Cellular Differentiation Research Articles

Volume-regulated anion channels conduct ATP in undifferentiated mammary cells and promote tumorigenesis in xenograft nude mouse

The high interstitial ATP concentration in the cancer microenvironment is a major source of adenosine, which acts as a strong immune suppressor. However, the source of ATP release has not been elucidated. We measured ATP release during hypotonic stress using a real-time ATP luminescence imaging system in breast cell lines and in primary cultured mammary cells. In breast cell lines, ATP was released with a slowly rising diffuse pattern, whereas in primary cultured cells, ATP was intermittently released with transient-sharp peaks. The diffuse ATP release pattern changed to a transient-sharp pattern by cholera toxin treatment and the reverse change was induced by transforming growth factor (TGF) β treatment. DCPIB, an inhibitor of volume-regulated anion channels (VRACs), suppressed the diffuse pattern. The inflammatory mediator sphingosine-1-phosphate (S1P) induced a diffuse ATP release pattern isovolumetrically. Knockdown of the A isoform of leucine-rich repeat-containing protein 8 (LRRC8A), the essential molecular entity of VRACs, using shRNA suppressed the diffuse pattern. In the nude mouse xenograft model, LRRC8A knockdown suppressed the tumorigenesis of subcutaneously implanted breast cancer cells. These results suggest that abundantly expressed VRACs are a conduit of ATP release in undifferentiated cells, including cancer cells.

Effect of introducing somatic mitochondria into an early embryo on zygotic gene activation†.

Unlike differentiated somatic cells, which possess elongated mitochondria, undifferentiated cells, such as those of preimplantation embryos, possess round, immature mitochondria. Mitochondrial morphology changes dynamically during cell differentiation in a process called mitochondrial maturation. The significance of the alignment between cell differentiation and mitochondrial maturity in preimplantation development remains unclear. In this study, we analyzed mouse embryos into which liver-derived somatic mitochondria were introduced (SM-embryos). Most SM-embryos were arrested at the two-cell stage. Some of the introduced somatic mitochondria became round, while others remained elongated and large. RNA-sequencing revealed a disruption of both minor and major zygotic gene activation (ZGA) in SM-embryos. Minor ZGA did not terminate before major ZGA, and the onset of major ZGA was inhibited, as shown by histone modification analyses of histone H3 lysine 4 trimethylation and histone H3 lysine 27 acetylation. Further analysis of metabolites involved in histone modification regulation in SM-embryos showed a significantly lower NAD+/NADH ratio in SM-embryos than in control embryos. Additionally, the mitochondrial membrane potential, an indicator of mitochondrial function, was lower in SM-embryos than in control embryos. Our results demonstrated that introducing somatic mitochondria into an embryo induces mitochondrial dysfunction, thereby disrupting metabolite production, leading to a disruption in ZGA and inducing developmental arrest. Our findings reveal that the alignment between cell differentiation and mitochondrial maturity is essential for early embryonic development.

PLUNC downregulates the expression of PD-L1 by inhibiting the interaction of DDX17/β-catenin in nasopharyngeal carcinoma

Background: Nasopharyngeal carcinoma (NPC) is characterized by high programmed death-ligand 1 (PD-L1) expression and abundant infiltration of non-malignant lymphocytes, which renders patients potentially suitable candidates for immune checkpoint blockade therapies. Palate, lung, and nasal epithelium clone (PLUNC) inhibit the growth of NPC cells and enhance cellular apoptosis and differentiation. Currently, the relationship between PLUNC (as a tumor-suppressor) and PD-L1 in NPC is unclear.Methods: We collected clinical samples of NPC to verify the relationship between PLUNC and PD-L1. PLUNC plasmid was transfected into NPC cells, and the variation of PD-L1 was verified by western blot and immunofluorescence. In NPC cells, we verified the relationship of PD-L1, activating transcription factor 3 (ATF3), and β-catenin by western blot and immunofluorescence. Later, we further verified that PLUNC regulates PD-L1 through β-catenin. Finally, the effect of PLUNC on β-catenin was verified by co-immunoprecipitation (Co-IP).Results: We found that PLUNC expression was lower in NPC tissues than in paracancer tissues. PD-L1 expression was opposite to that of PLUNC. Western blot and immunofluorescence showed that β-catenin could upregulate ATF3 and PD-L1, while PLUNC could downregulate ATF3/PD-L1 by inhibiting the expression of β-catenin. PLUNC inhibits the entry of β-catenin into the nucleus. Co-IP experiments demonstrated that PLUNC inhibited the interaction of DEAD-box helicase 17 (DDX17) and β-catenin.Conclusions: PLUNC downregulates the expression of PD-L1 by inhibiting the interaction of DDX17/β-catenin in NPC.

