The Acute Effects of a Preload Upper-Body Power Exercise on 50-m Freestyle Performance in Youth Swimmers.

ABSTRACT: Hill, V, Patterson, S, Buckthorpe, M, and Legg, HS. The acute effects of a preload upper-body power exercise on 50-m freestyle performance in youth swimmers. J Strength Cond Res 38(7): 1295-1299, 2024-This study aimed to investigate the acute effects of a medicine ball slam and the optimal recovery time required to induce a postactivation performance enhancement (PAPE) response on 50-meter freestyle swimming performance. Twenty-four (13 female, 11 male) competitive, adolescent swimmers (mean ± SD: age, 16.7 ± 1.2 years; height, 173.3 ± 6.7 cm; mass, 63.1 ± 6.4 kg) participated in a randomized crossover study. After the PAPE intervention (3 × 5 medicine ball slams), subjects had 1-minute, 4-minute, and 8-minute recovery periods before a 50-m maximal freestyle swim. A 1-way repeated-measures ANOVA revealed that different recovery times elicited changes in 50-m performance (F = 12.12, p < 0.0005). After 4 minutes of recovery, 50-m performance was 1.6% (0.47 seconds) faster (95% confidence interval [CI] [0.17-0.77], p < 0.001). When the data were split by sex, after 4 minutes of recovery, 50-m performance was 2% (0.64 seconds) faster for women (95% CI [0.279-0.998], p < 0.001). In conclusion, an upper-body power exercise, before performance, can induce a PAPE response and enhance 50-m freestyle performance after a 4-minute recovery period.

Providing a framework for seagrass mapping in United States coastal ecosystems using high spatial resolution satellite imagery.

Seagrasses have been widely recognized for their ecosystem services, but traditional seagrass monitoring approaches emphasizing ground and aerial observations are costly, time-consuming, and lack standardization across datasets. This study leveraged satellite imagery from Maxar's WorldView-2 and WorldView-3 high spatial resolution, commercial satellite platforms to provide a consistent classification approach for monitoring seagrass at eleven study areas across the continental United States, representing geographically, ecologically, and climatically diverse regions. A single satellite image was selected at each of the eleven study areas to correspond temporally to reference data representing seagrass coverage and was classified into four general classes: land, seagrass, no seagrass, and no data. Satellite-derived seagrass coverage was then compared to reference data using either balanced agreement, the Mann-Whitney U test, or the Kruskal-Wallis test, depending on the format of the reference data used for comparison. Balanced agreement ranged from 58% to 86%, with better agreement between reference- and satellite-indicated seagrass absence (specificity ranged from 88% to 100%) than between reference- and satellite-indicated seagrass presence (sensitivity ranged from 17% to 73%). Results of the Mann-Whitney U and Kruskal-Wallis tests demonstrated that satellite-indicated seagrass percentage cover had moderate to large correlations with reference-indicated seagrass percentage cover, indicative of moderate to strong agreement between datasets. Satellite classification performed best in areas of dense, continuous seagrass compared to areas of sparse, discontinuous seagrass and provided a suitable spatial representation of seagrass distribution within each study area. This study demonstrates that the same methods can be applied across scenes spanning varying seagrass bioregions, atmospheric conditions, and optical water types, which is a significant step toward developing a consistent, operational approach for mapping seagrass coverage at the national and global scales. Accompanying this manuscript are instructional videos describing the processing workflow, including data acquisition, data processing, and satellite image classification. These instructional videos may serve as a management tool to complement field- and aerial-based mapping efforts for monitoring seagrass ecosystems.

The influence of particle concentration and bulk characteristics on polarized oceanographic lidar measurements.

