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The world’s largest lakes, including the Laurentian Great Lakes, have experienced significant surface warming and loss of ice cover over the last several decades. Although changing surface conditions have received substantial research interest, changes below the surface remain largely unexplored, despite their importance for turbulent mixing, nutrient cycling, and primary production. In this study, we investigate changes in subsurface thermal structure and timing in Lake Michigan-Huron related to ongoing climate warming. This work utilizes atmospheric reanalysis data to drive the Great Lakes Finite Volume Community Ocean Model (GL-FVCOM), providing three-dimensional hydrodynamic and ice simulations between 1979 and 2021. Results are used to analyze trends in ice and temperature dynamics, revealing significant changes in annually averaged ice cover (− 2.1– − 5.2%/decade), ice thickness (− 0.68 – − 2.0 cm/decade), surface temperature (+ 0.47– + 0.51 $$^\circ{\rm C}$$ ∘ C /decade), and bottom temperature (+ 0.26– + 0.29 $$^\circ{\rm C}$$ ∘ C /decade) over the last 40 years, especially in ecologically important bays (e.g., Green Bay, Saginaw Bay). Significant warming was observed at all depth layers (0–270 m), with warming trends in the epilimnion and hypolimnion that agreed well with recent analysis of observational data in Lake Michigan. Shifting stratification dynamics led to dramatic changes in modelled overturning behavior, and earlier spring turnover dates (− 2.2– − 7.5 days/decade) and later fall turnover dates (+ 2.5– + 6.3 days/decade) led to a net lengthening of the stratified period. This study presents one of the most comprehensive analyses of changes in Great Lakes subsurface temperatures to date, providing important context for future climate modelling and coastal management efforts in the region.

Intra-operative electrophysiological testing is being increasingly used to determine device functionality. Impedance abnormalities (open or short circuits) measured at time of surgery pose a dilemma: is it likely to resolve or is it a permanent fault? There is little in the literature on how to manage these intraoperative finding and if, at time of surgery, the back-up device should be used.We routinely undertake impedance testing twice intraoperatively, as well as at switch on, 1 and 3 months postoperatively. Retrospective impedance thresholds were analysed for one surgeon's cases between January 2018 and December 2019.There were 235 cochlear implants performed for 217 patients (5,020 electrode contacts) analysed. Thirty-three electrodes had abnormal impedance thresholds on first intraoperative cycle of testing, 76% resolving with the second testing cycle electrode contacts that demonstrated abnormal impedance during both intraoperative test cycles were 16.54 times (95%CI 2.55-107.13, p = 0.003) more likely to be abnormal at three months. Fifty percent resolved by switch on. The intraoperative abnormalities made up 26% of electrode abnormalities seen at 3 months postoperatively.This study demonstrates the utility of 2 cycles of intraoperative impedance testing, with persistently abnormal electrodes having 16 times the likelihood of persistent abnormalities of impedance, and 50% resolution. These persistent intra-operative abnormal electrodes are responsibly for 26% of electrode abnormalities at 3 months. This information is useful for the surgeon when considering use of the backup cochlear implant device.

Application of lake models coupled within earth-system prediction models, especially for predictions from days to weeks, requires accurate initialization of lake temperatures. Commonly used methods to initialize lake temperatures include interpolation of global sea-surface temperature (SST) analyses to inland lakes, daily satellite-based observations, or model-based reanalyses. However, each of these methods have limitations in capturing the temporal characteristics of lake temperatures (e.g., effects of anomalously warm or cold weather) for all lakes within a geographic region and/or during extended cloudy periods. An alternative lake-initialization method was developed which uses two-way-coupled cycling of a small-lake model within an hourly data assimilation system of a weather prediction model. The lake model simulated lake temperatures were compared with other estimates from satellite and in situ observations and interpolated-SST data for a multi-month period in 2021. The lake cycling initialization, now applied to two operational US NOAA weather models, was found to decrease errors in lake surface temperature from as much as 5–10 K vs. interpolated-SST data to about 1–2 K compared to available in situ and satellite observations.

