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BACKGROUND: Bone marrow mesenchymal stem cells have been widely used to treat neurological diseases. However, due to limitations of the blood-brain barrier, low survival rate and differentiation rate of stem cells at damaged sites, the therapeutic effect is limited. OBJECTIVE: To investigate the effects of Shexiang Huangqi compound dripping pills on proliferation, migration and astrocyte differentiation of bone marrow mesenchymal stem cells. METHODS: Male SD rats were treated with Shexiang Huangqi compound dripping pills for 5 days after continuous gavage. Blood was collected from the abdominal aorta and serum was separated for later use. The effect of 5%, 10% and 20% drug-containing serum on the proliferation of bone marrow mesenchymal stem cells was detected by CCK-8 assay. The effect of 10% drug-containing serum on lateral migration of bone marrow mesenchymal stem cells was observed by scratch test. Bone marrow mesenchymal stem cells were cultured in Transwell cells. The effects of 10% drug-containing serum on longitudinal migration of bone marrow mesenchymal stem cells were observed by crystal violet staining and DAPI nuclear staining. Differentiation of bone marrow mesenchymal stem cells into astrocytes was observed by inducing solution with 10% drug-containing serum or co-culture with astrocytes. RESULTS AND CONCLUSION: (1) 10% and 20% drug-containing serum promoted cell proliferation more significantly on days 2 and 3, and there was no statistical difference between the two concentrations. (2) At 30 and 48 hours, bone marrow mesenchymal stem cell migration in 10% drug-containing serum group was significantly higher than that in the control group. (3) The number of bone marrow mesenchymal stem cells filtered through Transwell cells in 10% drug-containing serum group was higher than that in the control group. (4) 10% drug-containing serum might promote the differentiation of bone marrow mesenchymal stem cells to astrocytes, but the differentiation effect was weak, and astrocytes might further promote the differentiation of bone marrow mesenchymal stem cells into astrocytes induced by drug-containing serum. (5) The results exhibited that the 10% drug-containing serum could promote the proliferation and migration of bone marrow mesenchymal stem cells in vitro. Co-culture with astrocytes may promote the differentiation of bone marrow mesenchymal stem cells towards astrocytes. [ABSTRACT FROM AUTHOR]

A major challenge in therapeutic approaches applying hematopoietic stem cells (HSCs) is the cell quantity. The primary objective of this study was to predict the miRNAs and anti-miRNAs using bioinformatics tools and investigate their effects on the expression levels of key genes predicted in the improvement of proliferation, and the inhibition of differentiation in HSCs isolated from Human umbilical cord blood (HUCB). A network including genes related to the differentiation and proliferation stages of HSCs was constructed by enriching data of text (PubMed) and StemChecker server with KEGG signaling pathways, and was improved using GEO datasets. Bioinformatics tools predicted a profile from miRNAs containing miR-20a-5p, miR-423-5p, and chimeric anti-miRNA constructed from 5′-miR-340/3′-miR-524 for the high-score genes (RB1, SMAD4, STAT1, CALML4, GNG13, and CDKN1A/CDKN1B genes) in the network. The miRNAs and anti-miRNA were transferred into HSCs using polyethylenimine (PEI). The gene expression levels were estimated using the RT-qPCR technique in the PEI + (miRNA/anti-miRNA)-contained cell groups (n = 6). Furthermore, CD markers (90, 16, and 45) were evaluated using flow cytometry. Strong relationships were found between the high-score genes, miRNAs, and chimeric anti-miRNA. The RB1, SMAD4, and STAT1 gene expression levels were decreased by miR-20a-5p (P < 0.05). Additionally, the anti-miRNA increased the gene expression level of GNG13 (P < 0.05), whereas the miR-423-5p decreased the CDKN1A gene expression level (P < 0.01). The cellular count also increased significantly (P < 0.05) but the CD45 differentiation marker did not change in the cell groups. The study revealed the predicted miRNA/anti-miRNA profile expands HSCs isolated from HUCB. While miR-20a-5p suppressed the RB1, SMAD4, and STAT1 genes involved in cellular differentiation, the anti-miRNA promoted the GNG13 gene related to the proliferation process. Notably, the mixed miRNA/anti-miRNA group exhibited the highest cellular expansion. This approach could hold promise for enhancing the cell quantity in HSC therapy. [ABSTRACT FROM AUTHOR]

Shwachman–Diamond syndrome (SDS) is an inherited bone marrow failure disorder that often presents at infancy. Progress has been made in revealing causal mutated genes (SBDS and others), ribosome defects, and hematopoietic aberrations in SDS. However, the mechanism underlying the hematopoietic failure remained unknown, and treatment options are limited. Herein, we investigated the onset of SDS embryonic hematopoietic impairments. We generated SDS and control human‐derived induced pluripotent stem cells (iPSCs). SDS iPSCs recapitulated the SDS hematological phenotype. Detailed stepwise evaluation of definitive hematopoiesis revealed defects that started at the early emerging hematopoietic progenitor (EHP) stage after mesoderm and hemogenic endothelium were normally induced. Hematopoietic potential of EHPs was markedly reduced, and the introduction of SBDS in SDS iPSCs improved colony formation. Transcriptome analysis revealed reduced expression of ribosome and oxidative phosphorylation‐related genes in undifferentiated and differentiated iPSCs. However, certain pathways (e.g., DNA replication) and genes (e.g., CHCHD2) were exclusively or more severely dysregulated in EHPs compared with earlier and later stages. To our knowledge, this study offers for the first time an insight into the embryonic onset of human hematopoietic defects in an inherited bone marrow failure syndrome and reveals cellular and molecular aberrations at critical stages of hematopoietic development toward EHPs. [ABSTRACT FROM AUTHOR]

Phyllodes tumours or cystosarcoma phyllodes are fibroepithelial tumours of the breast and represent 1% of breast tumours. A 20-year-old nullipara presented with an enlarging left breast mass over 6 months. Although widely excised, it was reported to be a 12 × 10 × 5.5-cm borderline phyllodes tumour with involvement of the superior and inferior margins. Seven months later, she presented with a new ipsilateral breast lump measuring 8.5 × 7.5 × 4.6 cm. She underwent a left mastectomy, a three-rib resection with titanic rods for the thoracic cage reconstruction, and a latissimus dorsi flap wound closure. Histopathology revealed a high-grade malignant phyllodes tumour with features of osteoid differentiation with the nearest deep margin measuring 3 mm. She developed metastasis to the ipsilateral axillary lymph nodes and contralateral lung 2 months postoperatively. She was given palliative radiotherapy 60 Gy in 30 fractions to the left axilla. She developed sudden lower-limb weakness due to spinal metastases. The symptoms resolved with radiotherapy to the thoracic spine (T4–T8). As the lesion continued to grow rapidly from the anterior chest wall encircling towards the back, it was deemed unresectable. She was given palliative chemotherapy (doxorubicin six cycles, followed by ifosfamide one cycle) but had disease progression. She passed away 3 months later. The mainstay of treatment for phyllodes tumour is excision with a minimal margin of 1 cm. Although margins were involved after the first surgery, she was followed up as the pathology was a borderline phyllodes. When the lump recurred and had transformed, despite extensive surgery, it returned shortly and progressed. A borderline phyllodes should be excised to obtain a minimal margin of 1 cm, even if it means performing a mastectomy, to minimise recurrence. A recurrence may undergo malignant transformation which is largely chemotherapy and radiotherapy resistant. This will result in a poor outcome and decreased survival. [ABSTRACT FROM AUTHOR]

Mangrove sediments host a diverse array of microbial populations and are characterized by high heterogeneity along their vertical depths. However, the genetic diversity within these populations is largely unknown, hindering our understanding of their adaptive evolution across the sediment depths. To elucidate their genetic diversity, we utilized metagenome sequencing to identify 16 high‐frequency microbial populations comprised of two archaea and 14 bacteria from mangrove sediment cores (0–100 cm, with 10 depths) in Qi'ao Island, China. Our analysis of the genome‐wide genetic variation revealed extensive nucleotide diversity in the microbial populations. The genes involved in the transport and the energy metabolism displayed a high nucleotide diversity (HND; 0.0045–0.0195; an indicator of shared minor alleles with the microbial populations). By tracking the processes of homologous recombination, we found that each microbial population was subjected to different purification selection levels at different depths (44.12% genes). This selection resulted in significant differences in synonymous/non‐synonymous mutation ratio between 0–20 and 20–100 cm layers, indicating the adaptive evolutionary process of microbial populations. Furthermore, our assessment of differentiation in the allele frequencies between these two layers showed that the functional genes involved in the metabolic processes of amino acids or cofactors were highly differential in more than half of them. Together, we showed that the nucleotide diversity of microbial populations was shaped by homologous recombination and gene‐specific selection, finally resulting in the stratified differentiation occurring between 0–20 and 20–100 cm. These results enhance our cognition of the microbial adaptation mechanisms to vertical environmental changes during the sedimentation process of coastal blue carbon ecosystems. [ABSTRACT FROM AUTHOR]

