Initial Event Research Articles

Prevalence and Features of Post-stroke Urinary Incontinence: A Retrospective Cohort Study.

Long-term complications of stroke, persisting for more than 6 months after the initial event, substantially reduce the quality of life (QoL) in a significant percentage of stroke survivors. In this paper, we studied the prevalence of long-term urinary incontinence (UI) in post-stroke patients. In addition, we attempted to identify patient characteristics which were associated with higher UI prevalence, higher UI severity, and less UI-associated QoL. Medical records in a tertiary referral hospital were used to contact patients who had experienced a stroke between 6 to 32 months before the study date. The patients were given the International Consultation on Incontinence Questionnaire Short Form (ICIQ-UI-SF) questionnaire for determining the presence of UI and its severity. UI-positive patients were then given the I-QOL questionnaire to determine their QoL. The prevalence of UI in our study population (n=189) was 31%. Older age at the time of stroke was associated with higher UI severity (r=0.290) and lower QoL (r=-0.265). Furthermore, the presence of movement limitation was associated with higher UI prevalence (P<0.001, OR=3.89) and severity (P=0.002, d=1.05). Movement limitation also significantly impacted the psychological and social aspects of UI-associated QoL (P=0.035, d=-0.74). Conversely, higher body mass indices (BMIs) were associated with lower UI severity (r=-0.346) and higher QoL (r=0.281). In conclusion, UI continues to be prevalent in stroke survivors long after the cerebrovascular accident (CVA). As a result, these patients require continuous monitoring and UI prevention.

Novel methods for enhancing ECG visualization research

All Long QT syndrome (LQTS) patients are at elevated risk for channelopathy-induced delayed myocardial repolarization and consequently potentially life-threatening cardiac events with 90% of initial cardiac events occurring between preteen and 40 years old. Utilizing ECG and derived vectorcardiographic parameters, including T wave Vector Magnitude (TwVM) measurement data, this study attempts to determine whether TwVM from baseline ECGs is effectively predictive of future cardiac events for genotype-positive LQTS patients.Verified carriers of established LQTS disease-causing genotypes were selected from University of Minnesota patient encounters between 2010 and 2020 for inclusion in this retrospective study. Baseline and predictive ECG and derived vectorcardiographic parameter evaluation, clinical data, and statistical analysis were compared between patients with and patients without cardiac events. First recorded ECG was at presentation to our hospital and final ECG is defined as ECG just prior to cardiac event (event defined below in Methods) or the most final documented ECG before cut-off year of 2020 for the event-free group.Of 41 participants, 15 experienced cardiac events and 26 did not. While many baseline electrocardiographic parameter measurements did not show significant differences between patient groups, vectorcardiographic parameters at baseline, specifically the QRS vector magnitude (QRSVM) and azimuth of the spatial ventricular gradient, showed significance. Additionally, final vectorcardiographic parameters, particularly the QRSVM, TwVM, and azimuth of the spatial ventricular gradient showed significant differences between patient groups. Final T-wave frontal axis was significantly larger in those without cardiac events. Significant Kaplan-Meier curve separation between patient groups was noted based on a QRSVM of 1.43 mV or lower, with additional consideration to patient age, genotype, and beta blocker use.This study shows evidence of ECG and derived vectorcardiographic parameters, including TwVM, being effective in early prediction of cardiac events in genotype-positive LQTS patients.

Abstract #1405303: Common Adverse Events During Levoketoconazole Titration: An Integrated Analysis of Data from Two Prospective Cushing’s Syndrome (CS) Clinical Trials