Background Developing a noninvasive clinical test to accurately diagnose kidney allograft rejection is critical to improve allograft outcomes. Urinary exosomes, tiny vesicles released into the urine that carry parent cells' proteins and nucleic acids, reflect the biologic function of the parent cells within the kidney, including immune cells. Their stability in urine makes them a potentially powerful tool for liquid biopsy and a noninvasive diagnostic biomarker for kidney-transplant rejection. Methods Using 192 of 220 urine samples with matched biopsy samples from 175 patients who underwent a clinically indicated kidney-transplant biopsy, we isolated urinary exosomal mRNAs and developed rejection signatures on the basis of differential gene expression. We used crossvalidation to assess the performance of the signatures on multiple data subsets. Results An exosomal mRNA signature discriminated between biopsy samples from patients with all-cause rejection and those with no rejection, yielding an area under the curve (AUC) of 0.93 (95% CI, 0.87 to 0.98), which is significantly better than the current standard of care (increase in eGFR AUC of 0.57; 95% CI, 0.49 to 0.65). The exosome-based signature's negative predictive value was 93.3% and its positive predictive value was 86.2%. Using the same approach, we identified an additional gene signature that discriminated patients with T cell-mediated rejection from those with antibody-mediated rejection (with an AUC of 0.87; 95% CI, 0.76 to 0.97). This signature's negative predictive value was 90.6% and its positive predictive value was 77.8%. Conclusions Our findings show that mRNA signatures derived from urinary exosomes represent a powerful and noninvasive tool to screen for kidney allograft rejection. This finding has the potential to assist clinicians in therapeutic decision making.

Application of lake models coupled within earth-system prediction models, especially for short-term predictions from days to weeks, requires accurate initialization of lake temperatures. Here, we describe a lake initialization method by cycling within an hourly updated weather prediction model to constrain lake temperature evolution. We compare these simulated lake temperature values with other estimates from satellite and in situ and interpolated-SST data sets for a multi-month period in 2021. The lake cycling initialization, now applied to two operational US NOAA weather models, was found to decrease errors in lake temperature from as much as 5–10 K (using interpolated-SST data) to about 1–2 K (comparing with available in situ and satellite observations.

This study models the amount of contrast that could be conserved in computed tomographic examinations in the context of the current global shortage of iodinated contrast media.

Digital Image Correlation (DIC) is a known Non-Destructive Evaluation (NDE) method used in various industries. DIC requires a randomly produced pattern, known as speckle pattern, to track a...

