GC-MS-based metabonomic analysis of silkworm haemolymph reveals four-stage metabolic responses to nucleopolyhedrovirus infection.

Silkworm, Bombyx mori, an economically significant insect, plays a crucial role in silk production. However, silkworm breeding is highly susceptible to various pathogens, particularly the Bombyx mori nucleopolyhedrovirus (BmNPV), which poses a serious threat. Recent metabonomic studies have provided insights into the metabolic changes associated with BmNPV infection. BmNPV infection has obvious temporal characteristics. However, few studies have investigated the silkworms infected in different periods. This study employed gas chromatography-mass spectrometry (GC-MS) to perform a comprehensive analysis of haemolymph metabolites in silkworms at 48, 72, 96 and 120 h post-infection (h.p.i.). Through the integration of time-course analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment, the study revealed distinct four-stage metabolic characteristics in the silkworm's response to BmNPV infection. At Stage 1 (48 h.p.i.), silkworms activate antioxidant defence mechanisms, with significant enrichment in metabolic pathways involving key antioxidants such as glutathione, to mitigate oxidative stress induced by viral invasion. By Stage 2 (72 h.p.i.), pathways related to amino acid metabolism and protein synthesis become active, indicating an increase in protein synthesis. In Stage 3 (96 h.p.i.), energy metabolism and substance transport pathways are significantly upregulated to support the rapid viral replication and the enhanced locomotor behaviour of silkworm. Finally, at Stage 4 (120 h.p.i.), there is a further enhancement of pathways related to energy metabolism, nucleic acid synthesis, and substance transport, which align with peak viral assembly and release. These findings contribute to an in-depth understanding of the biochemical basis of silkworm resistance to NPV.

The 24-hour molecular landscape after exercise in humans reveals MYC is sufficient for muscle growth.

A detailed understanding of molecular responses to a hypertrophic stimulus in skeletal muscle leads to therapeutic advances aimed at promoting muscle mass. To decode the molecular factors regulating skeletal muscle mass, we utilized a 24-h time course of human muscle biopsies after a bout of resistance exercise. Our findings indicate: (1) the DNA methylome response at 30 min corresponds to upregulated genes at 3 h, (2) a burst of translation- and transcription-initiation factor-coding transcripts occurs between 3 and 8 h, (3) changes to global protein-coding gene expression peaks at 8 h, (4) ribosome-related genes dominate the mRNA landscape between 8 and 24 h, (5) methylation-regulated MYC is a highly influential transcription factor throughout recovery. To test whether MYC is sufficient for hypertrophy, we periodically pulse MYC in skeletal muscle over 4 weeks. Transient MYC increases muscle mass and fiber size in the soleus of adult mice. We present a temporally resolved resource for understanding molecular adaptations to resistance exercise in muscle ( http://data.myoanalytics.com ) and suggest that controlled MYC doses influence the exercise-related hypertrophic transcriptional landscape.

DEWNA: dynamic entropy weight network analysis and its application to the DNA-binding proteome in A549 cells with cisplatin-induced damage.

Cisplatin is one of the most commonly used chemotherapy drugs for treating solid tumors. As a genotoxic agent, cisplatin binds to DNA and forms platinum-DNA adducts that cause DNA damage and activate a series of signaling pathways mediated by various DNA-binding proteins (DBPs), ultimately leading to cell death. Therefore, DBPs play crucial roles in the cellular response to cisplatin and in determining cell fate. However, systematic studies of DBPs responding to cisplatin damage and their temporal dynamics are still lacking. To address this, we developed a novel and user-friendly stand-alone software, DEWNA, designed for dynamic entropy weight network analysis to reveal the dynamic changes of DBPs and their functions. DEWNA utilizes the entropy weight method, multiscale embedded gene co-expression network analysis and generalized reporter score-based analysis to process time-course proteome expression data, helping scientists identify protein hubs and pathway entropy profiles during disease progression. We applied DEWNA to a dataset of DBPs from A549 cells responding to cisplatin-induced damage across 8 time points, with data generated by data-independent acquisition mass spectrometry (DIA-MS). The results demonstrate that DEWNA can effectively identify protein hubs and associated pathways that are significantly altered in response to cisplatin-induced DNA damage, and offer a comprehensive view of how different pathways interact and respond dynamically over time to cisplatin treatment. Notably, we observed the dynamic activation of distinct DNA repair pathways and cell death mechanisms during the drug treatment time course, providing new insights into the molecular mechanisms underlying the cellular response to DNA damage.