In order to understand the coordinated proteome changes associated with differentiation of a cultured cell pluripotency model, protein expression changes induced by treatment of NT2 embryonal carcinoma cells with retinoic acid were monitored by mass spectrometry. The relative levels of over 5000 proteins were mapped across distinct cell fractions. Analysis of the chromatin fraction revealed major abundance changes among chromatin proteins and epigenetic pathways between the pluripotent and differentiated states. Protein complexes associated with epigenetic regulation of gene expression, chromatin remodelling (e.g., SWI/SNF, NuRD) and histone-modifying enzymes (e.g., Polycomb, MLL) were found to be extensively regulated. We therefore investigated histone modifications before and after differentiation, observing changes in the global levels of lysine acetylation and methylation across the four canonical histone protein families, as well as among variant histones. We identified the set of proteins with affinity to peptides housing the histone marks H3K4me3 and H3K27me3, and found increased levels of chromatin-associated histone H3 tail trimming following differentiation that correlated with increased expression levels of cathepsin proteases. We further found that inhibition of cathepsins B and D reduces histone H3 clipping. Overall, the work reveals a global reorganization of the cell proteome congruent with differentiation, highlighting the key role of multiple epigenetic pathways, and demonstrating a direct link between cathepsin B and D activity and histone modification. [ABSTRACT FROM AUTHOR]

Simple Summary: Simple Summary: Mesenchymal stem cells (MSCs) are adult stem cells capable of differentiating into various cell types, including osteoblasts, chondrocytes, and adipocytes. Renowned for their immunomodulatory and regenerative properties, MSCs hold great promise as tools for tissue repair and therapeutic applications. To facilitate the large-scale production of sheep umbilical cord MSCs (UCMSCs), the telomerase reverse transcriptase (TERT) gene was introduced into UCMSCs to promote immortalization. This study provides valuable insights into (a) the proliferation and anti-aging capabilities of transgenic MSCs compared to primary MSCs; (b) the absence of potential tumorigenicity exhibited in TERT-UCMSCs; and (c) the differentiation potential of TERT-UCMSCs. Immortalized MSCs exhibit enhanced proliferation and anti-aging abilities without tumorigenicity concerns, making them a safe immortalized cell line suitable for medical treatments. Mesenchymal stem cells (MSCs) possess significant differentiation potential, making them highly promising in medicine and immunotherapy due to their regenerative capabilities and exosome secretion. However, challenges such as limited cell divisions and complex testing hinder large-scale MSC production. In this study, we successfully established an immortalized MSC line by transfecting the human telomerase reverse transcriptase (TERT) gene into MSCs isolated from pregnant sheep umbilical cords. This approach effectively inhibits cell senescence and promotes cell proliferation, enabling the generation of umbilical cord mesenchymal stem cells (UCMSCs) on a larger scale. Our findings demonstrate that these transfected TERT-UCMSCs exhibit enhanced proliferative capacity and a reduced aging rate compared to regular UCMSCs while maintaining their stemness without tumorigenicity concerns. Consequently, they hold great potential for medical applications requiring large quantities of functional MSCs. [ABSTRACT FROM AUTHOR]

The current limitations of commercial microcarriers for mesenchymal stromal cell (MSC) expansion are driving the demand for novel tuneable microcarriers. Novel microcarriers are effective in improving MSC expansion under static and dynamic culture conditions without compromising their stemness and differentiation potential. Integrating microcarrier-based cultures within dynamic bioreactors provides an effective scalable approach for MSC expansion and harvest for downstream clinical translation. The development of good manufacturing practice (GMP)-grade systems, non-xenogenic culture media, and optimised bioreactors, which are compatible with novel microcarriers, is essential to improve cell yield and reproducibility. Microcarrier expansion systems show exciting potential to revolutionise mesenchymal stromal cell (MSC)-based clinical therapies by providing an opportunity for economical large-scale expansion of donor- and patient-derived cells. The poor reproducibility and efficiency of cell expansion on commercial polystyrene microcarriers have driven the development of novel microcarriers with tuneable physical, mechanical, and cell-instructive properties. These new microcarriers show innovation toward improving cell expansion outcomes, although their limited biological characterisation and compatibility with dynamic culture systems suggest the need to realign the microcarrier design pathway. Clear headway has been made toward developing infrastructure necessary for scaling up these technologies; however, key challenges remain in characterising the wholistic effects of microcarrier properties on the biological fate and function of expanded MSCs. [ABSTRACT FROM AUTHOR]

It has long been recognized that presence–absence, localisation, size, number and shape of fish scales can be important taxonomic features. Although there are some notes on the relationship between scale morphology and ecological needs, in the absence of a sufficiently large and detailed database, the morphological variability of fish scales and the factors responsible for this variability have not yet been explored in detail. For this reason, a database—which contains the shape and relative size data of 193 freshwater fish taxa of 14 orders, originated from five biogeographic realms—has been built. Database analyses showed that both the scale shape and relative size are proper taxonomic indicators. They can be used to separate higher taxonomic categories (e.g., orders), and by the simultaneous analysis of shape and size scale morphology, we showed increased sensitivity for species‐level detachments. Our results suggest that while both the shape and the size of the fish scales are genetically determined, they are also useful descriptors of the niche segregation (habitat use, flow preference) of close relative species. Scale morphology is a promising additional tool to specify the environmental preferences of lesser known or close relative recent and extinct fish species. And also can be of great help in such cases when only the scales are available for taxonomic identification, for example, in the research field of archaeology and palaeontology. [ABSTRACT FROM AUTHOR]

The proliferation of neural stem cells (NSCs) is precisely regulated by extracellular environmental factors. In situ hypoxia, one of the key factors involved in the regulation of NSC characteristics, has attracted increasing amounts of attention. Numerous studies have demonstrated that hypoxia can significantly promote the formation of neurospheres and the proliferation of NSCs in vitro and that intermittent hypoxia can promote the proliferation of endogenous NSCs in vivo. In this article, the effects of different concentrations of oxygen on NSC proliferation and differentiation both in vivo and in vitro are reviewed, and the potential applications of hypoxia-preconditioned NSCs, as well as research progress and challenges in the treatment of central nervous system diseases, are further summarized. Here, the critical role of oxygen in the neurogenesis of NSCs is emphasized, and insights into the use of hypoxia to regulate NSC characteristics are provided. [ABSTRACT FROM AUTHOR]

Oligodendrocytes are the myelinating cells in the CNS and multiple sclerosis (MS) is a demyelinating disorder that is characterized by progressive loss of myelin. Although oligodendroglial progenitor cells (OPCs) should be differentiated into oligodendrocytes, for multiple reasons, OPCs fail to differentiate into oligodendrocytes in MS. Therefore, increasing the maturation of OPCs to oligodendrocytes may be of therapeutic benefit for MS. The β‐hydroxy β‐methylbutyrate (HMB) is a muscle‐building supplement in humans and this study underlines the importance of HMB in stimulating the maturation of OPCs to oligodendrocytes. HMB treatment upregulated the expression of different maturation markers including PLP, MBP, and MOG in cultured OPCs. Double‐label immunofluorescence followed by immunoblot analyses confirmed the upregulation of OPC maturation by HMB. While investigating mechanisms, we found that HMB increased the maturation of OPCs isolated from peroxisome proliferator‐activated receptor β−/− (PPARβ−/−) mice, but not PPARα−/− mice. Similarly, GW6471 (an antagonist of PPARα), but not GSK0660 (an antagonist of PPARβ), inhibited HMB‐induced maturation of OPCs. GW9662, a specific inhibitor of PPARγ, also could not inhibit HMB‐mediated stimulation of OPC maturation. Furthermore, PPARα agonist GW7647, but neither PPARβ agonist GW0742 nor PPARγ agonist GW1929, alone increased the maturation of OPCs. Finally, HMB treatment of OPCs led to the recruitment of PPARα, but neither PPARβ nor PPARγ, to the PLP gene promoter. These results suggest that HMB stimulates the maturation of OPCs via PPARα and that HMB may have therapeutic prospects in remyelination. [ABSTRACT FROM AUTHOR]

