Cell Types Research Articles

Single cell transcriptomic analysis of SGLT2 expression supports an indirect or off-target role for the cardioprotective benefits of empagliflozin in heart failure

Sodium-glucose cotransporter 2 inhibitors (SGLT2i), such as empagliflozin, have shown remarkable benefits in reducing cardiovascular events and mortality in patients with heart failure irrespective of diabetes. Because of the magnitude of the benefits and broad application in both heart failure with reduced and preserved ejection fraction, there have been concerted efforts to identify a mechanism for the observed benefits. One hypothesis is that SGLT2i act directly on the heart. Given empagliflozin’s high specificity to SGLT2, we reasoned that SGLT2 expression would be a requirement for cells to respond to treatment via the expected drug target. Here, we present a comprehensive transcriptomic analysis of SLC5A2, which encodes SGLT2 at the single cell level in multiple datasets from healthy and HF donors, confirming its expression in a subset of kidney epithelial cells but minimal expression in other cell types. This was true irrespective of developmental stage, disease state, sequencing method or depth, and species. Therefore, it is likely that the cardioprotective benefits of SGLT2i cannot be explained by “canonical” interactions with SGLT2.

The Covert Side of Ascites in Cirrhosis: Cellular and Molecular Aspects

Ascites, a common complication of portal hypertension in cirrhosis, is characterized by the accumulation of fluid within the peritoneal cavity. While traditional theories focus on hemodynamic alterations and renin–angiotensin–aldosterone system (RAAS) activation, recent research highlights the intricate interplay of molecular and cellular mechanisms. Inflammation, mediated by cytokines (interleukin-1, interleukin-4, interleukin-6, tumor necrosis factor-α), chemokines (chemokine ligand 21, C-X-C motif chemokine ligand 12), and reactive oxygen species (ROS), plays a pivotal role. Besides pro-inflammatory cytokines, hepatic stellate cells (HSCs), sinusoidal endothelial cells (SECs), and smooth muscle cells (SMCs) contribute to the process through their activation and altered functions. Once activated, these cell types can worsen ascites accumulationthrough extracellular matrix (ECM) deposition and paracrine signals. Besides this, macrophages, both resident and infiltrating, through their plasticity, participate in this complex crosstalk by promoting inflammation and dysregulating lymphatic system reabsorption. Indeed, the lymphatic system and lymphangiogenesis, essential for fluid reabsorption, is dysregulated in cirrhosis, exacerbating ascites. The gut microbiota and intestinal barrier alterations which occur in cirrhosis and portal hypertension also play a role by inducing inflammation, creating a vicious circle which worsens portal hypertension and fluid accumulation. This review aims to gather these aspects of ascites pathophysiology which are usually less considered and to date have not been addressed using specific therapy. Nonetheless, it emphasizes the need for further research to understand the complex interactions among these mechanisms, ultimately leading to targeted interventions in specific molecular pathways, aiming towards the development of new therapeutic strategies.

The HOX code of human adult fibroblasts reflects their ectomesenchymal or mesodermal origin

Fibroblasts, the most abundant cell type in the human body, play crucial roles in biological processes such as inflammation and cancer progression. They originate from the mesoderm or neural-crest-derived ectomesenchyme. Ectomesenchyme-derived fibroblasts contribute to facial formation and do not express HOX genes during development. The expression and role of the HOX genes in adult fibroblasts is not known. We investigated whether the developmental pattern persists into adulthood and under pathological conditions, such as cancer. We collected adult fibroblasts of ectomesenchymal and mesodermal origins from distinct body parts. The isolated fibroblasts were characterised by immunocytochemistry, and their transcriptome was analysed by whole genome profiling. Significant differences were observed between normal fibroblasts from the face (ectomesenchyme) and upper limb (mesoderm), particularly in genes associated with limb development, including HOX genes, e.g., HOXA9 and HOXD9. Notably, the pattern of HOX gene expression remained consistent postnatally, even in fibroblasts from pathological tissues, including inflammatory states and cancer-associated fibroblasts from primary and metastatic tumours. Therefore, the distinctive HOX gene expression pattern can serve as an indicator of the topological origin of fibroblasts. The influence of cell position and HOX gene expression in fibroblasts on disease progression warrants further investigation.