Despite success in hematologic malignancies, chimeric antigen receptor (CAR) T cells have been less effective in solid tumors, in part because of the heterogeneous expression of the CAR-specific target antigen (1° Ag) found within the tumor mass. In combination with a CAR, bispecific T cell engagers (BiTEs) can target additional tumor antigens (2° Ag) while engaging CD3 signaling molecules on T cells, providing a unique way to address tumor heterogeneity, mitigate antigen escape and further potentiate durable effector function. However, in generating off-the-shelf universal CAR-T and NK cells, a combination strategy is not feasible as neither modified cell product is compatible to specifically engage with a BiTE. In developing allogeneic CAR-T cells, the T cell receptor (TCR) surface expression must be eliminated to prevent graft versus host disease, but the absence of surface TCR expression leads to loss of surface CD3 expression and BiTE compatibility. Similarly, NK cells naturally lack TCR expression and have no surface CD3 molecules for BiTE engagement. Here we discuss the development of a novel chimeric CD3ε fusion receptor (CD3-CFR) with a modified transmembrane and endodomain enabling surface expression in TCR-less T or NK cells, allowing for a novel combinatorial solution between universal CAR-T or NK cells with BiTEs in an allogeneic setting. CD3-CFR signal transduction was initially investigated in TRAC knockout NFAT reporter (T-KO) Jurkat cells engineered with CD3-CFR and co-cultured with EpCAM+ target cells. When soluble EpCAM-BiTE was added to the co-culture, the observed 6.9-fold increase in NFAT activity indicated successful engagement associated with the interaction of CD3-CFR, target cells and the BiTE. CD3-CFR T-KO Jurkat cells were further modified to secrete the EpCAM-BiTE and showed increased NFAT activity, demonstrating the feasibility of self-secreting BiTE-mediated targeting through CD3-CFR signaling. We next engineered CD3-CFR expressing iPSC-derived CAR-T (iT) cells to show preserved T cell differentiation kinetics and maintained CAR activity against 1° Ag positive targets (>90% cytolysis). Importantly, while CAR-iT cells showed only background lysis of 1° Ag negative tumor cells, CD3-CFR+ CAR-iT cells were able to lyse ~99% of 1° Ag negative tumor cells with the addition of soluble EpCAM-BiTE targeting the 2° Ag. When CD3-CFR+ CAR-iT cells were further engineered to secrete the EpCAM-BiTE, CD3-CFR+ BiTE+ CAR-iT cells demonstrated superior killing of heterogenous tumors compared to CD3-CFR+ BiTE- CAR-iT cells (70% vs. 5% cytolysis). Taken together these data demonstrate for the first time that CD3 expression and BiTE engagement can be combined in a universal iPSC-derived CAR-iT cell product to overcome antigen heterogeneity and enhance efficacy in solid tumor settings. Citation Format: Eigen Peralta, Dan Lu, Mark Landon, Hui-Yi Chu, Amit Mehta, Philip Chu, Alec Witty, Tom Lee, Bahram Valamehr. Chimeric CD3 fusion receptors expressed on iPSC-derived universal TCR-less CAR-T and -NK cells synergize with bispecific engagers to enhance antitumor activity and limit antigen escape [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5513.

New Onset Diabetes After Transplantation (NODAT) is a serious metabolic complication. While β-cell dysfunction is considered the main contributing factor in the development of NODAT, the precise pathogenesis is not well understood. Cytokines are thought to be involved in the inflammation of islet β-cells in diabetes; however, few studies have investigated this hypothesis in NODAT. A total of 309 kidney transplant recipients (KTRs) were included in this study. An association between kidney transplants, and the development of diabetes after transplant (NODAT) was investigated. Comparison was made between KTRs who develop diabetes (NODAT cases) or did not develop diabetes (control), using key cytokines, IL-6 G (− 174)C, macrophage mediator; IL-4 C (− 490)T, T helper (Th)-2 cytokine profile initiator; Th-1 cytokine profile initiator interferon-γ T (+ 874) A gene and TGF β1 C (+ 869) T gene polymorphisms were investigated. The genes were amplified using well-established polymerase chain reaction (PCR) techniques in our laboratory. Compared to the AA and AT genotypes of interferon gamma (IFNG), there was a strong association between the TT genotype of IFNG and NODAT kidney transplant recipients (KTRs) versus non-NODAT KTRs (p = 0.005). The AA genotype of IFNG was found to be predominant in the control group (p = 0.004). Also, significant variations of IL6 G (− 174) C, IL-4 C (− 590) T, interferon-γ T (+ 874) A gene and transforming growth factor β1 C (+ 869) T may contribute to NODAT. Our data is consistent with theTh-1/T-reg pathway of immunity. Further larger pan Arab studies are required to confirm our findings.

Ochronosis and alkaptonuria are manifestations of the same condition—a rare autosomal recessive disorder resulting from a constitutional lack of homogentisate 1,2-dioxygenase (HGD) with the consequent accumulation of homogentisic acid (HGA). In ochronosis, HGA undergoes autoxidation as well as enzymatic oxidation to form an ochronotic pigment that accumulates in cartilage and connective tissues. In the beginning, there is homogentisic aciduria and pigmentation of cartilages and other connective tissues. In later years, generalized osteoarthritis of the spine and large joints, termed ochronotic arthropathy, develops. The diagnosis is confirmed by quantitative measurement of HGA in urine and mutation analysis of the HGD gene. One of the differential diagnoses for the skin findings is exogenous ochronosis, a limited hyperpigmentation of skin caused by some chemicals. As for the lumbar spine findings, there can be radiographic similarities with ankylosing spondylitis (AS) including reduced intervertebral disc spaces and loss of lumbar lordosis; however, ochronosis will spare the sacroiliac joint, and the lumbar spine will show dense, wafer-like disk calcification with a vacuum disc phenomenon and broad syndesmophytes. Here, we present a case of a patient with probable ochronosis that was treated many years as ankylosing spondylitis without response, and we provide a review of the current literature on ochronosis pathogenesis, diagnosis, and treatment.