Stem cell status and prognostic applications of cuproptosis-associated lncRNAs in acute myeloid leukemia

IntroductionAcute myeloid leukemia (AML), a highly heterogeneous hematological malignancy, remains a major challenge in adult oncology. Stem cell research has highlighted the crucial role of long noncoding RNA (lncRNA) in regulating cellular differentiation and self-renewal processes, which are pivotal in AML pathogenesis and therapy resistance.MethodsThis study explores the relationship between cuproptosis-related lncRNAs and AML prognosis, providing novel insights into their impact on hematopoietic stem and progenitor cells.ResultsWe collected clinical information from 214 AML patients in our center and analyzed the association between granulocyte recovery after chemotherapy, cuproptosis, and prognosis. Additionally, we developed a prognostic model—the cuproptosis-associated long noncoding RNA prognostic model (CRLPM)—y analyzing data from The Cancer Genome Atlas (TCGA). Patients were stratified into high- and low-risk groups based on CRLPM, revealing significant survival differences. High-risk patients demonstrated lower sensitivity to chemotherapeutic agents such as Axitinib, GSK429286A, Navitoclax, and ZM-447439, underscoring the need for alternative therapeutic strategies.DiscussionCRLPM offers a promising framework for integrating stem cell-focused approaches into personalized treatment regimens, paving the way for precision medicine in AML management.

Relationship Between Pituitary Gland and Stem Cell in the Aspect of Hormone Production and Disease Prevention: A Narrative Review.

In the last two decades, scientists have gained a better understanding of several aspects of pituitary development. The signaling pathways that govern pituitary morphology and development have been identified, and the compensatory relationships among them are now known. This paper aims to emphasize the wide variety of relationships between Pituitary Gland and Stem cells in hormone Production and disease prevention. Based on many case reports and several types of research, a wide variety of relationships between the Pituitary Gland and Stem cells in the aspect of hormone Production and disease prevention are reviewed in this literature. In this paper, we focus on various roles and functions of the pituitary gland, the responsibilities of stem cells as a mode of hormone production, and disease prevention. Within this period, more was discovered concerning the contributions made to the transcription factors within the pituitary development, with factors such as Prop1, Pitx1, and Sox2 being defined as important in the development and action of hormone-secreting cells. They are also required in the appropriate specification of the cell types in the pituitary gland and the persistence of the progenitors. Manipulation of these factors causes developmental defects as well as tumors, thus the necessity of knowing the precise function and interaction of these factors. A closer look at these transcription factors could help expand treatment options for structural defect development or give rise to pituitary adenomas. It has been established that signaling pathways such as Sonic Hedgehog (Shh), Wnt, and Notch play a part in modulating pituitary development. These pathways are involved in regulating important processes such as cellular proliferation, differentiation, and organization of the pituitary gland tissues. Breaching any of these pathways has been correlated with the development of various pituitary-related conditions including adenomas and congenital hypopituitarism. Moving forward, further studies of these pathways and their associations with stem cells could provide a better understanding of disease processes and approaches to manage them. This way, there is a possibility of developing new approaches aimed at treating the cause of the dysfunction of the pituitary gland by modulating its specific signaling activities. Promising directions for the stimulation of hormone synthesis and restoration of normal pituitary function upon its disorders via tissue regrowth could be found in stem cell application. The fact that one can generate functional pituitary cells from iPSCs for instance provides new avenues both for the understanding of pituitary disease mechanisms as well as for personalized medicine. It is possible to utilize these stem cell- derived cells for modeling disease, drug discovery or even transplantation to restore the function of the damaged pituitary gland. In the future, however, the focus ought to be on the effective application of stem cell therapies that have been research during the development of better differentiation processes. The recent understanding of the system that carries the hypothalamic hormones to the pituitary gland, i.e., the hypophyseal portal vasculature, has had its implications too. This factorial consideration emphasizes the role of the vascular component in the control of pituitary activity - the release of hormones by the pituitary gland. Exploring stem cell-hypophyseal portal system interactions may open new avenues of treatment for diseases associated with deficient hormone transportation and/or pituitary dysfunction.

