Morphology Expression Research Articles

Morphological description of the anterior cerebral artery and its branches in cadavers’ samples

The diverse morphological expression of the anterior cerebral artery (ACA), described in several population groups, generates a complex spectrum of clinical signs when embolic or hemorrhagic strokes occur. This study evaluated the ACA and its branches in 162 cerebral hemispheres extracted from corpses at the Institute of Legal Medicine in Bucaramanga, Colombia. The proximal segments of the internal carotid and vertebral arteries were bilaterally perfused with semisynthetic resin dyed with red-colored minerals. The qualitative and morphometric characteristics of the ACA and its branches were recorded. The length of the A1 segment was 12.4±2.5 mm, with a diameter 2.4±0.6 mm. The medial orbitofrontal branch (MOF) originated from the A2 segment in 144 cases (93.5%), and in 41 specimens (26.6%), it formed common trunks with the frontopolar branch (FP) or with the anterior medial frontal branch (AMF). The length and diameter of the marginal callosal branch (MC) were 59.7±18.8 mm and 1.6±0.5 mm, respectively. The sixteen cases (10.4%) of observed medial frontal branches stemmed from the ACA and not from the MC. In thirty-three cases (21.4%), the branch of the paracentral lobule (PcL) emerged from the A3 segment; in 81 samples (52.6%) from the MC; and in 40 (26%) from the A4 segment. This study found great variability in the emergence of the ACA branches, especially the MC, the MF, and the PcL. Knowing the origin, unique or branched pathways, and territories irrigated by the ACA and its branches enriches the diagnosis, management, and interventions when these structures are compromised in clinical events.

Single-Larva RNA Sequencing Reveals That Red Sea Urchin Larvae Are Vulnerable to Co-Occurring Ocean Acidification and Hypoxia.

Anthropogenic carbon dioxide emissions have been increasing rapidly in recent years, driving pH and oxygen levels to record low concentrations in the oceans. Eastern boundary upwelling systems such as the California Current System (CCS) experience exacerbated ocean acidification and hypoxia (OAH) due to the physical and chemical properties of the transported deeper waters. Research efforts have significantly increased in recent years to investigate the deleterious effects of climate change on marine species, but have not focused on the impacts of simultaneous OAH stressor exposure. Additionally, few studies have explored the physiological impacts of these environmental stressors on the earliest life stages, which are more vulnerable and represent natural population bottlenecks in organismal life cycles. The physiological response of the ecologically and commercially important red sea urchin (Mesocentrotus franciscanus) was assessed by exposing larvae to a variety of OAH conditions, mimicking the range of ecologically relevant conditions encountered currently and in the near future along the CCS. Skeleton dissolution, larval development, and gene expression show a response with clearly delineated thresholds that were related to OAH severity. Skeletal dissolution and the induction of Acid-sensing Ion Channel 1A at pH 7.94/5.70 DO mg/L provide particularly sensitive markers of OAH, with dramatic shifts in larval morphology and gene expression detected at the pH/DO transition of 7.71/3.71-7.27/2.72 mg/L. Experimental simulations that describe physiological thresholds and establish molecular markers of OAH exposure will provide fishery management with the tools to predict patterns of larval recruitment and forecast population dynamics.

Lung Endothelial Cell Heterogeneity in Health and Pulmonary Vascular Disease.

Lung endothelial cells (ECs) are essential for maintaining organ function and homeostasis. Despite sharing some common features with ECs from organ systems, lung ECs exhibit significant heterogeneity in morphology, function, and gene expression. This heterogeneity is increasingly recognized as a key contributor to the development of pulmonary diseases like pulmonary hypertension (PH). In this mini-review, we explore the evolving understanding of lung EC heterogeneity, particularly through the lens of single-cell RNA sequencing (scRNA-seq) technologies. These advances have provided unprecedented insights into the diverse EC subpopulations, their specific roles, and the disturbances in their homeostatic functions that contribute to PH pathogenesis. In particular, these studies identified novel and functionally distinct cell types such as aerocytes and general capillary ECs that are critical for maintaining lung function in health and disease. In addition, multiple novel pathways and mechanisms have been identified that contribute to aberrant pulmonary vascular remodeling in PH. Emerging techniques like single-nucleus RNA sequencing and spatial transcriptomics have further pushed the field forward by discovering novel disease mediators. As research continues to leverage these advanced techniques, the field is poised to uncover novel EC subtypes and disease mechanisms, paving the way for new therapeutic targets in PH and other lung diseases.

