Cell Morphology Research Articles

Spraying KH2PO4 Alleviates the Damage of Spring Low-Temperature Stress by Improving the Physiological Characteristics of Wheat Flag Leaves.

The low-temperature stress (LTS) in spring results in tremendous yield loss in wheat production, and the application of potassium dihydrogen phosphate (KH2PO4) can alleviate stress-induced damage. However, the underlying effect of spraying KH2PO4 on the physiological characteristics of wheat flag leaves under spring LTS remains unclear. In this study, we investigated the effect of spraying KH2PO4 on flag leaf physiological traits and yield under spring LTS, including treatments at 15 °C and spraying H2O (CK), treatment at -4 °C and spraying H2O (LT1), and treatment at -4 °C and spraying KH2PO4 (LT2). The results showed that spraying KH2PO4 significantly increased the activities of the superoxide dismutase (SOD), the peroxidase (POD), and the catalase (CAT), and reduced malondialdehyde (MDA) content in the flag leaves. Compared to LT1, the SOD, POD, and CAT activities in the flag leaves of the Yangnong19 (YN19) and Xinmai26 (XM26) via LT2 increased by 5.5%, 10.9%, and 3.9%, and 5.4%, 9.2%, and 4.4%, respectively, and the MDA content of the YN19 and XM26 decreased by 10.5% and 9.1%, respectively, at 0-12 d after low temperature treatment (DALTT). Spraying KH2PO4 appreciably alleviated damage to the leaf cell morphology and tissue integrity, and increased the accumulation of proline and soluble protein, the chlorophyll content, and the activities of Ribulose-1,5-bisphosphate carboxylase and phosphoenolpyruvate carboxykinase. The net photosynthetic rate in the flag leaves of the YN19 and XM26 via LT2 increased by 37.9% and 35.9%, respectively, at 0-12 DALTT, compared to LT1. Moreover, spraying KH2PO4 reduced the yield loss rate of the YN19 and XM26 by 13.06% and 16.72%, respectively. The present study demonstrates that spraying KH2PO4 can enhance wheat resistance to spring LTS and maintain the photosynthetic capacity of flag leaves, alleviating the negative effects of LTS on grain yield.

Angiomatoid Fibrous Histiocytoma Initially Misdiagnosed as Elastofibroma Dorsi: A Case Report and Literature Review

Background and Objectives: Angiomatoid fibrous histiocytoma (AFH) is a rare soft-tissue tumor with a low-grade malignancy. It typically arises in superficial soft tissues of the extremities, head, neck and trunk in children or young adults. Because of its rare entity, it tends to be confused and misdiagnosed. Materials and Methods: A 12-year-old male presented with a painless mass located on his right upper back. The CT finding showed a 7.3 × 2.8 × 5.4 cm-sized, well-defined heterogeneous soft tissue mass in the right infrascapular area. We performed a complete excision, including the surrounding capsule. Result: The final pathology revealed an AFH of intermediate malignancy. On pathologic examination, the lesion was a 5.8 × 4.5 × 2.6 cm-sized mass with a mitotic count of 12/10 HPF, tumor necrosis of 0% and marked increased cellularity and spindle cell morphology. The immunohistochemical study showed negative for S100 and positive for SMA, focal positive for Ki-67, CD68 and positive for CD99, Desmin staining. During the five years of follow-up period, he did not show any evidence of recurrence. Conclusions: The result was satisfactory. We report a case of AFH of the back initially misdiagnosed as an elastofibroma dorsi (ED) with the review of the literature for this uncommon entity.

Effects of simulated microgravity on colorectal cancer organoids growth and drug response

Cellular and molecular dynamics of human cells are constantly affected by gravity. Alteration of the gravitational force disturbs the cellular equilibrium, which might modify physiological and molecular characteristics. Nevertheless, biological responses of cancer cells to reduced gravitational force remains obscure. Here, we aimed to comprehend not only transcriptomic patterns but drug responses of colorectal cancer (CRC) under simulated microgravity. We established four organoids directly from CRC patients, and organoids cultured in 3D clinostat were subjected to genome wide expression profiling and drug library screening. Our observations revealed changes in cell morphology and an increase in cell viability under simulated microgravity compared to their static controls. Transcriptomic analysis highlighted a significant dysregulation in the TBC1D3 family of genes. The upregulation of cell proliferation observed under simulated microgravity conditions was further supported by enriched cell cycle processes, as evidenced by the functional clustering of mRNA expressions using cancer hallmark and gene ontology terms. Our drug screening results indicated an enhanced response rate to 5-FU under conditions of simulated microgravity, suggesting potential implications for cancer treatment strategies in simulated microgravity.