Oceanographic lidar measurements of the linear depolarization ratio, δ, contain information on the bulk characteristics of marine particles that could improve our ability to study ocean biogeochemistry. However, a scarcity of information on the polarized light-scattering properties of marine particles and the lack of a framework for separating single and multiple scattering effects on δ have hindered the development of polarization-based retrievals of bulk particle properties. To address these knowledge gaps, we made single scattering measurements of δ for several compositionally and morphologically distinct marine particle assemblages. We then used a bio-optical model to explore the influence of multiple scattering and particle characteristics on lidar measurements of δ made during an expedition to sample a mesoscale coccolithophore bloom. Laboratory measurements of linear depolarization revealed a complex dependency on particle shape, size, and composition that were consistent with scattering simulations for idealized nonspherical particles. Model results suggested that the variability in δ measured during the field expedition was driven predominantly by shifts in particle concentration rather than their bulk characteristics. However, model estimates of δ improved when calcite particles were represented by a distinct particle class, highlighting the influence of bulk particle properties on δ. To advance polarized lidar retrievals of bulk particle properties and to constrain the uncertainty in satellite lidar retrievals of particulate backscattering, these results point to the need for future efforts to characterize the variability of particulate depolarization in the ocean and to quantify the sensitivity of operational ocean lidar systems to multiple scattering.

Polarized lidar and ocean particles: insights from a mesoscale coccolithophore bloom.

Oceanographic lidar can provide remote estimates of the vertical distribution of suspended particles in natural waters, potentially revolutionizing our ability to characterize marine ecosystems and properly represent them in models of upper ocean biogeochemistry. However, lidar signals exhibit complex dependencies on water column inherent optical properties (IOPs) and instrument characteristics, which complicate efforts to derive meaningful biogeochemical properties from lidar return signals. In this study, we used a ship-based system to measure the lidar attenuation coefficient (α) and linear depolarization ratio (δ) across a variety of optically and biogeochemically distinct water masses, including turbid coastal waters, clear oligotrophic waters, and calcite rich waters associated with a mesoscale coccolithophore bloom. Sea surface IOPs were measured continuously while underway to characterize the response of α and δ to changes in particle abundance and composition. The magnitude of α was consistent with the diffuse attenuation coefficient (Kd), though the α versus Kd relationship was nonlinear. δ was positively related to the scattering optical depth and the calcite fraction of backscattering. A statistical fit to these data suggests that the polarized scattering properties of calcified particles are distinct and contribute to measurable differences in the lidar depolarization ratio. A better understanding of the polarized scattering properties of coccolithophores and other marine particles will further our ability to interpret polarized oceanographic lidar measurements and may lead to new techniques for measuring the material properties of marine particles remotely.

Performance across WorldView-2 and RapidEye for reproducible seagrass mapping.

Satellite remote sensing offers an effective remedy to challenges in ground-based and aerial mapping that have previously impeded quantitative assessments of global seagrass extent. Commercial satellite platforms offer fine spatial resolution, an important consideration in patchy seagrass ecosystems. Currently, no consistent protocol exists for image processing of commercial data, limiting reproducibility and comparison across space and time. Additionally, the radiometric performance of commercial satellite sensors has not been assessed against the dark and variable targets characteristic of coastal waters. This study compared data products derived from two commercial satellites: DigitalGlobe's WorldView-2 and Planet's RapidEye. A single scene from each platform was obtained at St. Joseph Bay in Florida, USA, corresponding to a November 2010 field campaign. A reproducible processing regime was developed to transform imagery from basic products, as delivered from each company, into analysis-ready data usable for various scientific applications. Satellite-derived surface reflectances were compared against field measurements. WorldView-2 imagery exhibited high disagreement in the coastal blue and blue spectral bands, chronically overpredicting. RapidEye exhibited better agreement than WorldView-2, but overpredicted slightly across all spectral bands. A deep convolutional neural network was used to classify imagery into deep water, land, submerged sand, seagrass, and intertidal classes. Classification results were compared to seagrass maps derived from photointerpreted aerial imagery. This study offers the first radiometric assessment of WorldView-2 and RapidEye over a coastal system, revealing inherent calibration issues in shorter wavelengths of WorldView-2. Both platforms demonstrated as much as 97% agreement with aerial estimates, despite differing resolutions. Thus, calibration issues in WorldView-2 did not appear to interfere with classification accuracy, but could be problematic if estimating biomass. The image processing routine developed here offers a reproducible workflow for WorldView-2 and RapidEye imagery, which was tested in two additional coastal systems. This approach may become platform independent as more sensors become available.

