Simultaneous Assessment Research Articles

Multi-Trophic Species Diversity Contributes to the Restoration of Soil Multifunctionality in Degraded Karst Forests through Cascading Effects

The biodiversity–ecosystem function (BEF) relationship is the basis for studying the restoration of degraded ecosystems, and the simultaneous assessment of multi-trophic-level biodiversity and ecosystem multifunctionality relationship is more conducive to unravelling the restoration mechanism of degraded ecosystems, especially for degraded forest ecosystems with harsh habitats and infertile soils such as karst. In this study, we evaluated the biodiversity and soil multifunctionality (SMF) of degraded karst forests (scrub, SB; secondary growth forests, SG; old-growth forests, OG) in the Maolan National Nature Reserve, China, using 30 sample plots. Biodiversity and soil multifunctionality (SMF) at three trophic levels (plant–soil fauna–soil microorganisms), were assessed through vegetation surveys and soil sampling. One-way ANOVA showed that SMF increased with natural restoration, but multi-trophic level biodiversity showed different trends. Pearson’s correlation analysis showed a positive correlation between plant species diversity and SMF (p < 0.001), whereas soil fauna and soil microorganisms were negatively correlated with SMF. Structural equation modeling revealed a cascading effect of the multi-trophic level on the stimulation of the SMF during restoration. Only soil microorganisms exhibited a direct driving effect on SMF (p < 0.001), whereas plants indirectly influenced soil microorganisms through soil fauna, which subsequently affected the SMF. Although we observed the negative effects of increased plant diversity on soil fauna and soil microbial diversity in terms of quantitative relationships, the increase in soil fauna species and the evenness of soil microbial function still contributed to SMF restoration. This study revealed the cascading effects of multi-trophic diversity in promoting SMF restoration and emphasized that soil microbes are key to unraveling restoration mechanisms and processes, whereas soil fauna is an important intermediate link.

An Integrated Hepatocyte Stability Assay for Simultaneous Metabolic Stability Assessment and Metabolite Profiling.

The determination of metabolic stability is critical for drug discovery programs, allowing for the optimization of chemical entities and compound prioritization. As such, it is common to perform high-volume in vitro metabolic stability experiments early in the lead optimization process to understand metabolic liabilities. Additional metabolite identification experiments are subsequently performed for a more comprehensive understanding of the metabolic clearance routes to aid medicinal chemists in the structural design of compounds. Collectively, these experiments require extensive sample preparation and a substantial amount of time and resources. To overcome the challenges, a high-throughput integrated assay for simultaneous hepatocyte metabolic stability assessment and metabolite profiling was developed. This assay platform consists of four parts: 1) an automated liquid-handling system for sample preparation and incubation, 2) a liquid chromatography and high-resolution mass spectrometry-based system to simultaneously monitor the parent compound depletion and metabolite formation, 3) an automated data analysis and report system for hepatic clearance assessment; and 4) streamlined autobatch processing for software-based metabolite profiling. The assay platform was evaluated using eight control compounds with various metabolic rates and biotransformation routes in hepatocytes across three species. Multiple sample preparation and data analysis steps were evaluated and validated for accuracy, repeatability, and metabolite coverage. The combined utility of an automated liquid-handling instrument, a high-resolution mass spectrometer, and multiple streamlined data processing software improves the process of these highly demanding screening assays and allows for simultaneous determination of metabolic stability and metabolite profiles for more efficient lead optimization during early drug discovery. SIGNIFICANCE STATEMENT: Metabolic stability assessment and metabolite profiling are pivotal in drug discovery to fully comprehend metabolic liabilities for chemical entity optimization and lead selection. Process of these assays can be repetitive and resource demanding. Here, we developed an integrated hepatocyte stability assay that combines automation, high-resolution mass spectrometers, and batch-processing software to improve and combine the workflow of these assays. The integrated approach allows simultaneous metabolic stability assessment and metabolite profiling, significantly accelerating screening and lead optimization in a resource-effective manner.