Ethnopharmacological insights: hyperoside from Marsdenia latifolia (Benth.) K.Schum. mitigates reproductive toxicity in male Wistar rats induced by manganese exposure

This study provides novel findings on the ameliorative effect of hyperoside isolated from Marsdenia latifolia leaves (HIGLL) in reproductive health challenged by manganese toxicity. The study investigated the efficacy of HIGLL on male fertility in rats exposed to manganese chloride (MnCl2). The rats received either MnCl2 (30 mg/L) or in combination with HIGLL (100 mg/kg or 200 mg/kg). MnCl2 reduced fluid intake, organ-body weight, body weight; sperm count, sperm viability, sperm density, sperm motility, semen viscosity, daily sperm production, testicular sperm number; testosterone, follicle-stimulating and luteinising hormones, oestradiol, testosterone/oestradiol ratio; G6PDH, ALP, glucose, NO, acid phosphatase, sialic acid, 17-β-HSD, superoxide dismutase, CAT, GST, GSH, and T-SH without modifying the semen pH and volume, while it correspondingly raised the abnormalities associated with morphology of sperm cells in head, neck, and tail; H2O2 and lipid peroxidation levels. HIGLL abrogated the damaged histoarchitecture of the testes and epididymis caused by MnCl2. HIGLL can serve as a therapeutic agent in the management of male reproductive disorders related to oxidative stress and endocrine disruption.

Electroacupuncture preconditioning prevents urticaria by regulating inflammatory response in rats with urticaria

To observe the effect of electroacupuncture (EA) preconditioning at "Quchi" (LI11) and "Xuehai" (SP10) in prevention of urticaria. Twenty-four male SD rats were randomly divided into control, model and preconditioning of EA (Pre-EA) groups (8 rats/group). The urticaria model was established by intradermal injection of dilute allogeneic antioalbumin serum at the spots of the bilateral symmetry of the spine on the back, and followed by tail venous injection of mixture solution of egg albumin diluent, plus 0.5% Evans blue and normal saline. Ten days before the end of modeling, rats of the pre-EA group received EA stimulation of LI11 and SP10 for 20 min, once a day for 10 consecutive days. The times of rat's scratching the sensitized skin were recorded. HE staining method was used to observe the pathological changes of skin tissue, and toluidine blue staining method was used to observe the morphology of mast cells (MCs) in the skin, blood, mesentery, and peritoneal fluid, and calculate the degranulation rate. Immunohistochemical stainning was used to detect immunoglobulin E (IgE), histamine (HIS), and 5-hydroxytryptamine (5-HT) expressions in subcutaneous tissue. NOD like receptor thermal domain associated protein 3 (NLRP3) inflammasome, apoptosis related granule protein (ASC), and cysteine aspartate aminotransferase 1 (Caspase-1) protein expression levels in skin tissue were detected by Western blot. The contents of serum interleukin(IL)-1β and IL-18 were detected using ELISA method. Compared with the control group, the scratching times, amount of Evans blue exudation of the sensitized blue spots, degranulation rate of MCs in skin, blood, mesentery and peritoneal fluid, the expression levels of IgE, HIS, 5-HT in subcutaneous tissue, protein expression levels of NLRP3, ASC, Caspase-1 in skin tissue, and the contents of serum IL-1β and IL-18 were significantly increased (P<0.01, P<0.05) in the model group. In comparison with the model group, the scratching times, amount of Evans blue exudation of the sensitized blue spots, degranulation rate of MCs, the expression levels of IgE, HIS, 5-HT in subcutaneous tissue, protein expression levels of NLRP3, ASC, Caspase-1 in skin tissue, and the contents of serum IL-1β and IL-18 in EA group were significantly decreased (P<0.01, P<0.05). EA preconditioning at LI11 and SP10 can prevent and treat UR by inhibiting inflammatory response, which is related to the regulation of pyroptosis.