Illustrate common AEs reported during titration phases of the Phase 3 SONICS and LOGICS studies in adult patients with CS to inform the clinical use of levoketoconazole and enhance patient counseling. Treatment-emergent AE data with levoketoconazole were pooled from two prospective relatively similarly designed dose-titration phases from SONICS and LOGICS for integrated analysis. Titrations of up to 7 dose levels (from 150 mg BID to 600 mg BID) occurred no more frequently than once every 2 weeks as needed to normalize mean 24-hour urine free cortisol. AE preferred terms describing similar events were combined to form 24 mutually exclusive AE syndromic categories. Categories with a reported-patient frequency of ≥10% (n=14) were analyzed descriptively as to frequency, severity, action taken, times to onset and resolution, and recurrences during titration. Patient frequency of common AEs ranged between 10% (myalgia) and 30% (nausea/vomiting), with no apparent dose relationship among 13 of 14 AE categories (exception of upper respiratory infection). Drug-exposure duration differed by dose. Investigator-assessed AE severity was most often Grade 1 (mild, median patient frequency for 14 categories combined: 69%, range 41% to 90%). Overall, Grade 3 (severe) events were reported for 25% of patients with liver abnormality/injury, 15% with hypertension, and less than 10% for all other categories. Levoketoconazole dose reductions or interruptions were used for management of AEs most often for arrhythmia (typically asymptomatic QT interval prolongation; in 37% of arrhythmia events) and nausea/vomiting (19%), whereas withdrawal of levoketoconazole was most often used for management of liver abnormality/injury (29%). Median time to first onset for common AEs combined was 49.5 d, with fatigue the earliest onset (median 27 d) and arrhythmia (median 77 d) the latest. Median time to event resolution was 17 d, with headache (median 3d) the quickest to resolve and hypokalemia (median 64 d) the slowest. Among patients with a common AE, recurrence averaged 18% of patients across categories: nausea/vomiting (36%) recurring most often and hypertension (3.7%) least often. Median time to first recurrence following initial event resolution for combined categories was 25 d. Mean number of recurrences per patient was highest for hypokalemia (1.8±0.5). Detailed characterization of common AEs from an integrated dataset of 2 clinical studies populations during titration with levoketoconazole provides new knowledge relevant to levoketoconazole clinical use as chronic medical treatment in adult patients with CS.

Submarine landslide susceptibility assessment along the southern convergent margin of the Colombian Caribbean

Submarine landslides are a mixture of rock, sediment, and fluids moving downslope due to a slope's initial event of mechanical failure. Submarine landslides play a critical role in shaping the morphology of the seafloor and the transport of sediments from the continental shelf to the continental rise in the southern margin of the Colombian Caribbean. Two fundamental considerations can be highlighted: first, mass transport complexes produced by submarine landslides encompass significant portions of the stratigraphic record; second, these mass movements could affect underwater infrastructure. The mapping of the Southern Caribbean seafloor using 3D seismic surveys and multibeam bathymetry data in an area encompassing 59,471 km2 allowed the identification of 220 submarine landslides with areas ranging between 0.1 and 209 km2. Distinctive characteristics were found for submarine landslides associated with canyon walls, channel-levee systems, tectonically controlled ridges, and the continental shelf break. The analysis of the relationship between submarine landslides and seafloor morphological features made it possible to estimate a mass movement susceptibility map that suggests the following considerations: first, structural ridges and adjacent intraslope subbasins related to the South Caribbean Deformed Belt are more likely to be submarine landslide hazards; second, the continental shelf break and channelized systems produce a moderate submarine landslide hazard potential; and third, deep marine systems with a slope less than 5° show the lowest submarine landslide hazard potential. This work contributes to the understanding of submarine landslides in the study area through the presentation of conceptual diagrams that provide additional visual elements facilitating the level of abstraction necessary for visualizing bathymetric data. Likewise, the mass movement susceptibility map presented herein gives insights for future studies that seek to evaluate geohazards in the southern Colombian Caribbean margin.

Regulation of Hepatocytes in G0 and G1 Phases by NOTCH3 mRNA, miR-369-3p, and rno-Rmdn2_0006 during the Initial Stage of Rat Liver Regeneration.

The key event of liver regeneration initiation (LRI) is the switch of hepatocytes from the G0 phase to the G1 phase. This study aimed to use the data from large-scale quantitatively detecting and analyzing (LQDA) to reveal the regulation of hepatocytes in the G0 or G1 phase by competing endogenous RNAs (ceRNAs) during LRI. The hepatocytes of the rat liver right lobe were isolated 0, 6, and 24 h after partial hepatectomy. Their ceRNA expression level was measured using LQDA, and the correlation among their expression, interaction, and role was revealed by ceRNA comprehensive analysis. The expression of neurogenic loci notch homologous protein 3 (NOTCH3) mRNA was upregulated in 0 h, but the expression of miR-369-3p and rno-Rmdn2_0006 of hepatocytes did not change significantly. Meanwhile, the expression of the G0 phase-related gene CDKN1c was promoted by NOTCH3 upregulation, and the expression of the G1 phase-related gene PSEN2 was inhibited by NOTCH3 downregulation. On the contrary, the expression of NOTCH3 mRNA and rno-Rmdn2_0006 was upregulated at 6 h, but the expression of miR-136-3p was downregulated. The expression of the G1 phase-related genes CHUK, DDX24, HES1, NET1, and STAT3 was promoted by NOTCH3 upregulation, and the expression of the G0 phase-related gene CDKN1a was inhibited by NOTCH3 downregulation. These results suggested that the ceRNAs and the NOTCH3-regulated G0 phase- and G1 phase-related genes showed a correlation in expression, interaction, and role. They together regulated the hepatocytes in the G0 phase at 0 h and in the G1 phase at 6 h. These findings might help understand the mechanism by which ceRNA together regulated the hepatocytes in the G0 or G1 phase.

