Porcine epidemic diarrhea virus E protein induces unfolded protein response through activating both PERK and ATF6 rather than IRE1 signaling pathway.

Porcine epidemic diarrhea virus (PEDV) small envelope protein (E) plays important roles in virus budding, assembly, and release. Our previous study found that PEDV E protein localizes in the endoplasmic reticulum (ER) to trigger the unfolded protein response (UPR). However, how UPR is directly regulated by PEDV E protein remains elusive. Thus, in this study, we investigated the expression of ER chaperone glucose-regulated protein 78 (GRP78) and activations of the three main UPR signaling pathways to elucidate the underlying mechanisms of UPR triggered by PEDV E protein. The results showed that over-expression of PEDV E protein increased expression of GRP78 and induced stronger phosphorylation of both protein kinase RNA-like ER kinase (PERK) and eukaryotic initiation factor-2α (eIF2α), as well as caused the significant degradation of activating transcription factor 6 (ATF6), in both dose- and time-dependent manners. However, PEDV E protein did not induce UPR through the inositol-requiring enzyme 1 (IRE1) signaling pathway, as revealed by the splicing of XBP1 remaining unaffected and unchanged when PEDV E protein was overexpressed. Taken together, these results demonstrate that PEDV E protein induces UPR through activation of both PERK and ATF6 pathways rather than IRE1 signaling. This study not only provides mechanistic details of UPR induced by the PEDV E protein, but also provides insights into these new biologic functions to help us better understand the interactions between PEDV and host cells.

What University Students Learn In Visualization Classes.

As a step towards improving visualization literacy, this work investigates how students approach reading visualizations differently after taking a university-level visualization course. We asked students to verbally walk through their process of making sense of unfamiliar visualizations, and conducted a qualitative analysis of these walkthroughs. Our qualitative analysis found that after taking a visualization course, students engaged with visualizations in more sophisticated ways: they were more likely to exhibit design empathy by thinking critically about the tradeoffs behind why a chart was designed in a particular way, and were better able to deconstruct a chart to make sense of it. We also gave students a quantitative assessment of visualization literacy and found no evidence of scores improving after the class, likely because the test we used focused on a different set of skills than those emphasized in visualization classes. While current measurement instruments for visualization literacy are useful, we propose developing standardized assessments for additional aspects of visualization literacy, such as deconstruction and design empathy. We also suggest that these additional aspects could be incorporated more explicitly in visualization courses.

The Backstory to "Swaying the Public": A Design Chronicle of Election Forecast Visualizations.

A year ago, we submitted an IEEE VIS paper entitled "Swaying the Public? Impacts of Election Forecast Visualizations on Emotion, Trust, and Intention in the 2022 U.S. Midterms" [50], which was later bestowed with the honor of a best paper award. Yet, studying such a complex phenomenon required us to explore many more design paths than we could count, and certainly more than we could document in a single paper. This paper, then, is the unwritten prequel-the backstory. It chronicles our journey from a simple idea-to study visualizations for election forecasts-through obstacles such as developing meaningfully different, easy-to-understand forecast visualizations, crafting professional-looking forecasts, and grappling with how to study perceptions of the forecasts before, during, and after the 2022 U.S. midterm elections. This journey yielded a rich set of original knowledge. We formalized a design space for two-party election forecasts, navigating through dimensions like data transformations, visual channels, and types of animated narratives. Through qualitative evaluation of ten representative prototypes with 13 participants, we then identified six core insights into the interpretation of uncertainty visualizations in a U.S. election context. These insights informed our revisions to remove ambiguity in our visual encodings and to prepare a professional-looking forecasting website. As part of this story, we also distilled challenges faced and design lessons learned to inform both designers and practitioners. Ultimately, we hope our methodical approach could inspire others in the community to tackle the hard problems inherent to designing and evaluating visualizations for the general public.

Promises and Pitfalls: Using Large Language Models to Generate Visualization Items.