9 Randel Lecture: In Vivo investigation of pregnancy Physiology

Abstract Pregnancy in any mammal requires the integration of three distinct compartments; the maternal, placental and fetal compartments. Compromised function of any of the three compartments can result in suboptimal pregnancy outcomes, including increased morbidity and mortality at delivery, reduced livestock production efficiency and adult-onset of metabolically-related disease in humans. To accurately understand the progression of both normal and compromised pregnancies, therefore, requires the simultaneous assessment of all three compartments, which for most species is not technically feasible. Consequently, over the past 60 yr the pregnant sheep has been used extensively for in vivo investigation of pregnancy physiology. The ability to place indwelling catheters, within both maternal and fetal vessels, allows the simultaneous steady-state assessment of uterine and umbilical blood flows, hormone secretion, and nutrient uptake and utilization of nutrients by each of the three compartments, under non-stressed and non-anesthetized conditions. This has provided unrivalled insight into the physiology of pregnancy, and the realization that the placenta is a highly metabolic tissue, utilizing the majority of oxygen and glucose taken up by the uterus. However, the deficit in studying pregnant sheep is the lack of genetic models, readily available in rodents, that could help define specific gene function during pregnancy. Consequently, we developed in vivo lentiviral-mediated RNA interference (RNAi) methods to specifically alter the abundance of specific gene transcripts/proteins within the placenta, beginning at 9 d of gestation (dGA). Our lentiviral-mediated approach limits the RNAi to the trophectoderm derivatives of the placenta. Initially we used this approach to determine the importance of PRR15 and LIN28 during the establishment of pregnancies, but turned our efforts towards combining in vivo RNAi with steady-state physiological assessments during late gestation to examine the function of genes involved in placental nutrient transport (SLC2A3) and placenta derived hormones (CSH). By creating a placental deficiency in SLC2A3, not only was the fetus smaller and hypoglycemic at mid-gestation, but by interfering with placental glucose uptake, maternal hormone concentrations were also impacted, yet near-term, a distinctly different phenotype was present. For the placental hormone CSH (a.k.a. placental lactogen), CSH RNAi not only impacts placental and fetal growth, but also reductions in uterine blood flow, nutrient fluxes to the fetus, and produces an altered hormonal environment. Combining these methodological approaches provides new and often unexpected insights into the physiological integration of the maternal, placental and fetal compartments. Supported by NIH-NICHD grants HD093701 and HD094952.

Interaction effect between low birthweight and resistin gene rs1862513 variant on insulin resistance and type 2 diabetes mellitus in adulthood: Toon Genome Study.

Gene-environment interactions are considered to critically influence type 2 diabetes mellitus development; however, the underlying mechanisms and specific interactions remain unclear. Given the increasing prevalence of low birthweight (LBW) influenced by the intrauterine environment, we sought to investigate genetic factors related to type 2 diabetes development in individuals with LBW. The interaction between 20 reported type 2 diabetes susceptibility genes and the development of type 2 diabetes in LBW (<2,500 g) individuals in a population-based Japanese cohort (n = 1,021) was examined by logistic regression and stratified analyses. Logistic regression analyses showed that only the G/G genotype at the rs1862513 locus of the resistin gene (RETN), an established initiator of insulin resistance, was closely related to the prevalence of type 2 diabetes in individuals with LBW. Age, sex and current body mass index-adjusted stratified analyses showed a significant interaction effect of LBW and the RETN G/G genotype on fasting insulin, homeostatic model assessment 2-insulin resistance, Matsuda index and the prevalence of type 2 diabetes (all P-values for interaction <0.05). The adjusted odds ratio for type 2 diabetes in the LBW + G/G genotype group was 7.33 (95% confidence interval 2.43-22.11; P = 0.002) compared with the non-LBW + non-G/G genotype group. Similar results were obtained after excluding the influence of malnutrition due to World War II. Simultaneous assessment of LBW and the RETN G/G genotype can more accurately predict the risk of future type 2 diabetes than assessing each of these factors alone, and provide management strategies, including early lifestyle intervention in LBW population.

Quantitative and Visual Analysis of Data Augmentation and Hyperparameter Optimization in Deep Learning-Based Segmentation of Low-Grade Glioma Tumors Using Grad-CAM.

