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NKG2D (natural-killer group 2, member D) is a homodimeric transmembrane receptor that plays an important role in NK, γδ + , and CD8 + T cell-mediated immune responses to environmental stressors such as viral or bacterial infections and oxidative stress. However, aberrant NKG2D signaling has also been associated with chronic inflammatory and autoimmune diseases, and as such NKG2D is thought to be an attractive target for immune intervention. Here, we describe a comprehensive small-molecule hit identification strategy and two distinct series of protein–protein interaction inhibitors of NKG2D. Although the hits are chemically distinct, they share a unique allosteric mechanism of disrupting ligand binding by accessing a cryptic pocket and causing the two monomers of the NKG2D dimer to open apart and twist relative to one another. Leveraging a suite of biochemical and cell-based assays coupled with structure-based drug design, we established tractable structure–activity relationships with one of the chemical series and successfully improved both the potency and physicochemical properties. Together, we demonstrate that it is possible, albeit challenging, to disrupt the interaction between NKG2D and multiple protein ligands with a single molecule through allosteric modulation of the NKG2D receptor dimer/ligand interface.

As the total number of birds has declined in the billions over the last 50 years, an accurate method for classifying bird species is necessary for conservation efforts and population monitoring. One promising method is using machine learning models to classify birds by their sounds, which has emerged due to benefits such as being less affected by environmental factors (eg. habitat, time of day), and lower disturbances to bird species during the data collection process, contrary to other processes such as image classification. As audio processing may eventually become the main method of classifying birds and may be used as an important conservation tool, it is imperative to understand the challenges that must be overcome before it can be successfully applied. In this work, the programming language Python and the machine learning model Convolutional Neural Networks were used to process and classify audio recordings from over 150 different bird species. This study demonstrates that although audio classification is a promising method of classification, many challenges are still present in the field, such as the amount of variety in the different calls of a single bird, the presence of background noises in many audio recordings, and the difficulty in efficiently representing an audio signal with images, highlighting the importance of overcoming these challenges for conservation efforts.

Because of scarcity, vulnerability and heterogeneity in the population of circulating tumor cells (CTCs), the CTC isolation system relying on immunoaffinity interaction exhibits inconsistent efficiencies for all types of cancers...

Retrosynthetic deconstruction of a core aromatic ring is an especially simplifying retrosynthetic step, reducing the complexity of the precursor synthetic target. Moreover, when implemented to provide a penultimate intermediate, it enables late-stage divergent aryl introductions, permitting deep-seated core aryl modifications ordinarily accessible only by independent synthesis. Herein, we highlight the use of a ketone carbonyl group as the functionality to direct such late-stage divergent aryl introductions onto a penultimate intermediate with a projected application in the total synthesis of vinblastine and its presently inaccessible analogues containing indole replacements. Although the studies highlight this presently unconventional strategy with an especially challenging target in mind, the increase in molecular complexity (intricacy) established by the synthetic implementation of the powerful retrosynthetic disconnection, the use of a ketone as the precursor enabling functionality, and with adoption of either conventional or new wave (hetero)aromatic annulations combine to define a general and powerful strategy suited for widespread implementation with near limitless scope in target diversification.

Purpose We evaluated the reproducibility of a study characterizing newly‐diagnosed multiple myeloma (MM) patients within an electronic health records (EHR) database using different analytic tools. Methods We reproduced the findings of a descriptive cohort study using an iterative two‐phase approach. In Phase I, a common protocol and statistical analysis plan (SAP) were implemented by independent investigators using the Aetion Evidence Platform® (AEP), a rapid‐cycle analytics tool, and SAS statistical software as a gold standard for statistical analyses. Using the UK Clinical Practice Research Datalink (CPRD) dataset, the study included patients newly diagnosed with MM within primary care setting and assessed baseline demographics, conditions, drug exposure, and laboratory procedures. Phase II incorporated analysis revisions based on our initial comparison of the Phase I findings. Reproducibility of findings was evaluate by calculating the match rate and absolute difference in prevalence between the SAS and AEP study results. Results Phase I yielded slightly discrepant results, prompting amendments to SAP to add more clarity to operational decisions. After detailed specification of data and operational choices, exact concordance was achieved for the number of eligible patients (N = 2646), demographics, comorbidities (i.e., osteopenia, osteoporosis, cardiovascular disease [CVD], and hypertension), bone pain, skeletal‐related events, drug exposure, and laboratory investigations in the Phase II analyses. Conclusions In this reproducibility study, a rapid‐cycle analytics tool and traditional statistical software achieved near‐exact findings after detailed specification of data and operational choices. Transparency and communication of the study design, operational and analytical choices between independent investigators were critical to achieve this reproducibility.