Myosin heavy chain‐perinatal (MyHC‐perinatal) is one of two development‐specific myosin heavy chains expressed exclusively during skeletal muscle development and regeneration. The specific functions of MyHC‐perinatal are unclear, although mutations are known to lead to contracture syndromes such as Trismus‐pseudocamptodactyly syndrome. Here, we characterize the functions of MyHC‐perinatal during skeletal muscle differentiation and regeneration. Loss of MyHC‐perinatal function leads to enhanced differentiation characterized by increased expression of myogenic regulatory factors and differentiation index as well as reduced reserve cell numbers in vitro. Proteomic analysis revealed that loss of MyHC‐perinatal function results in a switch from oxidative to glycolytic metabolism in myofibers, suggesting a shift from slow type I to fast type IIb fiber type, also supported by reduced mitochondrial numbers. Paracrine signals mediate the effect of loss of MyHC‐perinatal function on myogenic differentiation, possibly mediated by non‐apoptotic caspase‐3 signaling along with enhanced levels of the pro‐survival apoptosis regulator Bcl2 and nuclear factor kappa‐B (NF‐κB). Knockdown of MyHC‐perinatal during muscle regeneration in vivo results in increased expression of the differentiation marker myogenin (MyoG) and impaired differentiation, evidenced by smaller myofibers, elevated fibrosis and reduction in the number of satellite cells. Thus, we find that MyHC‐perinatal is a crucial regulator of myogenic differentiation, myofiber oxidative phenotype and regeneration. [ABSTRACT FROM AUTHOR]

Allergen-stimulated basophils secrete leukotriene C4 (LTC4), acting on the cysteinyl leukotriene receptor (CysLTR2) in neurons, activating neuronal signaling that mediates acute itch exacerbation in atopic dermatitis (AD). CXCR2+ basophils recruited to the kidney release interleukin (IL)-6, which recruits type 17 T helper (T H 17) cells, contributing to the pathogenesis of renal fibrosis. Targeting CXCR2+ basophils might be considered a therapeutic strategy in renal fibrosis. In mouse bone marrow granulocyte–monocyte progenitors, high expression of E-cadherin on myeloid progenitors marks an early population of pro-basophil and mast cell progenitors, which is committed to the basophil and mast cell fates. Lung-resident basophils establish a lung-specific function influenced by IL-33 and granulocyte–macrophage colony-stimulating factor (GM-CSF), which are crucial for maintaining alveolar macrophage development and function. Basophils serve as multifunctional regulators in either maintaining homeostasis or contributing to disease pathogenesis. Recent findings highlight their diverse lineage-priming stages and involvement in disorders across organs such as the skin, lung, kidney, and heart, suggesting systemic influence and potential heterogeneous subpopulations. However, the underlying mechanisms of basophil differentiation and heterogeneity remain unclear, meriting further investigation, particularly when developing possible therapeutic strategies for relevant disorders. Basophils, rare granulocytes, have long been acknowledged for their roles in type 2 immune responses. However, the mechanisms by which basophils adapt their functions to diverse mammalian microenvironments remain unclear. Recent advancements in specific research tools and single-cell-based technologies have greatly enhanced our understanding of basophils. Several studies have shown that basophils play a role in maintaining homeostasis but can also contribute to pathology in various tissues and organs, including skin, lung, and others. Here, we provide an overview of recent basophil research, including cell development, characteristics, and functions. Based on an increasing understanding of basophil biology, we suggest that the precise targeting of basophil features might be beneficial in alleviating certain pathologies such as asthma, atopic dermatitis (AD), and others. [ABSTRACT FROM AUTHOR]

The clinical translation of induced pluripotent stem cells (iPSCs) holds great potential for personalized therapeutics. However, one of the main obstacles is that the current workflow to generate iPSCs is expensive, time‐consuming, and requires standardization. A simplified and cost‐effective microfluidic approach is presented for reprogramming fibroblasts into iPSCs and their subsequent differentiation into neural stem cells (NSCs). This method exploits microphysiological technology, providing a 100‐fold reduction in reagents for reprogramming and a ninefold reduction in number of input cells. The iPSCs generated from microfluidic reprogramming of fibroblasts show upregulation of pluripotency markers and downregulation of fibroblast markers, on par with those reprogrammed in standard well‐conditions. The NSCs differentiated in microfluidic chips show upregulation of neuroectodermal markers (ZIC1, PAX6, SOX1), highlighting their propensity for nervous system development. Cells obtained on conventional well plates and microfluidic chips are compared for reprogramming and neural induction by bulk RNA sequencing. Pathway enrichment analysis of NSCs from chip showed neural stem cell development enrichment and boosted commitment to neural stem cell lineage in initial phases of neural induction, attributed to a confined environment in a microfluidic chip. This method provides a cost‐effective pipeline to reprogram and differentiate iPSCs for therapeutics compliant with current good manufacturing practices. [ABSTRACT FROM AUTHOR]

Diagnosing cutaneous melanomas relies mainly on histopathological analysis, which, in selected cases, can be aided by immunohistochemical evaluation of conventional melanocytic markers. Nevertheless, these malignancies, particularly in metastatic settings, may display divergent differentiation with unusual histological and immunohistochemical features. In this context, we present the case of a 65-year-old male diagnosed with typical superficial spreading melanoma who developed recurrence and metastatic lesions featuring angiosarcomatous differentiation. The diagnosis of the initial tumour and the subsequently dedifferentiated lesions was confirmed by ample immunohistochemical analysis, which included several melanocytic markers, as well as mesenchymal and vascular markers. The recurrent tumour and lymph nodes metastases were completely negative for Melan-A and PRAME, and focally positive for SOX10. Additionally, they also displayed diffuse, intense positivity for CD10 and WT1 and focal positivity for CD99, ERB, and CD31. Thus, the diagnosis of primary cutaneous melanoma with recurrent and metastatic divergent angiosarcomatous differentiation was established. This occurrence is particularly rare and can pose important diagnostic challenges. Therefore, in addition to presenting this highly unusual case, we also performed a comprehensive review of the literature on divergent differentiation in melanomas. [ABSTRACT FROM AUTHOR]

S. boulardii, the probiotic variety of S. cerevisiae, is known for its different characteristics, even though the two strains share more than 99% genomic relatedness. Therefore, S. boulardii has been examined and evaluated for differential characteristics in many studies. This study aims to explain the differentiation of S. boulardii from well-identified S. cerevisiae strains by biochemical characteristics and the qPCR-HRM. Initially, S. cerevisiae isolates from the HUF Culture Collection, were screened for basic biochemical characteristics such as growth temperature, glucose, and galactose utilization tests. Out of all selected cultures, the viability at gastric pH was investigated by spectrophotometric readings. The results showed that one isolate was more resistant than other strains when exposed to gastric acids. After sequencing and phylogeny steps using LR0R universal primer, there was no significant differentiation for S. boulardii strains and therefore HRM analysis was performed to interrogate sequence variation and differentiate S. boulardii strains. HRM analysis has successfully distinguished even highly closely related yeast strains. [ABSTRACT FROM AUTHOR]

Yak (Bos grunniens) is a unique livestock animal originating from the Qinghai-Tibet Plateau in China. In the current study, we investigated the maternal genetic diversity, differentiation and phylogeny of wild yak population and four domestic yak breeds (Qinghai-Gaoyuan, Huanhu, Xueduo, and Yushu) in Qinghai, China by analyzing 166 mitochondrial cytochrome b (Cytb) gene sequence variations. The results showed that the five yak breeds/populations had high genetic diversity (Hd = 0.501 ± 0.088–0.770 ± 0.053) and each yak breed/population owned unique haplotypes. Estimates of FST values showed a moderate genetic differentiation between wild yak and Huanhu yak (FST = 0.0577) as well as that between Huanhu yak and Yushu yak breeds (FST = 0.0520), but a weak genetic differentiation was observed between the other yak breeds/populations (–0.0209 < FST < 0.0372). Additionally, the clustering analysis based on RST values showed that Xueduo yak and Huanhu yak were clustered into one group, and each of the other three yak breeds/populations was separated into one group, respectively. Overall, the clustering relationship between wild yak and Yushu yak was closer. Maternal phylogenetic analysis showed that wild yak and four domestic yak breeds/populations in Qinghai represented in three maternal lineages (Mt-I, Mt-II, and Mt-III), indicating three maternal origins in yak. Our study would provide valuable information for the conservation and utilization of wild yak and Qinghai domestic yak breeds. [ABSTRACT FROM AUTHOR]