Effect of cumulative uric acid to high-density lipoprotein cholesterol ratio on myocardial infarction in prospective cohorts.

OBJECTIVE: This study aimed to investigate the effect of the ratio of cumUHR on MI, based on the hypothesis that higher exposure to the ratio of cumUHR is associated with a higher risk of MI. METHODS: Participants who underwent three examinations between 2006 and 2010 were selected. The cumUHR from baseline to the third check was calculated, multiplying the mean between consecutive checks by the time interval between visits. The association between cumUHR and MI and its progression was evaluated by Cox proportional hazards regression model. The cumulative incidence of endpoint events between cumUHR groups was compared using a log-rank test. Stratification by age, sex, and BMI was further performed. RESULTS: A total of 53,697 people, with an average age of 53.08 years, 78 % of whom were male, with a median follow-up of 10.51 years and 744 myocardial infarction events, were enrolled. The highest cumUHR quartile, MI, had the highest cumulative incidence (log-rank P < 0.01). Multivariate COX regression analysis showed that in the fully adjusted model, there was a high level of concentration in the highest cumUHR quartile (HR, 1.52; 95 % CI, 1.20-1.92) and participants with longer duration of high UHR exposure (HR, 1.55; 95 % CI, 1.22-1.97). CONCLUSIONS: The risk of MI increases with cumUHR and is influenced by the time course of cumUHR. In particular, in people aged ≥ 60 years, males, and BMI < 28 kg/m2, the risk of MI is more affected by the level of UHR, and more attention should be paid to controlling the level of UHR.

Untargeted metabolomic analysis reveals a time-course hepatic metabolism disorder induced by short-term 6PPD exposure in rats.

The tire antioxidant 6PPD has garnered extensive attention due to its widespread presence in the environment and the harmful effects of its transformation products on aquatic organisms. 6PPD has been detected in airborne dust, and it can enter mammals through inhalation exposure. While the toxic effects of 6PPD exposure have been reported in mammals, its effects on hepatic metabolism still remain poorly understood. Here, we collected the serum and liver samples at 1, 6, and 72 h following a single oral exposure of 100 mg/kg body weight (bw) 6PPD, respectively. We also investigated changes in serum and hepatic physiological indicators and metabolites, correspondingly. Results indicated that single time oral exposure a high dose of 6PPD did not significantly affect the physiological indexes of rats within a short time frame. However, untargeted metabolomics analysis of the metabolites in the liver at 1, 6, and 72 h revealed that the number of differential expression metabolites gradually increased over time and the most affected substances were lipids and lipid-like molecules. Interestingly, the KEGG pathway enrichment analysis indicated that 6PPD disrupted the riboflavin metabolism, leading to a significant decrease in FMN levels at all time points. In addition, the hepatic glucose metabolism was significantly affected at 6 and 72 h after oral administration. Taken together, short-term exposure to 6PPD disturbed lipid and riboflavin metabolism and gradually affected glucose metabolism in the liver of rats. These findings revealed the impacts of 6PPD on the hepatic metabolism in animals, and also offered some important insights into its toxicology and health risk.

Sympathetic reinnervation in cardiac transplant recipients: Prevalence, time course and association with long-term survival.

BACKGROUND: Partial cardiac sympathetic reinnervation after cardiac transplant has been extensively investigated and evidenced. However, there have been no large-scale, long-term studies evaluating the prevalence, time-course, and association with long-term survival of sympathetic reinnervation of the heart. METHODS: Cardiac transplant recipients (n=232) were recruited from outpatient clinic at a single transplant centre in the United Kingdom. Participants were each tested once for the presence of sympathetic reinnervation of the sinus node using the low frequency component of power spectral analysis of heart rate variability, with a cut-off defined as 2 standard deviations above the mean for denervated participants (those tested <56 days post-transplant). Time-course was calculated based on the timing of testing post-transplant. Patients were then followed-up over a period of up to 27 years after transplant for survival analysis. RESULTS: The overall prevalence of cardiac sympathetic reinnervation in the 225 patients tested >56 days post-transplant was 64.9%. Sympathetic reinnervation primarily occurred in the first 18 months after transplant, with a plateau thereafter. The prevalence in participants tested >18 months post-transplant was 69.6%. In Kaplan-Meier survival analysis, sympathetic reinnervation was associated with significantly improved survival (Log-rank P=0.019), with a median survival time for reinnervated patients of 19.9 years compared to 14.4 years for the denervated group. CONCLUSIONS: Sympathetic reinnervation of the sinus node occurs mostly within 18 months of transplant, is found in 70% of cardiac transplant recipients tested >18 months post-transplant, and is associated with significantly improved long-term survival.

