Sampling Time Points Research Articles

P-714. Symptoms Associated with High Viral Loads of Common Cold Coronaviruses in Hematopoietic Cell Transplantation Recipients

Abstract Background Limited data exist on the association between specific symptoms and viral loads of common cold coronavirus (ccCOV). We investigated this potential association in allogeneic hematopoietic cell transplant (HCT) recipients.Figure 1.Proportion of sampling time points with each symptom Methods Four hundred seventy-one children and adults were prospectively followed with weekly symptom questionnaires through one year after allogeneic HCT at the Fred Hutchinson Cancer Center (12/2005-2/2010). Multiplex PCR for 11 respiratory viruses was performed on combined nasal wash and throat swab samples collected weekly through day 100 post-HCT, and then every three months or if symptoms were present through one-year post-HCT. Positive samples were evaluated for viral loads with quantitative real-time PCR (copies/ml). We included time points with the detection of ccCOV only and corresponding 15-point symptom survey data available post-HCT. To assess the association between specific symptoms and viral loads, the Mann-Whitney or Jonckheere-Terpstra tests were used to compare 2 or >2 groups for continuous variables, respectively.Figure 2.Symptoms associated with higher viral loads Results Among 6,276 samples tested, 251 samples (79 patients) were positive for ccCoV. One hundred thirty-six sampling time points (52 patients) met the inclusion criteria. The proportion of sample time points with each symptom present is shown in Figure 1. When we compared viral loads with the presence of each of the 15 symptoms, higher viral loads were associated with the presence of four respiratory tract symptoms (rhinorrhea, sneezing, sore throat, and coughing) and one systemic symptom (diarrhea) (p< 0.01 for each) (Figure 2). Viral loads appeared to increase with increasing number of respiratory tract symptoms (p< 0.001) (Figure 3).Figure 3.Viral loads compared with the number of respiratory tract symptoms Conclusion The presence of specific respiratory tract symptoms is associated with higher viral loads of ccCOV in the upper respiratory tract in allogeneic HCT recipients. The association with diarrhea suggests the possibility of fecal shedding, but this was not evaluated in our study. Patients with specific symptoms may be more likely to transmit virus given their higher viral loads. Studies of these associations for other respiratory viruses are warranted. Disclosures Chikara Ogimi, MD, PhD, AstraZeneca: Honoraria|bioMerieux Japan Ltd.: Honoraria|ELSEVIER: Honoraria|KYORIN: Honoraria|Miyarisan: Honoraria|MSD: Honoraria|NOVARTIS: Honoraria|Pfizer: Honoraria Alpana waghmare, MD, Allovir: Grant/Research Support|Ansun Biopharma: Grant/Research Support|GlaxoKlineSmith: Advisor/Consultant|GlaxoKlineSmith: Grant/Research Support|Pfizer: Grant/Research Support|Vir: Advisor/Consultant Janet A. Englund, MD, Abbvie: Advisor/Consultant|AstraZeneca: Advisor/Consultant|AstraZeneca: Grant/Research Support|GlaxoSmithKline: Advisor/Consultant|GlaxoSmithKline: Grant/Research Support|Meissa Vaccines: Advisor/Consultant|Merck: Advisor/Consultant|Pfizer: Board Member|Pfizer: Grant/Research Support|Pfizer: Speaker at meeting|SanofiPasteur: Advisor/Consultant|Shinogi: Advisor/Consultant Michael J. Boeckh, MD PhD, Allovir: Advisor/Consultant|Allovir: Grant/Research Support|AstraZeneca: Advisor/Consultant|AstraZeneca: Grant/Research Support|Merck: Advisor/Consultant|Merck: Grant/Research Support|Moderna: Advisor/Consultant|Moderna: Grant/Research Support|Symbio: Advisor/Consultant

The prevalence and antimicrobial resistance of respiratory pathogens isolated from feedlot cattle in Canada

