Dynamic Changes Research Articles

Application of time series analysis in predicting postpartum depression: integrating data from the hospitalization period and early postpartum weeks.

This study aimed to explore the dynamic changes in postpartum depressive symptoms from the hospitalization period to 4-8 weeks postpartum using time series analysis techniques. By integrating depressive scores from the hospital stay and the early postpartum weeks, we sought to develop a predictive model to enhance early identification and intervention strategies for Postpartum Depression (PPD). A longitudinal design was employed, analyzing Edinburgh Postnatal Depression Scale (EPDS) scores from 1,287 postpartum women during hospitalization and at 4, 6, and 8 weeks postpartum. Descriptive statistics summarized demographic characteristics. Time Series Analysis using the Auto-Regressive Integrated Moving Average (ARIMA) model explored temporal trends and seasonal variations in EPDS scores. Correlation analysis examined the relationships between EPDS scores and demographic characteristics. Model validation was conducted using a separate dataset. EPDS scores significantly increased from the hospitalization period to 4-8 weeks postpartum (p < .001). The ARIMA model revealed seasonal and trend variations, with higher depressive scores in the winter months. The model's fit indices (AIC = 765.47; BIC = 774.58) indicated a good fit. The Moving Average (MA) coefficient was - 0.69 (p < .001), suggesting significant negative impacts from previous periods' errors. Monitoring postpartum depressive symptoms dynamically was crucial, particularly during the 4-8 weeks postpartum. The seasonal trend of higher depressive scores in winter underscored the need for tailored interventions. Further research using longitudinal and multi-center designs was warranted to validate and extend these findings. Our predictive model aimed to enhance early identification and intervention strategies, contributing to better maternal and infant health outcomes.

7690 Acute hormonal signaling-induced 3D genome organization controls transcriptional dynamics during adipocyte thermogenesis

Abstract Disclosure: Y. Zhang: None. R. Zheng: None. T. Tsuji: None. Y. Xing: None. C. Wang: None. J. Darcy: None. K. Chen: None. Y. Tseng: None. Brown adipose tissue (BAT) maintains body temperature by dissipating energy as heat, presenting a promising target against obesity and its associated metabolic disorders. When exposed to cold, the β3-adrenergic receptor (β3-AR) in brown adipocytes is stimulated, initiating a sequential signaling cascade and activating a thermogenic gene program essential for heat generation. Despite remarkable progress has been made towards understanding the engagement of transcriptional and epigenetic regulators during thermogenesis in response to β3-AR stimulation, the precise impact of three-dimensional (3D) genome organization, an increasingly important modulator of gene expression, on the activation of thermogenic gene program remains elusive. In this study, we employed high-resolution Micro-C in murine brown adipocytes to interrogate the rapidity of 3D genome reorganization in response to β3-AR hormonal signaling and its impact on transcriptional dynamics during thermogenesis. Our findings unveiled substantial rewiring of chromatin loops within 4 hours of β3-AR stimulation. These dynamic changes in chromatin loops, particularly enhancer-promoter (E-P) interactions, were coupled with gene activation during thermogenesis. Furthermore, we uncovered the mechanisms by which β3-AR hormonal signaling mediates 3D chromatin reorganization to regulate the thermogenic gene program. We found that β3-AR hormonal signaling induced the phosphorylation of p18Hamlet, a subunit of the SRCAP complex, facilitating the deposition of the histone variant H2A.Z into regulatory DNA regions. This nucleosome incorporation of H2A.Z further mediated nucleosome rearrangement, favoring the formation of nucleosome-depleted regions (NDRs) and enhancing the accessibility of regulatory DNA regions. As a result, H2A.Z chromatin incorporation facilitated interactions between cis-regulatory regions and gene promoters, such as E-P interactions, thereby activating thermogenic gene expression in response to β3-AR stimulation. Loss of H2A.Z disrupted loop dynamics, impeding gene activation and BAT thermogenic capacity. The conserved role of H2A.Z in human thermogenesis was validated, and we identified adiposity-related SNPs in human homologous loop anchors that could potentially be targeted for treating obesity. Taken together, our results unveil a previously underappreciated aspect of acute hormonal signaling-induced chromatin dynamics in orchestrating the thermogenic program in brown adipocytes. These findings not only enhance our understanding of molecular regulation of thermogenesis but also shed light on the role of rapid and dynamic chromatin reconfiguration in regulating the cellular response to external stimuli. Our study bridges a critical gap between 3D genome organization and gene activation in the thermogenic program mediated by acute β3-AR hormonal signaling. Presentation: 6/2/2024