Intact Cohesion, Anaphase, and Chromosome Segregation in Human Cells Harboring Tumor-Derived Mutations in STAG2.

Somatic mutations of the cohesin complex subunit STAG2 are present in diverse tumor types. We and others have shown that STAG2 inactivation can lead to loss of sister chromatid cohesion and alterations in chromosome copy number in experimental systems. However, studies of naturally occurring human tumors have demonstrated little, if any, correlation between STAG2 mutational status and aneuploidy, and have further shown that STAG2-deficient tumors are often euploid. In an effort to provide insight into these discrepancies, here we analyze the effect of tumor-derived STAG2 mutations on the protein composition of cohesin and the expected mitotic phenotypes of STAG2 mutation. We find that many mutant STAG2 proteins retain their ability to interact with cohesin; however, the presence of mutant STAG2 resulted in a reduction in the ability of regulatory subunits WAPL, PDS5A, and PDS5B to interact with the core cohesin ring. Using AAV-mediated gene targeting, we then introduced nine tumor-derived mutations into the endogenous allele of STAG2 in cultured human cells. While all nonsense mutations led to defects in sister chromatid cohesion and a subset induced anaphase defects, missense mutations behaved like wild-type in these assays. Furthermore, only one of nine tumor-derived mutations tested induced overt alterations in chromosome counts. These data indicate that not all tumor-derived STAG2 mutations confer defects in cohesion, chromosome segregation, and ploidy, suggesting that there are likely to be other functional effects of STAG2 inactivation in human cancer cells that are relevant to cancer pathogenesis.

Cohesin mutations in human cancer.

Cohesin is a highly-conserved protein complex that plays important roles in sister chromatid cohesion, chromatin structure, gene expression, and DNA repair. In humans, cohesin is a ubiquitously expressed, multi-subunit protein complex composed of core subunits SMC1A, SMC3, RAD21, STAG1/2 and regulatory subunits WAPL, PDS5A/B, CDCA5, NIPBL, and MAU2. Recent studies have demonstrated that genes encoding cohesin subunits are somatically mutated in a wide range of human cancers. STAG2 is the most commonly mutated subunit, and in a recent analysis was identified as one of only 12 genes that are significantly mutated in four or more cancer types. In this review we summarize the findings reported to date and comment on potential functional implications of cohesin mutation in the pathogenesis of human cancer.

The genetics of melanoma: recent advances.

Cutaneous malignant melanoma results from the interplay of genetic, host, and environmental factors. Genetic factors implicated in melanoma etiology include inherited high-, intermediate-, and low-risk susceptibility genes as well as numerous somatic mutations in melanoma tumors. CDKN2A is the major high-risk melanoma susceptibility gene identified to date. Recent identification of low-risk loci has been accomplished predominantly through genome-wide association studies. Whole-exome and whole-genome studies have identified numerous genes somatically altered in melanoma tumors and highlighted a higher mutation load in melanoma tumors compared with those in other cancers. This higher load is believed to be attributable to the preponderance of cytosine-to-thymine nucleotide substitutions as a result of UV radiation exposure. Technological advances, particularly next-generation sequencing, have increased the opportunities for germline and somatic gene discovery in melanoma and are opening up new avenues for understanding melanoma pathogenesis as well as leading to new opportunities for treatment.

Stability of the CpG island methylator phenotype during glioma progression and identification of methylated loci in secondary glioblastomas.