Recently, the formation of three-dimensional (3D) cell aggregates known as embryoid bodies (EBs) grown in media supplemented with HSC-specific morphogens has been utilized for the directed differentiation of embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), into clinically relevant hematopoietic stem cells (HSCs). However, delivering growth factors and nutrients have become ineffective in inducing synchronous differentiation of cells due to their 3D conformation. Moreover, irregularly sized EBs often lead to the formation of necrotic cores in larger EBs, impairing differentiation. Here, we developed two gelatin microparticles (GelMPs) with different release patterns and two HSC-related growth factors conjugated to them. Slow and fast releasing GelMPs were conjugated with bone morphogenic factor-4 (BMP-4) and stem cell factor (SCF), respectively. The sequential presentation of BMP-4 and SCF in GelMPs resulted in efficient and effective hematopoietic differentiation, shown by the enhanced gene and protein expression of several mesoderm and HSC-related markers, and the increased concentration of released HSC-related cytokines. In the present study, we were able to generate CD34+, CD133+, and FLT3+ cells with similar cellular and molecular morphology as the naïve HSCs that can produce colony units of different blood cells, in vitro. [Display omitted] • Successful conjugation of BMP4 and SCF in fast and slow release gelatin microparticles. • BMP4/SCF conjugation in GelMPs promotes proliferation and reduces apoptosis. • Dual GelMP system promotes sequential differentiation of iPSC to mesoderm and HSC. • CD34+ iPS-derived HSCs (iHSCs) are capable of producing various blood cells, in-vitro. • Differentiated iHSCs were verified by RNA sequencing analysis. [ABSTRACT FROM AUTHOR]

Although the roles of E proteins and inhibitors of DNA‐binding (Id) in T follicular helper (TFH) and T follicular regulatory (TFR) cells have been previously reported, direct models demonstrating the impact of multiple E protein members have been lacking. To suppress all E proteins including E2A, HEB and E2‐2, we overexpressed Id1 in CD4 cells using a CD4‐Id1 mouse model, to observe any changes in TFH and TFR cell differentiation. Our objective was to gain better understanding of the roles that E proteins and Id molecules play in the differentiation of TFH and TFR cells. The CD4‐Id1 transgenic (TG) mice that we constructed overexpressed Id1 in CD4 cells, inhibiting E protein function. Our results showed an increase in the proportion and absolute numbers of Treg, TFH and TFR cells in the spleen of TG mice. Additionally, the expression of surface characterisation molecules PD‐1 and ICOS was significantly upregulated in TFH and TFR cells. The study also revealed a downregulation of the marginal zone B cell precursor and an increase in the activation and secretion of IgG1 in spleen B cells. Furthermore, the peripheral TFH cells of TG mice enhanced the function of assisting B cells. RNA sequencing results indicated that a variety of TFH‐related functional molecules were upregulated in TFH cells of Id1 TG mice. In conclusion, E proteins play a crucial role in regulating TFH/TFR cell differentiation and function and suppressing E protein activity promotes germinal centre humoral immunity, which has important implications for immune regulation and treating related diseases. [ABSTRACT FROM AUTHOR]

A pervasive view is that undifferentiated stem cells are alone responsible for generating all other cells and are the origins of cancer. However, emerging evidence demonstrates fully differentiated cells are plastic, can be coaxed to proliferate, and also play essential roles in tissue maintenance, regeneration, and tumorigenesis. Here, we review the mechanisms governing how differentiated cells become cancer cells. First, we examine the unique characteristics of differentiated cell division, focusing on why differentiated cells are more susceptible than stem cells to accumulating mutations. Next, we investigate why the evolution of multicellularity in animals likely required plastic differentiated cells that maintain the capacity to return to the cell cycle and required the tumor suppressor p53. Finally, we examine an example of an evolutionarily conserved program for the plasticity of differentiated cells, paligenosis, which helps explain the origins of cancers that arise in adults. Altogether, we highlight new perspectives for understanding the development of cancer and new strategies for preventing carcinogenic cellular transformations from occurring. This review summarises emerging views on how differentiated cells become cancer cells and contribute to malignancies due to accumulating mutations, aberrant tissue repair programs and hijacked cell state plasticity. [ABSTRACT FROM AUTHOR]

The number of immigrant and international multilingual learners enrolling in postsecondary education is on the rise. With this growth, there remain difficulties in identifying and supporting multilingual learners moving from K-12 to college due to demographic data collection procedures at the postsecondary level. Postsecondary institutions are challenged with appropriately identifying multilingual students in need of academic support and providing appropriate testing support and other academic supports. Our midWestern institution is set in a diverse urban setting with an increasing incidence of immigrants so we sought to meet both content and linguistic needs of these students at our institution. Through literature review research and stakeholder collaboration, we designed testing support guidelines and consultations for students and faculty to equitably support multilingual students taking course exams. [ABSTRACT FROM AUTHOR]

The full-length p200CUX1 protein encoded by the homology frame CUT-like protein (CUX1) plays an important role in tumors as a pro-oncogene or oncogene. However, its role and mechanism in acute myeloid leukemia remain unknown. p200CUX1 regulates several pathways, including the MAPK signaling pathway. Our data showed that p200CUX1 is lowly expressed in THP1 and U937 AML cell lines. Lentiviral overexpression of p200CUX1 reduced proliferation and promoted apoptosis and G0/G1 phase blockade, correlating with MAPK pathway suppression. Additionally, p200CUX1 regulated the expression of bone morphogenetic protein 8B (BMP8B), which is overexpressed in AML. Overexpression of p200CUX1 downregulated BMP8B expression and inhibited the MAPK pathway. Furthermore, BMP8B knockdown inhibited AML cell proliferation, enhanced apoptosis and the sensitivity of ATRA-induced cell differentiation, and blocked G0/G1 transition. Our findings demonstrate the pivotal function of the p200CUX1-BMP8B-MAPK axis in maintaining the viability of AML cells. Consequently, targeting p200CUX1 could represent a viable strategy in AML therapy. [ABSTRACT FROM AUTHOR]

Background: Differentiation of pluripotent stem cells into desired lineages is the key aspect of regenerative medicine and cell‐based therapy. Although RNA interference (RNAi) technology is exploited extensively for this, methods for long term silencing of the target genes leading to differentiation remain a challenge. Sustained knockdown of the target gene by RNAi is often inefficient as a result of low delivery efficiencies, protocol induced toxicity and safety concerns related to viral vectors. Earlier, we established octa‐arginine functionalized hydroxyapatite nano vehicles (R8HNPs) for delivery of small interfering RNA (siRNA) against a pluripotency marker gene in mouse embryonic stem cells. Although we demonstrated excellent knockdown efficiency of the target gene, sustained gene silencing leading to differentiation was yet to be achieved. Methods: To establish a sustained non‐viral gene silencing protocol using R8HNP, we investigated various methods of siRNA delivery: double delivery of adherent cells (Adh‐D), suspension delivery followed by adherent delivery (Susp + Adh), single delivery in suspension (Susp‐S) and multiple deliveries in suspension (Susp‐R). Sustained knockdown of a pluripotent marker gene followed by differentiation was analysed by reverse transcriptase‐PCR, fluoresence‐activated cell sorting and immunofluorescence techniques. Impact on cell viability as a result of repeated exposure of the R8HNP was also tested. Results: Amongst the protocols tested, the most efficient knockdown of the target gene for a prolonged period of time was obtained by repeated suspension delivery of the R8HNP‐siRNA conjugate. The long‐term silencing of a pluripotency marker gene resulted in differentiation of R1 ESCs predominantly towards the extra embryonic and ectodermal lineages. Cells displayed excellent tolerance to repeated exposures of R8HNPs. Conclusions: The results demonstrate that R8HNPs are promising, biocompatible, non‐viral alternatives for prolonged gene silencing and obtaining differentiated cells for therapeutics. [ABSTRACT FROM AUTHOR]