Epithelioid solitary fibrous tumors from CNS and soft tissues: an unusual morphologic variant

BackgroundSolitary fibroous tumors (SFTs) are distinctive soft tissue tumors characterized by rearrangements of NAB2-STAT6 gene, which are associated with thin-walled, branching, “staghorn”-shaped vessels. SFTs are originally classified as a type of hemangiopericytoma (HPC). Classical SFTs are composed of spindle to ovoid cells arranged haphazardly or in fascicles. Rarely, SFTs exhibit unusual morphological variants such as fat formation, giant cells, dedifferentiation, or epithelioid variant. The epithelioid cell variant, which is composed almost entirely of epithelioid cells and arranged in solid or nest patterns, is extremely rare and frequently malignant.Case presentationIn this study, we reported three cases of epithelioid SFTs (ESFTs) located in extrathoracic sites (right lateral ventricle, right lumbar, left pelvis). All the subjects in this study were elderly, with a predominance of female patients, accounting for two out of the three cases, and only one case involved a male patient. The tumor cells were entirely composed of epithelioid cells and exhibited positive for CD34 and STAT-6 markers. Ultimately, the majority of cases (two out of three) were diagnosed as malignant SFTs.ConclusionThis study aims to enhance the awareness of ESFTs. In these cases, irrespective of the onset location, the arrangement patterns of tumor cells, such as papillary structures and the morphology of epithelial-like cells, conspicuously lack the hallmark histological characteristics of Solitary Fibrous Tumors (SFTs). Consequently, it requires differential diagnosis from a plethora of malignant neoplasms. Moreover, the elevated malignancy level of this cohort of cases poses substantial diagnostic challenges to pathologists, compounding the complexity of accurate interpretation.

Cytologic features of gastric-type endocervical adenocarcinoma: Three cases report and literature review.

Cervical gastric-type adenocarcinoma shows gastric differentiation, and the tumor cell morphology appears benign and unrelated to human papillomavirus, which makes cervical gastric-type adenocarcinoma highly susceptible to misdiagnosis as normal glandular epithelium in cytologic diagnosis. We present 3 cases of gastric-type adenocarcinoma, with the first being a 57-year-old female with abnormal uterine bleeding and fluid drainage. The second patient was a 63-year-old female, and the third was a 59-year-old female with irregular vaginal bleeding after menopause. The 3 patients were diagnosed with cervical gastric-type adenocarcinoma by combining their history, clinical data, cytopathology, histopathology, and immunohistochemistry. Patient 1 underwent total hysterectomy and adnexectomy, but refused radiotherapy and chemotherapy. Patient 2 received a chemotherapeutic regimen, and patient 3 was treated with traditional Chinese medicine. Patient 1 was followed-up for 9 months and is currently in good general condition, while patients 2 and 3 were not followed-up. The "drunken honeycomb" cell arrangement is diagnostically important in liquid-based cytology, especially when three-dimensional spheroids are present, and may be a characteristic cytological finding of well-differentiated cervical gastric-type adenocarcinoma.

Impact of cannabinoids on synapse markers in an SH-SY5Y cell culture model

Patients suffering from schizophrenic psychosis show reduced synaptic connectivity compared to healthy individuals, and often, the use of cannabis precedes the onset of schizophrenic psychosis. Therefore, we investigated if different types of cannabinoids impact methylation patterns and expression of schizophrenia candidate genes concerned with the development and preservation of synapses and synaptic function in a SH-SY5Y cell culture model. For this purpose, SH-SY5Y cells were differentiated into a neuron-like cell type as previously described. Effects of the cannabinoids delta-9-THC, HU-210, and Anandamide were investigated by analysis of cell morphology and measurement of neurite/dendrite lengths as well as determination of methylation pattern, expression (real time-qPCR, western blot) and localization (immunocytochemistry) of different target molecules concerned with the formation of synapses. Regarding the global impression of morphology, cells, and neurites appeared to be a bit more blunted/roundish and to have more structures that could be described a bit boldly as resembling transport vesicles under the application of the three cannabinoids in comparison to a sole application of retinoic acid (RA). However, there were no obvious differences between the three cannabinoids. Concerning dendrites or branch lengths, there was a significant difference with longer dendrites and branches in RA-treated cells than in undifferentiated control cells (as shown previously), but there were no differences between cannabinoid treatment and exclusive RA application. Methylation rates in the promoter regions of synapse candidate genes in cannabinoid-treated cells were in between those of differentiated cells and untreated controls, even though findings were significant only in some of the investigated genes. In other targets, the methylation rates of cannabinoid-treated cells did not only approach those of undifferentiated cells but were also valued even beyond. mRNA levels also showed the same tendency of values approaching those of undifferentiated controls under the application of the three cannabinoids for most investigated targets except for the structural molecules (NEFH, MAPT). Likewise, the quantification of expression via western blot analysis revealed a higher expression of targets in RA-treated cells compared to undifferentiated controls and, again, lower expression under the additional application of THC in trend. In line with our earlier findings, the application of RA led to higher fluorescence intensity and/or a differential signal distribution in the cell in most of the investigated targets in ICC. Under treatment with THC, fluorescence intensity decreased, or the signal distribution became similar to the dispersion in the undifferentiated control condition. Our findings point to a decline of neuronal differentiation markers in our in vitro cell-culture system under the application of cannabinoids.