ObjectivesThe purpose of this study was to characterize the prevalence of antimicrobial resistance in Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni isolated from healthy feedlot cattle over 2 years, and investigate factors potentially associated with recovery of resistant isolates.MethodsDeep-guarded nasopharyngeal (NP) swabs were used to sample feedlot cattle in multiple randomly selected feedlots (2019 n = 21, 2020 n = 26) at 2 timepoints. NP swabs were collected from 16 animals in each enrolled group upon entry processing and later in the feeding period. Cattle from the same groups (not necessarily the same animals) were sampled at both timepoints. Susceptibility testing was performed using the broth microdilution.ResultsA total of 1,392 cattle within 47 housing groups were sampled over 2 years, providing 625 bacterial isolates for investigation. Pasteurella multocida (27.4%) was the most frequently isolated BRD organism, followed by H. somni (9%) and M. haemolytica (8.5%). Resistance to ≥3 antimicrobial classes was detected in 2.4% of M. haemolytica, 3.4% of H. somni, and 21.3% of P. multocida isolates. Potential associations were investigated between recovery of resistant organisms and time of year at sampling (quarter), sampling timepoint (arrival or second sample), days on feed (DOF) at sampling, animal age categories, and BRD risk categories. There was a significant (p < 0.05) increase in resistance prevalence after arrival for macrolide drugs in M. haemolytica, and for ampicillin, danofloxacin, enrofloxacin, spectinomycin, gamithromycin, tildipirosin, tulathromycin and tetracycline in P. multocida isolates. Resistance was higher in calves than in yearlings for tulathromycin in H. somni, and for gamithromycin, spectinomycin, tulathromycin, tildipirosin, and tetracycline for P. multocida (p < 0.05) Resistance to tetracycline, tildipirosin, and tulathromycin decreased between 61–80 DOF and 81–100 DOF when compared to 20–40 DOF, whereas for spectinomycin, resistance was lower in cattle sampled between 61–80 DOF than those sampled at 20–40 DOF for P. multocida.DiscussionThe diversity of AMR profiles and associated risk factors between the BRD pathogens studied, underscores the importance of including all three organisms in future AMR studies in beef cattle.

Transcriptional time-course analysis during ash dieback infection revealed different responses in tolerant and susceptible Fraxinus excelsior genotypes

Hymenoscyphus fraxineus, the causal agent of Ash Dieback (ADB), has been introduced to eastern Europe in the 1990s from where it spread causing decline in European ash populations. However, the genetic basis of the molecular response in tolerant and susceptible ash trees to this disease is still largely unknown. We performed RNA-sequencing to study the transcriptomic response to the disease in four ash genotypes (ADB-tolerant FAR3 and FS36, and ADB-susceptible UW1 and UW2), during a time-course of 7, 14, 21, and 28 days post-inoculation, including mock-inoculated trees as control samples for each sampling time point. The analysis yielded 395 and 500 Differentially Expressed Genes (DEGs) along the response for ADB-tolerant FAR3 and FS36, respectively, while ADB-susceptible UW1 and UW2 revealed 194 and 571 DEGs, respectively, with most DEGs found exclusively in just one of the genotypes. DEGs shared between tolerant genotypes FAR3 and FS36, included genes involved in the production of phytoalexins and other secondary metabolites with roles in plant defense. Moreover, we identified an earlier expression of genes involved in both pattern- and effector-triggered immunity (PTI and ETI) in ADB-tolerant genotypes, while in ADB-susceptible genotypes both responses were delayed (late response). Overall, these results revealed different transcriptomic expression patterns not only between ADB-tolerant and ADB-susceptible genotypes, but also within these two groups. This hints to individual responses in the natural tolerance to ADB, possibly revealing diversified strategies across ash genotypes.

A Novel Method Combining Radial Projection with Simultaneous Multislice Imaging for Measuring Cerebrovascular Pulse Wave Velocity

Magnetic resonance imaging (MRI) using a simultaneous multislice technique can measure dynamic vascular elasticity over time. However, conventional k-space undersampling can cause signal interference, owing to vertical projection between blood vessels within the same hemisphere. Here, we proposed a radial projection method that can reduce signal interference between the blood vessels and aimed to verify the theoretical and practical effects of this method. A dataset from the internal and common carotid arteries (ICA and CCA) was used for both projection methods. Pulse wave velocity (PWV) was calculated using the ICA and CCA time series, and the methods were compared using the mean absolute error of PWV. The feasibility of the radial projection method in an actual MRI environment was also evaluated. PWVs of the radial projection method were statistically indistinguishable from the ground truth. And the radial projection method was less sensitive to background noise levels and showed similar results to the ground truth. This method could effectively avoid signal interference between vessels and was feasible for use in real MRI environments, maintaining high temporal resolution even with fewer sampling timepoints. Therefore, it can contribute to the early diagnosis and treatment of cerebrovascular diseases through accurate and dynamic PWV measurements.

Bioavailability of rumen-protected histidine, lysine, and methionine assessed using different in vivo methods.