Groundwater historically has been a critical but understudied, underfunded, and underappreciated natural resource, although recent challenges associated with both groundwater quantity and quality have raised its profile. This is particularly true in the Laurentian Great Lakes (LGL) region, where the rich abundance of surface water results in the perception of an unlimited water supply but limited attention on groundwater resources. As a consequence, groundwater management recommendations in the LGL have been severely constrained by our lack of information. To address this information gap, a virtual summit was held in June 2021 that included invited participants from local, state, and federal government entities, universities, non-governmental organizations, and private firms in the region. Both technical (e.g., hydrologists, geologists, ecologists) and policy experts were included, and participants were assigned to an agricultural, urban, or coastal wetland breakout group in advance, based on their expertise. The overall goals of this groundwater summit were fourfold: (1) inventory the key (grand) challenges facing groundwater in Michigan; (2) identify the knowledge gaps and scientific needs, as well as policy recommendations, associated with these challenges; (3) construct a set of conceptual models that elucidate these challenges; and (4) develop a list of (tractable) next steps that can be taken to address these challenges. Absent this type of information, the sustainability of this critical resource is imperiled.

Solid organ transplantation is a lifesaving therapy for patients with end-organ disease. Current immunosuppression protocols are not designed to target antigen-specific alloimmunity and are uncapable of preventing chronic allograft injury. As myeloid-derived suppressor cells (MDSCs) are potent immunoregulatory cells, we tested whether donor-derived MDSCs can protect heart transplant allografts in an antigen-specific manner. C57BL/6 (H2Kb, I-Ab) recipients pre-treated with BALB/c MDSCs were transplanted with either donor-type (BALB/c, H2Kd, I-Ad) or third-party (C3H, H2Kk, I-Ak) cardiac grafts. Spleens and allografts from C57BL/6 recipients were harvested for immune phenotyping, transcriptomic profiling and functional assays. Single injection of donor-derived MDSCs significantly prolonged the fully MHC mismatched allogeneic cardiac graft survival in a donor-specific fashion. Transcriptomic analysis of allografts harvested from donor-derived MDSCs treated recipients showed down-regulated proinflammatory cytokines. Immune phenotyping showed that the donor MDSCs administration suppressed effector T cells in recipients. Interestingly, significant increase in recipient endogenous CD11b+Gr1+ MDSC population was observed in the group treated with donor-derived MDSCs compared to the control groups. Depletion of this endogenous MDSCs with anti-Gr1 antibody reversed donor MDSCs-mediated allograft protection. Furthermore, we observed that the allogeneic mixed lymphocytes reaction was suppressed in the presence of CD11b+Gr1+ MDSCs in a donor-specific manner. Donor-derived MDSCs prolong cardiac allograft survival in a donor-specific manner via induction of recipient’s endogenous MDSCs.

Summary During neonatal development, sensory cortices generate spontaneous activity patterns shaped by both sensory experience and intrinsic influences. How these patterns contribute to the assembly of neuronal circuits is not clearly understood. Using longitudinal in vivo calcium imaging in un-anesthetized mouse pups, we show that spatially segregated functional assemblies composed of interneurons and pyramidal cells are prominent in the somatosensory cortex by postnatal day (P) 7. Both reduction of GABA release and synaptic inputs onto pyramidal cells erode the emergence of functional topography, leading to increased network synchrony. This aberrant pattern effectively blocks interneuron apoptosis, causing increased survival of parvalbumin and somatostatin interneurons. Furthermore, the effect of GABA on apoptosis is mediated by inputs from medial ganglionic eminence (MGE)-derived but not caudal ganglionic eminence (CGE)-derived interneurons. These findings indicate that immature MGE interneurons are fundamental for shaping GABA-driven activity patterns that balance the number of interneurons integrating into maturing cortical networks.