This study executes a quantitative and visual investigation on the effectiveness of data augmentation and hyperparameter optimization on the accuracy of deep learning-based segmentation of LGG tumors. The study employed the MobileNetV2 and ResNet backbones with atrous convolution in DeepLabV3+ structure. The Grad-CAM tool was also used to interpret the effect of augmentation and network optimization on segmentation performance. A wide investigation was performed to optimize the network hyperparameters. In addition, the study examined 35 different models to evaluate different data augmentation techniques. The results of the study indicated that incorporating data augmentation techniques and optimization can improve the performance of segmenting brain LGG tumors up to 10%. Our extensive investigation of the data augmentation techniques indicated that enlargement of data from 90° and 225° rotated data,up to down and left to right flipping are the most effective techniques. MobilenetV2 as the backbone,"Focal Loss" as the loss function and "Adam" as the optimizer showed the superior results. The optimal model (DLG-Net) achieved an overall accuracy of 96.1% with a loss value of 0.006. Specifically, the segmentation performance for Whole Tumor (WT), Tumor Core (TC), and Enhanced Tumor (ET) reached a Dice Similarity Coefficient (DSC) of 89.4%, 70.1%, and 49.9%, respectively. Simultaneous visual and quantitative assessment of data augmentation and network optimization can lead to an optimal model with a reasonable performance in segmenting the LGG tumors.

Total-Body PET/CT Applications in Cardiovascular Diseases: A Perspective Document of the SNMMI Cardiovascular Council.

Digital PET/CT systems with a long axial field of view have become available and are emerging as the current state of the art. These new camera systems provide wider anatomic coverage, leading to major increases in system sensitivity. Preliminary results have demonstrated improvements in image quality and quantification, as well as substantial advantages in tracer kinetic modeling from dynamic imaging. These systems also potentially allow for low-dose examinations and major reductions in acquisition time. Thereby, they hold great promise to improve PET-based interrogation of cardiac physiology and biology. Additionally, the whole-body coverage enables simultaneous assessment of multiple organs and the large vascular structures of the body, opening new opportunities for imaging systemic mechanisms, disorders, or treatments and their interactions with the cardiovascular system as a whole. The aim of this perspective document is to debate the potential applications, challenges, opportunities, and remaining challenges of applying PET/CT with a long axial field of view to the field of cardiovascular disease.

Inception and development of aLC-MS/MS assay for the multiplexed quantitation of nine human drug transporter biomarkers.

Background: It has become common practice to assess solute carrier transporter (SLC)-mediated drug-drug interactions (DDIs) by quantitating various individual endogenous compounds as biomarkers in human plasma and urine. The goal of this work was to develop biomarker multiplex assays that could be utilized during first in human studies to support the simultaneous assessment of clinical DDI risk across various SLCs. Methodology: Hydrophilic interaction chromatography-MS/MS methods were developed, and validations were performed. Results: The multiplex assays were applied to a first in human study. Placebo/reference subject biomarker data were consistent with single assay in-house and published data. Conclusion: This work demonstrates the utility of these multiplex methods to support the concurrent evaluation of clinical DDI risk across various SLCs.

Large-scale smart bioreactor with fully integrated wireless multivariate sensors and electronics for long-term in situ monitoring of stem cell culture.

Achieving large-scale, cost-effective, and reproducible manufacturing of stem cells with the existing devices is challenging. Traditional single-use cell-bag bioreactors, limited by their rigid and single-point sensors, struggle with accuracy and scalability for high-quality cell manufacturing. Here, we introduce a smart bioreactor system that enables multi-spatial sensing for real-time, wireless culture monitoring. This scalable system includes a low-profile, label-free thin-film sensor array and electronics integrated with a flexible cell bag, allowing for simultaneous assessment of culture properties such as pH, dissolved oxygen, glucose, and temperature, to receive real-time feedback for up to 30 days. The experimental results show the accurate monitoring of time-dynamic and spatial variations of stem cells and myoblast cells with adjustable carriers from a plastic dish to a 2-liter cell bag. These advances open up the broad applicability of the smart sensing system for large-scale, lower-cost, reproducible, and high-quality engineered cell manufacturing for broad clinical use.

Spectral separation of evoked and spontaneous cortical activity, Part 1: Delta to high gamma band