The objective of this experiment was to estimate the bioavailability (BA) of rumen-protected (RP) His, RPLys, and 2 RPMet products using 3 in vivo methods: area under the curve (AUC), plasma dose-response (PDR), and fecal free AA (FFAA) methods. We used 8 rumen-cannulated cows in a replicated 4 × 4 Latin square experiment with 16-d periods. Treatments were (1) abomasal infusion of water (control), (2) abomasal infusion of free His, Lys, and Met (FAA), (3) administration of RPHis + RPLys + RPMet1 (rumen-protected methionine protected with ethyl cellulose; RPAA1), and (4) administration of RPHis + RPLys + RPMet2 (rumen-protected methionine protected with a pH-sensitive polymer; RPAA2). On d 7 of each experimental period, a pulse-dose of water (control) or FAA were infused into the abomasum of the cows, or RPAA were placed directly in the rumen, and blood samples were taken from the jugular vein through a catheter 11 times over a 24-h period for the AUC method. Following the AA pulse-dose, infusion lines were installed into the abomasum for continuous infusion of FAA for the PDR method, or RPAA were fed from d 12 to d 16 and cows were fitted with urinary catheters for total collection of feces for the FFAA method. Fecal collection and blood sampling were conducted from d 14 to 16. Due to technical issues likely leading to unrealistic BA estimates, data for the PDR method are reported in the supplemental material. Relative BA based on the AUC method (computed as AUC of RPAA treatment plasma AA concentration divided by AUC of FAA treatment plasma AA concentration) was lower for RPMet1 compared with RPMet2 (43% vs. 61%) and was 45% (SEM = 3.35) and 72% (SEM = 5.99), for RPHis and RPLys, respectively. Rumen escape fractions of RPAA, estimated in a previous study using an in situ method, and digestibility data from the current study were used for calculations of BA for the FFAA method. Bioavailability based on the FFAA method was lower for RPMet1 (67%) compared with RPMet2 (91%) and was 87% (SEM = 0.71) and 75% (SEM = 2.75) for RPHis and RPLys, respectively. The relative differences in estimated BA based on both the AUC and FFAA methods between the RPMet products were as expected, based on literature, and data for all 4 RPAA products corresponded well with previously estimated BA using the FFAA method. The unrealistic data for the PDR method were likely caused by technical deviations from the original method (e.g., once-daily dosing of RPAA and inability to capture representative plasma concentration data with the sampling time points). Therefore, comparison of the PDR method with the AUC and FFAA methods were not possible in this study. Further comparisons are needed without deviations from the original PDR method. Variability in BA data and differences in estimated BA between the in vivo methods highlight the current challenges for accurate measurements of relative in vivo BA of RPAA products. Different protection technologies may call for different methodology to be used for BA estimations. Further research and standardization of in vivo BA methods are warranted.

Evaluation of the covariation between leukotriene B4, prostaglandin E2, and hematologic inflammatory parameters in a canine pentylenetetrazole-induced seizure model.

Seizures can cause as well as result from neuroinflammation. This study was performed to identify the hematologic inflammatory parameters (HIPs) and inflammatory mediators that change after a single seizure in a canine pentylenetetrazole (PTZ)-induced seizure model. Five healthy Beagle dogs were used in this study. A 3% solution of PTZ was infused until the occurrence of generalized convulsion. Two separate experiments were conducted to observe changes in HIPs over short and long time periods. Blood sampling time points were divided into two periods as follows: short period (baseline, 30, 60, 90, and 120 min after seizure induction) and long period (baseline, 2, 6, 12, 24, and 48 h after seizure induction). The HIPs were calculated, and the serum prostaglandin E2 (PGE2) and leukotriene B4 (LTB4) concentrations were estimated using enzyme-linked immunosorbent assay. Significant changes (p < 0.05) in various HIPs were observed at different time point as follows: neutrophil × monocyte (90 min), neutrophil-to-lymphocyte ratio (60, 90, and 120 min), lymphocyte to monocyte ratio (60 min, 90 min, 120 min, 2 h, 12 h, and 24 h), platelet-to-albumin ratio (90 min), lymphocyte percentage × serum albumin concentration (LA; 60 min, 90 min, 120 min, 2 h), and neutrophil × platelet (6 h). LTB4 concentrations were significantly increased (p < 0.05) at 60 and 90 min, and 2, 6, and 48 h after seizure induction. PGE2 was significantly increased only 6 h after seizure induction (p < 0.05). LA was one of the HIPs that demonstrated a correlation with LTB4 concentration and showed significant changes that could be observed for a long-period (p < 0.05, r = -0.4194). The LA was the only HIP that reflected seizure-associated neuroinflammation. The 5-lipoxygenase pathway might be related to seizure-associated neuroinflammation.

Longitudinal Metabolomics Reveals Metabolic Dysregulation Dynamics in Patients with Severe COVID-19.