Effects of periodontal disease on the proteomic profile of the periodontal ligament.

Periodontal disease affects over 1 billion people globally. This study investigated how periodontitis affects the protein profile of the periodontal ligament (PDL) in rats. Eight Holtzman rats were divided into the control and experimental periodontitis groups. The PDL was isolated using laser capture microdissection, and protein extracts were analyzed by mass spectrometry. Data analysis utilized specialized software, and Gene Ontology enrichment analysis identified significant protein functions. The data are available via ProteomeXchange with identifier PXD055817. Proteins such as SerpinB1, C5, and Lgals3 were validated through immunohistochemistry, and their gene expression was examined in an in vitro human PDL cell line. This study identified 1326 proteins, with 156 unique to the control group, 294 unique to the periodontitis group, and 876 common to both groups. Enrichment analysis revealed that proteins associated with the regulation of enzyme activity and RNA binding were significantly represented in the periodontitis group. There were increased levels of SerpinB1, C5, and Lgals3 in the periodontitis group based on proteomic and immunohistochemical analyses. Furthermore, these targets showed increased gene expression in stimulated human PDL cells. This study provides insights into the periodontitis-related alterations in the protein composition of the PDL and PDL cells, identifying both novel and previously known disease-associated proteins. SIGNIFICANCE: The periodontal ligament plays a crucial role in oral functions by providing structural support to the tooth. Due to the presence of undifferentiated mesenchymal cells, research into its regenerative capacity is ongoing. Pathological conditions can affect these functions and protein composition. Currently, there is a lack of comprehensive research specifically focusing on evaluating the periodontal ligament in both healthy and diseased states. This pioneering study screened for protein alterations and the mechanisms related to periodontitis. The possibility of using proteomic analysis to evaluate the protein alterations that occur in periodontitis-a disease with a high global incidence-could provide therapeutic targets and new biomarkers for future clinical studies.

Scalable control of stem cell fate by riboswitch-regulated RNA viral vector without genomic integration.

Transgene expression in stem cells is a powerful means of regulating cellular properties and differentiation into various cell types. However, existing vectors for transgene expression in stem cells suffer from limitations such as the need for genomic integration, the transient nature of gene expression, and the inability to temporally regulate transgene expression, which hinder biomedical and clinical applications. Here we report a new class of RNA virus-based vectors for scalable and integration-free transgene expression in mouse embryonic stem cells (mESCs). The vector is equipped with a small molecule-regulated riboswitch and a drug selection marker that allow temporal regulation of transgene expression and stable maintenance of the vector in proliferating stem cells. We demonstrated the utility of the vector by maintaining the pluripotency of mESCs in a differentiation induction medium by expressing Nanog and inducing myogenic differentiation by triggering Myod1 expression, without altering the mESC genome.

Methylglyoxal compromises callus mineralization and impairs fracture healing through suppression of osteoblast terminal differentiation.