Visualization items-factual questions about visualizations that ask viewers to accomplish visualization tasks-are regularly used in the field of information visualization as educational and evaluative materials. For example, researchers of visualization literacy require large, diverse banks of items to conduct studies where the same skill is measured repeatedly on the same participants. Yet, generating a large number of high-quality, diverse items requires significant time and expertise. To address the critical need for a large number of diverse visualization items in education and research, this paper investigates the potential for large language models (LLMs) to automate the generation of multiple-choice visualization items. Through an iterative design process, we develop the VILA (Visualization Items Generated by Large LAnguage Models) pipeline, for efficiently generating visualization items that measure people's ability to accomplish visualization tasks. We use the VILA pipeline to generate 1,404 candidate items across 12 chart types and 13 visualization tasks. In collaboration with 11 visualization experts, we develop an evaluation rulebook which we then use to rate the quality of all candidate items. The result is the VILA bank of ∼1,100 items. From this evaluation, we also identify and classify current limitations of the VILA pipeline, and discuss the role of human oversight in ensuring quality. In addition, we demonstrate an application of our work by creating a visualization literacy test, VILA-VLAT, which measures people's ability to complete a diverse set of tasks on various types of visualizations; comparing it to the existing VLAT, VILA-VLAT shows moderate to high convergent validity (R = 0.70). Lastly, we discuss the application areas of the VILA pipeline and the VILA bank and provide practical recommendations for their use. All supplemental materials are available at https://osf.io/ysrhq/.

Sex differences in cardiac transcriptomic response to neonatal sleep apnea.

Pediatric obstructive sleep apnea poses a significant health risk, with potential long-term consequences on cardiovascular health. This study explores the dichotomous nature of neonatal cardiac response to chronic intermittent hypoxia (CIH) between males and females, aiming to fill a critical knowledge gap in the understanding of sex-specific cardiovascular consequences of sleep apnea in early life. Neonates were exposed to CIH until p28 and underwent comprehensive in vivo physiological assessments, including whole-body plethysmography, treadmill stress-tests, and echocardiography. Results indicated that male CIH rats weighed 13.7% less than age-matched control males (p = 0.0365), while females exhibited a mild yet significant increased respiratory drive during sleep (93.94 ± 0.84 vs. 95.31 ± 0.81;p = 0.02). Transcriptomic analysis of left ventricular tissue revealed a substantial sex-based difference in the cardiac response to CIH, with males demonstrating a more pronounced alteration in gene expression compared to females (5986 vs. 3174 genes). The dysregulated miRNAs in males target metabolic genes, potentially predisposing the heart to altered metabolism and substrate utilization. Furthermore, CIH in males was associated with thinner left ventricular walls and dysregulation of genes involved in the cardiac action potential, possibly predisposing males to CIH-related arrhythmia. These findings emphasize the importance of considering sex-specific responses in understanding the cardiovascular implications of pediatric sleep apnea.

Molecular and cellular neurocardiology in heart disease.

This paper updates and builds on a previous White Paper in this journal that some of us contributed to concerning the molecular and cellular basis of cardiac neurobiology of heart disease. Here we focus on recent findings that underpin cardiac autonomic development, novel intracellular pathways and neuroplasticity. Throughout we highlight unanswered questions and areas of controversy. Whilst some neurochemical pathways are already demonstrating prognostic viability in patients with heart failure, we also discuss the opportunity to better understand sympathetic impairment by using patient specific stem cells that provides pathophysiological contextualization to study 'disease in a dish'. Novel imaging techniques and spatial transcriptomics are also facilitating a road map for target discovery of molecular pathways that may form a therapeutic opportunity to treat cardiac dysautonomia.

Porcine epidemic diarrhea virus E protein induces formation of stress granules and attenuates protein translation through activation of the PERK/eIF2α signaling pathway.