The Origin of the Large Magellanic Cloud Globular Cluster NGC 2005

The ancient Large Magellanic Cloud (LMC) globular cluster NGC 2005 has recently been reported to have an ex situ origin, thus, setting precedents that the LMC could have partially formed from smaller merged dwarf galaxies. We here provide additional arguments from which we conclude that is also fairly plausible an in situ origin of NGC 2005, based on the abundance spread of a variety of chemical elements measured in dwarf galaxies, their minimum mass in order to form globular clusters, the globular cluster formation imprints kept in their kinematics, and the recent modeling showing that explosions of supernovae are responsible for the observed chemical abundance spread in dwarf galaxies. The present analysis points to the need for further development of numerical simulations and observational indices that can help us to differentiate between two mechanisms of galaxy formation for the LMC; namely, a primordial dwarf or an initial merging event of smaller dwarfs.

A critical role for altered red cell cation permeability in pathogenesis of sickle cell disease and other haemolytic anaemias.

The aetiology of sickle cell disease is well known, but pathogenesis is complicated and details remain uncertain. A thorough understanding may suggest novel ways for designing more effective therapies. One area of importance, covered here in Nader et al., is the altered cation permeability of sickle cells and how the co-ordinated operation of a number of membrane transport proteins contributes to disease progression, all driven by the initial event of HbS polymerisation. There are echoes here of the cation leaks of hereditary stomatocytosis. Nader et al. propose a central role for PIEZO1, a novel mechanosensitive channel found in red cells, which may be aberrantly activated in sickle cells following HbS polymerisation and which may have potential as a novel target for future chemotherapies. Commentary on: Nader et al. Piezo1 activation augments sickling propensity and the adhesive properties of sickle red blood cells in a calcium-dependent manner. Br J Haematol 2023;202:657-668.

Necessity of HuR/ELAVL1 for activation-induced cytidine deaminase-dependent decrease in topoisomerase 1 in antibody diversification.

Activation-induced cytidine deaminase (AID)-dependent DNA cleavage are the initial event of antibody gene-diversification processes such as class switch recombination (CSR) and somatic hypermutation (SHM). We previously reported the requirement of an AID-dependent decrease of topoisomerase 1 (Top1) for efficient DNA cleavage, but the underlying molecular mechanism has remained elusive. This study focuses on HuR/ELAVL1, a protein that binds to AU-rich elements in RNA. HuR-knockout (KO) CH12 cells derived from murine B lymphoma cells were found to have lower CSR and hypermutation efficiencies due to decreased AID-dependent DNA cleavage levels. The HuR-KO CH12 cells do not show impairment in cell cycles and Myc expression, which have been reported in HuR-reduced spleen B cells. Furthermore, drugs that scavenge reactive oxygen species (ROS) do not rescue the lower CSR in HuR-KO CH12 cells, meaning that ROS or decreased c-Myc protein amount is not the reason for the deficiencies of CSR and hypermutation in HuR-KO CH12 cells. We show that HuR binds to Top1 mRNA and that complete deletion of HuR abolishes AID-dependent repression of Top1 protein synthesis in CH12 cells. Additionally, reduction of CSR to IgG3 in HuR-KO cells is rescued by knockdown of Top1, indicating that elimination of the AID-dependent Top1 decrease is the cause of the inefficiency of DNA cleavage, CSR, and hypermutation in HuR-KO cells. These results show that HuR is required for initiation of antibody diversification and acquired immunity through the regulation of AID-dependent DNA cleavage by repressing Top1 protein synthesis.