Impaired fracture healing in diabetic patients leads to prolonged morbidity and increased healthcare costs. Methylglyoxal (MG), a reactive metabolite elevated in diabetes, is implicated in various complications, but its direct impact on bone healing remains unclear. Here, using a non-diabetic murine tibial fracture model, we demonstrate that MG directly impairs fracture healing. Micro-computed tomography revealed decreased volumetric bone mineral density in the callus, while callus volume remained unchanged, resulting in a brittle bone structure. This was accompanied by reduced expression of osteocalcin and bone sialoprotein, both critical for mineralization. Biomechanical analysis indicated that MG reduced the mechanical resilience of the fracture site without altering its elastic strength, suggesting that the impairment was not primarily due to the accumulation of advanced glycation end-products in the bone extracellular matrix. In vitro studies confirmed that non-cytotoxic concentrations of MG inhibited osteoblast maturation and mineralization. Transcriptomic analysis identified downregulation of Osterix, a key transcription factor for osteoblast maturation, without altering Runx2 levels, leading to decreased expression of key mineralization-related factors like osteocalcin. These findings align with clinical observations of reduced circulating osteocalcin levels in diabetic patients, suggesting that the detrimental effects of MG on osteoblasts may extend beyond bone metabolism. Our study highlights MG and MG-sensitive pathways as potential therapeutic targets for improving bone repair in individuals with diabetes and other conditions characterized by elevated MG levels.

Emerging Combinatorial Drug Delivery Strategies for Breast Cancer: A Comprehensive Review.

Breast cancer remains the second most prevalent cancer among women in the United States. Despite advancements in surgical, radiological, and chemotherapeutic techniques, multidrug resistance continues to pose significant challenges in effective treatment. Combination chemotherapy has emerged as a promising approach to address these limitations, allowing multiple drugs to target malignancies via distinct mechanisms of action. Increasingly, the use of phytoconstituents alongside chemotherapeutic agents has shown promise in enhancing treatment outcomes. This combination therapy acts on key signaling pathways such as Hedgehog, Notch, Wnt/β- catenin, tyrosine kinases, and phosphatidylinositol 3-kinase (PI3K), which play critical roles in cellular proliferation, apoptosis, angiogenesis, differentiation, invasion, and metastasis. This review explores various signaling pathways involved in breast cancer progression, discusses conventional treatment methods like surgery, adjuvant radiotherapy, hormonal therapy, and chemotherapy, and highlights emerging nanocarrier-based drug delivery systems (DDS). Liposomes, dendrimers, exosomes, polymeric micelles, and nanoparticles (organic, inorganic, gold, magnetic, carbon-based, and quantum dots) are examined as innovative strategies for enhancing drug delivery efficacy. Furthermore, stimuli-responsive DDSs, including reactive oxygen species (ROS), enzyme-, and hypoxia- responsive systems, are presented as cutting-edge approaches to overcoming drug resistance. Special emphasis is placed on the co-delivery of chemotherapeutic agents and plant-based compounds, particularly in estrogen receptor-positive (ER+) breast cancer. This review aims to provide a comprehensive overview of novel combinatorial strategies and advanced nanocarriers for the effective and targeted treatment of breast cancer.

Reconstructing Waddington Landscape from Cell Migration and Proliferation.

The Waddington landscape was initially proposed to depict cell differentiation, and has been extended to explain phenomena such as reprogramming. The landscape serves as a concrete representation of cellular differentiation potential, yet the precise representation of this potential remains an unsolved problem, posing significant challenges to reconstructing the Waddington landscape. The characterization of cellular differentiation potential relies on transcriptomic signatures of known markers typically. Numerous computational models based on various energy indicators, such as Shannon entropy, have been proposed. While these models can effectively characterize cellular differentiation potential, most of them lack corresponding dynamical interpretations, which are crucial for enhancing our understanding of cell fate transitions. Therefore, from the perspective of cell migration and proliferation, a feasible framework was developed for calculating the dynamically interpretable energy indicator to reconstruct Waddington landscape based on sparse autoencoders and the reaction diffusion advection equation. Within this framework, typical cellular developmental processes, such as hematopoiesis and reprogramming processes, were dynamically simulated and their corresponding Waddington landscapes were reconstructed. Furthermore, dynamic simulation and reconstruction were also conducted for special developmental processes, such as embryogenesis and Epithelial-Mesenchymal Transition process. Ultimately, these diverse cell fate transitions were amalgamated into a unified Waddington landscape.

Functional analysis of conserved C. elegans bHLH family members uncovers lifespan control by a peptidergic hub neuron.