The basolateral amygdala (BLA) plays a vital role in associating sensory stimuli with salient valence information. Excitatory principal neurons (PNs) undergo plastic changes to encode this association; however, local BLA inhibitory interneurons (INs) gate PN plasticity via feedforward inhibition (FFI). Despite literature implicating parvalbumin expressing (PV+) INs in FFI in cortex and hippocampus, prior anatomical experiments in BLA implicate somatostatin expressing (Sst+) INs. The lateral entorhinal cortex (LEC) projects to BLA where it drives FFI. In the present study, we explored the role of interneurons in this circuit. Using mice, we combined patch clamp electrophysiology, chemogenetics, unsupervised cluster analysis, and predictive modeling and found that a previously unreported subpopulation of fast-spiking Sst+ INs mediate LEC→BLA FFI.

BackgroundTransforming growth factor beta (TGFβ) is an immuno-suppressive cytokine present in the tumor microenvironment (TME) that creates considerable challenges for the treatment of solid tumors. Small molecule inhibitors targeting TGFβ exist, but the pleiotropic nature of TGFβ signaling suggests that a more targeted approach is preferential, especially in the context of cellular therapy. We hypothesized that primary T cells and iPSC-derived chimeric antigen receptor-T cells (CAR-iT cells) would benefit not only from blockade of TGFβ signaling, but also from re-direction of the signaling event toward specific cytokine pathways that activate cell function. Here we discuss novel synthetic TGFβ redirector constructs that overcome TME limitations and enhance CAR-iT cell function for improved efficacy in treating solid tumors.MethodsTo identify activation pathways for redirection of TGFβ signaling, we screened a panel of cytokines for their effect on the anti-tumor activity of CAR-iT cells. We then developed synthetic redirector receptors where a TGFBR2 ectodomain was fused to the top selected cytokine receptor endodomains. Redirection of TGFβ signaling was confirmed by phospho-flow of key signaling proteins. Anti-tumor activity of CAR-iT cells expressing these synthetic redirector constructs was tested in serial restimulation assays in the absence of cytokine support and in the presence of recombinant TGFβ (rTGFβ).ResultsA dose-dependent decrease in CAR-iT cell cytolytic capacity in the presence of rTGFβ was observed, with the activity of CAR-iT cells rescued in the presence of unique cytokines. We designed and tested synthetic TGFβ redirector constructs and demonstrated a rTGFβ-dependent increase in pSTAT5 positive cells (2.8-fold over control). The serial stimulation assay was then used to test CAR-iT cells engineered with synthetic TGFβ redirector receptors. After three rounds of restimulation, an increase in tumor cell numbers for non-engineered and dominant negative TGFBR2 CAR-iT cell controls was observed (41-fold and 32-fold increase over base input, respectively). In contrast, the synthetic TGFβ redirector receptor improved the ability of CAR-iT cells to control tumor cell growth with remarkable efficiency, limiting tumor growth to only 1.5-fold over three rounds of restimulation.ConclusionsThese studies demonstrate that a novel synthetic construct comprised of fusion of cytokine endodomains to a TGFBR2 ectodomain can be deployed to hijack the immuno-suppressive signal of TGFβ often found in the TME and activate CAR-iT cells for enhanced anti-tumor activity in solid tumors. Additional studies are underway to assess the temporal expression and activity of these synthetic redirector receptors in various preclinical models which will be further discussed.