GelMA as scaffold material for epithelial cells to emulate the small intestinal microenvironment

Host-microbe interactions in the intestine play a significant role in health and disease. Novel scaffolds for host cells, capable of potentially supporting these intricate interactions, are necessary to improve our current systems for mimicking host-microbiota interplay in vitro/ex vivo. In this research paper, we study the application of gelatin methacrylamide (GelMA) as scaffold material for intestinal epithelial cells in terms of permeability, mechanical strength, and biocompatibility. We investigated whether the degree of substitution (DS) of GelMA influences the permeability and found that both high and low DS GelMA show sufficient permeability of biorelevant transport markers. Additionally, we researched epithelial cell adherence and viability, as well as mechanical characteristics of different concentrations of GelMA. All concentrations of hydrogel show long-term biocompatibility for the mono- and co-cultures, despite the goblet-like cells (LS174T) showing lower performance than enterocyte-like cells (Caco-2). The mechanical strength of all hydrogel concentrations was in a physiologically relevant range to be used as scaffold material for intestinal cells. Lastly, we examined the effect of the two sterilization methods, ethylene oxide (EO) and 70% ethanol followed by UVC (EtOH/UVC). We found that the impact of the two methods on the mechanical characteristics was minimal, and we did not find a significant effect between the two methods on cell viability and confluency of Caco-2 cells seeded on the GelMA hydrogels. Based on these results, we conclude that GelMA is a suitable material as a scaffold for intestinal cell types in terms of permeability, mechanical strength and biocompatibility. These findings contribute to the growing field of in vitro modeling of the gut and moves the field further to ensuring more translatable research on host-microbe interactions.

Identification of potential drug targets for pelvic organ prolapse using a proteome-wide Mendelian randomization approach

Pelvic organ prolapse (POP) significantly impacts patients’ quality of life, and current treatment options remain limited due to high recurrence rates, making the exploration of new therapeutic targets essential. Using data from the FinnGen cohort, we performed a proteome-wide Mendelian randomization (PW-MR) analysis. Through PW-MR and Bayesian colocalization analyses, we identified EFEMP1 and MFAP4 as potential key drug targets, with EFEMP1 potentially exerting a protective effect, whereas MFAP4 may be associated with an increased risk of POP. To further support these findings, we analysed single-cell RNA sequencing data to evaluate the expression patterns of EFEMP1 and MFAP4 in different cell populations. The analysis revealed that EFEMP1 and MFAP4 are specifically enriched in cell types involved in tissue remodelling and fibrosis. Findings of phenome-wide association studies indicated that the risk of side effects for these targets may be low, suggesting the safety of treatment focused on these targets. Preliminary molecular docking analysis findings suggested that EFEMP1 and MFAP4 may have strong binding affinities with candidate drugs, further supporting the feasibility of EFEMP1 and MFAP4 as drug targets. In conclusion, our findings indicate that EFEMP1 and MFAP4 are promising therapeutic targets for POP, providing important insights for the development of safe and effective treatments.

The relationship mammalian p38 with human health and its homolog Hog1 in response to environmental stresses in Saccharomyces cerevisiae

The mammalian p38 MAPK pathway plays a vital role in transducing extracellular environmental stresses into numerous intracellular biological processes. The p38 MAPK have been linked to a variety of cellular processes including inflammation, cell cycle, apoptosis, development and tumorigenesis in specific cell types. The p38 MAPK pathway has been implicated in the development of many human diseases and become a target for treatment of cancer. Although MAPK p38 pathway has been extensively studied, many questions still await clarification. More comprehensive understanding of the MAPK p38 pathway will provide new possibilities for the treatment of human diseases. Hog1 in S. cerevisiae is the conserved homolog of p38 in mammalian cells and the HOG MAPK signaling pathway in S. cerevisiae has been extensively studied. The deep understanding of HOG MAPK signaling pathway will help provide clues for clarifying the p38 signaling pathway, thereby furthering our understanding of the relationship between p38 and disease. In this review, we elaborate the functions of p38 and the relationship between p38 and human disease. while also analyzing how Hog1 regulates cellular processes in response to environmental stresses. 1, p38 in response to various stresses in mammalian cells.2, The functions of mammalian p38 in human health.3, Hog1 as conserved homolog of p38 in response to environmental stresses in Saccharomyces cerevisiae. 1, p38 in response to various stresses in mammalian cells. 2, The functions of mammalian p38 in human health. 3, Hog1 as conserved homolog of p38 in response to environmental stresses in S. cerevisiae.