Pretreatment with tetramethylpyrazine alleviated the impairment of learning and memory induced by sevoflurane exposure in neonatal rats

Sevoflurane impairs learning and memory of the developing brain. However, strategies to mitigate these detrimental effects have been scarce. Herein, we investigated whether tetramethylpyrazine could alleviate the impairment of learning and memory and its underlying mechanisim in sevoflurane-exposed neonatal rats. Postnatal 7-day Sprague-Dawley (SD) rats or primary hippocampal neurons were pretreated with tetramethylpyrazine and then exposed to sevoflurane. The terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and lactate dehydrogenase (LDH) assays were used to detect neuronal injury. Learning and memory function were evaluated by novel object recognition and Morris water maze tests. Long-term potentiation (LTP) was recorded to evaluate synaptic plasticity electrophysiologically in the hippocampal slices. Golgi-Cox staining or PSD95 immunochemistry was used to detect the morphology of dendritic spines. Western blotting was employed to assess the expressions of cleaved Caspase-3, PSD95, NMDAR1, NMDAR2A and NMDAR2B in the hippocampus or cultured neurons. It was found that neonatal exposure of sevoflurane impaired learning and memory, increased neuronal apoptosis, altered the morphology of dendritic spine, upregulated the expressions of NMDAR2A and PSD95, and induced LTP deficits. Pretreatment with tetramethylpyrazine not only alleviated impairment of learning and memory, but also improved sevoflurane-induced changes in neuronal damage, dendritic spine morphology, NMDAR2A and PSD95 expressions, as well as LTP. These findings indicated that pretreatment with tetramethylpyrazine alleviated the impairment of learning and memory induced by sevoflurane through improvement of hippocampal synaptic plasticity in neonatal rats.

Influence of Multistrain Probiotics on Growth, Hematology, Gut and Liver Morphometry, and GH and IGFs Genes Expression in Rohu (Labeo Rohita) Fry

Probiotic is regarded as an effective feed additive to increase the growth, feeding efficiency, survivability, and overall productivity of fishes in aquaculture. The following study was intended to examine the effects of multistrain probiotics on growth, blood indices, histomorphometry of gastrointestinal tract and liver, and growth hormone (GH) and insulin‐like growth factors (IGFs) genes expression in rohu Labeo rohita. Probiotic containing Bacillus subtilis, B. thuringiencis, Lactobacillus plantarum, and L. buchneri was inoculated in water and three different doses, that is, 0 mL/L (control), 0.5 mL/L (lower dose, T1), and 1.0 mL/L (higher dose, T2) were used in this study. Each treatment had three replicates and fishes were reared in aquarium for 60 days. Results revealed that substantially higher weight gain (WG) and specific growth rate (SGR) along with least apparent feed conversion ratio (AFC) were documented in probiotic treated fish which was significantly different than that of control, however, higher dose (1.0 mL/L) showed best performance. Blood–biochemical indices such as red blood cell (RBC), white blood cell (WBC), hemoglobin (Hb), and glucose (Glu) level were observed statistically higher in probiotic additive culture water. Moreover, the inoculation of multispecies probiotic positively altered the intestinal histometry. Besides, immune response features of intestine were observed significantly higher in probiotic treated fish. The nucleus shape and distances between liver tissues (DLTs) were also improved with the upward doses of probiotic application. The mRNAs levels of GH in the pituitary and IGF‐1 and IGF‐2 in the liver were significantly higher in probiotic treated fish. In a nutshell, this study suggested that the inclusion of multistrain probiotics showed promising improvement in growth performance and feed conversion efficiency; additionally, uplifted the intestinal structure integrity, liver morphology, and GH and IGFs genes expression in rohu.