Although chimeric antigen receptor (CAR)-T cells have been transformational for several liquid tumors, their effectiveness in treating a variety of solid tumors has been limited in part because the heterogeneity of tumor-associated antigens found in many bulky tumors represents a major obstacle to the development and the delivery of effective CAR-T cell therapies in cancer. Here we demonstrate that iPSC-derived CAR-T (CAR-iT) cells are an ideal off-the-shelf approach that can be engineered to perform antibody-dependent cell-mediated cytotoxicity (ADCC) to facilitate multi-antigen targeting with the introduction of various monoclonal antibodies (mAbs). Under this strategy, while the CAR is equipped to predominantly target antigen 1, targeting additional tumor associated antigens through the use of various mAbs can be leveraged through ADCC. To this end, CAR-iT cells were genetically edited at the iPSC stage to uniformly express a novel high-affinity variant (158V), non-cleavable CD16A (hnCD16) Fc receptor to recognize and bind to the Fc fragment of mAbs and facilitate ADCC. When combined with therapeutic mAbs targeting HER2, EGFR, or PDL1, the cytolytic efficacy of hnCD16+ CAR-iT cells was significantly enhanced compared to parental CAR-iT cells in several tumor models, including ovarian, HER2+ breast, and triple-negative breast cancers, underscoring the flexibility of a combined CAR and ADCC – mediated targeting approach. Importantly, although both CD16A and CAR signaling share common CD3ζ pathways, simultaneous activation did not appreciably attenuate the activation and effector function of CAR-iT cells. Moreover, hnCD16+ CAR-iT cells demonstrated enhanced control in an established triple negative breast cancer solid tumor xenograft model. The control of antigenically heterogenous tumors by hnCD16+ CAR-iT cells was also significantly improved when compared to parental CAR-iT cells. To confirm selectivity and avoidance of immunoglobulin interference with functional output, various in vitro cellular cytotoxicity assays were conducted with IgG1 to confirm that hnCD16 requires crosslinking to elicit ADCC while maintaining full potentiation of CAR function. Additional in vitro and in vivo studies are ongoing and will be presented. Collectively, the data demonstrates that CAR-iT cells can be further modified with hnCD16 Fc receptor to elicit ADCC and the combination of CAR and hnCD16 is a unique approach to tackling tumor heterogeneity often found in bulky tumors. Citation Format: Martin Hosking, Soheila Shirinbak, Joy Grant, Wen-I Yeh, Yijia Pan, Mochtar Pribadi, Amit Mehta, Amanda Yzaguirre, Philip Chu, Shohreh Sikaroodi, Angela Gentile, Bruce Walcheck, Jianming Wu, Anhco Nguyen, Bahram Valamehr. A unique strategy to arm iPSC-derived CAR-T cells to perform antibody-directed cellular cytotoxicity to facilitate multi-antigen targeting [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1497.

The highly compact Linear Ion Trap Mass Spectrometer (LITMS) combines pyrolysis gas-chromatography/mass spectrometry (GCMS) and Mars-ambient laser desorption mass spectrometry (LDMS) through a single, miniaturized yet highly-capable linear ion trap mass analyzer. The LITMS instrument is based substantially on the Mars Organic Molecule Analyzer - Mass Spectrometer (MOMA-MS) for the 2020 ExoMars mission but features further analytical enhancements. In addition to the core MOMA capabilities, LITMS enhances the instrument performance by including negative ion detection, a dual frequency RF power supply to increase mass range, precision subsampling of drill cores at fine (≤ 1 mm) spatial scales, and pyrolysis of powdered sample for evolved gas analysis (EGA) of organic content and mineral assemblages. LITMS enables in situ characterization of inorganics and organics in individual rock core layers and features. This level of integrated analytical capability is critical to achieve advanced astrobiology objectives on Mars and other planets.

This paper describes the development and testing of the PlanetVac Xodiac sampler by Honeybee Robotics. This iteration of PlanetVac builds on Honeybee's heritage of pneumatic sampling systems and modifies it to function on an Entry, Descent, and Landing (EDL) test bed vehicle. PlanetVac Xodiac was flown on Masten Space Systems Xodiac vehicle in the Mojave Desert of California. The sampler was designed to withstand the high temperatures emitted by the propulsion system plume, as well as the vibration and impact stresses of takeoff and landing. PlanetVac Xodiac not only survived all three end-to-end field tests, it also collected over three times the expected 100g sample in each trial.