Expulsion of iron-rich ferritin via CD63-mediated exosome drives ferroptosis resistance in ovarian cancer cells

IntroductionFerroptosis is a promising new target for ovarian cancer (OVCA) treatment. However, some OVCA cell types resist the induction of ferroptosis by limiting the intracellular accumulation of the labile iron pool (LIP).MethodsHEY, COV318 and PEO4 were treated with erastin and assessed for cell viability by using PI flow cytometry assays. Erastin-affected iron metabolism was analysed by using FerroOrange assay, Western Blot (WB) analysis of ferritin heavy chain (FtH), transferrin receptor (CD71), and ferroportin (FPN). Mitochondrial reactive oxygen species (mitROS) and lipid peroxidation were quantified via MitoSOX and BODIPY-C11 flow cytometry assays, respectively. Exosomes (EVs) were collected from cell culture media through ultracentrifugation and then enumerated and analyzed by Nanoparticale Tracking Analysis (NTA) and transmission electron microscopy (TEM). CD63 protein expression in EVs was measured through WB by using CD9 as a loading control. Loss-of-function assays for FtH and CD63 were performed by using siRNA-mediated transient transfection.ResultsWe demonstrate that erastin treatment (8 µM, 8 h) is accompanied by the release of iron-rich ferritin via EV pathway in COV318 and PEO4 OVCA cells, thus failing to exert cytotoxic effects. Mechanistically, erastin causes the upregulation of CD63, a tetraspanin involved in forming multivesicular bodies (MVBs) and EVs, and the increase of MBVs assessed by transmission electron microscopy. Consistent with these findings, EV isolation followed by nanoparticle tracking analysis revealed a significant increase in EVs/cell in erastin-treated COV318 and PEO4 cells. Notably, EVs harvested from these cells contained CD63 and FtH, a major iron-storage protein. Inhibition of EV biogenesis with GW4869 prevented FtH release and restored LIP accumulation, lipid peroxidation, and ferroptosis sensitivity in COV318 and PEO4 cells.DiscussionOverall, our results indicate that OVCA cells can utilize CD63+ EVs to secrete iron-rich ferritin as a mechanism to evade erastin-induced ferroptosis. These findings suggest that combining erastin with EV inhibitors could offer promising strategy for overcoming ferroptosis resistance in OVCA.

Cladocroce caelum (Porifera) has a specialized growth region and a transcriptional regionalization across its body axis

Abstract It is widely acknowledged that sponges (Porifera) lack clear axes and regional specialization. Understanding whether sponges can have preferential growth regions and axial patterning is pivotal to comprehend the evolution of the body patterns in metazoans. We compared two body regions of the repent morph of Cladocroce caelum (Demospongiae): the proximal (with fully developed aquiferous system module) and distal regions (the hypothetical growth region). We found that specimens grew preferentially from the distal region, where the main cell type was archaeocytes (mean 65.3%), while the proximal region had aquiferous system modules comprising mainly choanocytes (47%). We also assembled a de novo reference transcriptome of C. caelum and recovered 99% of orthologous predicted genes for Eukarya and 94% for Metazoa. Our RNA-seq data revealed that most genes related to the germline, stem cell, and developmental signaling pathways are expressed in both proximal and distal regions. However, we retrieved 11,421 differentially expressed genes between the two regions (3,506 upregulated at the proximal region and 7,925 at the distal one). Some of these genes in the distal region (like WNT, TGF-β and MAPK pathway components) are related to development and cell proliferation. Our work compiles evidence for morphological and molecular differences between proximal and distal regions, suggesting that there is a preferential growth region in the distal end of C. caelum. Thus, our results indicate that despite the uniqueness of the sponge body plan, some poriferans may putatively share specialized growth regions with other animals.