BACKGROUND: Grade IV glioblastomas exist in two forms, primary (de novo) glioblastomas (pGBM) that arise without precursor lesions, and the less common secondary glioblastomas (sGBM) which develop from earlier lower grade lesions. Genetic heterogeneity between pGBM and sGBM has been documented as have differences in the methylation of individual genes. A hypermethylator phenotype in grade IV GBMs is now well documented however there has been little comparison between global methylation profiles of pGBM and sGBM samples or of methylation profiles between paired early and late sGBM samples. METHODS: We performed genome-wide methylation profiling of 20 matched pairs of early and late gliomas using the Infinium HumanMethylation450 BeadChips to assess methylation at >485,000 cytosine positions within the human genome. RESULTS: Clustering of our data demonstrated a frequent hypermethylator phenotype that associated with IDH1 mutation in sGBM tumors. In 80% of cases, the hypermethylator status was retained in both the early and late tumor of the same patient, indicating limited alterations to genome-wide methylation during progression and that the CIMP phenotype is an early event. Analysis of hypermethylated loci identified 218 genes frequently methylated across grade II, III and IV tumors indicating a possible role in sGBM tumorigenesis. Comparison of our sGBM data with TCGA pGBM data indicate that IDH1 mutated GBM samples have very similar hypermethylator phenotypes, however the methylation profiles of the majority of samples with WT IDH1 that do not demonstrate a hypermethylator phenotype cluster separately from sGBM samples, indicating underlying differences in methylation profiles. We also identified 180 genes that were methylated only in sGBM. Further analysis of these genes may lead to a better understanding of the pathology of sGBM vs pGBM. CONCLUSION: This is the first study to have documented genome-wide methylation changes within paired early/late astrocytic gliomas on such a large CpG probe set, revealing a number of genes that maybe relevant to secondary gliomagenesis.

Ganglion impar blockade: a review.

Since its original description in 1990, blockade of the ganglion impar (ganglion of Walther) has become a widely used intervention in the management of pelvic and perineal pain. We review the indications, techniques, and evidence for this pain intervention.

ß-lactam allergy: clinical implications and costs.

BACKGROUND: ß-lactam allergy is the most commonly reported medication allergy and it remains a key issue in antibiotic prescribing. A detailed and accurate history taking play a key role in preventing potentially serious clinical incidents and it may contribute in reducing costs. METHODS: Data were collected for patients with a documented penicillin allergy on their drug chart during a six month period. Sources included the inpatient drug charts and medical notes. Adherence to hospital guidelines was audited and costs of treatments were calculated. RESULTS: 94 patients with a history of penicillin allergy were included. Compliance with the hospital antibiotic policy was 81% and 52% of cases had a description of the reaction documented. The mean additional cost per patient was £89.29 (excluding VAT). CONCLUSIONS: It is important to maintain a high level of vigilance and constantly educate all healthcare professionals involved in prescribing and dispensing antibiotics in order to avoid the unnecessary use of non-penicillin-based antibiotics and associated cost implication.

Impact of Atmospheric Correction on Classification and Quantification of Seagrass Density from WorldView-2 Imagery.

Mapping the seagrass distribution and density in the underwater landscape can improve global Blue Carbon estimates. However, atmospheric absorption and scattering introduce errors in space-based sensors' retrieval of sea surface reflectance, affecting seagrass presence, density, and above-ground carbon (AGCseagrass) estimates. This study assessed atmospheric correction's impact on mapping seagrass using WorldView-2 satellite imagery from Saint Joseph Bay, Saint George Sound, and Keaton Beach in Florida, USA. Coincident in situ measurements of water-leaving radiance (LW), optical properties, and seagrass leaf area index (LAI) were collected. Seagrass classification and the retrieval of LAI were compared after empirical line height (ELH) and dark-object subtraction (DOS) methods were used for atmospheric correction. DOS left residual brightness in the blue and green bands but had minimal impact on the seagrass classification accuracy. However, the brighter reflectance values reduced LAI retrievals by up to 50% compared to ELH-corrected images and ground-based observations. This study offers a potential correction for LAI underestimation due to incomplete atmospheric correction, enhancing the retrieval of seagrass density and above-ground Blue Carbon from WorldView-2 imagery without in situ observations for accurate atmospheric interference correction.