8156 Exploring Gender-Specific Characteristics Of Diabetes Among Young Adults In Korea: The Korea National Health And Nutrition Examination Survey 2011-2012 And 2019-2020

Abstract Disclosure: C. Chung: None. Background: The escalating global incidence of diabetes among young adults could be linked to dynamic socio-environmental changes including pandemic and climate shifts. In this investigation, we aimed to discern the distinct features of diabetes among young adults in Korea. Methods: This cross-sectional study included 63,489,047 Korean young adults (32,173,248 male, 31,315,799 female) aged 20-39 years who participated in the Korean National Health and Nutrition Examination Survey 2011-2012 and 2020-2021. Diabetes cases were identified through hospital diagnoses or the prescription of insulin or non-insulinemic anti-diabetic medications. The significant insulin resistance was defined as HOMA-IR above 2.9. Complex sampling design and cross-tabulation analyses were used to examine both socioeconomic and laboratory factors. Results: The prevalence of diabetes was higher in men in both 2011-2012 and 2020-2021 (Men vs. Women, 2011-2012: 2.2 ± 0.5 vs. 1.2 ± 0.3 %, p&amp;lt;0.001; 2020-2021: 3.0 ± 0.5 vs. 1.6 ± 0.3 %, p&amp;lt;0.001). The use of insulin and oral hypoglycemic agent (OHA) was comparable between men and women in 2011-2012 (Insulin: 18.1 ± 8.2 vs. 5.2 ± 5.1 %, p=0.305; OHA, 18.2 ± 6.3 vs. 19.5 ± 6.5 %, p=0.239), but more women used insulin and more men used OHA in 2020-2021 (Insulin: 5.3 ± 3.7 vs. 28.7 ± 9.4 %, p=0.016; OHA: 31.4 ± 7.7 vs. 19.6 ± 7.3 %, p=0.009). Execution of fundus exam (25.7 ± 10.6 vs. 68.5 ± 14.1 %, p=0.021) and diagnosis of diabetic retinopathy (0 vs. 8.9 ± 3.3 %, p=0.001) was higher in women in 2020-2021. HbA1c was higher in men in 2020-2021 (8.1 ± 0.4 vs. 7.3 ± 0.3 %, p &amp;lt;0.001), but was similar between men and women in 2011-2012 (8.4 ± 0.6 vs. 8.1 ± 0.6 %, p=0.697). Obesity, one of the most common comorbidities of diabetes, was higher in women in 2011-2012 (24.8 ± 4.8 vs. 79.4 ± 9.3 %, p&amp;lt;0.001), but was higher in men in 2020-2021 (94.4 ± 2.9 vs. 67.8 ± 11.0 %, p&amp;lt;0.001). The factors associated with increase in HOMA-IR in 2020-2021 was different between men and women. Unexpectedly, men with low mental stress showed positive correlation with HOMA-IR (B = 3.9, 95% CI 0.1-7.7, p=0.047). For women, mental stress did not affect HOMA-IR, but those with regular anaerobic exercise (B = -10.5, 95% CI -18.3 - -2.6, p=0.011) and stable employment status (B = -11.9, 95% CI -19.4 - -4.8, p=0.002) presented negative correlation with HOMA-IR. Conclusion: In comparison to before the COVID-19 pandemic, men diabetic patients showed more obese tendency after COVID-19, and there was also a statistically significant difference in HbA1c disparity with women. The higher prevalence of diabetic retinopathy despite of lower HbA1c in women during pandemic era needs to be further investigated in well-designed cohort study. Presentation: 6/3/2024