Background/Objective: A dysregulated metabolism has been studied as a key aspect of the COVID-19 pathophysiology, but its longitudinal progression in severe cases remains unclear. In this study, we aimed to investigate metabolic dysregulation over time in patients with severe COVID-19 requiring mechanical ventilation (MV). Methods: In this single-center, prospective, observational study, we obtained 236 serum samples from 118 adult patients on MV in an ICU. The metabolite measurements were performed using capillary electrophoresis Fourier transform mass spectrometry, and we categorized the sampling time points into three time zones to align them with the disease progression: time zone 1 (T1) (the hyperacute phase, days 1-3 post-MV initiation), T2 (the acute phase, days 4-14), and T3 (the chronic phase, days 15-30). Using volcano plots and enrichment pathway analyses, we identified the differential metabolites (DMs) and enriched pathways (EPs) between the survivors and non-survivors for each time zone. The DMs and EPs were further grouped into early-stage, late-stage, and consistent groups based on the time zones in which they were detected. Results: With the 566 annotated metabolites, we identified 38 DMs and 17 EPs as the early-stage group, which indicated enhanced energy production in glucose, amino acid, and fatty acid metabolisms in non-survivors. As the late-stage group, 84 DMs and 10 EPs showed upregulated sphingolipid, taurine, and tryptophan-kynurenine metabolisms with downregulated steroid hormone synthesis in non-survivors. Three DMs and 23 EPs in the consistent group showed more pronounced dysregulation in the dopamine and arachidonic acid metabolisms across all three time zones in non-survivors. Conclusions: This study elucidated the temporal differences in metabolic dysregulation between survivors and non-survivors of severe COVID-19, offering insights into its longitudinal progression and disease mechanisms.

Application of an attention bias test after surgical castration in piglets

Surgical castration of male piglets is a routine procedure performed to improve meat quality. Prior studies have shown that pain due to castration can last for up to 4 days, negatively impacting animal welfare, however the impact on affect, such as anxiety, is unclear. The objective of this study was to test the application of a novel attention bias test to assess anxiety in piglets that underwent surgical castration with or without practical pain alleviation methods. Piglets were surgically castrated (n = 22), castrated with analgesics (n = 21), or sham-handled (n = 22) at 3 days of age. An attention bias test was performed in week 1 (n = 32, 10–11/treatment) and 12 (n = 29, 9–10/treatment) to assess anxiety (an affective state), with feed (positive stimulus), loud bangs, and flashing lights (negative stimuli) presented simultaneously. Latency to visit the feeder, behavioral responses, and activity were recorded during the test. Additional data on piglets’ activity, tails, and grimaces were collected at three timepoints, at 1, 6, and 24 h after castration to determine pain experience. Piglets’ increased activity (P = 0.065), the fact that fewer piglets visited the feeder (P = 0.029), and tended to have longer latencies to visit the feeder (P = 0.092) in the attention bias test in week 1 might suggest that pain caused by surgical castration increased anxiety. No differences were detected in week 12. Castration treatment and sampling timepoint impacted activity levels hours after treatments, however, other measures were not impacted. These results suggest that more research is needed to determine how affect is impacted by pain caused by surgical castration. Activity and behavioral results somewhat support previous findings that surgical castration causes pain in piglets. As this is the first study assessing the relationship between painful procedures and piglet anxiety, more research is needed to determine a valid method to understand the impacts of these procedures on pig affect.

Transcriptome-based analysis reveals a role for PpCDF5 in the promotion of anthocyanin accumulation at a low nighttime temperature

For red pear, the anthocyanin content is a crucial factor determining the fruit skin color, which affects consumer preferences. Low overnight temperatures promote anthocyanin accumulation, but the molecular mechanism responsible is unclear. In this study, ‘Hongzaosu’ pear (Pyrus pyrifolia × Pyrus communis) fruit were treated with a low nighttime temperature (LNT, 16 °C) or a warm nighttime temperature (WNT, 26 °C), with sampling conducted within two diurnal cycles. The results showed that LNT promoted anthocyanin accumulation in the fruit skin. The structural anthocyanin biosynthetic genes PpCHS, PpF3H, and PpUFGT exhibited a rhythmic increase in expression at night under LNT. To examine the underlying mechanism, RNA sequencing was conducted using pear calli exposed to LNT and WNT for different durations (24, 48, 72, or 96 h). Transcriptome analysis revealed 285 differentially expressed genes (DEGs) common to all pairwise comparisons of LNT- and WNT-treated calli of ‘Clapp's Favorite’ (P. communis) at the sampling time points. KEGG pathway and gene ontology enrichment analyses indicated that the common DEGs were enriched in secondary metabolic processes and phenylpropanoid metabolic processes, which are associated with anthocyanin biosynthesis. The transcription factor PpCDF5, which was responsive to LNT, was selected for further study. Dual-luciferase assays showed that PpCDF5 activated the transcription of anthocyanin biosynthetic genes PpMYB10, PpCHS, PpF3H, PpDFR, PpANS, and PpUFGT. The yeast one-hybrid and EMSA assays demonstrated that PpCDF5 directly binds to the PpF3H promoter, which contains an AAAG motif. Overexpression of PpCDF5 in pear calli and transient overexpression in pear fruit both increased anthocyanin accumulation. The results indicate that PpCDF5 is involved in LNT-induced anthocyanin biosynthesis in pear fruit and provide insights into the molecular regulation of commercial fruit coloration.

Acute and Chronic Resistance Training, Acute Endurance Exercise, nor Physiologically Plausible Lactate In Vitro Affect Skeletal Muscle Lactylation.