Dimensions of early life adversity and cognitive processing of emotion in youth.

BACKGROUND: Early life adversity (ELA) is a leading risk factor for psychopathology. The Dimensional Model of Adversity and Psychopathology (DMAP) elucidates processes altered by ELA and central to this association. Specifically, DMAP posits early experiences of deprivation alter cognitive and emotional processes in ways distinct from early experiences of threat. While evidence suggests that deprivation and threat predict alterations in cognitive and emotional processes, respectively, the influence of these dimensions on cognitive processing across emotionally valenced material remains unexamined. OBJECTIVE: This work is the first to investigate associations of deprivation versus threat on cognitive processing of multiple emotions (happy, sad, angry, and neutral facial expressions) and the time course of processing in a sample of youth. PARTICIPANTS AND SETTING: Eighty-two youth (48.80 % female, Mage = 12.85) were recruited from Vancouver. METHODS: Deprivation and threat were measured using the Traumatic Events Screening Inventory for Children (TESI-C), an interview-based measure assessing the instance and severity of 30+ experiences of ELA. Cognitive processing was measured using the Affective Posner Task, which assesses attentional biases and raw reaction times for happy, sad, angry, and neutral facial expressions. RESULTS: Interestingly, experiences of deprivation were associated with early attentional processing deficits regardless of valence, rs ≥ 0.22, ps ≤ 0.046, whereas experiences of threat were associated with late attentional biases for emotional material, Bs ≥ |4.15|, ps ≤ 0.036. CONCLUSIONS: Findings advance theoretical models of ELA by elucidating the nature and time course of cognitive and emotional alterations following deprivation and threat, and, if replicated, suggest the importance of cognitive processing of emotion in early interventions.

Time-course whole blood transcriptome profiling provides new insights into Microtus fortis natural resistance mechanism to Schistosoma japonicum.

Microtus fortis is known as a non-susceptible animal host of S. japonicum. A better understanding of this animal immune defense mechanism during the early stage of infection may offer an alternative route for vaccine development or therapy. Here, we analyzed the whole blood transcriptome of M. fortis using next-generation sequencing (NGS) to identify immune genes of biological relevance that might be involved in the mechanism of its resistance. The blood samples were collected from uninfected animals (control group) and infected animals at different time points (3, 7, 10 and 14 days post-infection). We identified 5310 sequences as unigenes and successfully annotated 4636 of them. The immune response was more intense at 10 dpi. The upregulated genes at this time point were mainly activated in the TNF and NF-kappa B signaling pathways, Th1, Th2and Th17 cell differentiation as well as cytokine-cytokine receptor interaction. Based on the differentially expressed genes analysis, we report that the IF27L2B, RETN, PGRP, IFI35, TYROBP, S100A8, S100A11, CD162, CD88, CYBA, and LBP could play important roles in the mechanism of M. fortis resistance.

TimeNorm: a novel normalization method for time course microbiome data.

Metagenomic time-course studies provide valuable insights into the dynamics of microbial systems and have become increasingly popular alongside the reduction in costs of next-generation sequencing technologies. Normalization is a common but critical preprocessing step before proceeding with downstream analysis. To the best of our knowledge, currently there is no reported method to appropriately normalize microbial time-series data. We propose TimeNorm, a novel normalization method that considers the compositional property and time dependency in time-course microbiome data. It is the first method designed for normalizing time-series data within the same time point (intra-time normalization) and across time points (bridge normalization), separately. Intra-time normalization normalizes microbial samples under the same condition based on common dominant features. Bridge normalization detects and utilizes a group of most stable features across two adjacent time points for normalization. Through comprehensive simulation studies and application to a real study, we demonstrate that TimeNorm outperforms existing normalization methods and boosts the power of downstream differential abundance analysis.