Despite IL-9 functioning as a pleiotropic cytokine in mucosal environments, the IL-9–responsive cell repertoire is still not well defined. Here, we found that IL-9 mediates proallergic activities in the lungs by targeting lung macrophages. IL-9 inhibits alveolar macrophage expansion and promotes recruitment of monocytes that develop into CD11c + and CD11c − interstitial macrophage populations. Interstitial macrophages were required for IL-9–dependent allergic responses. Mechanistically, IL-9 affected the function of lung macrophages by inducing Arg1 activity. Compared with Arg1-deficient lung macrophages, Arg1-expressing macrophages expressed greater amounts of CCL5. Adoptive transfer of Arg1 + lung macrophages but not Arg1 − lung macrophages promoted allergic inflammation that Il9r −/− mice were protected against. In parallel, the elevated expression of IL-9, IL-9R, Arg1, and CCL5 was correlated with disease in patients with asthma. Thus, our study uncovers an IL-9/macrophage/Arg1 axis as a potential therapeutic target for allergic airway inflammation.

The immune system is stratified into layers of specialized cells with distinct functions. Recently, Lin28b was shown to serve as a master regulator of fetal lymphopoiesis, programming the development of more innate-like lymphocytes in early life. However, it remains unclear whether Lin28b specifies innate functions in more conventional adaptive lymphocytes. In this report, we discovered that Lin28b promotes the development of a more innate-like lineage of CD8+ T cells that is capable of protecting the host against a wide variety of pathogens in the absence of TCR stimulation. Using RNA-seq and ATAC-seq, we found that Lin28b transcriptionally and epigenetically programs CD8+ T cells to be highly responsive to innate cytokines. We also performed scRNAseq and found that the shift from innate-like CD8+ T cells in early life to adaptive CD8+ T cells in adulthood is mediated by changes in the abundance of distinct subsets of cells. Remarkably, the innate CD8+ T cell subset predominates in early life but is also present in adult mice and humans. Collectively, our findings demonstrate that neonatal CD8+ T cells are a distinct lineage of lymphocytes that provide the host with innate defense in early life.One sentence SummaryHigh-dimensional analysis reveals how Lin28b programs neonatal CD8+ T cells for innate defense.

Neonates often develop poor immunity against intracellular pathogens. Because CD8+ T cells are essential for eliminating infectious agents, it is crucial to understand why they behave differently in early life. Previous studies in mice have demonstrated that neonatal CD8+ T cells fail to form memory because of an intrinsic propensity to differentiate into short-lived effectors. However, the underlying mechanisms remain undefined. We now show that neonatal CD8+ T cells exhibit higher glycolytic activity than adult CD8+ T cells postinfection, which may be due to age-related differences in Lin28b expression. Importantly, when glycolysis is pharmacologically inhibited, the impaired formation of neonatal memory CD8+ T cells can be restored. Collectively, these data suggest that neonatal CD8+ T cells are inherently biased toward undergoing glycolytic metabolism postinfection, which compromises their ability to develop into memory CD8+ T cells in early life.

Atopic dermatitis results in profound changes in the function of the skin that include diminished barrier function and altered production of antimicrobial peptides. Our previous work in a model of allergic skin inflammation identified a defect in the wound healing process that was dependent on IL-4. In this report, we show that allergic skin inflammation results in a dramatic decrease in the presence of the Vγ3+ dendritic epidermal T-cell (DETC) population of γδ T cells in the skin. In mice that express an active signal transducer and activator of transcription 6 in T cells, DETCs are lost early in life. The loss of DETCs is entirely dependent on IL-4 and is recovered with a genetic deficiency of IL-4. Moreover, injection of IL-4 into wild-type mice results in acute loss of the DETC population. A similar loss of DETCs was observed in mice treated topically with MC903. Wounding of skin from Stat6VT-transgenic or MC903-treated mice resulted in decreased production of DETC-dependent cytokines in the skin, coincident with diminished wound closure. Importantly, intradermal injection of the DETC-produced cytokine fibroblast GF 7 rescued the rate of wound closure in mice with allergic skin inflammation. Together, these results suggest that the atopic environment diminishes prohealing T-cell populations in the skin, resulting in attenuated wound healing responses.