We examined changes in skeletal muscle protein lactylation and acetylation in response to acute resistance exercise, chronic resistance training (RT), and a single endurance cycling bout. Additionally, we performed in vitro experiments to determine if different sodium lactate treatments affect myotube protein lactylation and acetylation. The acute and chronic RT study (12 college-aged participants) consisted of 10 weeks of unilateral leg extensor RT with vastus lateralis (VL) biopsies taken at baseline, 24 h following the first RT bout, and the morning of the last day of the RT bout. For the acute cycling study (9 college-aged participants), VL biopsies were obtained before, 2 h after, and 8 h after 60 min of cycling. For in vitro experiments, C2C12 myotubes were treated with varying levels of sodium lactate, including LOW (1 mM for 24 h), HIGH (10 mM for 24 h), and PULSE (10 mM for 30 min followed by 1 mM for 23.5-h). Neither acute nor chronic RT significantly affected nuclear or cytoplasmic protein lactylation. However, cytoplasmic protein acetylation was significantly reduced following one RT bout (-15%, p = 0.002) and chronic RT (-16%, p = 0.006). Cycling did not acutely alter post-exercise global protein lactylation or acetylation patterns. Lastly, varying 24 h lactate treatments did not alter nuclear or cytoplasmic protein lactylation or acetylation, cytoplasmic protein synthesis levels, or myotube diameters. These findings continue to support the idea that exercise induces more dynamic changes in skeletal muscle protein acetylation, but not lactylation. However, further human research with more sampling timepoints and a lactylomics approach are needed to determine if, at all, different exercise modalities affect skeletal muscle protein lactylation.

Abstract PR018: Plasma proteomic biomarkers reveal biological insights about the tumor microenvironment in melanoma patients after PD1 blockade

Abstract Most patients treated with immune checkpoint blockade (ICB) do not have durable treatment responses, stressing a critical need to identify early non-invasive biomarkers of response. Circulating biomarkers provide easy access for serial monitoring and can provide insight on the mechanisms of response to ICB. We performed plasma proteomics (2943 proteins) on 250 metastatic melanoma patients before and on ICB treatment and performed analysis to identify response- and time point-associated peripheral protein biomarkers. We next generated patient- matched peripheral blood lymphocyte (PBL) surface proteomic data to measure the proportions of immune cell subsets in circulation. We further obtained 13 tumor samples that were processed for single-cell RNA-sequencing (scRNA-seq). We fit linear mixed effects (LME) models to predict plasma protein abundances, using the sample time point, response status and interaction term between time point and response as covariates. We identified 397 proteins with a significant time point, response, or interaction effect. For proteins upregulated post-ICB, gene ontology analysis suggested these proteins were associated with inflammatory pathways involving the activation of effector immune functions. LME models fitted on the circulating immune cell proportions and parallel analysis of immune-related plasma proteins supported that post-ICB samples may be defined by increased immune activity, including T cell infiltration. Moreover, expression of genes corresponding to these post-ICB proteins in the tumor scRNA-seq data tended to be favored by immune subsets, namely those involved in immune cell recruitment and tumor reactivity, such as CXCL10+ macrophages and CXCL13+ T cells, respectively. On the other hand, expression of genes corresponding to plasma proteins upregulated in non-responders in the TME tended to be favored by suppressive myeloid subsets such as SPP1+ macrophages as well as malignant cells, suggesting the potential of immunosuppressive and pro-tumor biological processes to be involved in ICB resistance. Inference of cell-cell interactions in the TME further showed the involvement of these non-responder genes in potentially immunosuppressive and pro-tumor processes, including the PTPRC-MRC1 axis, involved in myeloid cell plasticity, the IGF1/2-IGF1R axis, involved in growth and survival of normal and neoplastic cells, and the GAS6-TYRO3 axis, associated with carcinogenetic mechanisms in multiple malignancies. We further identified a correlation between IL13 protein abundance in circulation and GAS6 expression in the tumor, suggesting that IL13 may reach the tumor from circulation to turn on the STAT6 pathway in receiving cells and upregulate GAS6 in turn to participate in this interaction with TYRO3. Together, these data represent one of the deepest peripheral biomarker studies using paired samples in melanoma patients treated with anti-PD1 therapy, and begin to elucidate the complex interplay between tumors and the systemic immune response within the host. Citation Format: Samuel J. Wright, Izabella Zamora, Milan Parikh, Deepika Yeramosu, Lynn Bi, Sarah Kang, Marijana Rucevic, Moshe Sade-Feldman, Baolin Liu, Ngan Nguyen, Jamey Guess, Alexis Schneider, David Lieb, Elliot Woods, William Michaud, Aleigha R. Lawless, Tatyana Sharova, Gyulnara Kasumova, Michelle S. Kim, Ryan J. Park, Russell W. Jenkins, Samuel J. Klempner, Ryan J. Sullivan, Keith T. Flaherty, Nir Hacohen, Genevieve M. Boland, Arnav Mehta. Plasma proteomic biomarkers reveal biological insights about the tumor microenvironment in melanoma patients after PD1 blockade [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr PR018.