Paving the way to a neural fate - RNA signatures in naive and trans-differentiating mesenchymal stem cells.

Mesenchymal Stem Cells (MSCs) derived from the embryonic mesoderm persist as a viable source of multipotent cells in adults and have a crucial role in tissue repair. One of the most promising aspects of MSCs is their ability to trans-differentiate into cell types outside of the mesodermal lineage, such as neurons. This characteristic positions MSCs as potential therapeutic tools for neurological disorders. However, the definition of a clear MSC signature is an ongoing topic of debate. Likewise, there is still a significant knowledge gap about functional alterations of MSCs during their transition to a neural fate. In this study, our focus is on the dynamic expression of RNA in MSCs as they undergo trans-differentiation compared to undifferentiated MSCs. To track and correlate changes in cellular signaling, we conducted high-throughput RNA expression profiling during the early time-course of human MSC neurogenic trans-differentiation. The expression of synapse maturation markers, including NLGN2 and NPTX1, increased during the first 24 h. The expression of neuron differentiation markers, such as GAP43 strongly increased during 48 h of trans-differentiation. Neural stem cell marker NES and neuron differentiation marker, including TUBB3 and ENO1, were highly expressed in mesenchymal stem cells and remained so during trans-differentiation. Pathways analyses revealed early changes in MSCs signaling that can be linked to the acquisition of neuronal features. Furthermore, we identified microRNAs (miRNAs) as potential drivers of the cellular trans-differentiation process. We also determined potential risk factors related to the neural trans-differentiation process. These factors include the persistence of stemness features and the expression of factors involved in neurofunctional abnormalities and tumorigenic processes. In conclusion, our findings contribute valuable insights into the intricate landscape of MSCs during neural trans-differentiation. These insights can pave the way for the development of safer treatments of neurological disorders.

Pupillary dilation response to the auditory food words in adolescents with obesity without binge eating disorder.

Childhood obesity is a growing global public health problem. Studies suggest that environmental cues contribute to developing and maintaining obesity. We aimed to evaluate pupillary changes to auditory food words vs. nonfood words and to conduct a dynamic temporal analysis of pupil size changes in adolescents with obesity without binge eating disorder by comparing healthy-weight adolescents. In this study, a total of 63 adolescents aged 12-18 years (n = 32, obesity group (OG); n = 31, control group (CG)) were included. In an auditory paradigm, participants were presented with a series of high and low-calorie food and nonfood words. A binocular remote eye-tracking device was used to measure pupil diameter. Generalized additive mixed models (GAMMs) were used for dynamic temporal analysis of pupillometry data. The results of GAMM analysis indicated that CG had larger pupil dilation than the OG while listening to auditory food words. CG had larger pupil dilation in food words than in nonfood words. However, the OG had a similar pupillary response in food and nonfood words. Pupil dilation response to higher-calorie foods was extended over the later stages of the time period (after 2000 ms) in the OG. In summary, our findings indicated that individuals with obesity had lower pupil dilation to auditory food words compared to normal-weight peers. Adolescents with obesity had prolonged pupillary dilation in higher calories of food words. The individual psychological factors affecting the dynamic changes of pupil responses to food cues in adolescents with obesity should be examined in further studies.

Investigation into temporal changes in the human bloodstain lipidome.

Bloodstains are crucial pieces of physical evidences found at violent crime scenes, providing valuable information for reconstructing forensic cases. However, there is limited data on how bloodstain lipidomes change over time after deposition. Hence, we deployed a high-throughput high-performance liquid chromatography-mass spectrometry (HPLC-MS) approach to construct lipidomic atlases of bloodstains, whole blood, plasma, and blood cells from 15 healthy adults. A time-course analysis was also performed on bloodstains deposited for up to 6 months at room temperature (~ 25°C). The molecular levels of 60 out of 400 detected lipid species differed dramatically between bloodstain and whole blood samples, with major disturbances observed in membrane glycerophospholipids. More than half of these lipids were prevalent in the cellular and plasmic fractions; approximately 27% and 10% of the identified lipids were uniquely derived from blood cells and plasma, respectively. Furthermore, a subset of 65 temporally dynamic lipid species arose across the 6-month room-temperature deposition period, with decreased triacylglycerols (TAGs) and increased lysophosphatidylcholines (LPCs) as representatives, accounting for approximately 8% of the total investigated lipids. The instability of lipids increased linearly with time, with the most variability observed in the first 10 days. This study sheds light on the impact of air-drying bloodstains on blood components at room temperature and provides a list of potential bloodstain lipid markers for determining the age of bloodstains.