Comparative therapeutic efficacy of Olea europaea L. leaf extract and bone marrow-derived mesenchymal stem cells in ameliorating streptozotocin-induced hepatic and splenic damage in pregnant Rats: Mechanisms of antioxidant and anti-apoptotic protection

Mesenchymal stem cells (MSCs) are multipotent stromal cells with remarkable plasticity, enabling them to differentiate into various tissue-specific cell types. MSCs play a pivotal role in tissue repair, hematopoiesis and immunomodulation. This study aimed to evaluate and compare the protective effects of olive leaf extract (OLE), bone marrow-derived mesenchymal stem cells (BM-MSCs) and their combination against hepatic and splenic toxicity in a rat model of gestational diabetes induced by streptozotocin (STZ). Histopathological and immunohistochemical analyses of liver and spleen tissues were conducted to assess these effects. Methods: Pregnant female rats were divided into five groups (n = 10 per group): Control group: Rats received STZ at a dose of 35 mg/kg body weight. OLE group: Rats were administered olive leaf extract (OLE) at 200 mg/kg body weight. GD + OLE group: Rats with STZ-induced gestational diabetes (GD) were treated with OLE (200 mg/kg body weight). GD + MSCs group: Rats with STZ-induced GD were treated with BM-MSCs. GD + OLE + MSCs group: Rats with STZ-induced GD were treated with both OLE and BM-MSCs. Results: STZ administration induced significant histopathological and immunohistochemical alterations in the liver and spleen tissues of pregnant femal rats. Treatment with OLE, BM-MSCs, or their combination markedly ameliorated these STZ-induced deteriorations, with the combined treatment showing the most pronounced protective effects. Conclusion: The findings demonstrate that both BM-MSCs and OLE exert protective effects against hepatotoxicity and splenic toxicity in a rat model of gestational diabetes. The combination of OLE and BM-MSCs exhibited synergistic benefits, highlighting their potential as therapeutic agents for mitigating organ damage in gestational diabetes.

Bone-on-a-Chip Systems for Hematological Cancers

Hematological malignancies originating from blood, bone marrow, and lymph nodes include leukemia, lymphoma, and myeloma, which necessitate the use of a distinct chemotherapeutic approach. Drug resistance frequently complicates their treatment, highlighting the need for predictive tools to guide therapeutic decisions. Conventional 2D/3D cell cultures do not fully encompass in vivo criteria, and translating disease models from mice to humans proves challenging. Organ-on-a-chip technology presents an avenue to surmount genetic disparities between species, offering precise design, concurrent manipulation of various cell types, and extrapolation of data to human physiology. The development of bone-on-a-chip (BoC) systems is crucial for accurately representing the in vivo bone microenvironment, predicting drug responses for hematological cancers, mitigating drug resistance, and facilitating personalized therapeutic interventions. BoC systems for modeling hematological cancers and drug research can encompass intricate designs and integrated platforms for analyzing drug response data to simulate disease scenarios. This review provides a comprehensive examination of BoC systems applicable to modeling hematological cancers and visualizing drug responses within the intricate context of bone. It thoroughly discusses the materials pertinent to BoC systems, suitable in vitro techniques, the predictive capabilities of BoC systems in clinical settings, and their potential for commercialization.

Proinflammatory Cytokines in Chronic Respiratory Diseases and Their Management

Pulmonary homeostasis can be agitated either by external environmental insults or endogenous factors produced during respiratory/pulmonary diseases. The lungs counter these insults by initiating mechanisms of inflammation as a localized, non-specific first-line defense response. Cytokines are small signaling glycoprotein molecules that control the immune response. They are formed by numerous categories of cell types and induce the movement, growth, differentiation, and death of cells. During respiratory diseases, multiple proinflammatory cytokines play a crucial role in orchestrating chronic inflammation and structural changes in the respiratory tract by recruiting inflammatory cells and maintaining the release of growth factors to maintain inflammation. The issue aggravates when the inflammatory response is exaggerated and/or cytokine production becomes dysregulated. In such instances, unresolving and chronic inflammatory reactions and cytokine production accelerate airway remodeling and maladaptive outcomes. Pro-inflammatory cytokines generate these deleterious consequences through interactions with receptors, which in turn initiate a signal in the cell, triggering a response. The cytokine profile and inflammatory cascade seen in different pulmonary diseases vary and have become fundamental targets for advancement in new therapeutic strategies for lung diseases. There are considerable therapeutic approaches that target cytokine-mediated inflammation in pulmonary diseases; however, blocking specific cytokines may not contribute to clinical benefit. Alternatively, broad-spectrum anti-inflammatory approaches are more likely to be clinically effective. Herein, this comprehensive review of the literature identifies various cytokines (e.g., interleukins, chemokines, and growth factors) involved in pulmonary inflammation and the pathogenesis of respiratory diseases (e.g., asthma, chronic obstructive pulmonary, lung cancer, pneumonia, and pulmonary fibrosis) and investigates targeted therapeutic treatment approaches.