Throughout the animal kingdom, several members of the basic helix-loop-helix (bHLH) family act as proneural genes during early steps of nervous system development. Roles of bHLH genes in specifying terminal differentiation of postmitotic neurons have been less extensively studied. We analyze here the function of 5 Caenorhabditis elegans bHLH genes, falling into 3 phylogenetically conserved subfamilies, which are continuously expressed in a very small number of postmitotic neurons in the central nervous system. We show (a) that 2 orthologs of the vertebrate bHLHe22/e23 genes, called hlh-17 and hlh-32, function redundantly to specify the identity of a single head interneuron class (AUA), as well as an individual motor neuron (VB2); (b) that the PTF1a ortholog hlh-13 acts as a terminal selector to control terminal differentiation and function of the sole octopaminergic neuron class in C. elegans, RIC; and (c) that the NHLH1/2 ortholog hlh-15 controls terminal differentiation and function of the peptidergic AVK head interneuron class, a known neuropeptidergic signaling hub in the animal. Strikingly, through null mutant analysis and cell-specific rescue experiments, we find that loss of hlh-15/NHLH in the peptidergic AVK neurons and the resulting abrogation of neuropeptide secretion from these neurons causes a substantially extended lifespan of the animal, which we propose to be akin to hypothalamic control of lifespan in vertebrates. Our functional analysis reveals themes of bHLH gene function during terminal differentiation that are complementary to the earlier lineage specification roles of other bHLH family members. However, such late functions are much more sparsely employed by members of the bHLH transcription factor family, compared to the function of the much more broadly employed homeodomain transcription factor family.

3D STED Imaging of Isolated Arabidopsis thaliana Nuclei.

Microscopy imaging of chromatin offers valuable insights into its spatial organization in the nucleus, a novel epigenetic dimension influencing the genome's functions. Particularly, visualization at the nanoscale in single cells is uniquely complementary to molecular profiling methods averaging chromatin configuration and composition over thousands of cells. How are chromatin and chromosomal domains distributed in relation to gene expression? How variable are these configurations? How do chromatin domains evolve in structure, composition, and distribution during cellular differentiation or cellular responses to environmental stimuli? Super-resolution microscopy techniques, like stimulated emission depletion (STED), are key in answering such questions. However, such imaging techniques are not often used in the field of plant cell biology compared to mammalian counterparts, which has greatly advanced our understanding of the 3D principles in genome organization. In an effort to bridge this gap, we provide a clear guide for isolating, embedding, immunostaining, and STED imaging intact leaf nuclei from Arabidopsis thaliana in 3D.

In vivo transplantation of intrahepatic cholangiocyte organoids with decellularized liver-derived hydrogels supports hepatic cellular proliferation and differentiation in chronic liver injury.

The limited replicative potential of primary hepatocytes (Hep) is a major hurdle for obtaining sufficient quantity and quality hepatocytes during cell therapy in patients with liver failure. Intrahepatic cholangiocyte organoids (ICOs) derived from intrahepatic bile ducts differentiate into both hepatocytes and cholangiocytes in vitro. Here, we studied in vivo effects of transplanting ICOs and Hep in chronic liver injury mice models. Well characterized primary mouse ICOs and Hep were mixed in decellularized liver (DCL) matrix hydrogels and injected into the subcapsular left lateral liver lobe of CCl4-induced liver injury models whereas mice given DCL alone were in the sham group. Two weeks post-transplantation, transplanted liver lobes were collected and studied by histology and RNA sequencing. Transplanted animals did not exhibit any tumors, mortality or morbidity. Mice livers transplanted with ICOs had increased cellular proliferation and vascularization as compared to Hep transplanted mice or sham. Collagen deposition in the liver was significantly reduced and serum albumin levels were significantly increased in transplanted groups compared to the sham group. Expression of genes associated with hepatocyte differentiation was highest in Hep transplanted livers among the three groups, but they were also upregulated in ICO transplanted livers compared to sham. Our study demonstrates that ICOs encapsulated in DCL hydrogels when transplanted in chronically injured mice livers engraft well and show hepatocyte differentiation and reduction of fibrosis, indicating that hydrogel transplanted cholangiocyte organoids may serve as an efficient cell source and therapy for renewal of hepatocytes, restoration of hepatocyte functions and resolution of liver injury.

Membrana preformativa: Unveiling the unexplored facets of dental development.