Porcine epidemic diarrhea virus (PEDV) envelope protein (E) has been characterized as an important structural protein that plays critical roles in the interplay with its host to affect the virus life cycle. Stress granules (SGs) are host translationally silent ribonucleoproteins, which are mainly induced by the phosphorylation of eIF2α in the PERK/eIF2α signaling pathway. Our previous study found that PEDV E protein caused endoplasmic reticulum stress response (ERS)-mediated suppression of antiviral proteins' translation. However, the link and the underlying mechanism by which PEDV induces SGs formation and suppresses host translation remain elusive. In this study, our results showed that PEDV E protein significantly elevated the expression of GRP78, CANX, and phosphorylation of PERK and eIF2α, indicating that the PERK/eIF2α branch of ERS was activated. PEDV E protein localized to the ER and aggregated into puncta to reconstruct ER structure, and further induced SGs formation, which has been caused through upregulating the G3BP1 expression level. In addition, a significant global translational stall and endogenous protein translation attenuation were detected in the presence of E protein overexpression, but the global mRNA transcriptional level remained unchanged, suggesting that the shutoff of protein translation was associated with the translation, not with the transcription process. Collectively, this study demonstrates that PERK/eIF2α activation is required for SGs formation and protein translation stall. This study is beneficial for us to better understand the mechanism by which PEDV E suppresses host protein synthesis, and provides us a new insight into the host translation regulation during virus infection.

Improvement of oxidized myofibrillar protein gel properties by black rice extract.

In order to investigate the effects of black rice extract (BE) on the composition of oxidized myofibrillar protein (MP) gel, different concentrations of BE (0, 10, 20, 50 mg g-1) were analyzed experimentally. Results revealed that the addition of small doses of BE significantly inhibited the formation of carbonyl groups in oxidized MP, and improved surface hydrophobicity and gel water holding capacity. Additionally, 10 and 20 mg g-1 BE increased the ordered structure of oxidized MP. Furthermore, dynamic rheometer results showed a significant increase in the storage modulus (G') of oxidized MP with 10 and 20 mg g-1 BE during heating. Scanning Electron Microscopy (SEM) showed that MP formed a denser network structure with addition of 10 and 20 mg g-1 BE. Low-Field Nuclear Magnetic Resonance (LF-NMR) and magnetic resonance imaging (MRI) showed that there is a significant increase in immobile water in MP gel and a decrease in free water within the 20 mg g-1 BE group. In conclusion, 20 mg g-1 supplemented BE significantly improved the structure order and hardness of oxidized MP gel, increased its structure density and water holding capacity, and it provides a theoretical basis for the application of antioxidants in meat products.

Risk of thrombosis and bleeding in gynecologic cancer surgery: systematic review and meta-analysis.

OBJECTIVE: This study aimed to provide procedure-specific estimates of the risk of symptomatic venous thromboembolism and major bleeding in the absence of thromboprophylaxis, following gynecologic cancer surgery. DATA SOURCES: We conducted comprehensive searches on Embase, MEDLINE, Web of Science, and Google Scholar for observational studies. We also reviewed reference lists of eligible studies and review articles. We performed separate searches for randomized trials addressing effects of thromboprophylaxis and conducted a web-based survey on thromboprophylaxis practice. STUDY ELIGIBILITY CRITERIA: Observational studies enrolling ≥50 adult patients undergoing gynecologic cancer surgery procedures reporting absolute incidence for at least 1 of the following were included: symptomatic pulmonary embolism, symptomatic deep vein thrombosis, symptomatic venous thromboembolism, bleeding requiring reintervention (including reexploration and angioembolization), bleeding leading to transfusion, or postoperative hemoglobin <70 g/L. METHODS: Two reviewers independently assessed eligibility, performed data extraction, and evaluated risk of bias of eligible articles. We adjusted the reported estimates for thromboprophylaxis and length of follow-up and used the median value from studies to determine cumulative incidence at 4 weeks postsurgery stratified by patient venous thromboembolism risk factors. The GRADE approach was applied to rate evidence certainty. RESULTS: We included 188 studies (398,167 patients) reporting on 37 gynecologic cancer surgery procedures. The evidence certainty was generally low to very low. Median symptomatic venous thromboembolism risk (in the absence of prophylaxis) was <1% in 13 of 37 (35%) procedures, 1% to 2% in 11 of 37 (30%), and >2.0% in 13 of 37 (35%). The risks of venous thromboembolism varied from 0.1% in low venous thromboembolism risk patients undergoing cervical conization to 33.5% in high venous thromboembolism risk patients undergoing pelvic exenteration. Estimates of bleeding requiring reintervention varied from <0.1% to 1.3%. Median risks of bleeding requiring reintervention were <1% in 22 of 29 (76%) and 1% to 2% in 7 of 29 (24%) procedures. CONCLUSION: Venous thromboembolism reduction with thromboprophylaxis likely outweighs the increase in bleeding requiring reintervention in many gynecologic cancer procedures (eg, open surgery for ovarian cancer and pelvic exenteration). In some procedures (eg, laparoscopic total hysterectomy without lymphadenectomy), thromboembolism and bleeding risks are similar, and decisions depend on individual risk prediction and values and preferences regarding venous thromboembolism and bleeding.