Methodological approaches to the comparative assessment of cellular reactions of innate immunity in mollusk &lt;I&gt;Pomacea&lt;/I&gt; sp. (&lt;I&gt;Caenogastropoda, Ampullariidae&lt;/I&gt;) in experimental aseptic inflammation

One of the significant methodological problems that arise in the study of cells of the innate immunity of invertebrates is the unsuitability of culture media and even salt solutions intended for mammals and humans. Another important methodological point is the significantly pronounced reactions of coagulation, which play an important protective role in all invertebrates, but manifest themselves in vitro in cellular suspensions prepared even with optimal saline solutions for this animal. To solve this problem, researchers widely use an anticoagulation buffer with ethylenediaminetetraacetic acid (EDTA) even in functional tests. From the standpoint of traditional approaches of mammalian immunology, the use of EDTA is possible only for the study of cell morphology, but not their functions. The aim of the work is to adapt methodological approaches to the comparative assessment of cellular reactions of innate immunity in Pomacea sp mollusks in experimental aseptic inflammation. To evaluate the functions of phagocytic cells in mollusk Pomacea sp., we used a complete salt solution (CSS) with optimal concentrations of glucose, calcium ions, magnesium with addition to prevent coagulation, cellular aggregation and degranulation instead of EDTA sodium salt of heparin at a concentration of 100 IU/mL (CSS-hep). As a result of a specially conducted experiment, it was shown that during incubation of hemolymph cells in this solution, their viability is preserved, evaluated by flow laser cytometry during incubation with annexin V-PE (PE Annexin V) and 7-aminoactinomycin D (7- AAD), as well as in a test with trypan blue. The possibility of studying the phagocytic activity and a phagosome acidification of leukocytes from hemolymph, hematopoiesis organ (kidney) and the focus of inflammation induced by intramuscular injection of sterile suspension of zymosan using CSS-hep is shown. Unlike the Limnaea stagnalis and Biomphalaria glabrata mollusks, in which the production of reactive oxygen species by phagocytes in the luminol-dependent chemiluminescence reaction is well documented, we were unable to induce this reaction in Pomacea sp. mollusks. In a specially conducted experiment, we did not reveal the effect of the sodium salt of heparin at a concentration of 100 IU/mL on the production of reactive oxygen species by rat phagocytes.

Bacterial Responses and Material-Cell Interplays With Novel MoAlB@MBene.

Developing efficient antibacterial nanomaterials has potential across diverse fields, but it requires a deeper understanding of material-bacteria interactions. In this study, a novel 2D core-shell MoAlB@MBene structure is synthesized using a mild wet-chemical etching approach. The growth of E. coli, S. aureus, and B. subtilis bacteria in the presence of MoAlB@MBene decreased in a concentration-dependent manner, with a prolonged lag phase in the initial 6h of incubation. Even under dark conditions, MoAlB@MBene triggered the formation of intercellular reactive oxygen species (ROS) and singlet oxygen (1O2) in bacteria, while the bacteria protected themselves by forming biofilm and altering cell morphology. The MoAlB@MBene shows consistent light absorption across the visible range, along with a distinctive UV absorption edge. Two types of band gaps are identified: direct (1.67eV) and indirect (0.74eV), which facilitate complex light interactions with MoAlB@MBene. Exposure to simulated white light led to decreased viability rates of E. coli (20.6%), S. aureus (22.9%), and B. subtilis (21.4%). Altogether, the presented study enhances the understanding of bacteria responses in the presence of light-activated 2D nanomaterials.