Dynamic changes of host immune response during Helicobacter pylori-induced gastric cancer development.

Helicobacter pylori infection is the main risk factor for gastric cancer. Chronic inflammation is usually induced by H. pylori infection and is accompanied by inherent immune disorders. However, the dynamic changes in the host immune response associated with the transition from normal to metaplasia, dysplasia, and gastric cancer are largely undefined. We established the H. pylori induced gastric cancer mice model. The gastric mucosa of H. pylori infected mice were subjected to RNA-sequencing analysis at different stages. We analyzed systemic immune disturbances in the spleen and changes in serum inflammatory cytokines during gastric cancer development, including gastritis, premalignant lesions (pre-gastric cancer), and gastric cancer stages. RNA-sequencing analysis of the gastric mucosa of H. pylori infected mice highlighted the important role of immune-associated pathways (especially inflammatory pathways) during gastric cancer development. Immune cell proportion analysis revealed the stage-dependent involvement of key immune cell types, including increased Th17 cells in early gastritis and pre-gastric cancer stages and decreased central memory CD4+ and CD8+ T cells during gastric cancer transition. Serum inflammatory cytokine analysis showed that IL-6 and IL-10 levels significantly increased, whereas IL-23 levels decreased during the gastric cancer stage. In summary, we illustrated systemic immune disturbances in the spleen and changes in serum inflammatory cytokines during gastric cancer development. Th17 cells were involved in early gastritis and premalignant processes, while central memory T cells participated in gastric cancer transition. Our findings provide valuable insights into identifying key inflection points and associated biomarkers for the early detection, diagnosis, and treatment of gastric cancer.

Antioxidant Properties of Aronia melanocarpa and Morinda citrifolia Juices and their Impact on Bladder Cancer Cell Lines.

BACKGROUND Aronia melanocarpa and Morinda citrifolia are valuable sources of bioactive compounds, mainly polyphenols, that provide health benefits. The antioxidants can scavenge free radicals that generate oxidative stress, resulting in various chronic diseases, including cancer. This research analyzed the in vitro effect of Aronia melanocarpa and Morinda citrifolia (Aronia and Noni) juices on urinary bladder cancer cells (T24) in comparison with normal uroepithelial cells (SV-HUC1). MATERIAL AND METHODS The study was preceded by analysis of the antioxidant properties of the tested juices - antioxidant capacity, total polyphenols, total flavonoids, and total anthocyanins. Moreover, the phenolic acids and selected flavonoids in the juices were determined through high-performance liquid chromatography. The cytotoxicity of the tested juices was analyzed in cell lines via 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay. RESULTS Aronia and Noni juices affected the viability of both tested cell types in a time- and concentration-dependent manner. The results showed that Aronia and Noni samples were significantly more cytotoxic to urinary bladder cancer cells than to normal cells. Furthermore, it was found that Aronia juice was more cytotoxic against the tested cells than Noni juice. Aronia melanocarpa juice affected T24 cancer cells already at a concentration of 1.56% while Noni juice needed higher concentrations, starting at 12.5%. The greater efficacy of Aronia could be the result of its higher antioxidant capacity (629.4 vs 77.3 mg Tx/100 mL) or its higher content of total polyphenols (1036.6 mg vs 235.7 mg gallic acid equivalents/100 mL). CONCLUSIONS The results are promising and could indicate that Aronia and Noni juices exert beneficial effects against bladder cancer cells. Further studies should be performed, especially on other cancer cell lines.

Novel function of TREK-1 in regulating adipocyte differentiation and lipid accumulation

K2P (two-pore domain potassium) channels, a diversified class of K+-selective ion channels, have been found to affect a wide range of physiological processes in the body. Despite their established significance in regulating proliferation and differentiation in multiple cell types, K2P channels’ specific role in adipogenic differentiation (adipogenesis) remains poorly understood. In this study, we investigated the engagement of K2P channels, specifically KCNK2 (also known as TREK-1), in adipogenesis using primary cultured adipocytes and TREK-1 knockout (KO) mice. Our findings showed that TREK-1 expression in adipocytes decreases substantially during adipogenesis. This typically causes an increased Ca2+ influx and alters the electrical potential of the cell membrane in 3T3-L1 cell lines. Furthermore, we observed an increase in differentiation and lipid accumulation in both 3T3-L1 cell lines and primary cultured adipocytes when the TREK-1 activity was blocked with Spadin, the specific inhibitors, and TREK-1 shRNA. Finally, our findings revealed that mice lacking TREK-1 gained more fat mass and had worse glucose tolerance when fed a high-fat diet (HFD) compared to the wild-type controls. The findings demonstrate that increase of the membrane potential at adipocytes through the downregulation of TREK-1 can influence the progression of adipogenesis.