Spectral analysis of repeatedly evoked potentials (EPs) is challenging since recordings contain a superposition of evoked signals and spontaneous activity. We developed a novel approach, N-interval Fourier Transform Analysis (N-FTA), which allows for reliable separation and simultaneous assessment of triggered and background spectral components. Median nerve stimulation data from a total of eleven volunteers recorded in two labs with different experimental settings were investigated. Consistently, short latency spectral components were mainly contained in the gamma and high gamma bands. In contrast, spontaneous activity displayed a 1/f spectral profile with distinct alpha and beta peaks. Spontaneous power spectral densities (PSDs) obtained for real and sham stimulation were highly comparable. The low frequency background PSD was more than two orders of magnitude above the spectral short latency peaks. Within the 30Hz to 90Hz band, the evoked peaks were -17dB to -4dB below the background suggesting that target band filtered short latency deflection might be extracted using less than 100 trials.SEPs following tibial nerve stimulation (3 subjects) displayed a narrower spectral band at about half the bandwidth as compared to median nerve stimulation. Evoked peaks were between 30Hz and 37Hz at PSD levels being -10dB to -4dB below the background activity. These spectral peaks were related to the short latency response of typical W-morphology.Cortical short latency responses are contained in distinct spectral target bands which are much narrower than the standard bandwidth recommendations for routine recordings. In particular, the high pass corner frequency may be selected about one order of magnitude above the current standard. This might render SEP recordings more robust since it eases the suppression of spontaneous activity and movement artifacts such as eye-blinks. Real-time zero-phase filters are required for translating these findings into improved recording systems.

Conformation of Pyroglutamated Amyloid β (3-40) and (11-40) Fibrils - Extended or Hairpin?

Amyloid β (Aβ) is a hallmark protein of Alzheimer's disease. One physiologically important Aβ variant is formed by initial N-terminal truncation at a glutamic acid position (either E3 or E11), which is subsequently cyclized to a pyroglutamate (either pE3 or pE11). Both forms have been found in high concentrations in the core of amyloid plaques and are likely of high importance in the pathology of Alzheimer's disease. However, the molecular structure of the fibrils of these variants is not entirely clear. Solid-state NMR spectroscopy studies have reported a molecular contact between Gly25 and Ile31, which would disagree with the conventional hairpin model of wildtype (WT-)Aβ1-40 fibrils, most often described in the literature. We investigated the conformation of the monomeric unit of pE3-Aβ3-40 and pE11-Aβ11-40 (and for comparison also wildtype (WT)-Aβ1-40) fibrils to find out whether the hairpin or a newly suggested extended structure dominates the structure of the Aβ monomers in these fibrils. To this end, solid-state NMR spectroscopy was applied probing the inter-residual contacts between Phe19/Leu34, Ala21/Leu34, and especially Gly25/Ile31 using suitable isotopic labeling schemes. In the second part, the flexible turn of the Aβ40 peptides was replaced by a (3-(3-aminomethyl)phenylazo)phenylacetic acid (AMPP)-based photoswitch, which can predefine the peptide conformation to either an extended (trans) or hairpin (cis) conformation. This enables simultaneous spectroscopic assessment of the conformation of the AMPP-photoswitch, allowing in situ structural investigations during fibrillation in contrast to structural techniques such as NMR spectroscopy or cryo-EM, which can only be applied to stable conformers. Both methods confirm an extended structure for the peptidic monomers in fibrils of all investigated Aβ variants. Especially the Gly25/Ile31 contact is a decisive indicator for the extended structure along with the characteristic absorption spectra of trans-AMPP-Aβ.

Simultaneous assessment of stress hyperglycemia ratio and glycemic variability to predict mortality in patients with coronary artery disease: a retrospective cohort study from the MIMIC-IV database

BackgroundStress hyperglycemia and glycemic variability (GV) can reflect dramatic increases and acute fluctuations in blood glucose, which are associated with adverse cardiovascular events. This study aimed to explore whether the combined assessment of the stress hyperglycemia ratio (SHR) and GV provides additional information for prognostic prediction in patients with coronary artery disease (CAD) hospitalized in the intensive care unit (ICU).MethodsPatients diagnosed with CAD from the Medical Information Mart for Intensive Care-IV database (version 2.2) between 2008 and 2019 were retrospectively included in the analysis. The primary endpoint was 1-year mortality, and the secondary endpoint was in-hospital mortality. Levels of SHR and GV were stratified into tertiles, with the highest tertile classified as high and the lower two tertiles classified as low. The associations of SHR, GV, and their combination with mortality were determined by logistic and Cox regression analyses.ResultsA total of 2789 patients were included, with a mean age of 69.6 years, and 30.1% were female. Overall, 138 (4.9%) patients died in the hospital, and 404 (14.5%) patients died at 1 year. The combination of SHR and GV was superior to SHR (in-hospital mortality: 0.710 vs. 0.689, p = 0.012; 1-year mortality: 0.644 vs. 0.615, p = 0.007) and GV (in-hospital mortality: 0.710 vs. 0.632, p = 0.004; 1-year mortality: 0.644 vs. 0.603, p < 0.001) alone for predicting mortality in the receiver operating characteristic analysis. In addition, nondiabetic patients with high SHR levels and high GV were associated with the greatest risk of both in-hospital mortality (odds ratio [OR] = 10.831, 95% confidence interval [CI] 4.494–26.105) and 1-year mortality (hazard ratio [HR] = 5.830, 95% CI 3.175–10.702). However, in the diabetic population, the highest risk of in-hospital mortality (OR = 4.221, 95% CI 1.542–11.558) and 1-year mortality (HR = 2.013, 95% CI 1.224–3.311) was observed in patients with high SHR levels but low GV.ConclusionsThe simultaneous evaluation of SHR and GV provides more information for risk stratification and prognostic prediction than SHR and GV alone, contributing to developing individualized strategies for glucose management in patients with CAD admitted to the ICU.