We develop robust models for earthquake preparedness by optimizing the number, location, and capacity of distribution centers (DCs). The goal is to minimize the total social costs, which include setup and initial supplies, as well as the deprivation costs associated with delayed access to supplies. The models incorporate various earthquake magnitude-specific uncertainties, such as facility damage, casualty by severity, and travel time. Examining the concept of social costs in light of an emerging concern in humanitarian logistics - the robustness of relief networks, we model two types of robustness: parameter uncertainty within a scenario and relative regret across scenarios. This unique approach reveals (1) the magnitude of social costs in the aftermath of an earthquake; (2) the hidden risks associated with inaccurate modeling of deprivation costs; and (3) the impact of budgetary constraints. We demonstrate the applicability of our approach via a case study featuring the Northridge region in California, which experienced two of the strongest earthquakes recorded in North America in 1971 and 1994.

T helper cell differentiation requires lineage-defining transcription factors and factors that have shared expression among multiple subsets. BATF is required for development of multiple Th subsets but functions in a lineage-specific manner. BATF is required for IL-9 production in Th9 cells but in contrast to its function as a pioneer factor in Th17 cells, BATF is neither sufficient nor required for accessibility at the Il9 locus. Here we show that STAT5 is the earliest factor binding and remodeling the Il9 locus to allow BATF binding in both mouse and human Th9 cultures. The ability of STAT5 to mediate accessibility for BATF is observed in other Th lineages and allows acquisition of the IL-9-secreting phenotype. STAT5 and BATF convert Th17 cells into cells that mediate IL-9-dependent effects in allergic airway inflammation and anti-tumor immunity. Thus, BATF requires the STAT5 signal to mediate plasticity at the Il9 locus., BATF is a transcription factor that is needed for IL-9 production by T helper 9 cells. Here the authors show that STAT5 is needed at the Il9 locus to enable BATF to function in this manner and that this interaction can reprogram other T helper subsets into IL-9 producing cells, thus regulating the immune response to disease.

ObjectivesPatients with multiple myeloma (MM) experience significant delays in diagnosis due to non-specific symptomatology. The aim of this study was to characterise the frequency and timing of clinical features in the primary care setting prior to MM diagnosis.DesignPopulation-based cohort study.SettingElectronic health records data of approximately 17 million patients (2006–2016) within the UK Clinical Practice Research Datalink.ParticipantsPatients aged ≥18 years with newly diagnosed MM (NDMM), no history of solid tumours and ≥2 years registration in a primary care practice prior to MM diagnosis.Main outcome measuresClinical features and symptoms including bone pain, skeletal-related events (SREs), investigation and confirmation of MM diagnostic CRAB criteria (hyperCalcaemia, Renal impairment, Anaemia, Bone lesions) during the 2 years prior to MM diagnosis; time between symptom manifestation and/or relevant investigation and diagnosis of MM.ResultsAmong 2646 patients with NDMM, 47.5% had a bone pain record during the 2-year period prior to MM diagnosis, mainly affecting the back. Regardless of baseline bone pain, investigations for serum calcium level were used in 36.4% of patients prior to MM diagnosis, followed by haemoglobin (65.6%) or renal function (74.1%). Median (Q1, Q3) time from first-recorded bone pain to MM diagnosis was 220 (80, 476) days. Median (Q1, Q3) time from first-recorded hypercalcaemia, renal impairment or anaemia to MM diagnosis was 23 (12, 46), 58 (17, 254) and 73 days (28, 232), respectively. An imaging investigation or referral for imaging was recorded for 60.0% of patients with bone pain/SRE and 32% without.ConclusionsNearly half of patients diagnosed with NDMM presented with bone pain approximately 7 months prior to MM diagnosis. Investigations to evaluate all CRAB criteria, including targeted imaging, were underused. Early recognition of myeloma clinical features and optimised use of investigations in primary care may potentially expedite MM diagnosis.