Risk of thrombosis and bleeding in gynecologic noncancer surgery: systematic review and meta-analysis.

OBJECTIVE: This study aimed to provide procedure-specific estimates of the risk for symptomatic venous thromboembolism and major bleeding in noncancer gynecologic surgeries. DATA SOURCES: We conducted comprehensive searches on Embase, MEDLINE, Web of Science, and Google Scholar. Furthermore, we performed separate searches for randomized trials that addressed the effects of thromboprophylaxis. STUDY ELIGIBILITY CRITERIA: Eligible studies were observational studies that enrolled ≥50 adult patients who underwent noncancer gynecologic surgery procedures and that reported the absolute incidence of at least 1 of the following: symptomatic pulmonary embolism, symptomatic deep vein thrombosis, symptomatic venous thromboembolism, bleeding that required reintervention (including re-exploration and angioembolization), bleeding that led to transfusion, or postoperative hemoglobin level <70 g/L. METHODS: A teams of 2 reviewers independently assessed eligibility, performed data extraction, and evaluated the risk of bias of the eligible articles. We adjusted the reported estimates for thromboprophylaxis and length of follow-up and used the median value from studies to determine the cumulative incidence at 4 weeks postsurgery stratified by patient venous thromboembolism risk factors and used the Grading of Recommendations Assessment, Development and Evaluation approach to rate the evidence certainty. RESULTS: We included 131 studies (1,741,519 patients) that reported venous thromboembolism risk estimates for 50 gynecologic noncancer procedures and bleeding requiring reintervention estimates for 35 procedures. The evidence certainty was generally moderate or low for venous thromboembolism and low or very low for bleeding requiring reintervention. The risk for symptomatic venous thromboembolism varied from a median of <0.1% for several procedures (eg, transvaginal oocyte retrieval) to 1.5% for others (eg, minimally invasive sacrocolpopexy with hysterectomy, 1.2%-4.6% across patient venous thromboembolism risk groups). Venous thromboembolism risk was <0.5% for 30 (60%) of the procedures; 0.5% to 1.0% for 10 (20%) procedures; and >1.0% for 10 (20%) procedures. The risk for bleeding the require reintervention varied from <0.1% (transvaginal oocyte retrieval) to 4.0% (open myomectomy). The bleeding requiring reintervention risk was <0.5% in 17 (49%) procedures, 0.5% to 1.0% for 12 (34%) procedures, and >1.0% in 6 (17%) procedures. CONCLUSION: The risk for venous thromboembolism in gynecologic noncancer surgery varied between procedures and patients. Venous thromboembolism risks exceeded the bleeding risks only among selected patients and procedures. Although most of the evidence is of low certainty, the results nevertheless provide a compelling rationale for restricting pharmacologic thromboprophylaxis to a minority of patients who undergo gynecologic noncancer procedures.

ggdist: Visualizations of Distributions and Uncertainty in the Grammar of Graphics.