Assessing the association between probable posttraumatic stress disorder symptoms and quality of life among emergency service workers using path analysis.

Emergency service workers have highly stressful occupations; the stressors encountered can contribute to the development of mental disorders such as depression, anxiety, and posttraumatic stress disorder (PTSD). The present study used a conceptual model and survey to identify variables influencing the association between probable PTSD and quality of life (QOL) in emergency service workers. PTSD was assessed using the Impact of Event Scale-Revised. QOL was assessed using the Medical Outcomes Study 36-Item Short-Form Health Survey. Path analysis was used to determine whether stress-coping (Brief Coping Orientation to Problems Experienced [Brief COPE] scores), social support (Multidimensional Scale of Perceived Social Support [MSPSS] scores), and resilience (Connor-Davidson Resilience Scale scores) explain the association between probable PTSD and QOL among 220 emergency service workers in Japan. Impact of Event Scale-Revised scores were significantly positively associated with Brief COPE Active coping scores and significantly negatively associated with MSPSS scores. Brief COPE Active coping and MSPSS scores were significantly positively associated with Connor-Davidson Resilience Scale scores, which were in turn significantly positively associated with Medical Outcomes Study 36-Item Short-Form Health Survey scores. Active coping in response to stressful situations is important for maintaining good mental health among emergency service workers. Active coping and social support may strengthen resilience, and resilience may improve QOL. Screening for mental health and QOL should include simultaneous assessment of stress-coping, social support, and resilience. Although the findings of this cross-sectional study are important, it could not confirm a causal relationship between PTSD and QOL.

The association of maternal-infant interactive behavior, dyadic frontal alpha asymmetry, and maternal anxiety in a smartphone-adapted still face paradigm

Mother-infant interactions form a strong basis for emotion regulation development in infants. These interactions can be affected by various factors, including maternal postnatal anxiety. Electroencephalography (EEG) hyperscanning allows for simultaneous assessment of mother-infant brain-to-behavior association during stressful events, such as the still-face paradigm (SFP). This study aimed at investigating dyadic interactive behavior and brain-to-behavior association across SFP and identifying neural correlates of mother-infant interactions in the context of maternal postnatal anxiety. We measured frontal alpha asymmetry (FAA), a physiological correlate of emotion regulation and a potential marker of risk for psychopathology. To emulate real-life interactions, EEG and behavioral data were collected from 38 mother-infant dyads during a smartphone-adapted dual-SFP. Although the behavioral data showed a clear still-face effect for the smartphone-adapted SFP, this was not reflected in the infant or maternal FAA. Brain-to-behavior data showed higher infant negative affect being associated with more infant leftward FAA during the still-face episodes. Finally, mothers with higher postnatal anxiety showed more right FAA during the first still-face episode, suggesting negative affectivity and a need to withdraw from the situation. Our results form a baseline for further research assessing the effects of maternal postnatal anxiety on infants’ FAA and dyadic interactive behavior.

Applications of Raman spectroscopy in the diagnosis and monitoring of neurodegenerative diseases.