Cell surface marker-based capture of neoantigen-reactive CD8+ T-cell receptors from metastatic tumor digests.

BACKGROUND: Cellular immunotherapies using autologous tumor-infiltrating lymphocytes (TIL) can induce durable regression of epithelial cancers in selected patients with treatment-refractory metastatic disease. As the genetic engineering of T cells with tumor-reactive T-cell receptors (TCRs) comes to the forefront of clinical investigation, the rapid, scalable, and cost-effective detection of patient-specific neoantigen-reactive TIL remains a top priority. METHODS: We analyzed the single-cell transcriptomic states of 31 neoantigen-specific T-cell clonotypes to identify cell surface dysfunction markers that best identified the metastatic transcriptional states enriched with antitumor TIL. We developed an efficient method to capture neoantigen-reactive TCRs directly from resected human tumors based on cell surface co-expression of CD39, programmed cell death protein-1, and TIGIT dysfunction markers (CD8+ TILTP). RESULTS: TILTP TCR isolation achieved a high degree of correlation with single-cell transcriptomic signatures that identify neoantigen-reactive TCRs, making it a cost-effective strategy using widely available resources. Reconstruction of additional TILTP TCRs from tumors identified known and novel antitumor TCRs, showing that at least 39.5% of TILTP TCRs are neoantigen-reactive or tumor-reactive. Despite their substantial enrichment for neoantigen-reactive TCR clonotypes, clonal dynamics of 24 unique antitumor TILTP clonotypes from four patients indicated that most in vitro expanded TILTP populations failed to demonstrate neoantigen reactivity, either by loss of neoantigen-reactive clones during TIL expansion, or through functional impairment during cognate neoantigen recognition. CONCLUSIONS: While direct usage of in vitro-expanded CD8+ TILTP as a source for cellular therapy might be precluded by profound TIL dysfunction, isolating TILTP represents a streamlined effective approach to rapidly identify neoantigen-reactive TCRs to design engineered cellular immunotherapies against cancer.

Specific recognition of an FGFR2 fusion by tumor infiltrating lymphocytes from a patient with metastatic cholangiocarcinoma.

BACKGROUND: Metastatic cholangiocarcinoma (CC), a form of gastrointestinal cancer that originates from the bile ducts, cannot be cured by currently available therapies, and is associated with dismal prognosis. In a previous case report, adoptive transfer of autologous tumor infiltrating lymphocytes (TILs), the majority of which recognized a tumor-specific point mutation, led to a profound and durable cancer regression in a patient with metastatic CC. Thus, more effective treatment for patients with this disease may be developed by using TILs that target cancer-specific mutations, but also other genetic aberrations such as gene fusions. In this context, fusions that involve fibroblast growth factor receptor 2 (FGFR2) and function as oncogenes in a subset of patients with intrahepatic CC (ICC) represent particularly attractive targets for adoptive cell therapy. However, no study to date has explored whether FGFR2 fusions can be recognized by patients' T cells. METHOD: To address whether FGFR2 fusions can be recognized by patients' T cells, we tested TILs from four patients with FGFR2 fusion-positive ICC for recognition of peptides and minigenes that represented the breakpoint regions of these fusions, which were unique to each of the four patients. RESULTS: We found that CD4+ TILs from one patient specifically recognized the breakpoint region of a unique FGFR2-TDRD1 (tudor domain-containing 1) fusion, and we isolated a T-cell receptor responsible for its recognition. CONCLUSIONS: This finding suggests that FGFR2 fusion-reactive TILs can be isolated from some patients with metastatic ICC, and thus provides a rationale for future exploration of T cell-based therapy targeting FGFR2 fusions in patients with cancer. Furthermore, it augments the rationale for extending such efforts to other types of solid tumors hallmarked by oncogenic gene fusions.