It is well known that males and females respond differently to intracellular pathogens. Females mount a more robust immune response than males, which decreases their susceptibility to infection but comes at the cost of increasing immunopathology. However, the underlying basis for sex-specific differences in the CD8+ T cell response to infection remains poorly understood. In this study, we show that female CD8+ T cells have an intrinsic propensity to become short-lived effectors, whereas male CD8+ T cells give rise to more memory precursor effector cells after murine infection with either a virus (vaccinia virus) or bacteria (Listeria monocytogenes). Interestingly, we found that the propensity of female CD8+ T cells to form short-lived effectors is not because they respond to lower amounts of cognate Ag but rather because they have an enhanced capacity to respond to IL-12, which facilitates more effector cell differentiation at each round of cell division. Our findings provide key insights into the sex-based immunological differences that underlie variations in the susceptibility to infection in males and females.

Neonates often develop poor immunity against intracellular pathogens. Since CD8+ T cells are essential for eliminating infectious agents, it is crucial to understand why they behave differently in early life. Previous studies in mice have demonstrated that neonatal CD8+ T cells fail to form memory because of an intrinsic propensity to differentiate into short-lived effectors. However, the underlying mechanisms remain undefined. We now show that neonatal CD8+ T cells exhibit higher glycolytic activity than adult CD8+ T cells after infection, which may be due to age-related differences in Lin28b expression. Importantly, when glycolysis is pharmacologically inhibited, the impaired formation of neonatal memory CD8+ T cells can be restored. Collectively, these data suggest that neonatal CD8+ T cells are inherently biased toward undergoing glycolytic metabolism after infection, which compromises their ability to develop into memory CD8+ T cells in early life.

Background: Diagnosing multiple myeloma (MM) is challenging, because MM can present with a myriad of non-specific signs and symptoms, such as bone pain, fatigue, fractures, infection, within an elderly patient population (median age of MM diagnosis is 70 years). Patients with MM therefore experience significant delays in diagnosis, resulting in increased morbidity and reduced survival. In the primary care setting, the presence of bone pain in combination with laboratory abnormalities, such as hypercalcaemia or unexplained renal impairment, have a critical diagnostic role in MM. However, the occurrence of these clinical features prior to MM diagnosis within a population-based incident cohort of patients with MM has not been previously well described. Aims: To describe the frequency and timing of the initial clinical features among patients with newly diagnosed MM (NDMM) in the UK primary care setting. Methods: We used Clinical Practice Research Datalink (CPRD), an electronic health records database, derived from routinely collected general practitioner (GP) data across the UK primary care, containing approximately 17.1 million patients at the time of analysis. We identified patients with NDMM (2006-2016), aged ≥18 years with no history of solid tumors, and ≥2 years of GP registration prior to MM diagnosis. Baseline clinical characteristics were assessed during the 2-year period before MM diagnosis. We assessed clinical features, including bone pain, skeletal-related events (SREs) and CRAB criteria (hyperCalcemia, Renal failure, Anemia, Bone lesions) for MM during the 2-year period before MM diagnosis and from the time of symptom presentation or relevant diagnostic CRAB criteria to the diagnosis of MM. Results: We identified 2,646 eligible patients with NDMM with a median [Q1, Q3] age of 70 [61, 77]; 53.3% of patients were male. During the 2-year period prior to MM diagnosis, 47.5% of patients had a record of bone pain, mainly affecting the back or other joints, and 4.8% had a record of SRE. The median time between the first record of bone pain and MM diagnosis was 220 days. Regardless of recorded bone pain, laboratory investigations for hypercalcaemia, renal impairment, and anaemia and were conducted in 36.4%, 74.2%, 65.6% and of patients prior to MM diagnosis, respectively. Approximately 5.5% of patients presented with hypercalcemia, 13.5% with renal impairment and 31.3% with anemia. The median time from the first-recorded hypercalcaemia, renal impairment or anaemia event to MM diagnosis was 23, 58 and 73 days, respectively. An imaging investigation or referral for an imaging investigation was recorded for approximately 60% of patients with bone pain/SRE and 32% without bone pain/SRE. One out of five patients with bone pain/SRE and imaging investigations had a record of magnetic resonance imaging or computed tomography scan (Table 1). Conclusions: Nearly half of patients with newly diagnosed MM presented with a bone pain symptom in primary care, approximately 7 months prior to MM diagnosis. Investigations for hypercalcaemia were not frequent in patients presenting with bone pain, and tests to explore CRAB criteria were underutilized. Underuse of targeted imaging in patients presenting with bone pain was observed. Early recognition and testing for clinical features of MM in primary care may potentially expedite the disease diagnosis and lead to timely medical care. Disclosures Seesaghur: Amgen (Europe) GmbH: Current Employment, Current equity holder in publicly-traded company. Petruski-Ivleva:Aetion: Current equity holder in private company, Ended employment in the past 24 months. Banks:Amgen (Europe) GmbH: Ended employment in the past 24 months, Other: Contract worker at Amgen during the conduct of the study. Wang:Aetion: Current Employment, Current equity holder in private company. Mattox:Aetion: Current Employment, Current equity holder in private company. Abbasi:Amgen (Europe) GmbH: Other: Contract worker at Amgen. Maskell:Amgen (Europe) GmbH: Current Employment, Current equity holder in publicly-traded company. Neasham:Amgen (Europe) GmbH: Current Employment, Current equity holder in publicly-traded company. Ramasamy:Takeda: Honoraria; Janssen: Honoraria; Janssen: Research Funding; Takeda: Research Funding; Sanofi: Honoraria; Amgen: Research Funding; Janssen: Membership on an entity's Board of Directors or advisory committees; Abbvie: Membership on an entity's Board of Directors or advisory committees; Amgen: Membership on an entity's Board of Directors or advisory committees; Takeda: Membership on an entity's Board of Directors or advisory committees; Bristol Myers Squibb: Honoraria; Oncopeptides: Honoraria; Takeda: Speakers Bureau; Bristol Myers squibb: Membership on an entity's Board of Directors or advisory committees; Bristol Myers Squibb: Research Funding; Amgen: Honoraria; GSK: Membership on an entity's Board of Directors or advisory committees; Sanofi: Membership on an entity's Board of Directors or advisory committees; Oncopeptides: Membership on an entity's Board of Directors or advisory committees.