Odontogenesis is a complex and highly regulated biological process that involves a range of molecular mechanisms. Among these, Ki67 and Cyclin D1 are crucial cell cycle regulators that play pivotal roles in controlling cell proliferation during tooth development. This study aims to provide detailed insights into the expression patterns and functional significance of Ki67 and Cyclin D1 in tooth development. Through rigorous analysis, we seek to elucidate the intricate mechanisms underlying tooth development, helping to advance our understanding of this vital biological process. The procurement of rabbit tooth germ was performed only after obtaining the requisite ethical clearance. Subsequently, the tissues were processed and subjected to Hematoxylin and Eosin staining to facilitate enhanced visualization of the overall tissue architecture and organization & immunohistochemical staining of Ki-67 and Cyclin D1 was performed. In the tooth germ, cyclin D1 demonstrated intense staining in the dental papilla, especially in the membrana preformativa, with this intensity decreasing following predentin formation. Odontoblasts showed mild staining as they transitioned from pre-odontoblasts, which further diminished after dentin formation. Both the dental papilla and differentiating odontoblasts were positive for Ki67, though Ki67 staining in the odontoblasts reduced after dentin formation. In conclusion, the membrana preformativa plays a key role in odontogenesis, as indicated by its involvement in cellular proliferation and differentiation during tooth development.

Reactive oxygen species favors Varicellovirus bovinealpha 5 (BoAHV-5) replication in neural cells.

Varicellovirus bovinealpha (BoAHV) 1 and 5 are closely related neurotropic alphaherpesviruses with distinct neuropathogenic potential. BoAHV-5 causes meningoencephalitis in calves whereas encephalitis by BoAHV-1 infection is sporadic. the mechanisms underlying the differences in tropism and clinical outcomes of the infections are not yet completely understood. Here, we used neuroblastoma SH-SY5Y cells as non-differentiated in comparison with the SH-SY5Y neuronal-like cells obtained after exposing SH-SY5Y undifferentiated cells to trans-retinoic acid. We aimed to establish whether there was a relationship between the production of reactive oxygen species (ROS) and the kinetics of virus replication. We demonstrated that ROS production after BoAHV infection was higher in differentiated cells. Generation of ROS was also dependent on the infecting BoAHV strain. Higher ROS levels were produced during BoAHV-5 infection concomitantly with enhanced viral replication. We propose that increased ROS production mechanistically contributes to the tissue damage and neuroinflammation induced by BoAHV-5 infection. Future studies will determine specific targets of ROS that mediate the effects on viral replication.

Tumor Shrinkage can be a Promising Diagnosis Toward Remission or can also be an Ominous Diagnosis Toward Fatality

The objective of this article is to rectify tumor shrinkage as a valid diagnosis of cancer therapy. Cancer incidence and cancer mortality keep on increasing, which are an indication that cancer has not been handled right. Perpetual proliferation of cancer cells (CCs) is the most outstanding feature of cancer. Elimination of CCs to shrink tumor became the commanding principle, which was wrong, because cancer evolved due to wound unhealing. Killing to create wound definitely is contra-indication of cancer therapy. The mistake was committed at a time when we did not have the full knowledge of cancer, which was excusable. Now we have better knowledge of cancer, but the mistake is carrying on to result in 10 million annual mortality worldwide in 2019 with an anticipated annual increment of 5%, which is not excusable. Cytotoxic therapies can only benefit a small minority of cancer patients in the early stage whose chem-surveillance have not yet fatally damaged, relying on the restoration of chemo-surveillance to subdue cancer stem cells (CSCs), which are resistant to cytotoxic agents. Under this circumstance, tumor shrinkage is a promising diagnosis toward remission. Cytotoxic therapies cause the death of a majority of cancer patients in the advanced stage whose chemo-surveillance have been fatally damaged. Under this circumstance, tumor shrinkage is an ominous diagnosis toward fatality. Tumor shrinkage should be used with discretion. Cancer is a disease created by multiple factors. The collapse of chemo-surveillance or immuno-surveillance, the evolution of cancer stem cells (CSCs) from progenitor stem cells (PSCs) due to wound unhealing, and the progression of CSCs to faster replicating CCs by the activation of oncogenes or the inactivation of suppressor genes all contribute significantly to the development of cancer. An effective cancer therapy must be able to rectify all these contributing factors. Focusing on a specific factor is insufficient. The obsession to eliminate CCs to achieve tumor shrinkage is a grave mistake of cancer establishments to result in ever-increasing cancer mortality. Actually, elimination of CSCs is more important than the elimination of CCs, because CSCs contribute most fatal effects of cancer such as metastasis, drug resistance, anti-apoptosis, angiogenesis and recurrence. Induction of terminal differentiation by cell differentiation agent (CDA) formulations is the only option to eliminate CSCs which are critically linked to wound unhealing at the primary site. Induction of terminal differentiation can also result in the elimination of CCs, which are not as tightly linked to wound unhealing as CSCs. Terminal differentiation of CSCs and CCs cannot make the tumor to disappear. Thus, CDAs violated the commanding principle of tumor shrinkage put up by the cancer establishments, and were blocked as acceptable cancer drugs. CDA formulations are obviously the only drugs that can save the lives of advanced cancer patients whose chemo-surveillance have been fatally damaged. Oncologists and advanced cancer patients must unite to push for the approval of CDA formulations to save advanced cancer patients.

