by the patient’s AML cells. Analysis of the recurrent AML revealed expression of the standard proteasome, but limited expression of the immunoproteasome, specifically the beta subunit 1i (β1i), which is required for presentation of WT1126–134. An analysis of a second patient treated with TTCR-C4demonstrated specific loss of AML cells coexpressing β1i and WT1. To determine whether the WT1 protein continued to be processed and presented in the absence of immunoproteasome processing, we identified and tested a TCR targeting an alternative, HLA-A2–restricted WT137–45epitope that was generated by immunoproteasome-deficient cells, including WT1-expressing solid tumor lines. T cells expressing this TCR (TTCR37–45) killed the first patients’ relapsed AML resistant to WT1126–134targeting, as well as other primary AML, in vitro. TTCR37–45controlled solid tumor lines lacking immunoproteasome subunits both in vitro and in an NSG mouse model. As proteasome composition can vary in AML, defining and preferentially targeting these proteasome-independent epitopes may maximize therapeutic efficacy and potentially circumvent AML immune evasion by proteasome-related immunoediting.

Human CD34+hematopoietic stem and progenitor cells (HSPCs) are a standard source of cells for clinical HSC transplantations as well as experimental xenotransplantation to generate “humanized mice”. To further extend the range of applications of these humanized mice, we developed a protocol to efficiently edit the genomes of human CD34+HSPCs before transplantation. In the past, manipulating HSPCs has been complicated by the fact that they are inherently difficult to transduce with lentivectors, and rapidly lose their stemness and engraftment potential duringin vitroculture. However, with optimized nucleofection of sgRNA:Cas9 ribonucleoprotein complexes, we are now able to edit a candidate gene in CD34+HSPCs with almost 100% efficiency, and without affecting their potential for engraftment and multilineage differentiation in mice. The result is a humanized mouse from which we knocked out a gene of interest from their human immune system.

Historically, some of the first cell-free protein expression systems studied in vitro translation in various human blood cells. However, because of limited knowledge of eukaryotic translation and the advancement of cell line development, interest in these systems decreased. Eukaryotic translation is a complex system of factors that contribute to the overall translation of mRNA to produce proteins. The intracellular translateome of a cell can be modified by various factors and disease states, but it is impossible to individually measure all factors involved when there is no comprehensive understanding of eukaryotic translation. The present work outlines the use of a coupled transcription and translation cell-free protein expression system to produce recombinant proteins derived from human donor peripheral blood mononuclear cells (PBMCs) activated with phytohemagglutinin-M (PHA-M). The methods outlined here could result in tools to aid immunology, gene therapy, cell therapy, and synthetic biology research and provide a convenient and holistic method to study and assess the intracellular translation environment of primary immune cells.

IntroductionHistorical and political factors underpin the disproportional burden of type 2 diabetes mellitus (T2DM) and gestational diabetes mellitus (GDM) in women, a harbinger of future T2DM, in Indigenous populations. There is a need for T2DM prevention strategies driven by the voices of Indigenous women. In this study, we aimed to understand the perspectives of Cree women with prior GDM living in northern Quebec, where over a quarter of pregnancies are complicated by GDM.Research design and methodsA local healthcare worker invited women with GDM in the prior 5 years to participate in semistructured interviews. A Cree-origin research partner and a researcher jointly conducted interviews in-person or by teleconference. Open-ended questions addressed GDM experience, maintaining a healthy lifestyle, and needs/preferences pertinent to designing a T2DM prevention program aimed at women affected by GDM. We adopted an inductive thematic analysis framework to categorize experiences and opinions.ResultsAmong the 13 mothers interviewed, some success with health behavior changes during pregnancy was reported but there were difficulties postpartum resulting from time constraints, costs of healthy foods, discomfort at the gym related to not being perceived as athletic, and safety concerns. They acknowledged the existence of programs addressing T2DM prevention in their community but did not participate. They endorsed preferences for group sessions, with family collaboration and childcare, that addressed healthy cooking and physical activity and incorporated traditional elements.ConclusionCree mothers with a history of GDM highlighted several barriers to diabetes prevention. We are working to address these barriers through the creation of a Cree-facilitator-led community-based intervention.