The grammar of graphics is ubiquitous, providing the foundation for a variety of popular visualization tools and toolkits. Yet support for uncertainty visualization in the grammar graphics-beyond simple variations of error bars, uncertainty bands, and density plots-remains rudimentary. Research in uncertainty visualization has developed a rich variety of improved uncertainty visualizations, most of which are difficult to create in existing grammar of graphics implementations. ggdist, an extension to the popular ggplot2 grammar of graphics toolkit, is an attempt to rectify this situation. ggdist unifies a variety of uncertainty visualization types through the lens of distributional visualization, allowing functions of distributions to be mapped to directly to visual channels (aesthetics), making it straightforward to express a variety of (sometimes weird!) uncertainty visualization types. This distributional lens also offers a way to unify Bayesian and frequentist uncertainty visualization by formalizing the latter with the help of confidence distributions. In this paper, I offer a description of this uncertainty visualization paradigm and lessons learned from its development and adoption: ggdist has existed in some form for about six years (originally as part of the tidybayes R package for post-processing Bayesian models), and it has evolved substantially over that time, with several rewrites and API re-organizations as it changed in response to user feedback and expanded to cover increasing varieties of uncertainty visualization types. Ultimately, given the huge expressive power of the grammar of graphics and the popularity of tools built on it, I hope a catalog of my experience with ggdist will provide a catalyst for further improvements to formalizations and implementations of uncertainty visualization in grammar of graphics ecosystems. A free copy of this paper is available at https://osf.io/2gsz6. All supplemental materials are available at https://github.com/mjskay/ggdist-paper and are archived on Zenodo at doi:10.5281/zenodo.7770984.

Swaying the Public? Impacts of Election Forecast Visualizations on Emotion, Trust, and Intention in the 2022 U.S. Midterms.

We conducted a longitudinal study during the 2022 U.S. midterm elections, investigating the real-world impacts of uncertainty visualizations. Using our forecast model of the governor elections in 33 states, we created a website and deployed four uncertainty visualizations for the election forecasts: single quantile dotplot (1-Dotplot), dual quantile dotplots (2-Dotplot), dual histogram intervals (2-Interval), and Plinko quantile dotplot (Plinko), an animated design with a physical and probabilistic analogy. Our online experiment ran from Oct. 18, 2022, to Nov. 23, 2022, involving 1,327 participants from 15 states. We use Bayesian multilevel modeling and post-stratification to produce demographically-representative estimates of people's emotions, trust in forecasts, and political participation intention. We find that election forecast visualizations can heighten emotions, increase trust, and slightly affect people's intentions to participate in elections. 2-Interval shows the strongest effects across all measures; 1-Dotplot increases trust the most after elections. Both visualizations create emotional and trust gaps between different partisan identities, especially when a Republican candidate is predicted to win. Our qualitative analysis uncovers the complex political and social contexts of election forecast visualizations, showcasing that visualizations may provoke polarization. This intriguing interplay between visualization types, partisanship, and trust exemplifies the fundamental challenge of disentangling visualization from its context, underscoring a need for deeper investigation into the real-world impacts of visualizations. Our preprint and supplements are available at https://doi.org/osf.io/ajq8f.

Mechanisms of reduced myocardial energetics of the dystrophic heart.

Heart disease is a leading cause of death in patients with Duchenne muscular dystrophy (DMD), characterized by the progressive replacement of contractile tissue with scar tissue. Effective therapies for dystrophic cardiomyopathy will require addressing the disease before the onset of fibrosis, however, the mechanisms of the early disease are poorly understood. To understand the pathophysiology of DMD, we perform a detailed functional assessment of cardiac function of the mdx mouse, a model of DMD. These studies use a combination of functional, metabolomic, and spectroscopic approaches to fully characterize the contractile, energetic, and mitochondrial function of beating hearts. Through these innovative approaches, we demonstrate that the dystrophic heart has reduced cardiac reserve and is energetically limited. We show that this limitation does not result from poor delivery of oxygen. Using spectroscopic approaches, we provide evidence that mitochondria in the dystrophic heart have attenuated mitochondrial membrane potential and deficits in the flow of electrons in complex IV of the electron transport chain. These studies provide evidence that poor myocardial energetics precede the onset of significant cardiac fibrosis and likely results from mitochondrial dysfunction centered around complex IV and reduced membrane potential. The multimodal approach used here implicates specific molecular components in the etiology of reduced energetics. Future studies focused on these targets may provide therapies that improve the energetics of the dystrophic heart leading to improved resiliency against damage and preservation of myocardial contractile tissue.NEW & NOTEWORTHY Dystrophic hearts have poor contractile reserve that is associated with a reduction in myocardial energetics. We demonstrate that oxygen delivery does not contribute to the limited energy production of the dystrophic heart even with increased workloads. Cytochrome optical spectroscopy of the contracting heart reveals alterations in complex IV and evidence of depolarized mitochondrial membranes. We show specific alterations in the electron transport chain of the dystrophic heart that may contribute to poor myocardial energetics.