Лексикограматичні засоби експлікації психоемоційних станів у сучасних українських медіатекстах воєнного періоду

The article comprehensively analyzes the lexical‑grammatical means of verbalizing psycho‑emotional states, elucidates their role in creating a negative emotogenic atmosphere of modern media discourse during the Russian‑Ukrainian war period. It was found that the linguistic means of expression for psycho‑emotional states should be analyzed within the close relationship of form and content. Lexical, phraseological, metaphorical units become maximally expressive in syntactic structures, and grammatical constructions acquire a bright pragmatic effect as a result of them being filled with emotional and evaluative vocabulary, synonymous series, phraseological units, tropes, etc. The most frequent and most expressive syntactic models are highlighted (monosyllabic impersonal sentences with the dative case in the function of the state bearer, two‑syllable personal sentences with the nominative subject case, two‑syllable with metaphorical or phraseological predicates and experiential in indirect cases), which strengthen the emotional and expressive background of the analyzed media texts due to the interaction of their syntactic semantics with the lexical meaning and morphological expression of the filler components, as well as the influence of context, intonation, and non‑linguistic factors.

Mesenchymal Traits as an Intrinsic Feature of Undifferentiated Cells

Since its first conceptualization over a century ago, the mesenchymal phenotype has traditionally been viewed as either a transient phase between successive epithelial stages or as a feature of cell types primarily devoted to structural support. However, recent findings in cancer research challenge this limited view, demonstrating that mesenchymal traits and hybrid mesenchymal/epithelial states can mark cancer cells with stem cell properties. By analyzing publicly available single-cell transcriptome datasets from early embryonic stages and adult tissues, this study aims to extend this concept beyond pathological contexts, suggesting that a partial or fully mesenchymal phenotype may represent the morphological expression of undifferentiated and multipotent states in both the developing embryo and adult organs.

Assessing the impact of airborne particulate pollution on human skin utilizing a novel human skin equivalent containing MUTZ‐3‐derived Langerhans cells

AbstractAir pollution is an exogenous stressor known to have a detrimental impact on skin health through the induction of inflammation; however, the direct effect of topical pollution exposure is still being elucidated. Human skin equivalents (HSE) aim to reproduce in vitro the structure and function of the native skin tissue. However, HSEs typically lack skin‐resident immune cells, which could play a key role in the inflammatory response induced by pollution exposure. We outline the development of a HSE‐containing MUTZ‐3‐derived Langerhans cells (MUTZ‐3‐LCs), which show dendritic morphology and Langerhans cell marker expression. We demonstrated that HSE‐containing MUTZ‐3‐LC have lower basal levels of proinflammatory cytokines, but topical stimulation with allergens and irritant compounds induced a greater inflammatory response in these models compared to HSE without immune cells. To study the effect of pollution, we created a technique to apply diesel particulate matter (DPM) to HSEs. Though our microscopic analysis demonstrated that DPM does not penetrate the stratum corneum, we showed that DPM did induce production of proinflammatory cytokines, but notably only in HSEs containing MUTZ‐3‐LCs. These data suggest that topical exposure to air pollution can induce cutaneous inflammation and that skin‐resident immune cells contribute to this response. This highlights the significance of immune‐competent HSEs to the study of exogenous stressors in vitro.

Genome Assembly and Winged fruit Gene Regulation of Chinese Wingnut: Insights from Genomic and Transcriptomic Analyses.

The genomic basis and biology of winged fruit are interesting issues in ecological and evolutionary biology. Chinese wingnut (Pterocarya stenoptera) is an important garden and economic tree species in China. The genomic resources of this hardwood tree could provide advanced genomic studies of Juglandaceae and their evolutionary relationships. Here, we reported a high-quality reference genome of P. stenoptera (N50 = 35.15 Mb) and provided a comparative analysis of Juglandaceae genomes. Paralogous relationships among the 16 chromosomes of the Chinese wingnut genome revealed eight main duplications representing the subgenome. Molecular dating suggested that the most recent common ancestor of P. stenopetera and Cyclocarya paliurus diverged from Juglans around 56.7 million years ago (Mya). The expanded and contracted gene families were associated with cutin, suberine, and wax biosynthesis, cytochrome P450, and anthocyanin biosynthesis. We identified large inversion blocks between the P. stenoptera genome and its relatives, which are enriched in genes related lipid biosynthesis and metabolism, and starch and sucrose metabolism. The twenty-eight individuals were clearly clustered into three groups responding to three species, namely Pterocarya macroptera, Pterocarya hupehensis, and P. stenoptera, based on whole genome resequencing data. Morphological and gene expression analysis showed that CAD, COMT, LOX, and MADS-box play important roles during the five developmental stages of wingnuts. Our study highlights the evolutionary history of the P. stenoptera genome and supports P. stenoptera as an appropriate Juglandaceae model for studying winged fruits. These results provide a theoretical basis for the evolution, development, and diversity of woody plant winged fruits.