Advanced understanding of T cell regulation has fueled cancer immunotherapy, but durable responses remain confined to a minority of patients. Poor responses can be linked to either suppressed T cell function in tumors or “cold” tumors that exclude immune cells. Identifying novel T cell regulators involved in these processes is a high priority. Our studies identified the protein YAP as a novel regulator of T cells. YAP mRNA and protein were rapidly expressed after T cell activation in mouse CD4+ and CD8+ T cells. T cell-specific Yap knockout (YapKO) generated CD4+ and CD8+ T cells that express elevated levels of activation markers (CD25, CD44, CD69) compared to WT cells. YapKO increased the sensitivity of T cells to anti-CD3 dose, and increased CD4+ T cell differentiation efficiency under Th1, Th2, Th17, and Treg skewing conditions. YapKO CD8+ T cells expanded >20-fold more than WT when cultured in engineered hydrogel matrices optimized for T cell expansions and adoptive transfer. Syngeneic tumors grown in T cell YapKO mice had impaired growth and increased immune infiltration compared to tumors in WT hosts. Co-adoptive transfer of YapKO and WT CD8+ T cells showed only YapKO cells infiltrated B16F10 tumors. RNAseq of CD4+ and CD8+ T cells from tumors and draining lymph nodes showed gene expression changes including upregulation of T cell receptor and cytokine signaling pathways in YapKO hosts. T cells from tumor-draining lymph nodes were not significantly different between YapKO and WT. YapKO gene signatures correlated with immune infiltration and survival in multiple human cancers in TCGA. Our data demonstrated YAP is a novel regulator of diverse T cell functions and may be a useful target for enhancing responses to cancer immunotherapy.

The article considers how the past and present tendency to focus on selecting the best projects based on the sole criterion of meritorious science may result in a sub-optimal portfolio. The authors argue that scientists need to proactively engage in the discussion over the need to improve the efficiency and effectiveness of societal investments to ensure that the next generation of the management and decision-making process for our science, technology and innovation system is rooted in sound principles. The classic peer review process tends to provide unintended overlap and allows for an ill fit between some of the pieces and unwanted gaps to occur. Areas of high risk and high return can be missed due to their controversial nature and split decisions typically resulting in negative funding decisions. In general, high risk and a high frequency of split decisions tend to be replaced with lower risk initiatives. The authors propose herein supplementing peer review with research portfolio evaluation approaches and decision-making tools that can better assess research uncertainties and other special features of the transformation of the resulting knowledge into improved social well-being. A coupling of research quality review by peers with more systematic portfolio meta-analysis of recommended projects is both possible and essential.

Microelectromechanical-systems (MEMS), microsystems technology (MST), and micromachines are roughly synonymous terms applied in the United States, Europe, and Japan respectively, to manufacturing technologies that are enabling the development of many exciting microscale or microminiaturized products. For convenience here, the terms are lumped together and labeled M3. In the parlance of management science, M3 technologies are: - exogenous (they originate from outside an existing or familiar frame of reference); and - disruptive (they are revolutionary rather than evolutionary). These technologies are creating a second micromanufacturing revolution - the first being microelectronics - as well as a fertile environment for discontinuous innovations that will revitalize existing industries and generate new markets.M3 technologies: - enable the manufacture of products that would otherwise be impossible toproduce; - improve manufacturing efficiencies by as much as an order of magnitude; - improve critical performance aspects of current products by as much as anorder of magnitude; and/or - improve product quality by reducing component size and number.

Background: Research regarding difficult mask ventilation (DMV) combined with difficult laryngoscopy (DL) is extremely limited even though each technique serves as a rescue for one another. Methods: Four tertiary care centers participating in the Multicenter Perioperative Outcomes Group used a consistent structured patient history and airway examination and airway outcome definition. DMV was defined as grade 3 or 4 mask ventilation, and DL was defined as grade 3 or 4 laryngoscopic view or four or more intubation attempts. The primary outcome was DMV combined with DL. Patients with the primary outcome were compared to those without the primary outcome to identify predictors of DMV combined with DL using a non-parsimonious logistic regression. Results: Of 492,239 cases performed at four institutions among adult patients, 176,679 included a documented face mask ventilation and laryngoscopy attempt. Six hundred ninety-eight patients experienced the primary outcome, an overall incidence of 0.40%. One patient required an emergent cricothyrotomy, 177 were intubated using direct laryngoscopy, 284 using direct laryngoscopy with bougie introducer, 163 using videolaryngoscopy, and 73 using other techniques. Independent predictors of the primary outcome included age 46 yr or more, body mass index 30 or more, male sex, Mallampati III or IV, neck mass or radiation, limited thyromental distance, sleep apnea, presence of teeth, beard, thick neck, limited cervical spine mobility, and limited jaw protrusion (c-statistic 0.84 [95% CI, 0.82–0.87]). Conclusion: DMV combined with DL is an infrequent but not rare phenomenon. Most patients can be managed with the use of direct or videolaryngoscopy. An easy to use unweighted risk scale has robust discriminating capacity.