The perturbation of parabens on the neuroendocrine system in zebrafish larvae

Parabens, as the synthetic preservatives, have caused universal environmental contamination and human exposure. Whether parabens could disturb neuroendocrine system was still ambiguous. In this study, the effects of four commonly-used parabens, i.e. methyl paraben (MeP), ethyl paraben (EtP), propyl paraben (PrP) and butyl paraben (BuP), were tested on the neuroendocrine system of zebrafish larvae by investigating the swimming behavior, the related hormones and biomarkers in the hypothalamic-pituitary-interrenal (HPI) axis. The results showed that all test chemicals significantly reduced the swimming distance and mean velocity of zebrafish larvae. The adrenocorticotropic hormone (ACTH) levels in zebrafish larvae were significantly increased, while the cortisol levels were obviously decreased by paraben exposure. The transcriptional analysis showed that the expressions of the target genes including gr, mr and crhr2 in the HPI axis were mostly down-regulated. The exploration of the initial molecular event showed that parabens could bind with the glucocorticoid receptor (GR) and trigger its transactivation, according to MDA-kb2 luciferase assay and molecular docking analysis. The interaction of parabens with the GR included the hydrogen bond and hydrophobic interaction. The findings herein revealed the potential deleterious effects of parabens on the neuroendocrine system of zebrafish larvae, thus accumulating the in vivo toxicological data on this kind of food preservatives.

Genome diversity of Leishmania aethiopica.

Leishmania aethiopica is a zoonotic Old World parasite transmitted by Phlebotomine sand flies and causing cutaneous leishmaniasis in Ethiopia and Kenya. Despite a range of clinical manifestations and a high prevalence of treatment failure, L. aethiopica is one of the most neglected species of the Leishmania genus in terms of scientific attention. Here, we explored the genome diversity of L. aethiopica by analyzing the genomes of twenty isolates from Ethiopia. Phylogenomic analyses identified two strains as interspecific hybrids involving L. aethiopica as one parent and L. donovani and L. tropica respectively as the other parent. High levels of genome-wide heterozygosity suggest that these two hybrids are equivalent to F1 progeny that propagated mitotically since the initial hybridization event. Analyses of allelic read depths further revealed that the L. aethiopica - L. tropica hybrid was diploid and the L. aethiopica - L. donovani hybrid was triploid, as has been described for other interspecific Leishmania hybrids. When focusing on L. aethiopica, we show that this species is genetically highly diverse and consists of both asexually evolving strains and groups of recombining parasites. A remarkable observation is that some L. aethiopica strains showed an extensive loss of heterozygosity across large regions of the nuclear genome, which likely arose from gene conversion/mitotic recombination. Hence, our prospection of L. aethiopica genomics revealed new insights into the genomic consequences of both meiotic and mitotic recombination in Leishmania.

Analysis of the Dynamics of Water Flow and Suspension Flow Discharge in Volcano Watershed with Settlement Land Use

Suspension flow into the upstream of volcano watershed is sensitive to land use. In Indonesia, a settlement is a form of land use in several volcanic landscapes. There is currently no detailed study on the suspension flow sediment from the settlement land use. The purpose of this study is to investigate the characteristics of the relationship between water and suspension flow discharge. The study was conducted through the measurements at a gully outlet that produced 747 suspension load data. For each rainfall event, suspension load measurements were made in the field, followed by laboratory analysis. Additionally, field surveys were used to determine the characteristics of settlement land use and the water flow into the gully system. According to the findings, the peak flow discharge corresponds to the peak suspension discharge, the peak flow discharge comes before the peak suspension discharge, and the peak flow discharge happens after the peak suspension discharge. The average time lag between initial rainfall events and suspension flow was 10.36 minutes, and the suspension peak content varied by an average of 2.22 gl-1. The grain size was also dominated by the clay fraction, averaging 67.86% on the ascending branch and 67.82% on the descending branch.&#x0D; Keywords: Erosion; Discharge; Settlements; Suspension; Watershed&#x0D; &#x0D; Copyright (c) 2023 Geosfera Indonesia and Department of Geography Education, University of Jember&#x0D; This work is licensed under a Creative Commons Attribution-Share A like 4.0 International License

Chronodisruption and Loss of Melatonin Rhythm, Associated with Alterations in Daily Motor Activity and Mitochondrial Dynamics in Parkinsonian Zebrafish, Are Corrected by Melatonin Treatment.