Raman scattering is an inelastic light scattering that occurs in a manner reflective of the molecular vibrations of molecular structures and chemical conditions in a given sample of interest. Energy changes in the scattered light can be assessed to determine the vibration mode and associated molecular and chemical conditions within the sample, providing a molecular fingerprint suitable for sample identification and characterization. Raman spectroscopy represents a particularly promising approach to the molecular analysis of many diseases owing to clinical advantages including its instantaneous nature and associated high degree of stability, as well as its ability to yield signal outputs corresponding to a single molecule type without any interference from other molecules as a result of its narrow peak width. This technology is thus ideally suited to the simultaneous assessment of multiple analytes. Neurodegenerative diseases represent an increasingly significant threat to global public health owing to progressive population aging, imposing a severe physical and social burden on affected patients who tend to develop cognitive and/or motor deficits beginning between the ages of 50 and 70. Owing to a relatively limited understanding of the etiological basis for these diseases, treatments are lacking for the most common neurodegenerative diseases, which include Alzheimer's disease, Parkinson's disease, Huntington's disease, and amyotrophic lateral sclerosis. The present review was formulated with the goal of briefly explaining the principle of Raman spectroscopy and discussing its potential applications in the diagnosis and evaluation of neurodegenerative diseases, with a particular emphasis on the research prospects of this novel technological platform.

Prognostic significance of left atrial strain combined with left ventricular strain using cardiac magnetic resonance feature tracking in patients with heart failure with preserved ejection fraction.

We aimed to evaluate the prognostic value of left ventricular global longitudinal strain (LVGLS) and left atrial strain (LAS) obtained from magnetic resonance imaging (MRI) feature tracking in patients with heart failure with preserved ejection fraction (HFpEF). We retrospectively enrolled consecutive patients with HFpEF admitted to our hospital who underwent cardiac MRI. LVGLS and LAS were obtained from cine MRI by feature tracking. The end point was defined as a composite of all-cause death, myocardial infarction, and hospitalization due to decompensated HF. One-hundred patients with HFpEF were enrolled. Mean LVGLS and LAS were -13.7 ± 3.7% and 22.5 ± 11.6%, respectively. During follow-up of 4.4 ± 1.9years, 24 events occurred. Multivariate Cox proportional hazards model analysis demonstrated LAS was independently associated with adverse cardiac events. Kaplan-Meier curve analysis revealed that the patients with both LVGLS and LAS worse than the median (LVGLS ≥ -12.2% and LAS ≤ 13.8%) had a significantly lower event-free rate compared to those with preserved strain (Log-rank P < 0.001). Simultaneous assessment of LVGLS and LAS using MRI was useful for risk stratification in the patients with HFpEF.

Abstract B032: Comprehensive analysis of clonality, segregation errors, chromosomal instability and heterogeneity in cancer cell lines and cancer samples reveals that cancer evolution is often promoted by highly aberrant cellular divisions

Abstract Chromosomal instability (CIN) is the most common form of genomic instability in cancer. CIN leads to structural and numerical chromosomal aberrations and to heterogeneity of such aberrations inside the same tumor. Together with other forms of genomic and epigenomic instability, CIN underlies the most dangerous feature of cancer cells, namely, ability to evolve, allowing cancer cells to evade immunologic surveillance, to metastasize, to become resistant to drug treatments. Studies of chromosomal instability are significantly impeded by the lack of comprehensive approaches for simultaneous detection of different forms of ongoing chromosomal instability and their outcomes. We developed an approach that allows to analyze the clonal composition and chromosomal heterogeneity in cell lines and cancer samples, as well as rates of different forms of ongoing chromosomal instability in cancer cell lines. At the heart our analysis is a multiplex interphase FISH (miFISH) with seven sequential hybridizations of color probe panels resulting in the assessment of a total of 35 probes per single cell. Each probe panel is designed to include the centromeric probe and probes on the p- and q-arm for a given chromosome that allows for the simultaneous assessment of centromeric probes and gene-specific arm probes. This approach was applied to interphase nuclei, metaphase spreads and bi-nucleated/dividing cancer cells in colorectal and cervical cell lines and cancer samples. miFISH allows for automated acquisition of nuclei and metaphase spreads on a BioView spotcounting system. A gallery provides detailed views for semi-automatic as well as manual analysis of the obtained images. A total of 10000-25000 cell images were collected from each slide. The simultaneous evaluation of our strategically chosen centromeric and gene-specific probes within the same cells allows unprecedented insights in the patterns of distribution of genetic material among cancer cells in cancer cell population, and, especially, in dividing cells. We evaluated clonality, clonal and non-clonal chromosomal heterogeneity, ploidy distributions, and occurrence of endoreplication. Assessment of dividing cells based on equal or unequal distribution of miFISH centromeric and gene-specific signals allowed to estimate and compare the rates and outcomes of different forms of ongoing chromosomal instability such as structural rearrangements and amplifications, loss of chromosomes or chromosomal fragments in micronuclei, non-disjunctions, grossly asymmetrical cell divisions (including multipolar mitoses). Conclusions: Gradual changes in chromosomal/genomic content of cancer cells occur due to structural rearrangements, losses of chromosomes and chromosomal fragments in micronuclei, and by relatively rare single chromosome non-disjunctions. Grossly abnormal cell divisions (massive non-disjunctions and structural rearrangements) is the main source for the high level of chromosomal abnormalities and aneuploidy in evolved cancer cell populations. Citation Format: Anna Roschke. Comprehensive analysis of clonality, segregation errors, chromosomal instability and heterogeneity in cancer cell lines and cancer samples reveals that cancer evolution is often promoted by highly aberrant cellular divisions [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Translating Cancer Evolution and Data Science: The Next Frontier; 2023 Dec 3-6; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(3 Suppl_2):Abstract nr B032.