This paper considers how and why the positive net effect of science-related activities substantially increases the value beyond what would be anticipated by the financial theory, which seems to work so well for other fields. A qualitative analysis of 25 small biotechnology R&D firms listed on a stock exchange illustrates that these firms do not follow the neo-classical expectation of Gaussian returns. To better understand this deviation from the expected Gaussian returns, the firms are compared to S&P 100 and the Thomson Reuters Global Innovator List. It is found that while these large firms have a higher than expected frequency of non-Gaussian events, the causes appear to be dominated by macro-economic or industrial events that impact a large number of firms. With the small R&D-intensive biotechnology firms, it is possible to identify specific events that appear to trigger a sudden increase or decrease in value. A better understanding of the nature and magnitude of these events allows policy makers, investors and managers to better comprehend the unusually large risks and new opportunities associated with biotechnology R&D. From this, a greater insight is afforded into the dynamic value of R&D in general.

Attempts have been made to compare the environmental impact or burden of different products or processes by equating different types of pollution. The results of these attempts are highly debated and time sensitive, due to the evolving understanding of the relative danger of different pollutants. In order to develop a method to better understand the magnitude of environmental impacts of a process or product, the relative impact on air pollution of electrical generation in forty-nine states is examined. Three pollutants (NOx, SOx, and CO2) are considered simultaneously. A State's electrical generation is ranked as efficient if its environmental burden is relatively less then at least one other state in one or more of the three categories. States that have a greater environmental burden for air pollution in all of the three categories of air pollutants are ranked as inefficient and the degree of inefficiency is calculated. Data envelopment analysis (DEA) is used for this analysis and is able to reduce a set of forty-nine options to one efficient option. The limitation of DEA for analysis of environmental burdens is also discussed.

Background Research regarding difficult mask ventilation (DMV) combined with difficult laryngoscopy (DL) is extremely limited even though each technique serves as a rescue for one another. Methods Four tertiary care centers participating in the Multicenter Perioperative Outcomes Group used a consistent structured patient history and airway examination and airway outcome definition. DMV was defined as grade 3 or 4 mask ventilation, and DL was defined as grade 3 or 4 laryngoscopic view or four or more intubation attempts. The primary outcome was DMV combined with DL. Patients with the primary outcome were compared to those without the primary outcome to identify predictors of DMV combined with DL using a non-parsimonious logistic regression. Results Of 492,239 cases performed at four institutions among adult patients, 176,679 included a documented face mask ventilation and laryngoscopy attempt. Six hundred ninety-eight patients experienced the primary outcome, an overall incidence of 0.40%. One patient required an emergent cricothyrotomy, 177 were intubated using direct laryngoscopy, 284 using direct laryngoscopy with bougie introducer, 163 using videolaryngoscopy, and 73 using other techniques. Independent predictors of the primary outcome included age 46 yr or more, body mass index 30 or more, male sex, Mallampati III or IV, neck mass or radiation, limited thyromental distance, sleep apnea, presence of teeth, beard, thick neck, limited cervical spine mobility, and limited jaw protrusion (c-statistic 0.84 [95% CI, 0.82-0.87]). Conclusion DMV combined with DL is an infrequent but not rare phenomenon. Most patients can be managed with the use of direct or videolaryngoscopy. An easy to use unweighted risk scale has robust discriminating capacity.

Research regarding difficult mask ventilation (DMV) combined with difficult laryngoscopy (DL) is extremely limited even though each technique serves as a rescue for one another.Four tertiary care centers participating in the Multicenter Perioperative Outcomes Group used a consistent structured patient history and airway examination and airway outcome definition. DMV was defined as grade 3 or 4 mask ventilation, and DL was defined as grade 3 or 4 laryngoscopic view or four or more intubation attempts. The primary outcome was DMV combined with DL. Patients with the primary outcome were compared to those without the primary outcome to identify predictors of DMV combined with DL using a non-parsimonious logistic regression.Of 492,239 cases performed at four institutions among adult patients, 176,679 included a documented face mask ventilation and laryngoscopy attempt. Six hundred ninety-eight patients experienced the primary outcome, an overall incidence of 0.40%. One patient required an emergent cricothyrotomy, 177 were intubated using direct laryngoscopy, 284 using direct laryngoscopy with bougie introducer, 163 using videolaryngoscopy, and 73 using other techniques. Independent predictors of the primary outcome included age 46 yr or more, body mass index 30 or more, male sex, Mallampati III or IV, neck mass or radiation, limited thyromental distance, sleep apnea, presence of teeth, beard, thick neck, limited cervical spine mobility, and limited jaw protrusion (c-statistic 0.84 [95% CI, 0.82-0.87]).DMV combined with DL is an infrequent but not rare phenomenon. Most patients can be managed with the use of direct or videolaryngoscopy. An easy to use unweighted risk scale has robust discriminating capacity.

Why is it that textbooks and prestigious publications such as Physics World persist in getting the definition of the fine-structure constant – a dimensionless number that describes the strength of the electromagnetic force – wrong (September p4)? The expression "e2/ch-" is clearly not "dimensionless" because it has the dimensions of charge squared on the top and mechanical units on the bottom, nor does it have the expected value of 1/137.