Identification of a novel small-molecule inhibitor of the HIV-1 reverse transcriptase activity with a non-nucleoside mode of action

BackgroundHuman immunodeficiency virus-1 (HIV-1) is the causative agent of acquired immunodeficiency syndrome, which is a major global health problem. Although combination antiretroviral therapy (cART) successfully expands the lifespan of HIV-1-infected patients, long-term cART often increases drug resistance and adverse effects. Therefore, efforts are ongoing to develop novel anti-HIV-1 drugs.MethodsThe anti-HIV-1 activities of compounds were investigated using TZM-bl reporter cell line, A3.01 T cell line, and peripheral blood mononuclear cells infected with several HIV-1 strains, including wild type and drug-resistance associated mutants. Next-generation sequencing analysis and in silico molecular docking studies were employed to determine the mode of action of the compound.ResultsWe identified a small-molecule inhibitor consisting of a thiadiazole core appended to two pyrazoles (BPPT), which exerted a highly potent inhibitory effect on HIV-1 infectivity, with a half-maximal effective concentration (EC50) of 60 nM, without causing cytotoxicity. In experiments with various HIV-1 strains and cell types, the potency of BPPT was found to be comparable to that of commercial antiretroviral agents (azidothymidine, nevirapine, and others). Further analysis of the mode of action demonstrated that BPPT is a novel type of HIV-1 non-nucleoside reverse transcriptase inhibitor (NNRTI). Analysis of viruses harboring drug-resistance-associated mutations showed that BPPT was potent against G190A (C or S) mutations in reverse transcriptase (RTase), exhibiting high-level resistance to other NNRTIs. Next-generation sequencing analysis of long-term treatment with BPPT displayed an RTase mutation profile different from that in the case of established NNRTIs. Given these data, in silico molecular docking studies demonstrated the molecular mechanism underlying the BPPT-mediated inhibition of RTase.ConclusionOur data suggest that BPPT is a novel small-molecule inhibitor of HIV-1 RTase and could serve as a promising chemical scaffold to complement or replace conventional treatments, particularly for overcoming resistance associated with the G190 mutation.

ENACT: End-to-end Analysis of Visium High Definition (HD) Data.

Spatial transcriptomics (ST) enables the study of gene expression within its spatial context in histopathology samples. To date, a limiting factor has been the resolution of sequencing based ST products. The introduction of the Visium High Definition (HD) technology opens the door to cell resolution ST studies. However, challenges remain in the ability to accurately map transcripts to cells and in assigning cell types based on the transcript data. We developed ENACT, a self-contained pipeline that integrates advanced cell segmentation with Visium HD transcriptomics data to infer cell types across whole tissue sections. Our pipeline incorporates novel bin-to-cell assignment methods, enhancing the accuracy of single-cell transcript estimates. Validated on diverse synthetic and real datasets, our approach is both scalableto samples with hundreds of thousands of cells and effective, offering a robust solution for spatially resolved transcriptomics analysis. ENACT source code is available at https://github.com/Sanofi-Public/enact-pipeline. Experimental data is available at https://zenodo.org/records/14748859. Supplementary data are available at Bioinformatics online.

Multi-Omics Analysis of the Anoikis Gene CASP8 in Prostate Cancer and Biochemical Recurrence (BCR)

Background: Prostate cancer, as an androgen-dependent malignant tumor in older men, has attracted the attention of a wide range of clinicians. BCR remains a significant challenge following early prostate cancer treatment. Methods: The specific expression pattern of the Anoikis gene set in prostate cancer cells was first explored by single-cell and spatial transcriptomics analysis. Genes causally associated with prostate cancer were screened using Summary-data-based Mendelian Randomization (SMR). Subsequently, we explored the role and mechanism of CASP8 in prostate cancer cells and defined a new cell type: the CASP8 T cell. We constructed a prediction model that can better predict the BCR of prostate cancer, and explored the differences in various aspects of clinical subgroups, tumor microenvironments, immune checkpoints, drug sensitivities, and tumor-immune circulations between high- and low-risk groups. The results of SMR analysis indicated that CASP8 could increase the risk of prostate cancer. Based on the differential genes of CASP8-positive and -negative T cells, we constructed a four-gene prognostic model with a 5-year AUC of 0.713. Results: The results revealed that high-risk prostate cancer BCR patients had various characteristics such as higher tumor purity, higher BCR rate, downregulated SIRPA immune checkpoints, and unique drug sensitivity. Conclusions: In summary, CASP8 may be a potential biomarker for prostate cancer.