Beyond sleep/wake, clock genes regulate the daily rhythms of melatonin production, motor activity, innate immunity, and mitochondrial dynamics, among others. All these rhythms are affected in Parkinson's disease (PD), suggesting that chronodisruption may be an early stage of the disease. The aim of this study was to evaluate the connection between clock genes and these rhythms in PD, and whether melatonin administration reestablished the normal clock function. Parkinsonism was induced with 600 μM MPTP (N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) in 24-120 h post fertilization (hpf) zebrafish embryos and melatonin was administered at a dose of 1 μM. Day-night melatonin rhythm disappeared in MPTP-treated embryos, which showed an advance in the activity phase in parallel with changes in the rhythm of clock genes. An alteration in the fission-to-fusion mitochondrial dynamics was also detected in parkinsonian embryos, increasing the former and leading to apoptosis. Melatonin administration to MPTP-treated embryos fully restored the circadian system, including the rhythms of clock genes, motor activity, melatonin rhythm, and mitochondrial dynamics, and decreasing apoptosis. Because clock-controlled rhythms such as sleep/wake alterations are early events in PD, the data here reported may point to chronodisruption as one initial pathophysiological event of the disease.

Fluorescent microspheres can affect in vitro fibrinolytic outcomes.

Hemostasis is the cessation of bleeding due to the formation of a blood clot. After the completion of wound healing, the blood clot is typically dissolved through the natural process of fibrinolysis, the enzymatic digestion by plasmin of the fibrin fibers that make up its structural scaffold. In vitro studies of fibrinolysis reveal mechanisms regulating these processes and often employ fluorescent microscopy to observe protein colocalization and fibrin digestion. In this study, we investigate the effects of labeling a fibrin network with 20 nm diameter fluorescent beads (fluorospheres) for the purpose of studying fibrinolysis. We observed fibers and 2-D fibrin networks labeled with fluorospheres during fibrinolysis. We found that the labeling of fibrin with fluorospheres can alter fibrinolytic mechanisms. In previous work, we showed that, during lysis, fibrin fibers are cleaved into two segments at a single location. Herein we demonstrate that fibrinolysis can be altered by the concentration of fluorospheres used to label the fibers, with high concentrations of fluorospheres leading to very minimal cleaving. Furthermore, fibers that are left uncleaved after the addition of plasmin often elongate, losing their inherent tension throughout the imaging process. Elongation was especially prominent among fibers that had bundled together due to other cleavage events and was dependent on the concentration of fluorophores used to label fibers. Of the fibers that do cleave, the site at which they cleave also shows a predictable trend dependent on fluorosphere concentration; low concentrations heavily favor cleavage locations at either end of fibrin fiber and high concentrations show no disparity between the fiber ends and other locations along the fiber. After the initial cleavage event bead concentration also affects further digestion, as higher bead concentrations exhibited a larger population of fibers that did not digest further. The results described in this paper indicate that fluorescent labeling strategies can impact fibrinolysis results.

Cascading disasters and mental health: The February 2021 winter storm and power crisis in Texas, USA

In February 2021, the state of Texas and large parts of the US were affected by a severe cold air outbreak and winter weather event. This event resulted in large-scale power outages and cascading impacts, including limited access to potable water, multiple days without electricity, and large-scale infrastructure damage. Little is known about the mental health implications of these events, as most research has predominantly focused on the mental health effects of exposure to hurricanes, wildfires, or other natural disasters that are more commonly found in the summer months. This study aimed to analyze the crisis responses from the 2021 winter weather event in Texas using Crisis Text Line, a text-based messaging service that provides confidential crisis counseling nationwide. To date, Crisis Text Line is the largest national crisis text service, with over 8 million crisis conversations since its inception in 2013. We employed multiple analytic techniques, including segmented regression, interrupted time series, autoregressive integrated moving average (ARIMA), and difference-in-difference (DID), to investigate distinct time periods of exposure for all crisis conversations. ARIMA and DID were further utilized to examine specific crisis outcomes, including depression, stress/anxiety, and thoughts of suicide. Results found increases in total crisis conversations and for thoughts of suicide after the initial winter weather event; however, crisis outcomes varied in time. Thoughts of suicide in high-impact regions were higher across multiple time periods (e.g., 4-weeks, 3-months, 6-months, 9-months and 11-months) compared to low-impact regions and were elevated compared to pre-event time periods for 6-months and 11-months from the initial event. Total crisis volume also remained elevated for high-impact regions compared to low-impact regions up to 11-months after the beginning of the winter event. Our work highlights that cascading winter weather events, like the Texas 2021 Winter storm, negatively impacted mental health. Future research is needed across different disaster types (e.g., cascading, concurrent events) and for specific crisis outcomes (e.g., depression, suicidal ideation) to understand the optimal timing of crisis intervention post-disaster.