12622 Molecular Physiology And Compensatory Response In Maladapted Pancreatic Islets Of Humanized Mice Model For Type 1 Diabetes

Abstract Disclosure: A. Hussain: None. A. Aldasouqi: None. S. Rafiqi: None. S.S. Khan: None. R. Paparodis: None. J.C. Jaume: None. S. Imam: None. Genetic predisposition linked with specific HLA (DR and DQ) alleles, environmental factors, and other uncharacterized physiological aberrations triggers an autoimmune attack and depletion of β cells in pancreatic islets, leading to decreased insulin secretion and diabetes. Our laboratory has generated humanized mice carrying the antigen-presenting HLA-DQ8 diabetes-linked haplotype and expressing the human autoantigen GAD65 in pancreatic beta cells. These humanized mice spontaneously develop type 1 diabetes (T1D) and represent the full spectrum of pathophysiological conditions experienced in T1D. Using humanized mice as a model of study, we studied the molecular physiology and compensatory response in maladapted pancreas under normal physiological conditions and hyperglycemic stress of T1D. We studied the differential expression of some important genes by quantitative PCR in maladapted islets to identify the physiological processes affected during the initial oxidative stress exerted by autoimmune conditions. Our results confirmed that levels of cyclin B, cMyc, Cyc D, Cyc E, PDGFRA, and NOTCH are reduced during autoimmune insult, reflecting the poor regenerative capacity of islet cells. However, Cyc A has an elevated expression, reflecting that islet cells, particularly the β cells of Type 1 diabetic mice, may have regenerative potential that may be explored. The genes for Desmin and Survivin showed decreased expression, whereas genes for Smad and Synuclein displayed increased expression. The genes for insulin and somatostatin showed elevated expression, whereas the gene for glucagon displayed reduced expression. The results suggest that during autoimmune insults, the β cells moderately whip up the synthesis of insulin-specific RNA to maintain the requirements. Among the endoplasmic reticulum-specific molecular chaperones, Calnexin showed an elevated expression, whereas calreticulin displayed a reduced expression. Among the genes involved with the architecture of the islet, Lamin and Integrin both display increased expression, reflecting the dynamic change in islet architecture during the autoimmune depletion of β cells. We also investigated the expression of ER-specific stress genes both at RNA and protein levels. Our preliminary results have confirmed that ER-specific stress genes showed an elevated expression during oxidative stress mediated by autoimmune T1D conditions. Presentation: 6/3/2024

Pseudomonas aeruginosa kills Staphylococcus aureus in a polyphosphate-dependent manner.

Due to their frequent coexistence in many polymicrobial infections, including in patients with cystic fibrosis or burn/chronic wounds, many studies have investigated the mechanistic details of the interaction between the opportunistic pathogens Pseudomonas aeruginosa and Staphylococcus aureus. P. aeruginosa rapidly outcompetes S. aureus under in vitro cocultivation conditions, which is mediated by several of P. aeruginosa's virulence factors. Here, we report that polyphosphate (polyP), an efficient stress defense system and virulence factor in P. aeruginosa, plays a role in the pathogen's ability to inhibit and kill S. aureus in a contact-independent manner. We show that P. aeruginosa cells characterized by low polyP levels are less detrimental to S. aureus growth and survival while the Gram-positive pathogen is significantly more compromised by the presence of P. aeruginosa cells that produce high levels of polyP. The polyP-dependent phenotype of P. aeruginosa-mediated killing of S. aureus could at least in part be direct, as polyP was detected in the spent media and causes significant damage to the S. aureus cell envelope. However, more likely is that polyP's effects are indirect through modulating the production of one of P. aeruginosa's virulence factors, pyocyanin. We show that pyocyanin production in P. aeruginosa occurs polyP-dependently and harms S. aureus through membrane damage and potentially the generation of reactive oxygen species, resulting in the increased expression of antioxidant enzymes. In summary, our study adds a new component to the list of biomolecules that the Gram-negative pathogen P. aeruginosa generates to compete with S. aureus for resources.IMPORTANCEHow do interactions between microorganisms shape the course of polymicrobial infections? Previous studies have provided evidence that the two opportunistic pathogens Pseudomonas aeruginosa and Staphylococcus aureus generate molecules that modulate their interaction with potentially significant impact on disease outcomes. Our study identified the biopolymer polyphosphate (polyP) as a new effector molecule that impacts P. aeruginosa's interaction with S. aureus. We show that P. aeruginosa kills S. aureus in a polyP-dependent manner, which occurs primarily through the polyP-dependent production of the P. aeruginosa virulence factor pyocyanin. Our findings add a new role for polyP to an already extensive list of functions. A more in-depth understanding of how polyP influences interspecies interactions is critical, as targeting polyP synthesis in bacteria such as P. aeruginosa may have a significant impact on other microorganisms and potentially result in dynamic changes in the microbial composition.