Phenotypic signatures of circulating neoantigen-reactive CD8+ T cells in patients with metastatic cancers.

Circulating T cells from peripheral blood (PBL) can provide a rich and noninvasive source for antitumor T cells. By single-cell transcriptomic profiling of 36 neoantigen-specific T cell clones from 6 metastatic cancer patients, we report the transcriptional and cell surface signatures of antitumor PBL-derived CD8+ T cells (NeoTCRPBL). Comparison of tumor-infiltrating lymphocyte (TIL)- and PBL-neoantigen-specific T cells revealed that NeoTCRPBL T cells are low in frequency and display less-dysfunctional memory phenotypes relative to their TIL counterparts. Analysis of 100 antitumor TCR clonotypes indicates that most NeoTCRPBL populations target the same neoantigens as TILs. However, NeoTCRPBL TCR repertoire is only partially shared with TIL. Prediction and testing of NeoTCRPBL signature-derived TCRs from PBL of 6 prospective patients demonstrate high enrichment of clonotypes targeting tumor mutations, a viral oncogene, and patient-derived tumor. Thus, the NeoTCRPBL signature provides an alternative source for identifying antitumor T cells from PBL of cancer patients, enabling immune monitoring and immunotherapies.

Photorespiration in eelgrass (Zostera marina L.): A photoprotection mechanism for survival in a CO2-limited world.

Photorespiration, commonly viewed as a loss in photosynthetic productivity of C3 plants, is expected to decline with increasing atmospheric CO2, even though photorespiration plays an important role in the oxidative stress responses. This study aimed to quantify the role of photorespiration and alternative photoprotection mechanisms in Zostera marina L. (eelgrass), a carbon-limited marine C3 plant, in response to ocean acidification. Plants were grown in controlled outdoor aquaria at different [CO2]aq ranging from ~55 (ambient) to ~2121 µM for 13 months and compared for differences in leaf photochemistry by simultaneous measurements of O2 flux and variable fluorescence. At ambient [CO2], photosynthesis was carbon limited and the excess photon absorption was diverted both to photorespiration and non-photochemical quenching (NPQ). The dynamic range of NPQ regulation in ambient grown plants, in response to instantaneous changes in [CO2]aq, suggested considerable tolerance for fluctuating environmental conditions. However, 60 to 80% of maximum photosynthetic capacity of ambient plants was diverted to photorespiration resulting in limited carbon fixation. The photosynthesis to respiration ratio (P E : R D) of ambient grown plants increased 6-fold when measured under high CO2 because photorespiration was virtually suppressed. Plants acclimated to high CO2 maintained 4-fold higher P E : R D than ambient grown plants as a result of a 60% reduction in photorespiration. The O2 production efficiency per unit chlorophyll was not affected by the CO2 environment in which the plants were grown. Yet, CO2 enrichment decreased the light level to initiate NPQ activity and downregulated the biomass specific pigment content by 50% and area specific pigment content by 30%. Thus, phenotypic acclimation to ocean carbonation in eelgrass, indicating the coupling between the regulation of photosynthetic structure and metabolic carbon demands, involved the downregulation of light harvesting by the photosynthetic apparatus, a reduction in the role of photorespiration and an increase in the role of NPQ in photoprotection. The quasi-mechanistic model developed in this study permits integration of photosynthetic and morphological acclimation to ocean carbonation into seagrass productivity models, by adjusting the limits of the photosynthetic parameters based on substrate availability and physiological capacity.

Vertical artifacts in high-resolution WorldView-2 and WorldView-3 satellite imagery of aquatic systems.