TaxaHFE: a machine learning approach to collapse microbiome datasets using taxonomic structure.

Motivation: Biologists increasingly turn to machine learning models not just to predict, but to explain. Feature reduction is a common approach to improve both the performance and interpretability of models. However, some biological datasets, such as microbiome data, are inherently organized in a taxonomy, but these hierarchical relationships are not leveraged during feature reduction. We sought to design a feature engineering algorithm to exploit relationships in hierarchically organized biological data. Results: We designed an algorithm, called TaxaHFE, to collapse information-poor features into their higher taxonomic levels. We applied TaxaHFE to six previously published datasets and found, on average, a 90% reduction in the number of features (SD = 5.1%) compared to using the most complete taxonomy. Using machine learning to compare the most resolved taxonomic level (i.e. species) against TaxaHFE-preprocessed features, models based on TaxaHFE features achieved an average increase of 3.47% in receiver operator curve area under the curve. Compared to other hierarchical feature engineering implementations, TaxaHFE introduces the novel ability to consider both categorical and continuous response variables to inform the feature set collapse. Importantly, we find TaxaHFE's ability to reduce hierarchically organized features to a more information-rich subset increases the interpretability of models. Availability and implementation: TaxaHFE is available as a Docker image and as R code at https://github.com/aoliver44/taxaHFE.

Outcomes of hypothalamic oxytocin neuron-driven cardioprotection after acute myocardial infarction.

Altered autonomic balance is a hallmark of numerous cardiovascular diseases, including myocardial infarction (MI). Although device-based vagal stimulation is cardioprotective during chronic disease, a non-invasive approach to selectively stimulate the cardiac parasympathetic system immediately after an infarction does not exist and is desperately needed. Cardiac vagal neurons (CVNs) in the brainstem receive powerful excitation from a population of neurons in the paraventricular nucleus (PVN) of the hypothalamus that co-release oxytocin (OXT) and glutamate to excite CVNs. We tested if chemogenetic activation of PVN-OXT neurons following MI would be cardioprotective. The PVN of neonatal rats was transfected with vectors to selectively express DREADDs within OXT neurons. At 6 weeks of age, an MI was induced and DREADDs were activated with clozapine-N-oxide. Seven days following MI, patch-clamp electrophysiology confirmed the augmented excitatory neurotransmission from PVN-OXT neurons to downstream nuclei critical for parasympathetic activity with treatment (43.7 ± 10 vs 86.9 ± 9 pA; MI vs. treatment), resulting in stark improvements in survival (85% vs. 95%; MI vs. treatment), inflammation, fibrosis assessed by trichrome blue staining, mitochondrial function assessed by Seahorse assays, and reduced incidence of arrhythmias (50% vs. 10% cumulative incidence of ventricular fibrillation; MI vs. treatment). Myocardial transcriptomic analysis provided molecular insight into potential cardioprotective mechanisms, which revealed the preservation of beneficial signaling pathways, including muscarinic receptor activation, in treated animals. These comprehensive results demonstrate that the PVN-OXT network could be a promising therapeutic target to quickly activate beneficial parasympathetic-mediated cellular pathways within the heart during the early stages of infarction.

Adaptive Assessment of Visualization Literacy.