Large expansion of plasma commensal viruses is associated with SIV pathogenesis in Macaca leonina.

Human immunodeficiency virus-1 (HIV-1) infection disrupts the homeostatic equilibrium between the host and commensal microbes. However, the dynamic changes of plasma commensal viruses and their role in HIV/simian immunodeficiency virus (SIV) pathogenesis are rarely reported. Here, we investigated the longitudinal changes of plasma virome, inflammation levels, and disease markers using an SIV-infected Macaca leonina model. Large expansions of plasma Anelloviridae, Parvoviridae, Circoviridae and other commensal viruses, and elevated levels of inflammation and D-dimer were observed since the chronic phase of SIV infection. Anelloviridae abundance appears to correlate positively with the CD4+ T cell count but negatively with SIV load especially at the acute phase, whereas other commensal viruses' abundances show opposite correlations with the two disease markers. Antiretroviral therapy slightly reduces but does not substantially reverse the expansion of commensal viruses. Furthermore, 1387 primate anellovirus open reading frame 1 sequences of more than 1500 nucleotides were annotated. The data reveal different roles of commensal viruses in SIV pathogenesis.

Dynamic changes of media prefrontal cortex astrocytic activity in response to negative stimuli in male mice

Astrocytes play significant roles in regulating the central stress response. Chronic stress impairs the structure and function of astrocytes in many brain regions such as media prefrontal cortex (mPFC) in multiple neuropsychiatric conditions, but the astrocytic dynamics on the timescale of behavior remains unclear. Here, we recorded mPFC astrocytic activity in freely behaving mice and found that astrocytes are activated immediately by different aversive stimuli. Astrocyte specific GCaMP6s calcium indicator were virally expressed in mPFC astrocytes and fiber photometry experiments revealed that astrocytes are activated by tail-restraint (TRT), foot shock (FS), open arm exploration, stressor of height, predator odor and social defeat (SD) stress. ΔF/F analyses demonstrated that an unpredictable stimulus such as elevated platform stress (EPS) at the initial encounter induced the most intense and rapid changes in astrocytic calcium activity, while a predictable 2,5-dihydro-2,4,5-trimethylthiazoline (TMT) stimulus resulted in the weakest response with a longer peak latency. In TRT, FS or SD test, a somatic stimulus led to higher average calcium activity level and faster average peak latency in repeated trails. Similar to TMT stimulus, astrocytic calcium activity in elevated plus maze (EPM) test exhibited a smaller average change in amplitude and the longest peak latency during open arm exploration. Moreover, astrocytic calcium activity exhibited different changes across behavioral states in SD tests. Our findings show that mPFC astrocytes exhibit distinct patterns of calcium activity in response to various negative stimuli, indicating that the dynamic activity of astrocytes may reflect the stress-related behavioral state under different stimulus conditions.

Survival strategies of microalgae in response to fluctuating brine environments in Saroma-ko Lagoon sea ice, Hokkaido, Japan.