MiRNA-455-5p regulates the growth and development of adipose tissue by targeting IGF-1R gene

LAY SUMMARY Adipose tissue is an important factor that influences porcine quality, and adipose tissue is also an important endocrine regulator of metabolic homeostasis. Adipose tissue development involves a series of highly synergistic processes, including clone amplification, proliferation arrest and terminal differentiation. In recent years, microRNAs (miRNAs) are a class of non-coding endogenous RNA molecules that have been considered as a class of novel regulators of adipogenesis. In this study, we used the dorsal subcutaneous adipose tissue of Guangxi Bama mini-pigs and Landrace pigs as experimental materials to study the molecular mechanism of adipogenic differentiation of preadipocytes, and the regulatory mechanism of miRNA-455-5p and its target gene IGF-1R involved in the development of adipose tissue. These findings may improve our understanding of mechanism of the molecular regulation of miRNA-455-5p and IGF-1R in pigs, and provide a theoretical basis for increasing pork production. TEASER TEXT The miRNA-455-5p was involved in the negative regulation of adipose tissue development by inhibiting the expression of its target gene IGF-1R. ABSTRACT Adipose tissue is an organ with metabolic, endocrine and immune functions, affecting the life span of animals and their susceptibility to diseases. Previous studies have reported the relationship between many miRNAs and the development of adipose tissue. However, the regulation of miR-455-5p on the development of porcine adipose has not been reported. In this study, we used the dorsal subcutaneous adipose tissue of Guangxi Bama mini-pigs and Landrace pigs as experimental materials to study the regulatory mechanism of miRNA-455-5p and its target gene IGF-1R involved in the development of adipose tissue. Bioinformatic analysis with Targetscan predicted that IGF-1R might be a target gene of miRNA-455-5p. Moreover, mouse 3T3-L1 preadipocytes were transfected with chemically modified mimics and inhibitors to observe the expression of miR-455-5p and IGF-1R genes from 0 to 8 days after transfection. And the expression of adipogenic marker genes PPARγ and C/EBPα was examined. Additionally, the miRNA-455-5p may be involved in the negative regulation of adipose tissue development by inhibiting the expression of its target gene IGF-1R. These results provide a theoretical basis for increasing pork production and provide a new potential target for the treatment of obesity and type 2 diabetes in humans.