Despite the fact that in the 2012 presidential election, two-thirds of voters waited less than 10 min and a mere 3% waited longer than an hour to cast their ballots, media accounts of excruciating waits have left a misleading impression on the general public. At the root of the problem is the allocation of voting machines based on efficiency as measured by average waiting time. This method does not account for the damaging consequences of the rare events that cause extremely long waits. We propose an extreme-value robust optimization model that can explicitly consider nominal and worst-case waiting times beyond the single-point estimate commonly seen in the literature. We benchmark the robust model against the published deterministic model using a real case from the 2008 presidential election in Franklin County, Ohio. The results demonstrate that the proposed robust model is superior in accounting for uncertainties in voter turnout and machine availability, reducing the number of voters experiencing waits that exceed two hours by 61%.

The U.S. Department of Agriculture (USDA) currently uses a bidding system to determine carriers and suppliers that would partner in providing food aid annually in response to global emergencies and famine. We mimic the USDA approach via a robust optimization model featuring box and ellipsoid uncertainty frameworks to account for uncertainties in demand, supplier and carrier bid prices. Through a case study utilizing historical invoice data, we demonstrate our model applicability in improving ocean carrier and food supplier bid pricing strategy and similar supply chain network optimization problems. Through a validation algorithm we demonstrate the value of our robust models.

Heterogeneity is a hallmark feature of the adaptive immune system in vertebrates. Following infection, naive T cells differentiate into various subsets of effector and memory T cells, which help to eliminate pathogens and maintain long-term immunity. The current model suggests there is a single lineage of naive T cells that give rise to different populations of effector and memory T cells depending on the type and amounts of stimulation they encounter during infection. Here, we have discovered that multiple sub-populations of cells exist in the naive CD8

During the ontogeny of the mammalian immune system, distinct lineages of cells arise from fetal and adult hematopoietic stem cells (HSCs) during specific stages of development. However, in some cases, the same immune cell type is produced by both HSC populations, resulting in the generation of phenotypically similar cells with distinct origins and divergent functional properties. In this report, we demonstrate that neonatal CD8+ T cells preferentially become short-lived effectors and adult CD8+ T cells selectively form long-lived memory cells after infection because they are derived from distinct progenitor cells. Notably, we find that naive neonatal CD8+ T cells originate from a progenitor cell that is distinguished by expression of Lin28b. Remarkably, ectopic expression of Lin28b enables adult progenitors to give rise to CD8+ T cells that are phenotypically and functionally analogous to those found in neonates. These findings suggest that neonatal and adult CD8+ T cells belong to separate lineages of CD8+ T cells, and potentially explain why it is challenging to elicit memory CD8+ T cells in early life.