We consider the role of personalized pricing (PP) on product differentiation when PP is costly to implement. Using a stylized yet commonly used formulation, we find that when firms decide on positioning before deciding on PP implementation, PP implementation cost affects not only the amount of differentiation firms choose in their positioning, firm profits, consumer surplus, and social welfare, but also whether firms implement PP. When PP implementation cost is low, firms cannot help but to implement PP and engage in direct price competition. Moreover, firms implementing PP reduce their differentiation, further intensifying price competition, and are worse off. When PP implementation cost is moderate, firms position to reduce their differentiation to commit to not implementing PP, again aggravating price competition. In contrast, when PP implementation cost is higher, firms increase their differentiation due to the threat of PP but do not implement PP. As a result, the availability of PP improves firm profits, even though firms do not implement PP. However, if differentiation is restricted, then PP availability cannot improve firm profits. If an information seller sets the PP implementation cost, then it sets the cost low. Consequently, firms implement PP and are worse off. We also find that when firms decide whether to implement PP before deciding on positioning, they never implement PP. This is the case when PP implementation is complex, and differentiation can be affected by short-run advertising and promotion. Finally, we show that banning PP can benefit consumers when accounting for changes in firm positioning. This paper was accepted by D. J. Wu, information systems. Funding: This work was supported by the Social Sciences and Humanities Research Council of Canada and the Hong Kong Research Grants Council [Grant 21500920]. Supplemental Material: The online appendix is available at https://doi.org/10.1287/mnsc.2021.02740. [ABSTRACT FROM AUTHOR]

Backgrounds: The direct and indirect interactions between multiple myeloma (MM) cells and bone marrow mesenchymal stromal cells (MSCs) play crucial roles in the formation of the bone marrow environment, disease progression, and drug resistance development. However, it remains unclear how MM cells and MSCs individually influence each other to induce these phenomena. Objective: In this study, we focused on observing changes in MSCs induced by MM cells. Changes in MSCs due to exposure to MM cells were observed by assessing cell proliferation, apoptosis, cell cycle, and morphology. Furthermore, the unique abilities of MSCs were confirmed through differentiation potential and MSC marker expression, along with the demonstration of senescence. Gene profiling was performed to elucidate the mechanisms underlying these changes. Results: Co-culturing MM cells with MSCs did not alter the morphology or proliferation of MSCs but increased apoptosis. As apoptosis increased, damaged deoxyribonucleic acid (DNA) was repaired, leading to the activation of the cell cycle with an increase in the S phase, resulting in no significant changes in cell proliferation and morphology. Osteogenesis and adipogenesis generally decreased by co-culturing with MM cells, and senescence increased. Significant differences were observed in the expression of MSC marker genes. Gene profiling revealed changes in gene expression following osteogenic differentiation. Conclusion: Based on these results, MSCs exposed to MM cells exhibited an increase in the S phase of the cell cycle, leading to the recovery of cells undergoing apoptosis. Osteogenesis and adipogenesis decreased, whereas senescence increased, suggesting that these changes were attributed to the overall MSC characteristics and genetic mechanisms. [ABSTRACT FROM AUTHOR]

Objective: To investigate the role of ginsenoside Rd (GRd) in acute myeloid leukemia (AML) cell differentiation. Methods: AML cells were treated with GRd (25, 50, 100 and 200 µg/mL), retinoic acid (RA, 0.1g/L) and PD98059 (20 mg/mL) for 72 h, cell survival was detected by methylthiazolyldiphenyl-tetrazolium bromide and colony formation assays, and cell cycle was detected by flow cytometry. Cell morphology and differentiation were observed by Wright-Giemsa staining, peroxidase chemical staining and cellular immunochemistry assay, respectively. The protein expression levels of GATA binding protein 1 (GATA-1), purine rich Box-1 (PU.1), phosphorylated-extracellular signal-related kinase (p-ERK), ERK, phosphorylated-glycogen synthase kinase-3β (p-GSK3β), GSK3β and signal transducer and activator of transcription 1 (STAT1) were detected by Western blot. Thirty-six mice were randomly divided into 3 groups using a random number table: model control group (non-treated), GRd group [treated with 200 mg/(kg·d) GRd] and homoharringtonine (HTT) group [treated with 1 mg/(kg·d) HTT]. A tumor-bearing nude mouse model was established, and tumor weight and volume were recorded. Changes of subcutaneous tumor tissue were observed after hematoxylin and eosin staining. WT1 and GATA-1 expressions were detected by immunohistochemical staining. Results: The cell survival was inhibited by GRd in a dose-dependent manner and GRd caused G0/G1 cell arrest (p<0.05). GRd treatment induced leukemia cell differentiation, showing increased expressions of peroxidase and specific proteins concerning erythrogenic or granulocytic differentiation (p<0.05). GRd treatment elicited upregulation of p-ERK, p-GSK-3β and STAT1 expressions in cells, and reversed the effects of PD98059 on inhibiting the expressions of peroxidase, GATA-1 and PU.1 (P<0.05). After GRd treatment, tumor weight and volume of mice were decreased, and tumor cells underwent massive apoptosis and necrosis (P<0.05). WT1 level was decreased, and GATA-1 level was significantly increased in subcutaneous tumor tissues (P<0.05 or P<0.01). Conclusion: GRd might induce the differentiation of AML cells via regulating the ERK/GSK-3β signaling pathway. [ABSTRACT FROM AUTHOR]

Cultivated meat refers to edible meat obtained by proliferating cells without killing livestock in a laboratory. The selection of donor animals is a crucial factor for efficient cell culture production. Hanwoo is a native Korean taurine cattle breed raised as livestock in Korea since before 2000 B.C. Cells isolated from Hanwoo, which has little genetic diversity, are expected to be advantageous in cell culture because of the existence of fewer individual differences. However, cells collected from Hanwoo are in a state where efficient culture conditions have not been established. Therefore, in this study, we investigated the effects of mimicking an in vivo environment on the proliferation and differentiation of Hanwoo muscle satellite cells. The culture conditions consisted of CON (37°C/20% O2), T1 (37°C/2% O2), T2 (39°C/20% O2), and T3 (39°C/2% O2). Cell numbers decreased and expression levels of PAX7 and MYF5 decreased at a temperature of 39°C (p < 0.05). Conversely, 2% oxygen increased the number of cells and expression levels of PAX7 and MYF5 (p < 0.05). A temperature of 39°C inhibited the proliferation of Hanwoo muscle satellite cells by reducing the expression of PAX7 and MYF5 (p < 0.05). Conversely, 2% oxygen promoted the proliferation of Hanwoo muscle satellite cells by enhancing the expression of PAX7 and MYF5 (p < 0.05). During differentiation, a temperature of 39°C improved the myotube area and fusion index (p < 0.05). The RT-qPCR and Western blotting results revealed that a culture temperature of 39°C increased expression levels of the MYH2 gene and DES and MYOG proteins (p < 0.05). Additionally, an interactive condition increased expression levels of MYOD1, DES, and MYOG genes (p < 0.05). These results indicated that a temperature of 39°C promoted the differentiation of Hanwoo muscle satellite cells by increasing DES and MYOG protein expression. Thus, the production of cultivated meat using Hanwoo muscle satellite cells is expected to be efficient under 2% oxygen for proliferation and 39°C for differentiation. [ABSTRACT FROM AUTHOR]

Cortactin (CTTN), a cytoskeletal protein and substrate of Src kinase, is implicated in tumor aggressiveness. However, its role in bone cell differentiation remains unknown. The current study revealed that CTTN was upregulated during osteoblast and adipocyte differentiation. Functional experiments demonstrated that CTTN promoted the in vitro differentiation of mesenchymal stem/progenitor cells into osteogenic and adipogenic lineages. Mechanistically, CTTN was able to stabilize the protein level of mechanistic target of rapamycin kinase (mTOR), leading to the activation of mTOR signaling. In-depth investigation revealed that CTTN could bind with casitas B lineage lymphoma-c (c-CBL) and counteract the function of c-CBL, a known E3 ubiquitin ligase responsible for the proteasomal degradation of mTOR. Silencing c-Cbl alleviated the impaired differentiation of osteoblasts and adipocytes caused by CTTN siRNA, while silencing mTOR mitigated the stimulation of osteoblast and adipocyte differentiation induced by CTTN overexpression. Notably, transplantation of CTTN-silenced bone marrow stromal cells (BMSCs) into the marrow of mice led to a reduction in trabecular bone mass, accompanied by a decrease in osteoblasts and an increase in osteoclasts. Furthermore, CTTN-silenced BMSCs expressed higher levels of receptor activator of nuclear factor κB ligand (RANKL) than control BMSCs did and promoted osteoclast differentiation when cocultured with bone marrow-derived osteoclast precursor cells. This study provides evidence that CTTN favors osteoblast differentiation by counteracting the c-CBL-induced degradation of mTOR and inhibits osteoclast differentiation by downregulating the expression of RANKL. It also suggests that maintaining an appropriate level of CTTN expression may be advantageous for maintaining bone homeostasis. [ABSTRACT FROM AUTHOR]