This study investigated the changes in sea ice temperature, microalgae species distribution, shape changes, and photosynthetic activity observed in the first-year ice that forms in winter in Saroma-ko Lagoon, Hokkaido, Japan. Temperatures at the bottom of the ice remained constant at -1.7°C, near the freezing point, while they varied between -6 and -1°C with diel fluctuations at the surface layer. Carefully collected algal samples showed high photosynthetic quantum yield and acclimation to the light intensities of individual ice layers; this indicates that the algal photosynthetic activity responds to dynamic changes in the ice environment, such as variations in temperature, salinity, and brine space. The algal communities consisted of more than 95% diatoms. Smaller algal cells were distributed in the upper layer of the sea ice compared to the lower layers. Chaetoceros sp., the dominant small-cell species, was evenly distributed throughout the layers. In contrast, Detonula confervacea, the dominant large-cell species, was unevenly distributed in the lower layer, with smaller colony size and cell volume in the upper layer. The shape differences observed in this species were thought to be a response to the changing environment within the first-year sea ice.

Plasma EGFR mutation ctDNA dynamics in patients with advanced EGFR-mutated NSCLC treated with Icotinib: phase 2 multicenter trial result

Plasma epidermal growth factor receptor mutation (EGFRm) circulating tumor DNA (ctDNA) dynamics exhibit promise in predicting outcomes in patients with EGFRm-advanced non-small cell lung cancer (NSCLC). However, there remains limited trial-level data on integrating ctDNA monitoring into clinical practice. We performed a prospective, multicenter trial to investigate the relationship between EGFRm ctDNA dynamic changes and clinical outcomes in NSCLC patients with EGFRm. Ninety-eight treatment-naive EGFRm-advanced NSCLC patients were recruited and administered icotinib until disease progression. Plasma samples were collected at baseline and four weeks after icotinib administration. ctDNA was analyzed using a droplet-digital polymerase chain reaction. At baseline, 71.4% of patients had detectable EGFRm ctDNA. Among them, 45.9% of patients’ ctDNA became undetectable within four weeks of treatment. These patients demonstrated significantly longer progression-free survival (PFS) and overall survival (OS) than those with detectable ctDNA after treatment (P = 0.004 and < 0.001, respectively) and were comparable to those with undetectable ctDNA at both baseline and four weeks. ctDNA detectable at four weeks emerged as a poor independent risk factor for PFS and OS. Patients whose ctDNA became undetectable after treatment had outcomes similar to those with initially undetectable ctDNA, underscoring the predictive value of ctDNA dynamics in treatment efficacy.Registry and the Registration No. of the study/trial: ChiCTR-DDD-17013131. Date of registration: 2017-10-27.

Team dynamics after implementation of guidelines for anaesthesia for children with CHD.

Guidelines were created at our single centrer institution for which anesthesiology team should care for pediatric cardiac patients for noncardiac surgery. The goal of the survey was to assess inter-team dynamics after the implementation of guidelines and revealed that practice behaviour can quickly change but a sustained change in team dynamics and workplace culture takes time.

Serial single-cell RNA sequencing unveils drug resistance and metastatic traits in stage IV breast cancer

Metastasis is a complex process that remains poorly understood at the molecular levels. We profiled single-cell transcriptomic, genomic, and epigenomic changes associated with cancer cell progression, chemotherapy resistance, and metastasis from a Stage IV breast cancer patient. Pretreatment- and posttreatment-specimens from the primary tumor and distant metastases were collected for single-cell RNA sequencing and subsequent cell clustering, copy number variation (CNV) estimation, transcriptomic factor estimation, and pseudotime analyses. CNV analysis revealed that a small population of pretreatment cancer cells resisted chemotherapy and expanded. New clones including Metastatic Precursor Cells (MPCs), emerged in the posttreatment primary tumors in CNV similar to metastatic cells. MPCs exhibited expression profiles indicative of epithelial–mesenchymal transition. Comparison of MPCs with metastatic cancer cells also revealed dynamic changes in transcription factors and calcitonin pathway gene expression. These findings demonstrate the utility of single-patient clinical sample analysis for understanding tumor drug resistance, regrowth, and metastasis.

Aglycone Analysis and Quantitative Tracking of Correlated Terminal Glycan Pair on Spermatozoa.