This paper uses the concept of social cost, comprised of private and externality costs, to capture the time-elapsed economic value of losses inflicted on users of a service under cyberattack. Our approach offers a holistic treatment of cybersecurity that not only employs prevention as a first defense but also uses detection & containment safeguards to mitigate the damage of successful attacks. It examines the optimal balance between these two safeguards under three sources of uncertainty through a robust optimization model with the help of distribution-free ellipsoidal uncertainty sets to ease the challenge of providing exact estimates for uncertain input. Our method is more appropriate than stochastic programming and other competing RO methods in addressing cybersecurity parameter uncertainty. Tested on a case study, results from 25 deterministic scenarios first reveal a strong resource-allocation preference for the prevention safeguard, but when the budget constraint is relaxed, preference shifts toward the containment & detection safeguard. Results from 54 robust test instances indicate that, for the three sources of uncertainty, the adjusted effectiveness of the prevention safeguard has the greatest impact on both social cost and the optimal configuration of safeguards. Our analysis points to some serious flaws in the existing cybersecurity framework's reliance on prevention and provides decisionmakers with urgently needed guidelines.

Atopic dermatitis (AD) is a common Th2-biased chronic inflammatory skin disease. Major clinical manifestations include erythema, severe itchiness, and dry skin that impose a burden on patients and impairs their quality of life. Extensive research has been conducted to identify possible underlying mechanisms and develop therapeutic approaches. γδT cells, an understudied T cell population, play important roles in maintaining skin integrity in both human and mice. They are among the first responders to epithelial wounding and are a source of keratinocyte/fibroblast growth factors that promote wound repair. Hence, we wanted to investigate their behavior during wounding in an AD-like environment. Using a mouse model, we observed that γδT cell numbers are drastically decreased in atopic skin. Multiple factors contribute to this decrease, including insufficient activation, less proliferative capacity and increased cell death. These changes in γδT cell function are IL-4 dependent. Administration of IL-4 to wild type (WT) mice resulted in significant loss of γδT cells, and Il4−/− mice in the AD model showed numbers of γδT cells similar to WT controls. We further observed that the levels of keratinocyte/fibroblast growth factors are lower in AD animals, suggesting that loss of γδT cells could compromise re-epithelialization and repair following wounding. Collectively, our data indicate that in an atopic environment, γδT cells decrease drastically in an IL-4 dependent fashion, which may contribute to the aberrant wound healing process in AD animals.

International audience; Mirror mirror on the wall: By taking advantage of the unique structural features of L-DNA, the first examples of left-helical enantioselective induction in the field of DNA-based asymmetric catalysis were realized. Most importantly, this approach is the only one that allows a reliable and predictable access to both enantiomers for any given reaction.

A new strategy for enantioselective synthesis of azacyclic molecules in which dynamic kinetic equilibration of diastereomeric iminium ions precedes a stereochemistry-determining sigmatropic rearrangement is reported. The method is illustrated by the synthesis, in high enantiomeric purity (generally 95-99% ee), of a variety of 1-azabicyclic molecules containing angular allyl or 3-substituted 2-propenyl side chains adjacent to nitrogen and up to three stereogenic centers. In these products, the size of the carbocyclic ring is varied widely (5-12 membered); however, useful yields are obtained in forming 1-azabicyclic products containing only fused pyrrolidine and piperidine rings. Chirality transfer from substituents at carbons 1 and 2 of the 3-butenylamine fragment of the starting material is investigated, with methyl and phenyl substituents at the allylic position shown to provide exquisite stereocontrol (generally 98-99% chirality transfer). An attractive feature of the method is the ability to carry out the key transformation in the absence of solvent. Illustrated also is the high yielding conversion of four such products to a new family of bicyclic β-amino acids of high enantiomeric purity.

A critical issue when solving the share-of-choice product design problem is the reliability of the optimal solution in the presence of partworth uncertainty. Existing approaches use point estimates of an individual's partworth utilities as input to the product optimization stage, ignoring within-person variability in estimates. Post-optimality sensitivity analysis is occasionally performed to assess the degree to which a solution is negatively impacted by partworth uncertainty. We propose a robust optimization model that explicitly captures variation in partworth estimates during the optimization process. Using a large, commercial dataset, we benchmark our model's performance against its deterministic counterpart. We also present inferential theory to guide the selection of model parameters controlled by the analyst. Results reveal that the new approach produces robust solutions in the face of measurement error. Out-of-sample coverage for individuals drawn from the target population is significantly higher than corresponding solutions from published methods.