Research Summary: We study product digitization as an impetus for firm strategy change. Product digitization can erode a firm's ability to differentiate through physical product attributes and prompts them to increase emphasis on nonphysical product attributes to sustain their competitive advantage. We expect this effect is pronounced among firms that have pursued a physical differentiation strategy prior to the digital age. However, we expect that countervailing forces exist such that the internal supply cost of and the external market demand for the nonphysical differentiator mitigate this effect. Evidence from publishers in the Amazon Kindle e‐book ecosystem supports our hypotheses. This study bridges the growing digital strategy literature and the classical competitive strategy literature. We discuss how our findings are relevant to a range of industries. Managerial Summary: Many companies invest in superior physical product attributes to increase a customer's willingness‐to‐pay and entice purchase. However, when products become digitized—such as music streaming displacing CDs or e‐books replacing printed books—the effectiveness of physical product differentiation is diminished. We examine how book publishers manage this challenge at the advent of Amazon's Kindle e‐book technology. We find that publishers that have emphasized physical product attributes turn to emphasizing nonphysical product differentiation, namely, through offering more content. Yet, the extent of the strategy change is limited by the internal development cost of and the external demand for the nonphysical differentiator. We discuss how our findings are relevant to a range of settings, including both product and service companies. [ABSTRACT FROM AUTHOR]

Everyday ventures are important engines of economic and social development around the world. These types of ventures tend to be resource constrained, focused on regional service areas, and rely almost exclusively on replicative approaches to business. Despite their prevalence, these types of ventures receive far less attention in the literature than their innovative, growth-focused counterparts. Thus, there is still much we do not know about how the entrepreneurs behind these types of ventures approach strategy and measure their success. In this qualitative study, we utilize self-determination theory to interpret interview data and illustrate how venture launch and strategic choice allow everyday entrepreneurs to self-fulfill their psychological needs through their business. Doing so, our research answers questions related to: 1) everyday venture launch; 2) the strategies everyday entrepreneurs pursue to position their venture in the market; and, 3) the metrics by which everyday entrepreneurs measure the success of their venture. We find that everyday entrepreneurs self-fulfill their needs for autonomy, relatedness, and competence by launching their ventures and making specific strategic choices regarding competition. More specifically, we find that everyday entrepreneurs primarily rely on differentiation strategies focused on high-quality offerings coupled with a premium customer service experience, while measuring success using an approach similar to the balanced scorecard. We conclude with a discussion of the implications for theory and practice. [ABSTRACT FROM AUTHOR]

This study addressed a gap in the literature by investigating the link between corporate strategy and board composition and how it connects to firm value. Most studies have explored whether board structure predicts firm outcomes rather than the other way around even though firm strategy plays a significant role in structuring the board. The sample covered the period between 2013 and 2018 and included 20,677 firm‐year observations, on which we executed country‐industry‐year fixed‐effects regression analysis. We found that, first, cost leadership strategy was positively associated with board size, board independence, board gender diversity, and board tenure, whereas it was negatively associated with board skills. Second, differentiation strategy was positively associated with board size and board gender diversity. Third, the moderating effects revealed that while board size and board tenure negatively moderated between cost leadership and firm value, board skills positively moderated between cost leadership and firm value. On the other hand, while board size, board gender diversity, and board tenure positively moderated between differentiation and firm value, board skills negatively moderated between differentiation and firm value. The results were robust to an alternative firm value proxy and endogeneity concerns. These findings provide firms with the opportunity to better shape their board structure in line with their corporate strategies and shareholder expectations. [ABSTRACT FROM AUTHOR]

Neuroblastoma is a type of peripheral neuroblastic tumor and is a common malignant solid tumor in children with significant biological heterogeneity and rapid development. Determining the type of differentiation is important in predicting the prognosis of neuroblastoma and making early judgments on postoperative treatment plans. The whole slide images of neuroblastoma offer high resolution that greatly facilitates clinical reading. However, the high resolution of these images makes computer-aided diagnosis quite challenging. To address the challenge, we propose in this paper a network model based on the traditional encoder-decoder structure of U-Net. The model uses DenseNet as the encoder to extract features of image, initializes network weights by transfer learning, and builds the decoder based on the up-sampling module and dense block to extract different types of cells. K-means algorithm is used to cluster and count the poorly differentiated cells and differentiated cells, identifying the histopathological types and the prognostic effects. Our experimental results demonstrate that our model performs excellently, having achieved 97.55% of segmentation accuracy, 92.10% and 89.02% counting accuracy for poor differentiated cells and differentiated cells, respectively. [ABSTRACT FROM AUTHOR]

The use of loose parts can be an effective teaching method to improve and enhance student understanding of Next Generation Science Standards (NGSS). Loose parts can inspire children to use their creativity and critical thinking skills in the classroom, which encourages them to embrace science curriculum and connections to standards at any age. Students learn most readily and positively in a laboratory type environment, where they can experiment, create, invent, appreciate, and find out things for themselves. Children should be encouraged to be part of the experiment, not just listening at their desk, or watching on the sidelines. This article outlines the theory, associated research, principles, benefits, and developing a loose parts mindset, with examples of light exploration, magnetic games, forces loose parts learning lab, and nest building using natural materials. Loose parts can be applied to most educational topics and content, making it a universal learning tool, putting students at the center of their education. [ABSTRACT FROM AUTHOR]

Background: The classical concept of brain sex differentiation suggests that steroid hormones released from the gonads program male and female brains differently. However, several studies indicate that steroid hormones are not the only determinant of brain sex differentiation and that genetic differences could also be involved. Methods: In this study, we have performed RNA sequencing of rat brains at embryonic days 12 (E12), E13, and E14. The aim was to identify differentially expressed genes between male and female rat brains during early development. Results: Analysis of genes expressed with the highest sex differences showed that Xist was highly expressed in females having XX genotype with an increasing expression over time. Analysis of genes expressed with the highest male expression identified three early genes, Sry2, Eif2s3y, and Ddx3y. Discussion: The observed sex-specific expression of genes at early development confirms that the rat brain is sexually dimorphic prior to gonadal action on the brain and identifies Sry2 and Eif2s3y as early genes contributing to male brain development. [ABSTRACT FROM AUTHOR]

Social enterprises are organisations comprised of two conflicting logics: social and commercial. However, simultaneous commitment to both logics often leads to tensions, making social enterprises fragile organisations at the risk of mission drift. The extant literature on hybridity highlights two important overall strategies – integration and differentiation - for managing tensions. However, integration or differentiation alone can create additional problems, which is especially true in the case of differentiation, creating a situation comparable to black and white, with a salient faultline and heated conflicts. In this inductive qualitative case study, responding to the recent call in the literature on how to mitigate such challenges, we will show that social enterprises could employ two strategies called people circulation and weaving warp and weft to achieve a reintegrated, embedded situation referred as yin and yang with a mitigated faultline and lessened tensions. [ABSTRACT FROM AUTHOR]

Cancer cells often overexpress CD47, which triggers the inhibitory receptor SIRPα expressed on macrophages, to elude phagocytosis and antitumor immunity. Pharmacological blockade of CD47 or SIRPα is showing promise as anticancer therapy, although CD47 blockade has been associated with hematological toxicities that may reflect its broad expression pattern on normal cells. Here we found that, in addition to triggering SIRPα, CD47 suppressed phagocytosis by a SIRPα-independent mechanism. This mechanism prevented phagocytosis initiated by the pro-phagocytic ligand, SLAMF7, on tumor cells, due to a cis interaction between CD47 and SLAMF7. The CD47-SLAMF7 interaction was disrupted by CD47 blockade and by a first-in-class agonist SLAMF7 antibody, but not by SIRPα blockade, thereby promoting antitumor immunity. Hence, CD47 suppresses phagocytosis not only by engaging SIRPα, but also by masking cell-intrinsic pro-phagocytic ligands on tumor cells and knowledge of this mechanism may influence the decision between CD47 blockade or SIRPα blockade for therapeutic purposes.