How a Cosmology of a Nonlocally Unified, Meaningfully in-formed, Holographically Manifested and Purposefully Evolving Universe Engenders Planetary Systems as Gravitational Environments for the Emergence of Biological and Biomechanical Complexity

The emergent cosmology of a nonlocally unified, meaningfully in-formed, holographically manifested and purposefully evolving Universe, treats gravity as an emergent consequence of the in-formational and holographic nature of space-time and describes it as the consequence of the informational content, or intropy, associated with positions in space-time of massive bodies. In doing so, the evolutionary impulse inherently embodied in the Universe’s finite lifecycle engenders solar systems and planetary homes for the eventual evolution and complexity of biological organisms. Showing that planetary gravitational fields are required to enable cellular differentiation and subsequent development of a range of biomechanical strategies for building complex morphology, describes how such complex organisms such as human beings could eventually develop

Rare Histological Types of Malignant Gastric Neoplasms: Features of Histomorphological and Visual Structure

Diagnosing rare histological types of malignant gastric neoplasms causes difficulty due to the nature of tumor growth, the lack of clear cellular differentiation and an extremely rare incidence. The aim of the study is to increase the accuracy of diagnosing rare types of malignant gastric neoplasms using modern methods of radiation diagnosis, depending on their histological characteristics. Materials and methods. The authors examined the data of patients with malignant gastric neoplasms of epithelial (N1, n = 385) and non-epithelial origin – gastrointestinal stromal tumor of the stomach (N2, n = 27), who were diagnosed and (or) treated at the BI «Republican Clinical Oncological Dispensary» under the Health Ministry of Chuvashia and the BI «City Clinical Hospital No. 1» under the Health Ministry of Chuvashia in 2022-2023. The control group is N3 (n = 40). A comparative analysis of data obtained by instrumental diagnostic methods (computed tomography, ultrasound) with a histomorphological description was carried out. Results. Rare histological types of gastric malignancies were identified: gastrointestinal stromal tumor in 6.6% (n = 27), neuroendocrine neoplasias in 3.6% (n = 15), undifferentiated cancer in 3.4% (n = 14), squamous cell carcinoma of the gastric cardia made 2.4% (n = 10). Gastrointestinal stromal tumor, neuroendocrine neoplasias and undifferentiated cancer were predominantly found in the body of the stomach (70.4%, 73.3% and 35.7%, respectively). Squamous cell carcinoma was found exclusively in the gastric cardia, involving the cardioesophageal junction (below the Z-line by 2 cm). G astrointestinal stromal tumor in 55.5% of cases was diagnosed as having a low risk of the disease progression. Neuroendocrine gastric neoplasias in 60% of cases had a low degree of tumor malignancy, and squamous-cell carcinoma of gastric cardia in 60% of cases had an intermediate degree of malignancy. In undifferentiated gastric cancer, cellular differentiation of the tumor was lost. A combined use of radiation diagnostic methods increased their informative value in identifying rare histological types of gastric malignancies: sensitivity – 95.5%, specificity – 98.5%, accuracy – 98.1%. Conclusions. A complex use of radiation examination methods increased the diagnostic accuracy of rare histological types of malignant gastric neoplasms.

Alternative splicing controls pan-neuronal homeobox gene expression.

The pan-neuronally expressed and phylogenetically conserved CUT homeobox gene ceh-44/CUX orchestrates pan-neuronal gene expression throughout the nervous system of Caenorhabditis elegans. As in many other species, including humans, ceh-44/CUX is encoded by a complex locus that also codes for a Golgi-localized protein, called CASP (Cux1 alternatively spliced product) in humans and CONE-1 ("CASP of nematodes") in C. elegans How gene expression from this complex locus is controlled-and, in C. elegans, directed to all cells of the nervous system-has not been investigated. We show here that pan-neuronal expression of CEH-44/CUX is controlled by a pan-neuronal RNA splicing factor, UNC-75, the C. elegans homolog of vertebrate CELF proteins. During embryogenesis, the cone-1&ceh-44 locus exclusively produces the Golgi-localized CONE-1/CASP protein in all tissues, but upon the onset of postmitotic terminal differentiation of neurons, UNC-75/CELF induces the production of the alternative CEH-44/CUX CUT homeobox gene-encoding transcript exclusively in the nervous system. Hence, UNC-75/CELF-mediated alternative splicing not only directs pan-neuronal gene expression but also excludes a phylogenetically deeply conserved golgin from the nervous system, paralleling surprising spatial specificities of another golgin that we describe here as well. Our findings provide novel insights into how all cells in a nervous system acquire pan-neuronal identity features and reveal unanticipated cellular specificities in Golgi apparatus composition.