Satellite image artefacts are features that appear in an image but not in the original imaged object and can negatively impact the interpretation of satellite data. Vertical artefacts are linear features oriented in the along-track direction of an image system and can present as either banding or striping; banding are features with a consistent width, and striping are features with inconsistent widths. This study used high-resolution data from DigitalGlobe's (now Maxar) WorldView-3 satellite collected at Lake Okeechobee, Florida (FL), on 30 August 2017. This study investigated the impact of vertical artefacts on both at-sensor radiance and a spectral index for an aquatic target as WorldView-3 was primarily designed as a land sensor. At-sensor radiance measured by six of WorldView-3's eight spectral bands exhibited banding, more specifically referred to as non-uniformity, at a width corresponding to the multispectral detector sub-arrays that comprise the WorldView-3 focal plane. At-sensor radiance measured by the remaining two spectral bands, red and near-infrared (NIR) #1, exhibited striping. Striping in these spectral bands can be attributed to their time delay integration (TDI) settings at the time of image acquisition, which were optimized for land. The impact of vertical striping on a spectral index leveraging the red, red edge, and NIR spectral bands-referred to here as the NIR maximum chlorophyll index (MCINIR)-was investigated. Temporally similar imagery from the European Space Agency's Sentinel-3 and Sentinel-2 satellites were used as baseline references of expected chlorophyll values across Lake Okeechobee as neither Sentinel-3 nor Sentinel-2 imagery showed striping. Striping was highly prominent in the MCINIR product generated using WorldView-3 imagery, as noise in the at-sensor radiance exceeded any signal of chlorophyll in the image. Adjusting the image acquisition parameters for future tasking of WorldView-3 or the functionally similar WorldView-2 satellite may alleviate these artefacts. To test this, an additional WorldView-3 image was acquired at Lake Okeechobee, FL, on 26 May 2021 in which the TDI settings and scan line rate were adjusted to improve the signal-to-noise ratio. While some evidence of non-uniformity remained, striping was no longer noticeable in the MCINIR product. Future image tasking over aquatic targets should employ these updated image acquisition parameters. Since the red and NIR #1 spectral bands are critical for inland and coastal water applications, archived images not collected using these updated settings may be limited in their potential for analysis of aquatic variables that require these two spectral bands to derive.

Temporal Stability of Seagrass Extent, Leaf Area, and Carbon Storage in St. Joseph Bay, Florida: a Semi-automated Remote Sensing Analysis.

Seagrasses are globally recognized for their contribution to blue carbon sequestration. However, accurate quantification of their carbon storage capacity remains uncertain due, in part, to an incomplete inventory of global seagrass extent and assessment of its temporal variability. Furthermore, seagrasses are undergoing significant decline globally, which highlights the urgent need to develop change detection techniques applicable to both the scale of loss and the spatial complexity of coastal environments. This study applied a deep learning algorithmto a 30-year time series of Landsat 5 through 8 imagery to quantify seagrass extent, leaf area index (LAI), and belowground organic carbon (BGC) in St. Joseph Bay, Florida, between 1990 and 2020. Consistent with previous field-based observations regarding stability of seagrass extent throughout St. Joseph Bay, there was no temporal trend in seagrass extent (23 ± 3 km2, τ = 0.09, p = 0.59, n = 31), LAI (1.6 ± 0.2, τ = -0.13, p = 0.42, n = 31), or BGC (165 ± 19 g C m-2, τ = - 0.01, p = 0.1, n = 31) over the 30-year study period. There were, however, six brief declines in seagrass extent between the years 2004 and 2019 following tropical cyclones, from which seagrasses recovered rapidly. Fine-scale interannual variability in seagrass extent, LAI, and BGC was unrelated to sea surface temperature or to climate variability associated with the El Niño-Southern Oscillation or the North Atlantic Oscillation. Although our temporal assessment showed that seagrass and its belowground carbon were stable in St. Joseph Bay from 1990 to 2020, forecasts suggest that environmental and climate pressures are ongoing, which highlights the importance of the method and time series presented here as a valuable tool to quantify decadal-scale variability in seagrass dynamics. Perhaps more importantly, our results can serve as a baseline against which we can monitor future change in seagrass communities and their blue carbon.