Mammalian sperm glycans directly mediate several key life events. However, previous studies have not focused on two key factors that regulate these processes, the terminal glycan pattern and the anchoring sites. Herein, we group the capping monosaccharide sialic acid (Sia) and its capping substrates galactose/N-acetylgalactosamine (Gal/GalNAc) into a "correlated terminal glycan pair" (glycopair) and, for the first time, reveal the differences in the aglycone pattern of this pair on spermatozoa using glyco-selective in situ covalent labeling techniques. Sia is mainly found in glycoproteins, whereas terminal Gal/GalNAc is mainly found in glycolipids. We quantitatively track the dynamic changes of the glycopair during sperm epididymal migration and find that the Sia capping ratio decreases with the increased expression of the glycopair; caudal upswim spermatozoa also show a lower Sia capping ratio than down spermatozoa. We thus propose two new parameters reflecting the terminal glycoforms of spermatozoa, which can well distinguish the maturity of spermatozoa. By fluorescence imaging of the glycopair in different regions of the sperm, we find that different parts of the sperm contribute differently to the overall glycan changes.

Summer daily dynamics of the vertical structure of atmospheric aerosol over Baikal coastal area from ground-based lidar measurements

ABSTRACT The spatial structure of atmospheric aerosol over Baikal in summer is studied on a regular basis with the use of ground-based LOSA-M2 lidar measurements at the Boyarsky scientific station of the Institute of Physical Material Science, Siberian Branch, Russian Academy of Sciences (51.83°N, 106.06°E, 456 m a.s.l.), Republic of Buryatia, Russia. The lake is located in a basin surrounded by mountain ranges on all side; its water volume is large. The lidar data are analysed along with the regional meteorological situation and meteorological parameters of the atmosphere. The daily dynamics of aerosol distribution over the atmospheric is examined for the period from 2015 to 2023 accounting the effect of different air masses in the region. Three main typical air circulation types are identified for the coastal zone of southern Baikal in summer, which determine the atmospheric aerosol generation and transport. The first and most common type relates to breeze circulation, where the main changes in the spatial structure of aerosol occur in the 2–3-km layer and are determined by changes in the transport direction within a breeze cell during the day. The main feature of this type is a decrease in the altitude of an air layer which contains most of aerosol in the lower atmosphere up to 1 km. The second type refers to southwestern transport under the presence of an anticyclone over the region. The maximal altitude of the aerosol layers attains 7 km in this case. The most complex and dynamic changes in the daily spatial structure of atmospheric aerosol occurred under transport of cyclones and associated atmospheric fronts over Lake Baikal. Under this third type, the altitude of the aerosol layers decreases throughout the day within the altitude range from the surface to 3–4 km.

A Method for Constructing an Urban Waterlogging Emergency Knowledge Graph Based on Spatiotemporal Processes

Urban waterlogging is one of the major “diseases” faced by cities, posing a great challenge to the healthy and sustainable development of cities. The traditional geographic knowledge graph struggles to capture dynamic changes in urban waterlogging over time. Therefore, the objective of this study is to analyze the time, events, properties, geographic objects, and activities associated with urban waterlogging emergency responses from the geographic spatial and temporal processes perspective and to construct an urban waterlogging emergency knowledge graph by combining top-down and bottom-up approaches. We propose a conceptual model of urban waterlogging emergency response ontology based on spatiotemporal processes by analyzing the basic laws and influencing factors of urban waterlogging occurrence and development. Secondly, we describe the construction process of the urban waterlogging emergency response knowledge graph from knowledge extraction, knowledge fusion, and knowledge storage. Finally, the knowledge graph was visualized using 159 urban waterlogging events in China from 2020–2022, with a quality assessment indicating 81% correctness, 65.5% completeness, and 95% data conciseness. The results show that this method can effectively express the spatiotemporal process of an urban waterlogging emergency response and can provide a reference for the spatiotemporal modeling of the knowledge graph.