Ferrocenyl β-Diketonate Compounds: Extended Ring Systems for Improved Anticancer Activity.

A library of ferrocenyl β-diketonate compounds with varying degrees of aromatic functionality have been synthesized and fully characterized. This includes cyclic voltammetry and the analysis of four new structures by single crystal X-ray diffraction. The compounds cytotoxic potential has been determined by MTT screening against pancreatic carcinoma (MIA PaCa-2), ovarian adenocarcinoma (A2780), breast adenocarcinomas (MDA-MB-231 and MCF-7) and normal epithelial retinal (ARPE-19). The compounds show a general trend, where increasing the number of aromatic rings in the molecule yields an increase in cytotoxicity and follows the trend anthracenyl > naphthyl > phenyl > methyl. The compounds are particularly sensitive to the triple negative cancer cell line MDA-MB-231, and the potential modes of action have been studied by production of reactive oxygen species using fluorescence microscopy and cell morphology using Scanning Electron Microscopy. All assays highlight the ferrocenyl β-diketonate with an anthracenyl substituent to be the lead compound in this library. The decomposition was also observed within cells, to give a cytotoxic fluorescent molecule, which has been visualized by confocal microscopy.

Unfractionated Heparin Enhances Sepsis Prognosis Through Inhibiting Drp1‐Mediated Mitochondrial Quality Imbalance

AbstractUnfractionated heparin (UFH) is commonly used as an anticoagulant in sepsis treatment and has recently been found to have non‐anticoagulant effects, but underlying mechanisms remain unclear. This retrospective clinical data showed that UFH has significant protective effects in sepsis compared to low‐molecular‐weight heparin and enoxaparin, indicating potential benefits of its non‐anticoagulant properties. Recombinant protein chip screening, surface plasmon resonance, and molecular docking data demonstrated that UFH specifically bound to the cytoplasmic Drp1 protein through its zone 2 non‐anticoagulant segment. In‐vitro experiments verified that UFH's specific binding to Drp1 suppressed Drp1 translocation to mitochondria following “sepsis” challenge, thereby improving mitochondrial morphology, function and metabolism in vascular endothelial cells. Consequently, UHF comprehensively protected mitochondrial quality, thus reducing vascular leakage and improving prognosis in a sepsis rat model. These findings highlight the potential of UFH as a sepsis treatment strategy targeting non‐anticoagulation mechanism.

An innovative 4D printing approach for fabrication of anisotropic collagen scaffolds

Collagen anisotropy is known to provide the essential topographical cues to guide tissue-specific cell function. Recent work has shown that extrusion-based printing using collagenous inks yield 3D scaffolds with high geometric precision and print fidelity. However, these scaffolds lack collagen anisotropy. In this study, extrusion-based 3D printing was combined with a magnetic alignment approach in an innovative 4D printing scheme to generate 3D collagen scaffolds with high degree of collagen anisotropy. Specifically, the 4D printing process parameters-collagen (Col):xanthan gum (XG) ratio (Col:XG; 1:1, 4:1, 9:1 v/v), streptavidin-coated magnetic particle concentration (SMP; 0, 0.2, 0.4 mg ml-1), and print flow speed (2, 3 mm s-1)-were modulated and the effects of these parameters on rheological properties, print fidelity, and collagen alignment were assessed. Further, the effects of collagen anisotropy on human mesenchymal stem cell (hMSC) morphology, orientation, metabolic activity, and ligamentous differentiation were investigated. Results showed that increasing the XG composition (Col:XG 1:1) enhanced ink viscosity and yielded scaffolds with good print fidelity but poor collagen alignment. On the other hand, use of inks with lower XG composition (Col:XG 4:1 and 9:1) together with 0.4 mg ml-1SMP concentration yielded scaffolds with high degree of collagen alignment albeit with suboptimal print fidelity. Modulating the print flow speed conditions (2 mm s-1) with 4:1 Col:XG inks and 0.4 mg ml-1SMP resulted in improved print fidelity of the collagen scaffolds while retaining high level of collagen anisotropy. Cell studies revealed hMSCs orient uniformly on aligned collagen scaffolds. More importantly, collagen anisotropy was found to trigger tendon or ligament-like differentiation of hMSCs. Together, these results suggest that 4D printing is a viable strategy to generate anisotropic collagen scaffolds with significant potential for use in tendon and ligament tissue engineering applications.