Mutations in the microexon splicing regulator srrm4 have minor phenotypic effects on zebrafish neural development.

Achieving a diversity of neuronal cell types and circuits during brain development requires alternative splicing of developmentally regulated mRNA transcripts. Microexons are a type of alternatively spliced exon that are 3-27 nucleotides in length and are predominantly expressed in neuronal tissues. A key regulator of microexon splicing is the RNA-binding protein Serine/arginine repetitive matrix 4 (Srrm4). Srrm4 is a highly conserved, vertebrate splicing factor that is part of an ancient family of splicing proteins. To better understand the function of Srrm4 during brain development, we examined neural expression of zebrafish srrm4 from days 1-5 of development using fluorescence in situ hybridization. We found that srrm4 has a dynamically changing expression pattern, with expression in diverse cell types and stages during development. We then used CRISPR-based mutagenesis to generate zebrafish srrm4 mutants. Unlike previously described morphant phenotypes, srrm4 mutants did not show overt morphological defects. Whole brain morphometric analysis revealed a reduction in optic tectum neuropil in G0 crispants that, unexpectedly, was also not replicated in stable mutants. Sequencing of wild-type and mutant transcriptomes revealed only minor changes in splicing and did not support a hypothesis of transcriptional adaptation, suggesting that another, as yet, unidentified mechanism of compensation is occurring. srrm4 thus appears to have a limited role in zebrafish neural development.

Catestatin in Cardiovascular Diseases

Cardiovascular diseases are one of the leading causes of mortality and morbidity worldwide. The pathogenesis of this group of disorders is highly complex and involves interactions between various cell types and substances, among others, catestatin (CTS). In recent years, numerous researchers have expanded our knowledge about CTS’s role in development and its potential for the treatment of a variety of diseases. In this review, the authors discuss CTS’s importance in the pathogenesis of arterial hypertension, coronary artery disease, and heart failure. Moreover, we present CTS’s influence on heart and vessel function.

Optimizing deep learning for accurate blood cell classification: A study on stain normalization and fine-tuning techniques

Abstract: BACKGROUND: Deep learning’s role in blood film screening is expanding, with recent advancements including algorithms for the automated detection of sickle cell anemia, malaria, and leukemia using smartphone images. OBJECTIVES: This study aims to build the artificial intelligence (AI) models and assess their performance in classifying blood film cells as normal or abnormal. MATERIALS AND METHODS: The dataset included 171,374 images from 961 patients which were classified by experts. These images were resized, denoized, normalized, augmented, and classified into two categories, normal and abnormal cells. Two stain normalization techniques were used in this study; Reinhard and Mackenko techniques. The data were split into training and testing sets with a ratio of (8:2). The model was built through transfer learning by using the pretrained model Inception-Resnet v2 as a backbone. Three different fine-tuning techniques were tested in this study. The training was done using Python with Keras library on Google Colab for 10 epochs. The model was tested for accurately classifying individual blood cells whether normal or abnormal and evaluated using accuracy and area under receiver operator characteristic curve. RESULTS: The counts of the three most common cell types were as follows: Segmented neutrophils: 29,424; erythroblasts: 27,395; and lymphocytes: 26,242. The Reinhard stain normalization had better accuracy than Mackenko, the best AI model achieved the highest accuracy of 96.7%%, the area under the curve (AUC) of 99.87%, while the second technique achieved an accuracy of 91.46% and an AUC of 97.23% in classifying normal from abnormal cells. CONCLUSION: In conclusion, AI can effectively classify the blood cells as either normal or abnormal, yielding accurate results in a time-effective manner, especially with the use of transfer learning of pretrained models and fine-tuning. In this study, Inception-Resnet V2 showed good accuracy in differentiating normal from abnormal cells.