T cells undergo extensive chromatin remodeling over several days following stimulation through the T cell receptor. However, the kinetics and gene loci targeted by early remodeling events within the first 24 hours of T cell priming to orchestrate effector differentiation have not been well described. We identified that chromatin accessibility is rapidly and extensively remodeled within 1 hour of stimulation of naïve CD8+ T cells, leading to increased global chromatin accessibility at many effector T cell-associated genes that are enriched for AP-1, early growth response (EGR), and nuclear factor of activated T cells (NFAT) binding sites, but this short duration of stimulation is insufficient for commitment to clonal expansion in vivo. Sustained 24-hour stimulation led to further chromatin remodeling and was sufficient to enable clonal expansion. These data suggest that the duration of antigen receptor signaling is intimately coupled to chromatin remodeling and activation of genes involved in effector cell differentiation and highlight a potential mechanism that helps CD8+ T cells discriminate between foreign- and self-antigens.

BackgroundApproximately 50% of patients who receive anti-CD19 CAR-T cells relapse, and new immunotherapeutic targets are urgently needed. We recently described CD72 as a promising target in B-cell malignancies and developed nanobody-based CAR-T cells (nanoCARs) against it. This cellular therapy design is understudied compared with scFv-based CAR-T cells, but has recently become of significant interest given the first regulatory approval of a nanoCAR in multiple myeloma.MethodsWe humanized our previous nanobody framework regions, derived from llama, to generate a series of humanized anti-CD72 nanobodies. These nanobody binders were inserted into second-generation CD72 CAR-T cells and were evaluated against preclinical models of B cell acute lymphoblastic leukemia and B cell non-Hodgkin's lymphoma in vitro and in vivo. Humanized CD72 nanoCARs were compared with parental ("NbD4") CD72 nanoCARs and the clinically approved CD19-directed CAR-T construct tisangenlecleucel. RNA-sequencing, flow cytometry, and cytokine secretion profiling were used to determine differences between the different CAR constructs. We then used affinity maturation on the parental NbD4 construct to generate high affinity binders against CD72 to test if higher affinity to CD72 improved antitumor potency.ResultsToward clinical translation, here we humanize our previous nanobody framework regions, derived from llama, and surprisingly discover a clone ("H24") with enhanced potency against B-cell tumors, including patient-derived samples after CD19 CAR-T relapse. Potentially underpinning improved potency, H24 has moderately higher binding affinity to CD72 compared with a fully llama framework. However, further affinity maturation (KD

Abstract Spermatogonial stem cells (SSCs) play a crucial role in the male reproductive system, responsible for maintaining continuous spermatogenesis. The microenvironment or niche of SSCs is a key factor in regulating their self-renewal, differentiation and spermatogenesis. This microenvironment consists of multiple cell types, extracellular matrix, growth factors, hormones and other molecular signals that interact to form a complex regulatory network. This review aims to provide an overview of the main components of the SSCs microenvironment, explore how they regulate the fate decisions of SSCs, and discuss the potential impact of microenvironmental abnormalities on male reproductive health. Graphical Abstract

Abstract Background Cardiovascular progenitor cells (CPCs) derived from human embryonic stem cells (hESCs) are considered valuable cell sources for investigating cardiovascular physiology in vitro. Meeting the diverse needs of this application requires the large-scale production of CPCs in an in vitro environment. This study aimed to use an effective culture system utilizing signaling factors for the large-scale expansion of hESC-derived CPCs with the potential to differentiate into functional cardiac lineage cells. Methods and results Initially, CPCs were generated from hESCs using a 4-day differentiation protocol with a combination of four small molecules (CHIR99021, IWP2, SB-431542, and purmorphamine). These CPCs were then expanded and maintained in a medium containing three factors (bFGF, CHIR, and A83-01), resulting in a > 6,000-fold increase after 8 passages. These CPCs were successfully cryopreserved for an extended period in late passages. The expanded CPCs maintained their gene and protein expression signatures as well as their differentiation capacity through eight passages. Additionally, these CPCs could differentiate into four types of cardiac lineage cells: cardiomyocytes, endothelial cells, smooth muscle cells, and fibroblasts, demonstrating appropriate functionality. Furthermore, the coculture of these CPC-derived cardiovascular lineage cells in rat tail collagen resulted in cardiac microtissue formation, highlighting the potential of this 3D platform for studying cardiovascular physiology in vitro. Conclusion In conclusion, expandable hESC-derived CPCs demonstrated the ability to self-renewal and differentiation into functional cardiovascular lineage cells consistently across passages, which may apply as potential cell sources for in vitro cardiovascular studies. Graphical Abstract

Abstract Low back pain (LBP) is largely attributed to intervertebral disc degeneration (IVDD), of which the endplate changes are an important component. However, the alterations in cell fate and properties within the endplates during degeneration remain unknown. Here, we firstly performed the single-cell RNA-sequencing analysis (scRNA-seq) of the cells focusing on degenerative human endplates. By unsupervised clustering of the 8,534 single-cell based on the gene expression, we identified nine distinct cell types. We employed Gene ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways analysis, and the single-cell regulatory network inference and clustering (SCENIC) to determine the enriched pathways and transcriptional activities across seven chondrocyte subpopulations. Furthermore, two cell fates of chondrocyte differentiation were found by trajectory analysis, one was enriched in inflammation-related genes, and the other was related to extracellular matrix (ECM). Additionally, the intercellular interactions of macrophages (MA) and chondrocytes, T cells/natural killer cells (T/NK) and chondrocytes were examined by ligand-receptor pairs analysis, showing the important regulative function of FN1 from MA and CD74 from T/NK during endplate degeneration. Overall, our findings provide novel perspectives on the endplate degeneration at the single-cell level and a whole-transcriptome size.

Background and aim: A surplus of glucocorticoids (GC) is a main cause of non-traumatic osteonecrosis of the femoral head (ONFH), and Jintiange (JTG), as one of the traditional Chinese medicines (TCM), also plays an instrumental role in the alleviation of bone loss simultaneously. Therefore, JTG was thought to be able to reverse GC-induced ONFH (GC-ONFH) to a certain extent. Experimental procedure: In vivo, the effect of JTG on trabeculae in the subchondral bone of the femoral head was investigated using micro-computed tomography (micro-CT), TdT-mediated dUTP nick end labeling (TUNEL) and histological staining; in vitro, proliferation, viability, apoptosis, and senescence of purified bone mesenchymal stem cells (BMSCs) were examined to demonstrate the direct impact of JTG on these cells. Meanwhile after using a series of interventions, the function of JTG on BMSC differentiation could be assessed by measuring of osteogenic and adipogenic markers at levels of protein and mRNA. Results: Our final results demonstrated that with the involvement of Wnt/β-catenin pathway, JTG was able to significantly promote osteogenesis, restrain adipogenesis, delay senescence in BMSCs, reduce osteoclast number, weaken apoptosis, and enhance proliferation of osteocytes, all of which could mitigate the progression of subchondral osteonecrosis. Conclusion: According to the results of experiments in vitro and vivo, JTG was deemed to relieve the early GC-ONFH using the prevention of destruction of subchondral bone, which was contributed to regulating the differentiation of BMSCs and the number of osteoclasts.

Infectious diseases caused by ubiquitous bacteria with variability of virulence factors and multiple antibiotic resistance reliably often represent the structure of the general pathology of poultry. The aim of the study was to analyze dynamics of morph functional parameters of immune system of chickens in experimental pseudomonas infection. The initiation, development, and outcome of the experimental infectious process were considered using bacteriological and immunological methods with vital dye. Dissemination ofP. aeruginosabacteria revealed significant increase in extravasation of lung blood vessels, signs of hemodynamic disorders, and development of dystrophic and compensatory-adaptive processes. Pathogenic mechanisms are mediated by the dynamics of changes in reticuloendothelial and lymph epithelial systems: accidental transformation of thymus, atrophy of bursa of Fabricius, hyperplasia of esophageal and cecum lymphoid follicles, Meckels diverticulum, and spleen.