Caspase-9 and p53 Protein Levels in Cancer Patients after Different Anesthesia Techniques

The aim of this study was to investigate the changes in caspase-9 and p53 levels as biomarkers of pro- and anti-apoptotic pathways in the early postoperative period in patients who underwent lung surgery for malig-nant tumors under different types of multimodal or inhalation-intravenous anesthesia. Material and Methods. A single-center prospective study of 22 patients aged 45–64 years was conducted at the Omsk Clinical Oncology Early Treatment and Prevention Center from January to April 2020. The participants were divided into two groups. Group 1 patients received multimodal anesthesia, which included sympathetic nerve block and prolonged epidural analgesia in the postoperative period. Group 2 patients received inhalational and intravenous anesthesia followed by systemic morphine analgesia. Serum caspase-9 and p53 protein levels were measured at four time points: before anesthesia, one, twelve, and twenty-four hours after surgery. Statistical hypotheses were tested using nonparametric (rank) analysis methods. Friedman's ANOVA was used to compare multiple time points, while the Wilcoxon test was used to compare variables between two time points in dependent samples. The Mann-Whitney test was used to assess differences between groups in independent samples. P-values &lt; 0.05 were considered statistically significant. Results are expressed as median ± half interquartile range (Me ± (LQ – UQ) / 2). Results. At time point 2, caspase-9 levels were significantly higher in group 2 patients than in group 1 (P = 0.045). There were no significant differences between the groups at any other time points. Conclusion. The lack of a significant difference in serum levels of caspase-9 and p53 protein at most time points between the groups demonstrates the efficacy of the anesthesia and analgesia methods used. Mean-while, a significantly higher level of caspase-9 one hour after surgery demonstrates a greater susceptibility of patients without sympathetic blockade to activation of the apoptotic cell death program.

Simulation of spatially varying non-stationary seismic waves using random frequencies and non-uniform fast fourier transform

The simulation of spatially varying non-stationary seismic waves is a crucial consideration for the seismic analysis of engineering structures. The traditional approach often incurs substantial computational expenses and suffers from low efficiency due to the extensive implementation of large-scale matrix factorizations at numerous sampling points in both time and frequency domains. In this study, an enhanced approach is developed for the efficient simulation of non-stationary seismic waves. The core of this approach lies in the utilization of random frequency sampling coupled with time-domain interpolation to reduce matrix factorizations, and non-uniform fast Fourier transform for expediting the summation of trigonometric functions. A parametric analysis is conducted to investigate the effect of vital parameters, including the decoupling strategy of time-frequency spectrum, the number of random frequencies, and the number of interpolation knots over time, on both the efficiency and accuracy of the enhanced approach. Consequently, the preferred values for these parameters are determined. By considering the simulation of seismic waves acting on a group of bridges, the enhanced approach is validated in the view of correlation functions. The results demonstrate a high level of fidelity of the simulated seismic waves and proves the effectiveness of the enhanced approach in the efficient simulation of spatially varying non-stationary earthquake ground motions. The developed approach can be used for the simulation of non-stationary seismic waves for a wide range of engineering structures.

Physiological effects of research handling on the northern elephant seal (Mirounga angustirostris)

Wildlife researchers must balance the need to safely capture and handle their study animals to sample tissues, collect morphological measurements, and attach dataloggers while ensuring their results are not confounded by stress artifacts caused by handling. To determine the physiological effects of research activities including chemical immobilization, transport, instrumentation with biologgers, and overnight holding on a model marine mammal species, we collected hormone, blood chemistry, hematology, and heart rate data from 19 juvenile northern elephant seals (Mirounga angustirostris) throughout a translocation experiment. Across our six sampling timepoints, cortisol and aldosterone data revealed a moderate hormonal stress response to handling accompanied by minor changes in hematocrit and blood glucose, but not ketone bodies or erythrocyte sedimentation rate. We also examined heart rate as a stress indicator and found that interval heart rate, standard deviation of heart rate, and apnea-eupnea cycles were influenced by handling. However, when seals were recaptured after several days at sea, all hormonal and hematological parameters had returned to baseline levels. Furthermore, 100 % of study animals were resighted in the wild post-translocation, with some individuals observed over four years later. Together, these findings suggest that while northern elephant seals exhibit measurable physiological stress in response to handling, they recover rapidly and show no observable long-term deleterious effects, making them a robust species for ecological and physiological research.