Visualization literacy is an essential skill for accurately interpreting data to inform critical decisions. Consequently, it is vital to understand the evolution of this ability and devise targeted interventions to enhance it, requiring concise and repeatable assessments of visualization literacy for individuals. However, current assessments, such as the Visualization Literacy Assessment Test (VLAT), are time-consuming due to their fixed, lengthy format. To address this limitation, we develop two streamlined computerized adaptive tests (CATs) for visualization literacy, A-VLAT and A-CALVI, which measure the same set of skills as their original versions in half the number of questions. Specifically, we (1) employ item response theory (IRT) and non-psychometric constraints to construct adaptive versions of the assessments, (2) finalize the configurations of adaptation through simulation, (3) refine the composition of test items of A-CALVI via a qualitative study, and (4) demonstrate the test-retest reliability (ICC: 0.98 and 0.98) and convergent validity (correlation: 0.81 and 0.66) of both CATs via four online studies. We discuss practical recommendations for using our CATs and opportunities for further customization to leverage the full potential of adaptive assessments. All supplemental materials are available at https://osf.io/a6258/.

A Pilot Study of F-18 Fluciclovine-PET/CT as a Diagnostic Tool for Bone Metastases in Patients With Castrate Resistant Prostate Adenocarcinoma and Correlative Analysis of Blood and Bone Molecular Testing (The FACT Study).

BACKGROUND: Suspicious F-18 fluciclovine PET/CT findings for osseous metastases from prostate cancer (PC) were targeted for core needle biopsy. We correlated the maximum standardized uptake value (SUVmax) of biopsied lesions, with biopsy results, other diagnostic outcomes, and blood and tissue molecular analysis (TMA). MATERIAL AND METHODS: Patients with castrate resistant prostate cancer (CRPC) were recruited from a university oncology clinic. SUVmax, histology, blood, and TMA were correlated. RESULTS: Fifteen patients were enrolled and 12 underwent bone biopsies. Fifty percent of bone biopsies demonstrated malignancy. Higher SUVmax was associated with positive biopsies for adenocarcinoma (P = .003), and lesions with SUVmax ≥ 5.1 were all positive for malignancy. Significant correlation between blood and somatic TMA (P = .002) was also found. CONCLUSION: Higher uptake of F-18 fluciclovine was associated with higher predictive value for osseous metastasis on biopsy. There was a significant correlation between blood and TMA.

High-throughput optical sensing of peri-cellular oxygen in cardiac cells: system characterization, calibration, and testing.

Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) represent a scalable experimental model relevant to human physiology. Oxygen consumption of hiPSC-CMs has not been studied in high-throughput (HT) format plates used in pre-clinical studies. Here, we provide comprehensive characterization and validation of a system for HT long-term optical measurements of peri-cellular oxygen in cardiac syncytia (human iPSC-CM and human cardiac fibroblasts), grown in glass-bottom 96-well plates. Laser-cut oxygen sensors having a ruthenium dye and an oxygen-insensitive reference dye were used. Ratiometric measurements (409 nm excitation) reflected dynamic changes in oxygen, as validated with simultaneous Clark electrode measurements. Emission ratios (653 nm vs. 510 nm) were calibrated for percent oxygen using two-point calibration. Time-dependent changes in the Stern-Volmer parameter, ksv, were observed during the initial 40-90 min of incubation, likely temperature-related. Effects of pH on oxygen measurements were negligible in the pH range of 4-8, with a small ratio reduction for pH > 10. Time-dependent calibration was implemented, and light exposure time was optimized (0.6-0.8 s) for oxygen measurements inside an incubator. Peri-cellular oxygen dropped to levels <5% within 3-10 h for densely-plated hiPSC-CMs in glass-bottom 96-well plates. After the initial oxygen decrease, samples either settled to low steady-state or exhibited intermittent peri-cellular oxygen dynamics. Cardiac fibroblasts showed slower oxygen depletion and higher steady-state levels without oscillations, compared to hiPSC-CMs. Overall, the system has great utility for long-term HT monitoring of peri-cellular oxygen dynamics in vitro for tracking cellular oxygen consumption, metabolic perturbations, and characterization of the maturation of hiPSC-CMs.