Categorial Semantic Contours Semi-Phraseologized Sentences

The article is devoted to the analysis of the categorical meanings of the semi-phraseologized sentences and characterisation of their semantic types. The study is based on the interpretation of semi-phraseologized sentences as colloquial syntactic constructions not only in Ukrainian but also in other languages. The categorical meanings of the semi-phraseological sentences “affirmation”, “negation”, “asessment” and “expression of will” make it possible to distinguish certain semantic types of semi-phraseologized sentences with corresponding categorical meanings. The classification feature is the phraseosyntactic meaning fixed to a stable structural scheme. The phrase-syntactic meaning forms the content of the phrase scheme components: morphological expression of permanent and variable components. Semi-phraseologized sentences with the categorical meaning of “affirmation” formed three semantic types: 1) with the actual affirmation meaning; 2) with the meaning of categorical affirmation; 3) with the meaning of conditional affirmation. Semi-phraseologized sentences have two manifestations of the categorical meaning of “negation”: 1) with actual negation; 2) with categorical negation. Semi-phraseologized sentences with categorical meaning of the “assessment” formed the two semantic types: with undifferentiated assessment and with negative assessment. Semi-phraseologized sentences with categorical meaning of the “expression of will” do not form semantic types due to the chaotic selection components of the colloquial type of phrase schemes.The proposed algorithm will help to organize specific non-segmented syntactic constructions in the syntax oral speech of speakers of different linguistic cultures.

Autofluorescence Virtual Staining System for H&E Histology and Multiplex Immunofluorescence Applied to Immuno-Oncology Biomarkers in Lung Cancer.

Virtual staining for digital pathology has great potential to enable spatial biology research, improve efficiency and reliability in the clinical workflow, as well as conserve tissue samples in a non-destructive manner. In this study, we demonstrate the feasibility of generating virtual stains for hematoxylin and eosin (H&E) and a multiplex immunofluorescence (mIF) immuno-oncology panel (DAPI, PanCK, PD-L1, CD3, CD8) from autofluorescence images of unstained non-small cell lung cancer tissue by combining high-throughput hyperspectral fluorescence microscopy and machine learning. Using domain-specific computational methods, we evaluated the accuracy of virtual H&E for histologic subtyping and virtual mIF for cell segmentation-based measurements, including clinically-relevant measurements such as tumor area, T cell density, and PD-L1 expression (tumor proportion score and combined positive score). The virtual stains reproduce key morphologic features and protein biomarker expressions at both tissue and cell levels compared to real stains, enable the identification of key immune phenotypes important for immuno-oncology, and show moderate to good performance across various evaluation metrics. This study extends our previous work on virtual staining from autofluorescence in liver disease and prostate cancer, further demonstrating the generalizability of this deep learning technique to a different disease (lung cancer) and stain modality (mIF).

Morphological characteristics, developmental dynamics, and gene temporal expressions across various development stages of Aphis gossypii sexual female

BackgroundAphis gossypii (Hemiptera: Aphididae) is a worldwide polyphagous phloem-feeding agricultural pest, and it can produce offspring by sexual or asexual reproduction. Compared with dozens of generations by parthenogenesis, sexual reproduction is performed in only one generation within one year, and little is known about the sexual reproduction of A. gossypii. In this study, sexual females of A. gossypii were successfully obtained through a previously established induction platform, and the morphological characteristics, developmental dynamics, and temporal gene expression were examined. Subsequently, signaling pathways potentially involved in regulating the growth, development, and reproduction of sexual females were investigated.ResultsThe morphological observation showed that from the 1st instar nymph to adult, sexual females exhibited a gradually deepened body color, an enlarged body size, longer antennae with a blackened end, and obviously protruding cauda (in adulthood). The anatomy found that the ovaries of sexual females developed rapidly from the 2nd instar nymph, and the embedded oocytes matured in adulthood. In addition, time-course transcriptome analysis revealed that gene expression profiles across the development of sexual females fell into 9 clusters with distinct patterns, in which gene expression levels in clusters 1, 5, and 8 peaked at the 2nd instar nymphal stage with the largest number of up-regulated genes, suggesting that the 2nd instar nymph was an important ovary development period. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis revealed that a large number of genes in the sexual female adult were enriched in the TGF-beta signaling pathway and Forkhead box O (FoxO) signaling pathway, highlighting their important role in sexual female adult development and reproduction.ConclusionThe morphological changes of the sexual female at each developmental stage were revealed for the first time. In addition, time-course transcriptomic analyses suggest genes enriched in the TGF-beta signaling pathway and FoxO signaling pathway probably contribute to regulating the development and oocyte maturation of sexual females. Overall, these findings will facilitate the regulating mechanism research in the growth and development of sexual females by providing candidate genes.