Research on coastline extraction and dynamic change from remote sensing images based on deep learning

Accurate coastline extraction is crucial for the scientific management and protection of coastal zones. Due to the diversity of ground object details and the complexity of terrain in remote sensing images, the segmentation of sea and land faces challenges such as unclear segmentation boundaries and discontinuous coastline contours. To address these issues, this study improve the accuracy and efficiency of coastline extraction by improving the DeepLabv3+ model. Specifically, this study constructs a sea-land segmentation network, DeepSA-Net, based on strip pooling and coordinate attention mechanisms. By introducing dynamic feature connections and strip pooling, the connection between different branches is enhanced, capturing a broader context. The introduction of coordinate attention allows the model to integrate coordinate information during feature extraction, thereby allowing the model to capture longer-distance spatial dependencies. Experimental results has shown that the model can achieves a land-sea segmentation mean intersection over union (mIoU) ration and Recall of over 99% on all datasets. Visual assessment results show more complete edge details of sea-land segmentation, confirming the model’s effectiveness in complex coastal environments. Finally, using remote sensing data from a coastal area in China as an application instance, coastline extraction and dynamic change analysis were implemented, providing new methods for the scientific management and protection of coastal zones.

Time-course characterization of whole-transcriptome dynamics of HepG2/C3A spheroids and its toxicological implications

Physiologically relevant in vitro models are a priority in predictive toxicology to replace and/or reduce animal experiments. The compromised toxicant metabolism of many immortalized human liver cell lines grown as monolayers as compared to in vivo metabolism limits their physiological relevance. However, recent efforts to culture liver cells in a 3D environment, such as spheroids, to better mimic the in vivo conditions, may enhance the toxicant metabolism of human liver cell lines. In this study, we characterized the dynamic changes in the transcriptome of HepG2/C3A hepatocarcinoma cell spheroids maintained in a clinostat system (CelVivo) to gain insight into the metabolic capacity of this model as a function of spheroid size and culture time. We assessed morphological changes (size, necrotic core), cell health, and proliferation rate from initial spheroid seeding to 35 days of continuous culture in conjunction with a time-course (0, 3, 7, 10, 14, 21, 28 days) of the transcriptome (TempO-Seq, BioSpyder). The phenotypic characteristics of HepG2/C3A growing in spheroids were comparable to monolayer growth until ∼Day 12 (Day 10–14) when a significant decrease in cell doubling rate was noted which was concurrent with down-regulation of cell proliferation and cell cycle pathways over this time period. Principal component analysis of the transcriptome data suggests that the Day 3, 7, and 10 spheroids are pronouncedly different from the Day 14, 21, and 28 spheroids in support of a biological transition time point during the long-term 3D spheroid cultures. The expression of genes encoding cellular components involved in toxicant metabolism and transport rapidly increased during the early time points of spheroids to peak at Day 7 or Day 10 as compared to monolayer cultures with a gradual decrease in expression with further culture, suggesting the most metabolically responsive time window for exposure studies. Overall, we provide baseline information on the cellular and molecular characterization, with a particular focus on toxicant metabolic capacity dynamics and cell growth, of HepG2/C3A 3D spheroid cultures over time.

Dynamic Transition of Regulatory T Cells to Cytotoxic Phenotype Amid Systemic Inflammation in Graves' Ophthalmopathy.

Graves' disease (GD) is an autoimmune condition that can progress to Graves' Ophthalmopathy (GO), leading to irreversible damage to orbital tissues and potential blindness. The pathogenic mechanism is not fully understood. In this study, we conducted single-cell multi-omics analyses on healthy individuals, GD patients without GO, newly diagnosed GO patients, and treated GO patients. Our findings revealed gradual systemic inflammation during GO progression, marked by overactivation of cytotoxic effector T cell subsets, and expansion of specific T cell receptor clones. Importantly, we observed a decline in the immunosuppressive function of activated regulatory T cells (aTreg) accompanied by a cytotoxic phenotypic transition. In vitro experiments revealed that dysfunction and transition of GO-autoreactive Treg were regulated by the yinyang1 (YY1) upon secondary stimulation of thyroid stimulating hormone receptor (TSHR) under inflammatory conditions. Furthermore, adoptive transfer experiments of GO mouse model confirmed infiltration of these cytotoxic Treg into the orbital lesion tissues. Notably, these cells were found to upregulate inflammation and promote pathogenic fibrosis of orbital fibroblasts (OFs). Our results revealed the dynamic changes in immune landscape during GO progression and provided novel insights into the instability and phenotypic transition of Treg, offering potential targets for therapeutic intervention and prevention of autoimmune diseases.