Description of a non-competitive ELISA based on time course analysis of ligand binding at saturation, and a direct method for calculating the affinity of monoclonal antibodies.

We present a time-course saturation ELISA for measuring the equilibrium constant of the monoclonal antibody (mAb) SIM 28 against horse radish peroxidase (HRP). The curves of HRP binding to a series of fixed mAb dilutions were plotted to completion, and the Kt (= Ks) value (time to occupy 50 % of the mAb paratopes) was determined for each mAb dilution and HRP concentration. Analysis of the kinetic mechanism of the reaction by Lineweaver-Burk and Hanes plots showed that the slope and y-intercept were affected, indicating that mAb ligand saturation follows non-competitive inhibition kinetics in this assay format. In this kinetics, the inhibition constant Ki (= Kd) is the time required to double the slope or halve the Vmax of the Lineweaver-Burk plot. The Kt values of the time courses were doubled (2 x Kt) and normalized by dividing by the total reaction time to obtain a unitless factor which, when multiplied by the concentration of HRP, gives the Ki. The affinity constant of mAb SIM 28 was determined from ELISA data (n = 16) by three methods: i) doubling of Kt, ii) Beatty equation (Kaff = (n-1)/2 (n [HRP']t - [HRP]t), and iii) SPR (Biacore) analysis. The calculated affinities (mean ± 95 % confidence limits) were i) 4.6 ± 0.67 × 10-9 M, ii) Kaff = 0.23 ± 0.03 × 109 M-1 (Kd = 4.8 ± 0.81 × 10-9 M), and iii) 4.3 ± 0.57 × 10-9 M, respectively. The similar results obtained with the three different techniques indicate that this time-course saturation ELISA, combined with the double Kt method, is a repeatable and direct approach to mAb affinity determination.

Time course of remnant cholesterol and the risk of heart failure.

BACKGROUND AND AIM: Previous studies have shown that remnant cholesterol (RC) was associated with heart failure (HF). However, lack of evidence regarding the long-term trend of RC with HF risk. We aimed to investigate the association between cumulative RC exposure with incident HF and to further explore the modulating effects of the time course of RC accumulation. METHODS AND RESULTS: We enrolled 41,168 participants free of CVD from the Kailuan Study who completed the first three health examinations from 2006 to 2010. Cumulative RC exposure included cumulative RC and time-weighted cumulative RC. The combination of cumulative RC and RC slope over time was characterized as the time course of RC accumulation. Multivariable adjusted Cox proportional hazards models were used to calculate hazard ratios (HRs) and 95% confidence intervals (CIs) for HF risk. We also considered non-HF-related death as a competing event by performing competing risk models as a sensitivity analysis. During 8.84 years, 839 participants developed HF events. Cumulative RC exposure increased the HF risk, with HRs for cumulative RC of 1.72 (1.41-2.10) and for time-averaged cumulative RC of 1.54 (1.25-1.89). There was a nonlinear relationship between cumulative RC exposure and HF risk. Participants with higher cumulative RC and negative slope had the highest HF risk (HR, 1.46; 95% CI, 1.16-1.83). CONCLUSIONS: Both cumulative long-term exposure and the time course of RC accumulation were associated with HF risk. Early RC accumulation resulted in a greater increase in risk compared to later accumulation. This finding suggests that long-term exposure to RC may be useful in identifying individuals at high risk of developing HF and highlights the need for early initiation of appropriate RC control to prevent or reduce incident HF.

Time course and neural locus of the Flashed Face Distortion Effect.