Tendon-targeted knockout of collagen XI disrupts patellar and Achilles tendon structure and mechanical properties during murine postnatal development

ABSTRACT Background Collagen XI is a fibril-forming collagen typically associated with type II collagen tissues but is also expressed in type I collagen-rich tendons, especially during development. We previously showed that tendon-targeted (Scx-Cre) Col11a1 knockout mice have smaller tendons in adulthood with aberrant fibril structure and impaired mechanical properties. However, the manifestation of this phenotype is not clearly understood. Therefore, our objective is to define the spatiotemporal roles of collagen XI in tendon structure-function during postnatal development. Given the high expression of collagen XI during embryonic development, we hypothesized that collagen XI knockout leads to the deposition of weakened extracellular matrix during early postnatal timepoints, disrupting the establishment of tendon structure and function. Methods Patellar and Achilles tendons from postnatal (P) days 0, 10, 20, and 30 tendon-targeted scleraxis-Cre heterozygous and homozygous Col11a1 knockout mice were evaluated for morphology, nuclear organization, fibril morphology, mechanical properties, and gene expression. Results At P0, there were no differences in tendon length or fibril diameter of either tendon. By P10, striking structural and functional differences emerged, with collagen XI deficiency resulting in increased tendon length, a heterogeneous and larger diameter population of fibrils, and inferior mechanical properties in both patellar and Achilles tendons. Differences increased in magnitude through P30, supporting our hypothesis that impaired structure-function during postnatal development may drive tendon lengthening and reduced mechanical properties. Conclusions Though collagen XI is a quantitatively minor component of the tendon extracellular matrix, these results highlight the critical role of collagen XI in the acquisition of tendon structure-function.

Morphological and physiological root traits and expression of the gene relationship withnitrogen uptake in Wheat (Triticum aestivum L.)

Morphological and physiological root traits and expression of the gene relationship withnitrogen uptake in Wheat (Triticum aestivum L.)

GLP-1R/NPY2R regulate gene expression, ovarian and adrenal morphology in HFD mice.

Glucagon-like peptide-1 receptor (GLP-1R) and neuropeptide Y receptors (NPYRs) are expressed in reproductive tissues contributing to the regulation of gonadal function. This exploratory study examines the potential impact of their modulation by assessing effects of exendin-4 (Ex-4) and peptide YY (PYY) (3-36) on endocrine ovaries and adrenals, in high-fat diet (HFD) mice. Ex-4 and PYY(3-36) reduced blood glucose and energy intake, with no effects on body weight. While HFD did not impact estrous cycle, Ex-4 increased metestrus frequency and decreased diestrus frequency resulting in 0% mice experiencing repeated diestrus or becoming acyclic. LH levels were significantly higher in the Ex-4 and PYY(3-36) groups compared to ND and HFD controls. In the adrenals, reduced capsule and zona glomerulosa thickness cause by HFD was reversed after peptide treatments. Within the ovaries, HFD increased the number of atretic follicles, an effect that disappeared after Ex-4 and PYY(3-36) treatments. Ex-4 also increased the number of corpora lutea owing to the prolonged metestrus phase. Gene expression analysis within the adrenals revealed upregulation of Insr and downregulation of Prgtr in HFD mice, while Ex-4 downregulated the expression of Gipr. Ovarian gene expression of Gipr, Npy1r and Prgtr were downregulated by Ex-4 treatment, while PYY(3-36) significantly downregulated Prgtr expression compared to HFD mice. These data indicate that manipulating GLP-1R and NPY2R lead to changes in the reproductive physiology of mice. Additionally, the observed alterations in the morphology and gene expression in the adrenals and ovaries imply a direct impact of these peptides on female reproductive function.