Gastric metastasis of small cell lung carcinoma: A rare but noteworthy entity to consider.

Small cell lung carcinoma (SCLC) is an aggressive malignancy known for its propensity for early and extensive metastatic spread. Gastric metastasis, where cancer cells disseminate from the lung to the stomach, is a rare but increasingly recognized complication of SCLC. This review provides a comprehensive overview of gastric metastasis in SCLC, addressing its clinical significance, diagnostic challenges, management strategies, and prognosis. Additionally, it examines the broader metastatic patterns of SCLC and compares them with other malignancies known for gastric metastasis. Gastric metastasis in SCLC, though infrequent, is clinically significant and often indicates advanced disease with a poor prognosis. SCLC typically metastasizes to the liver, brain, bones, and adrenal glands, with the stomach being an unusual site. The incidence of gastric metastasis ranges from 1% to 5% in autopsy studies, although this may be underestimated due to diagnostic difficulties and asymptomatic early lesions. Diagnosing gastric metastasis presents several challenges, including the asymptomatic nature of many cases, limitations of conventional imaging techniques, and difficulties in distinguishing metastatic lesions from primary gastric cancer via endoscopy. Histopathological diagnosis requires careful examination to identify SCLC cells through their characteristic small cell morphology and neuroendocrine markers. Management of gastric metastasis in SCLC typically involves a multidisciplinary approach. Systemic therapy, primarily chemotherapy, remains the cornerstone of treatment, with palliative care addressing symptoms and complications. Surgical intervention is usually reserved for specific cases requiring symptomatic relief. The prognosis for patients with gastric metastasis from SCLC is generally poor, reflecting the advanced stage of the disease. Median survival is significantly reduced compared to patients without gastric metastasis. This review emphasizes the need for enhanced awareness and early detection to improve patient outcomes and highlights the importance of ongoing research into better diagnostic and therapeutic strategies.

Performance analysis of CsFAMAPbIBr perovskite cells compared with MAPbI3 perovskite cells

The experimental preparation of Cesium Methylammonium Lead Iodide-Bromide (CsFAMAPbIBr) perovskite cell, and for the two kinds of perovskite cell morphology, optical properties, cell performance parameters and stability were compared, found that CsFAMAPbIBr perovskite cell in the above aspects have more outstanding performance. The CsFAMAPbIBr cell shows the best power conversion efficiency (PCE = 22.37 %) is higher than the MAPbI3 cell’s best power conversion efficiency (PCE = 18.46 %), and has high repeatability and long-term storage stability (air, >40 %RH, 500 h, 82 % of initial PCE).

Cytotoxicity of Orthodontic Archwires Used in Clinical Practice: In Vitro Study

This study investigates the cytotoxicity of various orthodontic archwires, which are essential in directing tooth movement through biomechanical forces. With advancements in material science, different archwire materials have been developed to balance mechanical performance with aesthetic and biological considerations. The study focuses on evaluating the biocompatibility and mechanical properties of stainless steel, nickel–titanium, and chromium–cobalt archwires, particularly their cytotoxic effects on oral cavity cells. In vitro cell culture experiments with fibroblasts, combined with scanning electron microscopy (SEM) analysis, were conducted to assess cell viability and morphology. The results revealed significant differences in cytotoxicity, with copper wires showing high toxicity and causing extensive cell death, while nickel–titanium and chromium–cobalt wires supported better cell viability and healthier cell morphology. These findings highlight the importance of selecting archwire materials that ensure mechanical efficiency without compromising cellular health, emphasizing the need for ongoing assessment of material biocompatibility in the oral environment.

3D Micropatterned Traction Force Microscopy: A Technique to Control 3D Cell Shape While Measuring Cell-Substrate Force Transmission.