We previously found that neonatal and adult CD8+ T cells adopt different fates after infection with specific pathogens because they are derived from a distinct progenitor cell that is distinguished by expression of Lin28b. In this report, we investigated if CD8+ T cells respond differently to unrelated pathogens in early life. To start, we compared gene expression profiles of neonatal and adult CD8+ T cells after stimulation with innate cytokines (IL-12 + IL-18). Interestingly, we found that neonatal CD8+ T cells are more responsive and express a distinct cytokine profile compared to their adult counterparts. To examine the biological significance of these age-related differences, we next adoptively transferred monoclonal populations of neonatal or adult CD8+ T cells into T cell-deficient recipients and compared their ability to mediate immune protection against an irrelevant strain of Listeria monocytogenes. Surprisingly, we found that neonatal donor cell displayed increased proliferation and effector functions, and the pathogen burden was significantly reduced when compared to recipients containing adult donor cells. We also found that Lin28b may be responsible for the enhanced responsiveness to innate cytokines in early life, as ectopic expression of Lin28b in adult cells enabled them to initiate a program of bystander activation that was analogous to neonates. Collectively, these findings indicate that neonatal CD8+ T cells possess innate-like functions during infections at the cost of generating memory, and these unique behaviors may be driven by high levels of Lin28b. We currently are employing ATAC-seq to determine whether Lin28b changes the program of bystander activation by altering the chromatin landscape.

It is well known that males and females respond differently to intracellular pathogens. In general, females mount more robust immune responses than males, which decreases susceptibility to infection but comes at the cost of increasing auto-immunopathology. However, the underlying basis for these sex-related differences is poorly understood. In this study, we investigated cell-intrinsic differences between male and female CD8+ T cells. First, we compared the ability of male and female CD8+ T cells to proliferate following in vitro stimulation. Interestingly, we found that female CD8+ T cells proliferate sooner and faster than their male counterparts. Next, we asked whether cell-intrinsic differences between male and female CD8+ T cells contribute to changes in the CD8+ T cell response to infection in vivo. We co-transferred equal numbers of male and female donor CD8+ T cells from TCR transgenic mice into the same host and compared their ability to respond to Listeria monocytogenes. The data indicated that female CD8+ T cells preferentially become short-lived effectors (SLECs), while male donor CD8+ T cells give rise to more memory precursor cells (MPECs), even when placed in the same environment. To gain insight into why female CD8+ T cells are inherently more responsive to infection, we performed a series of in vitro and in vivo experiments, whereby either antigen (signal 1) or IL-12 (signal 3) was titrated. These studies showed that male and female CD8+ T cells respond similarly to low amounts of cognate antigen, but female CD8+ T cells appear to be more sensitive to IL-12. Collectively, our data demonstrates that female CD8+ T cells are biased towards forming SLECs after infection because of their enhanced capacity to respond to IL-12.

Neonates often generate incomplete immunity against intracellular pathogens, although the mechanism of this defect is poorly understood. An important question is whether the impaired development of memory CD8+ T cells in neonates is due to an immature priming environment or lymphocyte-intrinsic defects. In this article, we show that neonatal and adult CD8+ T cells adopted different fates when responding to equal amounts of stimulation in the same host. Whereas adult CD8+ T cells differentiated into a heterogeneous pool of effector and memory cells, neonatal CD8+ T cells preferentially gave rise to short-lived effector cells and exhibited a distinct gene expression profile. Surprisingly, impaired neonatal memory formation was not due to a lack of responsiveness, but instead because neonatal CD8+ T cells expanded more rapidly than adult cells and quickly became terminally differentiated. Collectively, these findings demonstrate that neonatal CD8+ T cells exhibit an imbalance in effector and memory CD8+ T cell differentiation, which impairs the formation of memory CD8+ T cells in early life.

This method starts from substrates with high enantiomeric excesses and proceeds with excellent chirality transfers (usually ≥ 96%) yielding products with up to three stereocenters.