Simulated response of St. Joseph Bay, Florida, seagrass meadows and their belowground carbon to anthropogenic and climate impacts.

Seagrass meadows are degraded globally and continue to decline in areal extent due to human pressures and climate change. This study used the bio-optical model GrassLight to explore the impact of climate change and anthropogenic stressors on seagrass extent, leaf area index (LAI) and belowground organic carbon (BGC) in St. Joseph Bay, Florida, using water quality data and remotely-sensed sea surface temperature (SST) from 2002 to 2020. Model predictions were compared with satellite-derived measurements of seagrass extent and shoot density from the Landsat images for the same period. The GrassLight-derived area of potential seagrass habitat ranged from 36.2 km2 to 39.2 km2, averaging 38.0 ± 0.8 km2 compared to an observed seagrass extent of 23.0 ± 3.0 km2 derived from Landsat (range = 17.9-27.4 km2). GrassLight predicted a mean seagrass LAI of 2.7 m2 leaf m-2 seabed, compared to a mean LAI of 1.9 m2 m-2 estimated from Landsat, indicating that seagrass density in St. Joseph Bay may have been below its light-limited ecological potential. Climate and anthropogenic change simulations using GrassLight predicted the impact of changes in temperature, pH, chlorophyll a, chromophoric dissolved organic matter and turbidity on seagrass meadows. Simulations predicted a 2-8% decline in seagrass extent with rising temperatures that was offset by a 3-11% expansion in seagrass extent in response to ocean acidification when compared to present conditions. Simulations of water quality impacts showed that a doubling of turbidity would reduce seagrass extent by 18% and total leaf area by 21%. Combining climate and water quality scenarios showed that ocean acidification may increase seagrass productivity to offset the negative effects of both thermal stress and declining water quality on the seagrasses growing in St. Joseph Bay. This research highlights the importance of considering multiple limiting factors in understanding the effects of environmental change on seagrass ecosystems.

Breast Cancers Are Immunogenic: Immunologic Analyses and a Phase II Pilot Clinical Trial Using Mutation-Reactive Autologous Lymphocytes.

PURPOSE: Metastatic breast cancer (mBrCa) is most often an incurable disease with only modest responses to available immunotherapies. This study investigates the immunogenicity of somatic mutations in breast cancer and explores the therapeutic efficacy in a pilot trial of mutation-reactive tumor-infiltrating lymphocytes (TILs) in patients with metastatic disease. PATIENTS AND METHODS: Forty-two patients with mBrCa refractory to previous lines of treatment underwent surgical resection of a metastatic lesion(s), isolation of TIL cultures, identification of exomic nonsynonymous tumor mutations, and immunologic screening for neoantigen reactivity. Clinically eligible patients with appropriate reactivity were enrolled into one cohort of an ongoing phase II pilot trial of adoptive cell transfer of selected neoantigen-reactive TIL, with a short course of pembrolizumab (ClinicalTrials.gov identifier: NCT01174121). RESULTS: TILs were isolated and grown in culture from the resected lesions of all 42 patients with mBrCa, and a median number of 112 (range: 6-563) nonsynonymous mutations per patient were identified. Twenty-eight of 42 (67%) patients contained TIL that recognized at least one immunogenic somatic mutation (median: 3 neoantigens per patient, range: 1-11), and 13 patients demonstrated robust reactivity appropriate for adoptive transfer. Eight patients remained clinically eligible for treatment, and six patients were enrolled on a protocol of adoptive cell transfer of enriched neoantigen-specific TIL, in combination with pembrolizumab (≤ 4 doses). Objective tumor regression was noted in three patients, including one complete response (now ongoing over 5.5 years) and two partial responses (6 and 10 months). CONCLUSION: Most patients with breast cancer generated a natural immune response targeting the expressed products of their cancer mutations. Adoptive transfer of TIL is a highly personalized experimental option for patients with mBrCa shown to be capable of mediating objective responses in this pilot trial and deserves further study.