Viewing a rapid sequence of face images shown in the periphery can lead to large caricature-like distortions in the perceived images, a phenomenon known as the Flashed Face Distortion Effect (FFDE). The mechanisms underlying FFDE are poorly understood. Here we examined the timing and sites of the adaptation processes giving rise to the FFDE. To investigate the effects of presentation rate, we maintained consistent trial lengths while assessing how variations in the temporal frequencies of face presentation influenced the magnitude of face distortion and the averaging of facial expressions. Over a wide range of temporal frequencies (1.2-60 Hz) tested, we observed a decrease in FFDE strength as the presentation rate increased. To probe the neural sites of FFDE, we varied whether successive faces were presented to the same or different eyes using a dichoptic display. Distortion effects were comparable for monocular, binocular, and interocular conditions, yet much larger than a control condition where faces were presented with a temporal interval between successive images, suggesting a cortical locus for FFDE.

Effect of Phase Angle on Change in Quadriceps Muscle Strength 1 Year After Total Knee Arthroplasty: The Screening for People Suffering Sarcopenia in Orthopedic Cohort of Kobe Study.

BACKGROUND: Phase angle (PhA) has been reported to be associated with quadriceps strength in patients who have knee osteoarthritis and were scheduled for total knee arthroplasty (TKA). The PhA can also be expected to predict the time course of quadriceps muscle strength loss and recovery. We aimed to investigate the relationship between the preoperative PhA and the time course of quadriceps muscle strength change in patients undergoing TKA. METHODS: A prospective cohort study was conducted on patients scheduled for primary unilateral TKA. A total of 855 patents were included in the analysis. The PhA and quadriceps muscle strength of the operated knee were measured preoperatively and at 3, 6, and 12 months postoperatively. To analyze the effect of the preoperative PhA on the change in postoperative quadriceps muscle strength, a linear mixed model with the quadriceps muscle strength as a dependent variable with the preoperative PhA, evaluation period (dummy variable), and their product terms as independent variables was conducted after adjusting for preoperative covariates. RESULTS: A statistically significant negative effect was present for a higher PhA, resulting in a greater decrease in quadriceps muscle strength between preoperative and 3 months postoperative (P = 0.012). In contrast, the effect was not statistically significant between 3 and 6 months postoperatively (P = 0.17). However, a statistically significant positive effect for a higher PhA resulting in a greater increase in quadriceps muscle strength was present between 6 and 12 months postoperatively (P = 0.027). CONCLUSIONS: Preoperative PhA is a useful predictor of the quadriceps muscle strength change after TKA. These findings suggest that evaluating the preoperative PhA could aid in the development of targeted rehabilitation programs aimed at optimizing quadriceps muscle function in patients undergoing TKA.

Integrating multiregulatory analysis reveals the negative regulatory function of miR482a in the response of poplar to canker pathogen infection.

Canker disease caused by the bacterium Lonsdalea populi is one of the most destructive diseases affecting poplar stems. However, the detailed stress response mechanisms of poplar have not been widely characterized. To explore the diverse regulatory RNA landscape and the function of key regulators in poplar subjected to L. populi stress, we integrated time-course experiment with mock-inoculation (CK) and inoculation (IN) with L. populi at the first, third, and sixth day (IN1, IN3, IN6) on Populus × euramericana cv. '74/76' (107), small RNA-seq, whole transcriptome-wide analysis, degradome analysis and transgenic experiments. A total of 98 differentially expressed (DE) miRNA, 17 974 DEmRNA, and 807 DElncRNA were identified in poplar infected by L. populi, presenting dynamic changes over the infection course. Regulatory networks among RNAs were further constructed. Notably, a network centered on ptc-miR482a in CK-vs-IN3 contained most DEGs. We show that miR482a and miR1448 are located in one transcript as a polycistron. Overexpression of pre-miR482a-miR1448 (OX482-1448) and pre-miR482a (OX482) increased poplar susceptibility to canker pathogen with reduced accumulation of reactive oxygen species, while the suppression of miR482a (STTM482) conferred poplar disease resistance. PHA7 was validated as the target of miR482a with degradome sequencing and tobacco transient co-transformation, its expression being downregulated in OX482-1448 and OX482 lines. Additionally, a series of phasiRNAs were triggered by miR482a targeting PHA7, forming regulatory cascades with more RLP, NBS-LRR, and PK genes, further verifying the defense function of miR482a. These findings provide insights for understanding the roles of ncRNAs and regulatory networks involved in poplar immunity.