Transportation increases circulating corticosterone levels and decreases central serotonergic activity in a sex dependent manner in Pekin ducks

Previous studies from our lab suggest that transportation of early adulthood ducks can have long lasting physiological effects. To better understand how transportation affects the ducks’ physiology, we evaluated several central and peripheral parameters. Thirty-six, 23-week-old ducks were collected at a commercial breeder facility and randomly assigned to one of three treatment groups (n = 6/sex/treatment): 1) caught and euthanized (control), 2) caught and put in a crated in the pen for 90 min (crate), or 3) caught, crated, and transported in a truck for 90 min (transport) to simulate actual transportation. Blood was collected for serum corticosterone and blood smear analyses. Brains were hemisected and each half was dissected into three brain areas: caudal mesencephalon (CM), rostral mesencephalon (RM), and diencephalon (DI). Mass spectrometry was run on the right half of the brain, and gene expression of TPH1, TPH2, TH, CRH, and NPY were measured on the left half of brain using qRT-PCR. Serum corticosterone levels were increased (p = 0.01) in crated hens and in transported hens and drakes (p = 0.0084) when compared to control. HLR was increased (p = 0.035) in crated hens and transported hens and drakes compared to control. No differences in serotonin turnover were observed in drakes but increased in hens within the CM and RM from control to crate (p = 0.01) and crate to transport (p = 0.016). There were no differences in DA turnover or in gene expression for all brain areas for drakes and CM and RM for hens. Within the DI, hens showed a decrease (p = 0.03) in TPH1 for transport compared to crate. Overall, transportation elicits an acutely stressful event that increases corticosterone and HLR in a sex dependent manner where hens appear to be more reactive to the stressor than drakes. Our data supports that when assessing a stress response, care must be given to the sex of the bird and to the relative timepoint of sampling compared to the perceived onset of the stressor.

Constrained Pseudo-time Ordering for Clinical Transcriptomics Data.

Time series RNASeq studies can enable understanding of the dynamics of disease progression and treatment response in patients. They also provide information on biomarkers, activated and repressed pathways, and more. While useful, data from multiple patients is challenging to integrate due to the heterogeneity in treatment response among patients, and the small number of timepoints that are usually profiled. Due to the heterogeneity among patients, relying on the sampled time points to integrate data across individuals is challenging and does not lead to correct reconstruction of the response patterns. To address these challenges, we developed a new constrained based pseudotime ordering method for analyzing transcriptomics data in clinical and response studies. Our method allows the assignment of samples to their correct placement on the response curve while respecting the individual patient order. We use polynomials to represent gene expression over the duration of the study and an EM algorithm to determine parameters and locations. Application to three treatment response datasets shows that our method improves on prior methods and leads to accurate orderings that provide new biological insight on the disease and response. Code for the method is available at https://github.com/Sanofi-Public/ RDCS-bulkRNASeq-pseudo ordering.

Defining the optimal setting for transcriptomic analyses on blood samples for response prediction in immunotherapy-treated NSCLC patients

Transcriptomic profiling of blood immune cells offers a promising alternative to invasive, sampling bias-prone tissue-based biomarker assays for predicting immune checkpoint inhibitor (ICI) therapy response in non-small cell lung cancer (NSCLC) patients. However, the optimal analytical approach to identify systemic correlates of response still needs to be explored. We collected peripheral blood mononuclear cells and whole blood (WB) samples from 33 ICI-treated NSCLC patients before ICI treatment and at the first response evaluation. After bulk polyadenylated RNA-sequencing, we assessed differences in gene expression profiles between non-responders and responders using differential expression analysis, single sample gene set enrichment analysis (ssGSEA), and cell type deconvolution. We evaluated gene expression values, ssGSEA scores, and deconvolved cell type proportions to distinguish non-responders from responders via ROC curve (AUC) analysis, training a logistic regression classification model. Gene expression values and deconvolved proportions yielded the best results with WB samples after treatment (AUC = 0.87 and 0.85, respectively). Overall, ssGSEA scores showed superior classification performance across all sample types and timepoints (AUC > 0.7). In conclusion, transcriptomic analysis through ssGSEA demonstrated the best performance as a non-invasive biomarker for predicting clinical benefit in ICI-treated NSCLC patients, with gene expression and deconvolution on post-treatment WB samples also showing promising results.

Calcium propionate is an alkalizing agent in exercising Standardbreds

The objective of this study was to determine if calcium propionate is an alkalizing agent in exercising Standardbreds and if it alters plasma glucose and serum insulin concentrations. This study used a randomized crossover design to test the hypotheses that calcium propionate alters total CO2 (tCO2), Ca++, pH, strong ion difference (SID), glucose, and insulin in Standardbreds completing a simulated race test (SRT) on a high-speed equine treadmill. Blood was collected from eight horses (mean age ± SD = 16 ± 2.7 years; range = 13 - 21 years) at 10 and 5 min prior to treatment or control administration, just prior to the subsequent SRT, directly after the SRT, and at 60- and 90-min post SRT. Plasma pH and plasma concentrations of tCO2, glucose, Ca++, and Na+, K+, Cl-, Lac- (for SID calculation) were measured in duplicate by blood gas analyzer and serum insulin by radioimmunoassay. Data were analyzed by two-way repeated-measures ANOVA. For plasma pH, Na+, K+, Cl-, Lac-, SID, insulin and glucose, no effects of treatment or treatment by sampling-timepoint interaction were found (p>0.05). Plasma tCO2, Ca++, pH, Na+, SID, and glucose concentrations were significantly lower, and Lac- was significantly higher, directly after the SRT compared to all other timepoints (p<0.05). Plasma tCO2, HCO3- and Ca++ were significantly higher in treated than in control horses at multiple sampling timepoints after treatment/control administration (p<0.05). Serum insulin concentration, measured only pre and 30 min post treatment/control administration, was unaffected by treatment. Calcium propionate is an alkalizing agent in horses.