ЗНАЧЕНИЕ ОФЭКТ/КТ ДЛЯ ОДНОВРЕМЕННОЙ ОЦЕНКИ КАЛЬЦИНОЗА КОРОНАРНЫХ АРТЕРИЙ, ПЕРФУЗИИ И КОНТРАКТИЛЬНОЙ ФУНКЦИИ МИОКАРДА У ЖЕНЩИН С ИШЕМИЧЕСКОЙ БОЛЕЗНЬЮ СЕРДЦА

Purpose: To study the frequency of calcinosis of coronary arteries and its effect on myocardial perfusion and contractile function among females with coronary heart disease (CHD). Material and methods: A survey was conducted among 141 females with coronary heart disease (CHD): simultaneous assessment of coronary artery calcinosis, perfusion and contractile function with 99m Tc-technetril SPECT/CT with ECG-synchronization and X-ray radiation correction, as well as comparison of data with the results of electrocardiography (ECG), echocardiography (ECHO-CG), clinical and biochemical blood tests. Results: Coronary artery calcinosis was detected in 33 (23 %) females with coronary heart disease (CHD) out of all patients identified: over the age of 55 years (M = 67 years with variations from 58 to 83 years) ‒ in 25 (17.7 %), aged 41‒55 years (M = 50 years with variations from 46 to 54 years) ‒ in 7 (4.6 %), at the age of 25‒40 years ‒ in one female 35 years (0.7 %). The Agatston calcium score interval was set as follows: at the maximum degree &gt; 400 ‒ for 6 females (4.2 % of all surveyed); at 101‒400 ‒ in 9 (6.4 %); at 11‒100 ‒ in 17 (12.0 %); at 1‒10 ‒ in one female (0.7 %); with a minimum degree of 0 ‒ in 108 females (76.7 % of all surveyed). In the presence of maximum degree of calcinosis (&gt; 400 units), a slight significant decrease in perfusion, an increase in ejection fraction and diastole duration was revealed. Conclusion: The use of combined SPECT/CT with ECG synchronization and CT radiation correction technology in 141 females with coronary artery disease made it possible to identify coronary artery calcinosis in 23 % of patiets.

P313 Panenteric capsule endoscopy in the investigation and assessment of Crohn’s disease at a tertiary centre