Abstract The significant advances in the differentiation of human pluripotent stem (hPS) cells into pancreatic endocrine cells, including functional β-cells, have been based on a detailed understanding of the underlying developmental mechanisms. However, the final differentiation steps, leading from endocrine progenitors to mono-hormonal and mature pancreatic endocrine cells, remain to be fully understood and this is reflected in the remaining shortcomings of the hPS cell-derived islet cells (SC-islet cells), which include a lack of β-cell maturation and variability among different cell lines. Additional signals and modifications of the final differentiation steps will have to be assessed in a combinatorial manner to address the remaining issues and appropriate reporter lines would be useful in this undertaking. Here we report the generation and functional validation of hPS cell reporter lines that can monitor the generation of INS+ and GCG+ cells and their resolution into mono-hormonal cells (INS eGFP , INS eGFP /GCG mCHERRY ) as well as β-cell maturation (INS eGFP /MAFA mCHERRY ) and function (INS GCaMP6 ). The reporter hPS cell lines maintained strong and widespread expression of pluripotency markers and differentiated efficiently into definitive endoderm and pancreatic progenitor (PP) cells. PP cells from all lines differentiated efficiently into islet cell clusters that robustly expressed the corresponding reporters and contained glucose-responsive, insulin-producing cells. To demonstrate the applicability of these hPS cell reporter lines in a high-content live imaging approach for the identification of optimal differentiation conditions, we adapted our differentiation procedure to generate SC-islet clusters in microwells. This allowed the live confocal imaging of multiple SC-islets for a single condition and, using this approach, we found that the use of the N21 supplement in the last stage of the differentiation increased the number of monohormonal β-cells without affecting the number of α-cells in the SC-islets. The hPS cell reporter lines and the high-content live imaging approach described here will enable the efficient assessment of multiple conditions for the optimal differentiation and maturation of SC-islets.

Abstract Pregestational diabetes mellitus (PGDM) has an impact on fetal bone formation, but the underlying mechanism is still obscure. Although miRNAs have been extensively investigated throughout bone formation, their effects on fetal bone development caused by PGDM still need clarification. This study intends to examine the mechanism by which hyperglycemia impairs the bone formation of offspring via miR-322-5p (miR-322). In this study, miR-322 was selected by systemically screening utilizing bioinformatics and subsequent validation experiments. Using streptozotocin (STZ)-induced diabetic mice and ATDC5 cell lines, we found that miR-322 was abundantly expressed in the proliferative and hypertrophic zones of the growth plate, and its expression pattern was disturbed in the presence of hyperglycemia, suggesting that miR-322 is involved in the chondrocyte proliferation and differentiation in absence/presence of hyperglycemia. This observation was proved by manipulating miR-322 expression in ATDC5 cells by transfecting mimic and inhibitor of miR-322. Furthermore, Adamts5, Col12a1, and Cbx6 were identified as the potential target genes of miR-322, verified by the co-transfection of miR-322 inhibitor and the siRNAs, respectively. The evaluation criteria are the chondrocyte proliferation and differentiation and their relevant key gene expressions (proliferation: Sox9 and PthIh; differentiation: Runx2 and Col10a1) after manipulating the gene expressions in ATDC5 cells. This study revealed the regulative role miR-322 on chondrocyte proliferation and differentiation of growth plate by targeting Adamts5, Col12a1, and Cbx6 in hyperglycemia during pregnancy. This translational potential represents a promising avenue for advancing our understanding of bone-related complications in diabetic pregnancy and mitigating bone deficiencies in diabetic pregnant individuals, improving maternal and fetal outcomes.

Abstract Acute myeloid leukemia (AML) is a hematologic neoplasm, characterized by a blockage of differentiation and an unconstrained proliferation of immature myeloid cells. Recently, the survival of leukemia patients has increased thanks to the use of differentiating agents, though these may cause serious side effects. Hence, the search for safer differentiating compounds is necessary. Our aim was to assess the pro-differentiating effects of a flavonoid-rich extract of bergamot juice (BJe) in human monocytic leukemia THP-1 cells, an in vitro AML model. For the first time, we showed that treatment with BJe induced differentiation of THP-1 cells, changes in cell morphology and increased expression of differentiation-associated surface antigens CD68, CD11b and CD14. Moreover, BJe enhanced protein levels of autophagy-associated markers, such as Beclin-1 and LC3, as well as induced the phosphorylation of the MAPKs JNK, ERK and p38, hence suggesting a potential mechanism underlying its antiproliferative effects. Indeed, parallel experiments highlighted that BJe was able to hamper THP-1 cell growth. In conclusion, our study suggests that BJe induces the differentiation of THP-1 cells and reduces their proliferation, highlighting its potential in differentiation therapy of AML.

The research examines the dynamism of health tourism, with an emphasis on the contribution of segmentation, differentiation, and communication of entrepreneurs. It highlights the convergence of health tourism entrepreneurs' behavior into the best opportunities to gain profit and create a brand in the healthcare market. Organizational culture, worldwide trends and changes, and an open mind for a collaborative relationship entrepreneur-employer are significant driving forces in this field. The necessity for deliberate support in listening to customers' voices is a goal of market researchers in health tourism acknowledgment.

The aim of this study was the effectiveness of systematic differentiation training based on systemic family therapy on marital boredom and communication patterns of women referring to counseling centers in Najafabad in 1399. The research method was quasi-experimental pre-test-post-test. The statistical population of the study was all women referred to counseling centers in Najafabad. The sampling method used in this study was purposeful available sampling method that finally 30 people were selected. 15 people in the experimental group and 15 people in the control group were randomly assigned. The tools used for data collection included the Marital Boredom Questionnaire (Pines 1996), the Communication Patterns Questionnaire (Christensen and Salawi 1984) and the Differentiation Therapy Training Package (based on Bowen's system theory). Data analysis was performed using 24SPSS software at two levels of descriptive and inferential statistics. At the inferential level, analysis of covariance was used. The results of analysis of covariance showed that systematic differentiation training based on systemic family therapy has an effect on marital boredom (P

Abstract Background Using natural killer (NK) cells to treat hematopoietic and solid tumors has great promise. Despite their availability from peripheral blood and cord blood, stem cell-derived NK cells provide an “off-the-shelf” solution. Methods In this study, we developed two CAR-NK cells targeting PD-L1 derived from lentiviral transduction of human umbilical cord blood (UCB)-CD34+ cells and UCB-CD34+-derived NK cells. The transduction efficiencies and in vitro cytotoxic functions including degranulation, cytokine production, and cancer cell necrosis of both resultants PD-L1 CAR-NK cells were tested in vitro on two different PD-L1 low and high-expressing solid tumor cell lines. Results Differentiated CAR‑modified UCB-CD34+ cells exhibited enhanced transduction efficiency. The expression of anti-PD-L1 CAR significantly (P

Abstract Background The mechanisms enabling dynamic shifts between drug-resistant and drug-sensitive states in cancer cells are still underexplored. This study investigated the role of targeted autophagic protein degradation in regulating ovarian cancer stem cell (CSC) fate decisions and chemo-resistance. Methods Autophagy levels were compared between CSC-enriched side population (SP) and non-SP cells (NSP) in multiple ovarian cancer cell lines using immunoblotting, immunofluorescence, and transmission electron microscopy. The impact of autophagy modulation on CSC markers and differentiation was assessed by flow cytometry, immunoblotting and qRT-PCR. In silico modeling and co-immunoprecipitation identified ID1 interacting proteins. Pharmacological and genetic approaches along with Annexin-PI assay, ChIP assay, western blotting, qRT-PCR and ICP-MS were used to evaluate effects on cisplatin sensitivity, apoptosis, SLC31A1 expression, promoter binding, and intracellular platinum accumulation in ID1 depleted backdrop. Patient-derived tumor spheroids were analyzed for autophagy and SLC31A1 levels. Results Ovarian CSCs exhibited increased basal autophagy compared to non-CSCs. Further autophagy stimulation by serum-starvation and chemical modes triggered proteolysis of the stemness regulator ID1, driving the differentiation of chemo-resistant CSCs into chemo-sensitive non-CSCs. In silico modeling predicted TCF12 as a potent ID1 interactor, which was validated by co-immunoprecipitation. ID1 depletion freed TCF12 to transactivate the cisplatin influx transporter SLC31A1, increasing intracellular cisplatin levels and cytotoxicity. Patient-derived tumor spheroids exhibited a functional association between autophagy, ID1, SLC31A1, and platinum sensitivity. Conclusions This study reveals a novel autophagy-ID1-TCF12-SLC31A1 axis where targeted autophagic degradation of ID1 enables rapid remodeling of CSCs to reverse chemo-resistance. Modulating this pathway could counter drug resistance in ovarian cancer.