A Multiscale Model for Shear-Mediated Platelet Adhesion Dynamics: Correlating In Silico with In Vitro Results.

Platelet adhesion to blood vessel walls is a key initial event in thrombus formation in both vascular disease processes and prosthetic cardiovascular devices. We extended a deformable multiscale model (MSM) of flowing platelets, incorporating Dissipative Particle Dynamics (DPD) and Coarse-Grained Molecular Dynamics (CGMD) describing molecular-scale intraplatelet constituents and their interaction with surrounding flow, to predict platelet adhesion dynamics under physiological flow shear stresses. Binding of platelet glycoprotein receptor Ibα (GPIbα) to von Willebrand factor (vWF) on the blood vessel wall was modeled by a molecular-level hybrid force field and validated with in vitro microchannel experiments of flowing platelets at 30 dyne/cm2. High frame rate videos of flipping platelets were analyzed with a Semi-Unsupervised Learning System (SULS) machine learning-guided imaging approach to segment platelet geometries and quantify adhesion dynamics parameters. In silico flipping dynamics followed in vitro measurements at 15 and 45dyne/cm2 with high fidelity, predicting GPIbα-vWF bonding and debonding processes, distribution of bonds strength, and providing a biomechanical insight into initiation of the complex platelet adhesion process. The adhesion model and simulation framework can be further integrated with our established MSMs of platelet activation and aggregation to simulate initial mural thrombus formation on blood vessel walls.

Abstract 4907: Atovaquone-induced increase in reactive oxygen species leads to impaired glycolytic functions in ovarian cancer cells

Abstract Atovaquone (ATO) is an FDA-approved anti-malarial drug, which has also shown potent anti-cancer effects. We have previously demonstrated ATO as an effective chemotherapy against high-grade serous ovarian cancer in cell lines and mouse model. In the present study, we aim to determine if ATO is asserting its anti-cancer properties by altering cellular metabolism. Glycolysis and oxidative phosphorylation (OXPHOS) are the two major sources of intracellular energy. ATO reduces OXPHOS energy production by inhibiting mitochondrial complex III. We first determined whether there is a shift in energy production when OXPHOS is inhibited by assessing glycolytic function in various human (OVCAR3 and OVCAR5) and mouse (ID8) ovarian cancer cell lines. Mitochondrial oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) were assessed by the Seahorse XF Analyzer. First, we pretreated cells with ATO at IC70 concentrations for 1 hour prior to metabolic analysis. As expected, pretreatment with ATO reduces OCR in all cell lines. However, ATO treatment increased basal glycolysis but decreased glycolytic capacity and glycolytic reserve. This suggests that ATO-treated cells have an impaired ability to shift energy production towards glycolysis when there is an increase in energy demand. One of the initial events that occur in ATO-treated cells is a surge in reactive oxygen species (ROS) as measured by H2DCFDA ROS assay. In the human cell lines, we found that ATO significantly increased ROS by approximately 100-fold (MFI). Whereas, in the mouse cell line, the ATO-induced increase in ROS were comparatively attenuated. We hypothesized that the increase in ROS leads to inhibition of glycolysis. We tested this by pre-treating cells with 2mM N-acetylcysteine (NAC), an oxygen radical scavenger, for 1 hour before measuring the metabolic responses to ATO. We found that NAC attenuated the inhibition of glycolytic processes in human cell lines but not in the mouse cell line. Our lab has previously shown that ATO induced activation and increased expression of p53. Studies have shown that ROS activate p53, which in turn upregulates the TP53 induced glycolysis and apoptosis regulatory phosphatase (TIGAR), inhibiting the conversion of F6P to F-2,6-BP and ultimately leads to inhibition of glycolysis. Increases in ROS also shuttle G6P towards the phosphate pentose pathways through G6P dehydrogenase for antioxidant production. We found that long-term ATO treatment for 24 hours also resulted in increases in both TIGAR and G6PD mRNA expression. Taken together, these results suggest that ATO decreased plasticity of metabolic capacity in ovarian cancer cells through ROS and ROS associated glycolytic pathways. Further, this study provides mechanistic insights into ATO’s potential as a chemotherapeutic agent. Citation Format: Nicha Boonpattrawong, Sejal Sharma, Rob Schultz, Mayra A. Betancourt Ponce, Lisa M. Barroilhet, Manish Patankar. Atovaquone-induced increase in reactive oxygen species leads to impaired glycolytic functions in ovarian cancer cells. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4907.