Abstract Background Small bowel &amp; colon capsule endoscopy have been established in their use to visualise the gastrointestinal (GI) tract. The Panenteric Crohn’s Capsule (PCC) is an evolution of these capsules enabling simultaneous assessment of both the small &amp; large bowel with dual cameras for expanded mucosal coverage. This study investigates the use of PCC in patients suspected for inflammatory bowel disease (IBD), specifically Crohn's disease (CD), and in reassessing patients with established CD. The primary objective is to evaluate the impact of PCC results on patients’ management. Methods All consecutive patients who had PCC for suspected or established CD were included in the study. Those at risk of capsule retention (previous surgery, obstructive symptoms, stricturing disease) underwent a patency capsule, unless MR enterography was done within six months of PCC. Data was collected prospectively and included demographics, indication, procedural information such as completion rates, bowel cleansing &amp; need for subsequent colonoscopy. PCC findings were recorded and the clinical outcome based on those was assessed at the subsequent clinic follow up. Results In this prospective single-centre study, 95 patients (average age 34 years, range 14-84) underwent PCC from November 2020 to May 2023, 61 were females (64%). 43 procedures were conducted remotely, 52 in-hospital, &amp; 5 as inpatients. The primary indications were suspicion of IBD (n=54, 57%) and reassessment of established CD (n=41, 43%). Among the 95 patients,68 (72%) had successful PCC procedures. 18% (17/95) required a subsequent colonoscopy, primarily for biopsy (7,7.4%). PCC findings revealed inflammation in 41 patients (43.2%), diverticular disease in 13 (13.7%), &amp; polyps in 15 (15.8%). The data demonstrated a 51% change in management post-PCC, including treatment escalation (15.8%), treatment de-escalation due to IBD remission (10.5%), exclusion of IBD with redirection for other pathologies or IBS diagnoses (15.8%), new IBD diagnoses (5.3%), &amp; discharged due to normal investigation without need of further investigation (3.2%). Particularly in patients undergoing PCC for CD reassessment, 66% experienced a change in the treatment plan (14 escalated treatment, 7 de-escalated treatment due to remission, 3 achieved IBD treatment response &amp; 3 treated for symptoms found to be caused by other pathologies). Conclusion PCC is a single, one-stop, non-invasive GI examination, ideal for suspected or established CD patients requiring pan-gut assessment. Despite suboptimal completion rates, PCC spared a colonoscopy in 82% of patients and resulted in change of management in over half of the patients and in 2/3 of those with established CD, suggesting a promising future role in diagnosing and monitoring CD.

Adaptive Bayesian information borrowing methods for finding and optimizing subgroup-specific doses.

In precision oncology, integrating multiple cancer patient subgroups into a single master protocol allows for the simultaneous assessment of treatment effects in these subgroups and promotes the sharing of information between them, ultimately reducing sample sizes and costs and enhancing scientific validity. However, the safety and efficacy of these therapies may vary across different subgroups, resulting in heterogeneous outcomes. Therefore, identifying subgroup-specific optimal doses in early-phase clinical trials is crucial for the development of future trials. In this article, we review various innovative Bayesian information-borrowing strategies that aim to determine and optimize subgroup-specific doses. Specifically, we discuss Bayesian hierarchical modeling, Bayesian clustering, Bayesian model averaging or selection, pairwise borrowing, and other relevant approaches. By employing these Bayesian information-borrowing methods, investigators can gain a better understanding of the intricate relationships between dose, toxicity, and efficacy in each subgroup. This increased understanding significantly improves the chances of identifying an optimal dose tailored to each specific subgroup. Furthermore, we present several practical recommendations to guide the design of future early-phase oncology trials involving multiple subgroups when using the Bayesian information-borrowing methods.

Recreational and Medical Cannabis Legalization and Opioid Prescriptions and Mortality

While some have argued that cannabis legalization has helped to reduce opioid-related morbidity and mortality in the US, evidence has been mixed. Moreover, existing studies did not account for biases that could arise when policy effects vary over time or across states or when multiple policies are assessed at the same time, as in the case of recreational and medical cannabis legalization. To quantify changes in opioid prescriptions and opioid overdose deaths associated with recreational and medical cannabis legalization in the US. This quasiexperimental, generalized difference-in-differences analysis used annual state-level data between January 2006 and December 2020 to compare states that legalized recreational or medical cannabis vs those that did not. Recreational and medical cannabis law implementation (proxied by recreational and medical cannabis dispensary openings) between 2006 and 2020 across US states. Opioid prescription rates per 100 persons and opioid overdose deaths per 100 000 population based on data from the US Centers for Disease Control and Prevention. Between 2006 and 2020, 13 states legalized recreational cannabis and 23 states legalized medical cannabis. There was no statistically significant association of recreational or medical cannabis laws with opioid prescriptions or overall opioid overdose mortality across the 15-year study period, although the results also suggested a potential reduction in synthetic opioid deaths associated with recreational cannabis laws (4.9 fewer deaths per 100 000 population; 95% CI, -9.49 to -0.30; P = .04). Sensitivity analyses excluding state economic indicators, accounting for additional opioid laws and using alternative ways to code treatment dates yielded substantively similar results, suggesting the absence of statistically significant associations between cannabis laws and the outcomes of interest during the full study period. The results of this study suggest that, after accounting for biases due to possible heterogeneous effects and simultaneous assessment of recreational and medical cannabis legalization, the implementation of recreational or medical cannabis laws was not associated with opioid prescriptions or opioid mortality, with the exception of a possible reduction in synthetic opioid deaths associated with recreational cannabis law implementation.