Key features of the CD8+ T cell response to infection are the robust clonal expansion and phenotypic diversification of naïve precursors into effector and memory cells. However, mechanisms that regulate the fate of naïve CD8+ T cells after microbial challenge remain a subject of controversy. Previous studies have suggested heterogeneity in the effector pool is driven by differences in TCR avidity, asymmetric cell division, and environmental cues. However, one variable that has not been closely examined is the developmental origin of naïve CD8+ T cells. Notably, the pool of naïve CD8+ T cells in the adult is comprised of clonotypes that were produced during various stages of life. To determine whether the developmental origin of naïve CD8+ T cells in adults plays a deterministic role in their fate after infection, we developed a system to ‘timestamp’ CD8+ T cells from various stages of development (1 day, 1 week, and 4 weeks) and examined their phenotype and behavior in 8 week-old adult mice. Interestingly, we found naïve CD8+ T cells made early in life predominantly display a CD44hi CD122hi memory phenotype whereas cells made later in life are CD44lo CD122lo. In vitro stimulation of stamped populations showed that fetal- and neonatal-derived CD8+ T cells are hypersensitive to IL12/IL18 and proliferate more rapidly than cells made in older animals. In response to infection in adulthood, both polyclonal and monoclonal CD8+ T cells produced in early life skew towards a terminally differentiated short-lived effector phenotype (KLRG1hiCD127lo), indicating this phenomenon is TCR independent. These data indicate that the spectrum of CD8+ T cell differentiation observed after infection is influenced by when the responding cells were initially made.

Neonates often generate incomplete immunity against intracellular pathogens. As CD8+ T cells are essential in clearing intracellular infections, it is crucial to understand how neonatal CD8+ T cells respond to infection. We showed that neonatal CD8+ T cells fail to form memory. However, the underlying basis for this is unclear. Here, we investigated whether neonatal CD8+ T cells fail to form memory cells because (i) they have undergone more extensive homeostatic proliferation (HP) prior to infection (proliferation model) or (ii) they are created from a distinct hematopoietic stem cell (HSC) population (origin model). We first compared the behavior and gene expression profiles of different aged CD8+ T cells that had undergone similar amounts of HP. Interestingly, neonatal CD8+ T cells still possessed unique gene expression profiles and failed to form memory post-infection. Hence, the proliferation model cannot fully explain the differences. We tested the origin model by comparing neonatal and adult CD8+ T cells at the same stage of thymic development. Again, we observed distinct gene expression profiles, indicating that CD8+ T cells are created differently in neonates. To examine whether different HSCs underlie these differences, we adoptively transferred fetal and adult progenitors into adult thymii and found that fetal progenitor derived CD8+ T cells exhibited a memory phenotype and selectively formed short-lived effectors after infection. The adult progenitor derived cells exhibited a more naïve phenotype and preferentially formed memory precursors after infection. Collectively, our data extends the layered immune hypothesis to CD8+ T cells and suggests that neonatal and adult CD8+ T cells are derived from different HSCs.

A useful enantioselective synthesis of angularly substituted 1-azabicyclic molecules that delivers the bicyclic amine products in good yield and 99% ee is reported. Angularly substituted cis-octahydrocyclopenta[b]pyrroles, cis-octahydroindoles, cis-decahydrocyclohepta[b]pyrroles, and cis-octahydrocyclopenta[b]pyridine are formed in good yields and 99% ee exclusively as the cis stereoisomers. Decahydroquinolines are generated as mixtures of cis and trans stereoisomers in similarly high ee. The starting materials for this cascade transformation are assembled in five steps from cycloalkanones and enantiomerically pure (R)-2-phenyl-3-butenamine. This enantioselective synthesis introduces a new strategy for dynamic kinetic resolution in which a rapid tautomeric equilibration of diastereomeric iminium cations is combined with a diastereoselective sigmatropic rearrangement.

A new synthesis of 3,3′-bioxindoles is reported that is well suited for the preparation of unsymmetrical structures. In the key step, 3-hydroxy-3,3′-bioxindoles are constructed by Mukaiyama aldol reaction of 2-siloxyindoles with isatins These tertiary carb inols are formed in high diastereoselectivities, with substitution at various positions of the isatin and the 2-siloxyindole being tolerated.

A new synthesis of 3,3′-bioxindoles is reported that is well suited for the preparation of unsymmetrical structures. In the key step, 3-hydroxy-3,3′-bioxindoles are constructed by Mukaiyama aldol reaction of 2-siloxyindoles with isatins These tertiary carb inols are formed in high diastereoselectivities, with substitution at various positions of the isatin and the 2-siloxyindole being tolerated.