The timing of the mouse hind paw incision does not influence postsurgical pain.

Chronobiological approaches have emerged as tools to study pain and inflammation. Although time-of-day effects on the expression of pain after injury have been studied, it remains unaddressed whether the timing of the injury itself can alter subsequent pain behaviors. The aim of this study was to assess postsurgical pain behaviors in a mouse hind paw incision assay in a circadian-dependent manner. Incisions were made at one of four equally spaced time points over a 24-hour period, with evoked and spontaneous pain behaviors measured using the von Frey mechanical sensitivity test, Hargreaves' radiant heat paw-withdrawal test, and the Mouse Grimace Scale. Algesiometric testing was performed in C57BL/6 mice prior to and at multiple time points after incision injury, at the same time of day, until pain resolution. No statistically significant differences were observed between groups. This study adds to the literature on circadian rhythms and their influence on pain in the pursuit of more biologically informed pre- and postoperative care.

Investigating the kinetics of marine and terrestrial organic carbon incorporation and degradation in coastal bulk sediment and water settings through isotopic lenses.

Coastal sediments are the main deposition center for allochthonous and autochthonous organic carbon (OC). The discharge of terrestrial biomass, anthropogenic activities, oceanic primary productivity, and natural events contribute to this carbon pool. The OC buried in sediments undergoes alteration through physical, biological and chemical processes, becoming progressively refractory and more likely to be preserved on geological time scales. However, little is known about the rate of bulk OC alteration post weathering and bloom. We incubated coastal sediment slurries with isotopically distinct spikes of C4 corn leaves and cultured phytoplankton, individually and in 1:1 mixture. OC isotopic values and concentrations were probed at different time points to track degradation and incorporation in solid and liquid phases. Both amendments were composed of fresh OC with a high proportion of labile biochemicals (e.g. polysaccharides and proteins). Despite the small differences in their lability, corn leaves were incorporated into the sediments at a slower rate compared to phytoplankton. Following combined spiking of the terrestrial and marine amendments, no sign of synergistic effects was observed in system's response. Despite sediment sensitivity to OC input and the rapid alterations in its properties within the initial days of incubation, swiftly transitioning to a state of minimal change is indicative of a relatively stable system that retained the isotopic imprint of the OC spike for a long time (> 32 days). This isotopic remanence is likely due to heterotrophic bacteria that degrade OC to synthesize their biomass (food stock for successive generations) and incorporate its stable isotope characteristics. Hence, our work sheds light on the kinetics of biogeochemical changes, and recovery time of the system for returning to its pre-perturbation state.

Time-Course Analysis and Transcriptomic Identification of a Group III ERF CmTINY2 Involved in Waterlogging Tolerance in Chrysanthemums × morifolium Ramat.

'Hangju' is a variety of Chrysanthemum × morifolium Ramat. with both edible and medicinal value, cultivated as a traditional Chinese medicine for four centuries. The cultivation of 'Hangju' is currently at risk due to waterlogging, yet there is a lack of comprehensive understanding regarding its response to waterlogging stress. This study compared the waterlogging-tolerant 'Hangju' variety Enhanced Waterlogging Tolerance (EWT) with the waterlogging-sensitive variety CK ('zaoxiaoyangju'). EWT exhibited a more developed aeration tissue structure and demonstrated rapid growth regarding the adventitious roots following waterlogging. The time-course transcriptome analysis indicated that EWT could swiftly adjust the expression of the genes involved in the energy metabolism signaling pathways to acclimate to the waterlogged environment. Through WGCNA analysis, we identified Integrase-Type DNA-Binding Protein (CmTINY2) as a key factor in regulating the waterlogging tolerance in EWT. CmTINY2, a transcription factor belonging to the ethylene-responsive factor (ERF) subfamily III, operated within the nucleus and activated downstream gene expression. Its role in enhancing the waterlogging tolerance might be linked to the control of the stomatal aperture via the Ethylene-Responsive Element (ERE) gene. In summary, our research elucidated that the waterlogging tolerance displayed by EWT is a result of a combination of the morphological structure and molecular regulatory mechanisms. Furthermore, the study of the functions of CmTINY2 from ERF subfamily III also broadened our knowledge of the role of the ERF genes in the waterlogging signaling pathways.