Converting suspension-type human primary hepatocytes to be plateable-type for use as an in vitro pharmacokinetics model

Preclinical animal models cannot precisely predict medication failures in clinical trials. Human primary hepatocytes (HPHs) are the gold standard for in vitro culture systems that recapitulate in vivo human liver characteristics. Consequently, demand for HPHs is rising despite their high cost, batch-to-batch variability, and inadequate supply. Existing commercial HPHs are plateable (pHPHs) and suspension (sHPHs) using same isolation and cryopreservation technique. The sHPHs cannot be cultured on the plate surface or cannot efficiently relocalize efflux transporters. They can also have a life of maximum of 2–4 hours. Therefore, they are unsuitable for assays requiring well-organized efflux transporters or long-term studies, such as enzyme induction or metabolic assays for low-turnover drugs. We aim to develop a novel pharmacokinetic model of converted sHPHs to be attached like pHPHs. We successfully cultured sHPHs as 2D sandwich-cultured sHPHs for use as a promising, reproducible, and affordable in vitro model for efflux trans- porter assays, cytochrome P450 induction, and metabolic studies. The sHPHs successfully attached, converted to be plateable, similar morphology, and mRNA expression as pHPHs, as well as detectable metabolic activity and extended lifespan. Overall, this study is the first to culture functional 2D sandwich sHPHs as a novel in vitro pharmacokinetic model.

Proliferative potential and angiogenic characteristics of blood outgrowth endothelial cells derived from middle-aged and older adults.

Autologous blood outgrowth endothelial cells (BOECs) have been proposed to induce therapeutic angiogenesis for treating cardiovascular diseases (CVDs). The aim of the present study was to investigate the proliferative potential and angiogenic characteristics of BOECs among middle-aged and older adults, the population particularly susceptible to CVDs. BOECs were isolated from 48 peripheral blood samples of subjects aged 56 ± 4 years. The cells were then distinguished based on their proliferative abilities, and their phenotype, tube formation capacity, and migratory activity were compared using immunofluorescence staining, flow cytometry, tube formation assay, and wound healing assay, respectively. Correlations between demographic, clinical, and dietary parameters with the number of BOECs were also assessed. A total of 132 BOEC colonies with different proliferative potentials were obtained, including colonies lost proliferative ability before passage 3 (named LPA), stopped proliferating during passage 3-8 (HPA (3-8)), and proliferated after passage 8 (HPA (> 8)). LPA cells appeared later and displayed abnormal morphology, while HPA (3-8) cells exhibited alterations in von Willebrand factor morphology and lower KDR expression. HPA (> 8) cells obtained higher branching intervals and individual cell migration velocity compared with those of HPA (3-8) cells. Correlation analysis showed that the number of both LPA and HPA colonies were positively associated with several CVD risk factors. Additionally, the number of LPA colonies was positively associated with servings of meats and alternatives, fruits, fruits and vegetables, as well as the protein intake. Our findings provide evidence that the middle-aged and older populations possess BOECs with different proliferative and angiogenic potentials, exhibiting distinctions in cell morphology, appearance dates, VWF morphology, and KDR expression. Strikingly, a higher number of BOECs is likely associated with an increased risk of CVDs, while the number of BOECs with low proliferative ability may be regulated by diet.

Development and use of two Xenopus laevis spleen stromal cell lines to study the role of splenic stromal cells in anuran immune processes

The spleen is an important immune organ in adult Xenopus laevis, supporting the differentiation of B cells and acting as the main peripheral lymphoid organ. Key to these processes are the supporting non-hematopoietic cells, or stromal cells, within the spleen tissue. Despite the importance of the spleen to frog immunity, few frog cell lines originating from spleen tissue have been reported. In this study, we report on the establishment and characterization of two cell lines originating from X. laevis spleen tissue, Xela S5F and Xela S5E. Morphological observations and gene expression profiling suggest that Xela S5F is fibroblast-like and Xela S5E is epithelial-like. Both cell lines express transcripts corresponding to a variety of hematopoietic growth factors, suggesting their potential utility as a feeder cell line to support ex vivo myelopoietic cell differentiation. Xela S5F and Xela S5E produce transcripts for a diversity of pattern recognition receptors including toll-like receptors, scavenger receptors, and cytosolic nucleic acid sensors, suggesting anuran spleen stromal cells may be important cellular sensors of pathogens filtered through the spleen. This idea is supported by the increase in transcript levels for antiviral and proinflammatory genes in both cell lines in response to treatment with the commercially available toll-like receptor ligands, flagellin and poly(I:C). However, despite the ability to sense extracellular synthetic analogues of viral nucleic acids [i.e. poly(I:C)] and susceptibility and permissibility of both cell lines to frog virus 3 (FV3), a large double-stranded DNA virus that infects amphibians, neither cell line upregulates key antiviral or proinflammatory transcripts when challenged with FV3. The establishment of Xela S5F and S5E cell lines expands the current X. laevis invitrome and provides new in vitro cell model systems to investigate the role of splenic stromal cells in anuran immune functions.