Veklury®(remdesivir) formulations inhibit initial membrane-coupled events of SARS-CoV-2 infection due to their sulfobutylether-β-cyclodextrin content.

Despite its contradictory clinical performance, remdesivir (Veklury®) has a pivotal role in COVID-19 therapy. Possible contributions of the vehicle, sulfobutylether-β-cyclodextrin (SBECD) toVeklury® effects have been overlooked. The powder and solution formulations of Veklury® are treated equivalently despite their different vehicle content. Our objective was to study Veklury® effects on initial membrane-coupled events of SARS-CoV-2 infection focusing on the cholesterol depletion-mediated role of SBECD. Using time-correlated flow cytometry and quantitative three-dimensional confocal microscopy, we studied early molecular events of SARS-CoV-2-host cell membrane interactions. Veklury® and different cholesterol-depleting cyclodextrins (CDs) reduced binding of the spike receptor-binding domain (RBD) to ACE2 and spike trimer internalization for Wuhan-Hu-1, Delta and Omicron variants. Correlations of these effects with cholesterol-dependent changes in membrane structure and decreased lipid raft-dependent ACE2-TMPRSS2 interaction establish that SBECD is not simply a vehicle but also an effector along with remdesivir due to its cholesterol-depleting potential. Veklury® solution inhibited RBD binding more efficiently due to its twice higher SBECD content. The CD-induced inhibitory effects were more prominent at lower RBD concentrations and in cells with lower endogenous ACE2 expression, indicating that the supportive CD actions can be even more pronounced during in vivo infection when viral load and ACE expression are typically low. Our findings call for the differentiation of Veklury® formulations in meta-analyses of clinical trials, potentially revealing neglected benefits of the solution formulation, and also raise the possibility of adjuvant cyclodextrin (CD) therapy, even at higher doses, in COVID-19.

Study protocol for a prospective, multi-centered randomized controlled trial comparing pelvic fascia-sparing radical prostatectomy with conventional robotic-assisted prostatectomy: The PARTIAL trial

BackgroundPelvic fascia-sparing robotic-assisted radical prostatectomy (PFS-RARP) is a novel approach that spares the endopelvic fascia ventral to the prostate. The preservation of more native structures compared to conventional robotic-assisted radical prostatectomy (RARP) may lead to faster recovery of urinary function, fewer penile changes, and decreased inguinal hernia sequelae, but may have a higher risk for positive surgical margins and poorer cancer control. However, high-level evidence is absent. The PARTIAL trial is a surgical randomized controlled trial (RCT) aiming to bridge this evidence gap (NCT05155501). MethodsWe describe a prospective RCT with a projected enrollment of 600 men randomized to PFS-RARP vs. RARP. The primary outcome is cancer control (positive surgical margins and prostate-specific antigen failure) and secondary outcomes include health-related quality of life pertaining to urinary and sexual function, decision regret, and adverse events (30-day complications, inguinal hernias, penile shortening, and Peyronie's disease). The anticipated duration of trial participation is 24 months. Study participation is incentivized with the use of innovative methodologies such as a novel, two-stage informed consent and a validated web-based interface to monitor patient-reported symptoms and empower individuals to improve their recovery. ConclusionIf PFS-RARP is non-inferior to RARP in terms of cancer control and has better functional outcomes, it should be the surgical standard of care for men with localized prostate cancer. Using the innovative two-stage consent process, completion of the trial will not only provide much needed evidence on one of the most common cancer surgeries but also insight on improving surgical RCT methodology.Trial statusThis trial is registered at ClinicalTrials.gov (NCT05155501; first posted on December 13, 2021); Institutional approval number: WCM IRB # 21-07023781, BRANY's initial approval event ID # 186333. The trial is not yet recruiting.