P38α kinase governs muscle strength through PGC1α in mice.

Skeletal muscle, with its remarkable plasticity and dynamic adaptation, serves as a cornerstone of locomotion and metabolic homeostasis in the human body. Muscle tissue, with its extraordinary capacity for force generation and energy expenditure, plays a fundamental role in the movement, metabolism, and overall health. In this context, we sought to determine the role of p38α in mitochondrial metabolism since mitochondrial dynamics play a crucial role in the development of muscle-related diseases that result in muscle weakness. We conducted our study using male mice (MCK-cre, p38αMCK-KO and PGC1α MCK-KO) and mouse primary myoblasts. We analyzed mitochondrial metabolic, physiological parameters as well as proteomics, western blot, RNA-seq analysis from muscle samples. Our findings highlight the critical involvement of muscle p38α in the regulation of mitochondrial function, a key determinant of muscle strength. The absence of p38α triggers changes in mitochondrial dynamics through the activation of PGC1α, a central regulator of mitochondrial biogenesis. These results have substantial implications for understanding the complex interplay between p38α kinase, PGC1α activation, and mitochondrial content, thereby enhancing our knowledge in the control of muscle biology. This knowledge holds relevance for conditions associated with muscle weakness, where disruptions in these molecular pathways are frequently implicated in diminishing physical strength. Our research underscores the potential importance of targeting the p38α and PGC1α pathways within muscle, offering promising avenues for the advancement of innovative treatments. Such interventions hold the potential to improve the quality of life for individuals affected by muscle-related diseases.

Impact of waitlist weight change on outcomes in heart transplant recipients: a UNOS database analysis.

While the effect of pre-transplant weight on patient outcomes following heart transplantation (HTx) has previously been studied, data regarding the impact of dynamic weight change prior to HTx are extremely limited. We sought to elucidate the interaction between HTx listing weight and weight change while waitlisted, and explore how that interaction impacts post-HTx survival in a continuous manner. Adult patients listed for HTx from 1987 to 2020 were identified from UNOS database. Three-dimensional restricted cubic spline analysis explored post-HTx survival relative to both changes in BMI/weight and BMI at time of HTx listing. Continuous predictor variables were analyzed with Cox proportional hazards method. 9,628 included patients underwent HTx. Median recipient age was 55 [IQR 46-62] years, and 21% were females. 53% of patients lost while 47% gained weight on the waitlist. Median BMI (27.6kg/m2 [24.3-31.3] vs. 27.4kg/m2 [24.2-30.9], paired p < 0.001) and weight (84.8kg [73.0-98.0] kg vs. 84.4kg [72.6-96.6], p < 0.001) were similar at listing and transplant. One-year survival was 89.3%. Weight loss over 3 BMI points or 10kg was associated with higher hazard of death irrespective of listing BMI. In non-obese patients, some weight gain (1-4 BMI points or 5-15kg) was associated with improved survival. In cachectic patients (BMI < 18.5), failure to gain weight was associated with worse survival. Impact of weight change varies depending on listing BMI. While a survival benefit is seen in non-obese patients who gain some weight, significant weight loss is associated with poorer survival.

Real-time visualisation of developing chick embryos cultured in transparent plastic films from the blastoderm stage until hatching

Real-time visualisation of chick embryo development occurring inside an opaque eggshell is a major goal of developmental biology. This goal was partially achieved when 3-day-old embryos recovered from fertilised shelled eggs, preincubated in a conventional incubator, hatched after the egg contents had been transferred into shell-less embryo culture systems constructed of transparent plastic film placed in a plastic cup. However, the hatchable shell-less embryo culture systems described thus far cannot be used to visualise the dynamic morphological changes associated with the emergence of major organ systems during the first 3 days of embryo development, as the blastoderm area needs to be covered with an opaque membrane. Here, we present the development of a chick embryo culture system that enables omnidirectional real-time visualisation of the developing embryo, beginning from the newly laid blastoderm stage embryo until hatching.