A systematic evidence map protocol for mapping global exposure to bisphenols and their alternatives and social and environmental justice implications

BackgroundBisphenol A (BPA) is one of the highest-volume chemicals produced worldwide, and human exposure to BPA is thought to be ubiquitous. Stricter regulations around the use of BPA have led many manufacturers to switch to other bisphenol chemicals with similar functions such as bisphenol S and F. Even though exposure to BPA, other bisphenol chemicals and bisphenol alternatives poses a health risk for humans, very little is known about the granular exposure levels of different populations around the world. AimThis systematic evidence map (SEM) will identify human studies reporting concentrations of bisphenols and their alternatives measured in human bio-samples with the aim to chart the global human exposure levels by country and population characteristics to identify research gaps and discuss any social and environmental injustice implications. Search strategy and eligibility criteriaMEDLINE®, Embase and Web of Science (WoS) databases as well as grey literature sources will be searched using predefined search strings. The database search results will be supplemented by backward and forward citation tracking on included studies. A scoping exercise conducted during planning identified 90 bisphenol chemicals and alternatives used in plastics. These include BPA, other bisphenol chemicals with/without similar functions to BPA as well as alternatives with similar functions to bisphenol chemicals. Eligible studies must measure concentrations of at least one relevant bisphenol chemical/alternative in human bio-samples. Study selectionOnly primary studies published in English since 2010 will be considered. The title, abstract and keywords will be screened by the DistillerAI tool and two independent reviewers. Grey literature will be screened by two reviewers for inclusion and exclusion. The full text of the included studies will then be screened by two independent reviewers. Study appraisalStudy quality will not be evaluated in this SEM. Data extraction and codingData extraction and coding will be performed by two independent reviewers. Parameters of interest will include the following: study characteristics (e.g., year of publication, sampling timepoints and study design), population information (e.g., country, age, sex, ethnicity, number of participants) and exposure information (sources of exposure, bio-sample analyzed, chemical name, concentration, and detection frequencies). Synthesis and visualizationThe results will be presented using a narrative summary, tables, bar plots and color-coded maps. The interactive database will be available on a dedicated freely accessible website. Systematic map protocol registry and registration numberThis protocol has been registered on Open Science Framework (OSF) and is available at https://doi.org/10.17605/OSF.IO/MNWTD.

Comparative Preclinical Pharmacokinetics and Disposition of Favipiravir Following Pulmonary and Oral Administration as Potential Adjunct Therapy Against Airborne RNA Viruses.

Favipiravir is administered orally, even against airborne RNA viruses, in a loading-dose/maintenance dose regimen. We investigated whether-(a) pulmonary delivery of favipiravir would generate high concentrations in the luminal side of the respiratory tract; and (b) avoiding first-pass metabolism by the liver by inhaled drug would generate comparable pharmacokinetics (PK) with doses significantly smaller than the oral maintenance dose. A dry powder inhalation (DPI) of favipiravir formulated by mixing with Inhalac 400® was prepared and characterized. Inhalations of ~ 120µg dose strength, with or without a prior oral loading dose were administered to mice. Comparator mice received human-equivalent oral doses (3mg). Three mice per sampling time point were sacrificed and favipiravir concentrations in the blood plasma, bronchio-alveolar lavage fluid (BALF) and lung tissue homogenate determined by HPLC. One-compartment PK modeling of concentration-time data indicated that the area under the curve (AUC0-24h) generated in the BALF recovered from mice receiving inhalations of ~ 1/25th of the oral dose subsequent to an oral loading dose was 86.72 ± 4.48µg⋅mL-1⋅h. This was consistently higher than the AUC observed in the BALF of orally-dosed mice (56.71 ± 53.89µgmL-1⋅h). In blood serum, the respective values of AUC were 321.55 ± 124.91 and 354.71 ± 99.60µg⋅mL-1⋅h. Pulmonary delivery of significantly smaller doses of favipiravir generates meaningful drug disposition and pharmacokinetics at the site of respiratory viral infections. We provide the rationale for designing a self-administered, non-invasive, low-cost, targeted drug delivery system against airborne RNA virus infection.