Lipophagy Enhances Fat Graft Retention by Stimulating Dedifferentiation of Mature Adipocytes and Modulating Inflammation.

Maintaining lipid metabolic homeostasis is essential for cells to respond to environmental stresses, ensure survival, and preserve functionality. Lipophagy, a form of autophagy in which intracellular lysosomes target and degrade lipid droplets. The authors hypothesize that pharmacologically enhancing or disrupting lipophagy could either improve or impair the retention rates of fat grafts. Human fat graft nodules, retained for six months, were collected through surgical procedures for analysis. Additionally, a mouse fat graft model was established by mixing fragmented adipose tissue with the autophagy inducer rapamycin (Rapa), the autophagy inhibitor 3-methyladenine (3-MA), or PBS, and subcutaneously implanting into the bilateral lower back of mice for localized activation or inhibition of lipophagy. Samples were collected at various time points for morphological, histological, transcriptomic, and gene expression analysis. A significant increase in lipophagy was demonstrated in both human and mouse fat grafts. Rapa-induced upregulation of lipophagy promoted long-term retention of mouse fat grafts and resulted in fewer cysts and reduced fibrosis. Lipophagy activation facilitated the dedifferentiation of mature adipocytes and adipogenesis. Moreover, lipophagy enhanced the metabolism of oil droplets in macrophages. Activation of lipophagy also reduced long-term macrophage infiltration and collagen deposition in fat grafts while simultaneously increasing the proportion of M2 macrophages. Lipophagy was activated following fat grafting. Lipophagy improves fat graft retention and quality by stimulating the dedifferentiation of mature adipocytes, promoting oil metabolism, and reducing inflammation. Thus, lipophagy serves as a crucial integrative point connecting lipid metabolism, regeneration, and immunity in fat grafts.

Induction of neuronal differentiation in glioma cells by histone deacetylase inhibitors based on Connectivity Map discovery.

Neuron conversion leads to proliferation inhibition of glioma cells and may be an effective strategy to combat glioma and prevent recurrence. In this study, drug repositioning based on Connectivity Map (CMap) was conducted to discover drugs that could induce the differentiation of glioma cells into neuron-like cells, complemented by in vitro experimental validation. Downregulated neuronal genes in glioma were identified by the Human Protein Atlas database and the GeneCards database, and enrichment analysis and Gene Expression Profiling Interactive Analysis (GEPIA) were performed to ensure their reliability before they were uploaded to CMap for drug screening. The potential drug targets were screened through GEPIA and validated by the Chinese Glioma Genome Atlas database. Cell morphology, proliferation, and neuronal marker expression were detected to evaluate the differentiation-inducing effect of the selected drugs. The bioinformatics analysis identified histone deacetylase (HDAC) inhibitors as potential drugs. HDAC1/3/7 showed the relationship with neuronal genes, and HDAC1 showed the highest level of inverse correlation with neuronal gene expression and had the highest hazard ratio. In vitro study showed that both the pan-HDAC inhibitor belinostat, class I and class IIa HDAC inhibitor valproic acid, and selective HDAC1 inhibitor parthenolide induce morphology alteration, proliferation inhibition, expression of neuronal markers including microtubule-associated protein 2, neuronal nuclei antigen, and synaptophysin in U87 cells. This study suggests that the HDAC inhibitors belinostat, valproic acid, and parthenolide can induce glioma cells to differentiate into neuron-like cells, with HDAC1/3/7 being the likely drug targets and HDAC1 potentially playing an important role in this.