Clinical and Genetic Characteristics of Patients with Moyamoya Disease who Experienced Both Ischemic and Hemorrhagic Events.

Cerebrovascular events in moyamoya disease are mainly classified into ischemic or hemorrhagic onset. It is rare for one patient to develop both ischemia and hemorrhage in moyamoya disease; detailed clinical course and genetic characteristics of such patients have not been elucidated. We aimed to clarify the clinical features of patients with both ischemic and hemorrhagic cerebrovascular events. We analyzed the background factors, radiological features, and genotype of ring finger protein 213 c.14429G>A (p.Arg4810Lys) of patients with moyamoya disease who visited our hospital between 1996 and 2020, and experienced both ischemic and hemorrhagic cerebrovascular events. Additionally, we analyzed factors that caused subsequent hemorrhage in adult-onset ischemic moyamoya disease. Of 262 patients, 12 presented with both ischemia and hemorrhage, of which, 4 exhibited pediatric onset and 8 had adult onset. In pediatric-onset subjects, ischemia was the initial event in all cases. Hemorrhagic events occurred at a median of 24.7years postoperatively in patients who had undergone bypass surgery. In adult-onset subjects, ischemia preceded hemorrhage in 7 patients. In males, the interval to subsequent hemorrhage was significantly shorter for adult-onset ischemic moyamoya disease, and the hazard ratio for hemorrhagic events was 5.45. The ring finger protein 213 p.Arg4810Lys heterozygous variant was present in 9 patients. A majority of patients with moyamoya disease with both ischemia and hemorrhage experience an ischemic event first. Patients who developed ischemia in childhood may develop subsequent hemorrhage in approximately 20-25years after bypass surgery. Male sex is a risk factor for a subsequent hemorrhagic event in adult-onset ischemic moyamoya disease.

Association between Organophosphate Ester Exposure and Insulin Resistance with Glycometabolic Disorders among Older Chinese Adults 60-69 Years of Age: Evidence from the China BAPE Study.

Organophosphate esters (OPEs) are common endocrine-disrupting chemicals, and OPE exposure may be associated with type 2 diabetes (T2D). However, greater knowledge regarding the biomolecular intermediators underlying the impact of OPEs on T2D in humans are needed to understand biological etiology. We explored the associations between OPE exposure and glycometabolic markers among older Chinese adults 60-69 years of age to elucidate the underlying mechanisms using a multi-omics approach. This was a longitudinal panel study comprising 76 healthy participants 60-69 years of age who lived in Jinan city of northern China. The study was conducted once every month for 5 months, from September 2018 to January 2019. We measured a total of 17 OPEs in the blood, 11 OPE metabolites in urine, and 4 glycometabolic markers (fasting plasma glucose, glycated serum protein, fasting insulin, and homeostatic model assessment for insulin resistance). The blood transcriptome and serum/urine metabolome were also evaluated. The associations between individual OPEs and glycometabolic markers were explored. An adverse outcome pathway (AOP) was established to determine the biomolecules mediating the associations. Exposure to five OPEs and OPE metabolites (trimethylolpropane phosphate, triphenyl phosphate, tri-iso-butyl phosphate, dibutyl phosphate, and diphenyl phosphate) was associated with increased levels of glycometabolic markers. The mixture effect analysis further indicated the adverse effect of OPE mixtures. Multi-omics analyses revealed that the endogenous changes in the transcriptional and metabolic levels were associated with OPE exposure. The putative AOPs model suggested that triggers of molecular initiation events (e.g., insulin receptor and glucose transporter type 4) with subsequent key events, including disruptions in signal transduction pathways (e.g., phosphatidylinositol 3-kinase/protein kinase B and insulin secretion signaling) and biological functions (glucose uptake and insulin secretion), may constitute the diabetogenic effects of OPEs. OPEs are associated with the elevated risk of T2D among older Chinese adults 60-69 years of age. Implementing OPE exposure reduction strategies may help reduce the T2D burden among these individuals, if the relationship is causal. https://doi.org/10.1289/EHP11896.