Cell shape and function are intimately linked, in a way that is mediated by the forces exerted between cells and their environment. The relationship between cell shape and forces has been extensively studied for cells seeded on flat 2D substrates, but not for cells in more physiological 3D settings. Here, a technique called 3D micropatterned traction force microscopy (3D-µTFM) to confine cells in 3D wells of defined shape, while simultaneously measuring the forces transmitted between cells and their microenvironment is demonstrated. This technique is based on the 3D micropatterning of polyacrylamide wells and on the calculation of 3D traction force from their deformation. With 3D-µTFM, it is shown that MCF10A breast epithelial cells exert defined, reproducible patterns of forces on their microenvironment, which can be both contractile and extensile. Cells switch from a global contractile to extensile behavior as their volume is reduced are further shown. The technique enables the quantitative study of cell mechanobiology with full access to 3D cellular forces while having accurate control over cell morphology and the mechanical conditions of the microenvironment.

Revisiting Virchow’s triad: exploring the cellular and molecular alterations in cerebral venous congestion

BackgroundCerebral venous thrombosis (CVT) is a rare but serious condition that can lead to significant morbidity and mortality. Virchow’s triad elucidates the role of blood hypercoagulability, blood flow dynamics, and endothelial damage in the pathogenesis of CVT. Cerebral venous congestion (CVC) increases the risk of cerebral venous sinus thrombosis and can lead to recurrent episodes and residual symptoms. However, the precise mechanism by which blood congestion leads to thrombosis remains unclear. Our objective was to investigate the cellular and molecular alterations linked to CVC through analysis of the pathological morphology of venous sinus endothelial cells and transcriptomic profiling.ResultsThis study demonstrated a remarkable correlation between CVC and the phenotypic transformation of endothelial cells from an anticoagulant to a procoagulant state. The findings revealed that cerebral venous stasis results in tortuous dilatation of the venous sinuses, with slow blood flow and elevated pressure in the sinuses and damaged endothelial cells of the retroglenoid and internal jugular vein ligation (JVL) rat model. Mechanistically, analysis of transcriptomic results of cerebral venous sinus endothelial cells showed significant activation of platelet activation, complement and coagulation cascades pathway in the JVL rats. Furthermore, the expression of von Willebrand factor (vWF) and coagulation factor VIII (F8) in the complement and coagulation cascades and Fgg and F2 in the platelet activation was increased in the cerebral venous sinuses of JVL rats than in sham rats, suggesting that endothelial cell injury in the venous sinus induced by CVC has a prothrombotic effect. In addition, endothelial cell damage accelerates coagulation and promotes platelet activation. Significantly, the concentrations of vWF, F2 and F8 in venous sinus blood of patients with internal jugular vein stenosis were higher than in their peripheral blood.ConclusionCollectively, our data suggest that CVC can induce endothelial cell damage, which then exhibits a procoagulant phenotype and ultimately increases the risk of CVT. This research contributes to our understanding of the pathophysiology of CVC associated with procoagulant factors and reexamines the components of Virchow’s triad in the context of CVC.

Assessing CellaVision peripheral blood and advanced RBC application software's impact on hematologic morphology reporting: Real-world implementation experience in a large Veterans Affairs hospital.

This quality improvement study, conducted at the Kansas City Veterans Affairs Medical Center in Kansas City, Missouri, examined the change in patient reporting after transitioning from manual to CellaVision software-guided differentials and slide reviews. The primary focus was blasts and schistocytes, given their clinical significance. Three months of manual and CellaVision patient data were examined between May 2022 and February 2023. Blast and schistocyte patients were standardized to the total specimens performed and compared using 2-sample proportion testing. Other red blood cell (RBC) morphologies were also compared using this statistical analysis. Blast and schistocyte reporting after technologist review statistically increased after implementing the CellaVision software. This finding was most prevalent for low-percentage blasts. In addition, both morphologies experienced varying degrees of false-positive reporting, with 33% for low-percentage blasts and 91.7% for schistocytes. Other RBC morphologies displayed different levels of change, which could be clinically significant. The CellaVision software's increased sensitivity to blasts and schistocytes may benefit patient care, especially those with hematologic disorders. The software's high false-positive rate can be reduced by implementing quality metrics that prioritize clinically significant cells. This can be accomplished by implementing a CellaVision Champion to monitor reporting changes, perform patient lookbacks through the software, and provide technologist education. In addition, adopting a more stringent grading scale for schistocytes could also improve the high false-positive rate. Overall, CellaVision provides the ability to enhance hematology quality metrics by providing access to how patient cells are categorized and offering prompt education.

3D cultivation of non-small-cell lung cancer cell lines using four different methods

PurposeThe aim of this study was to set up reliable and reproducible culture conditions for 3D tumoroids derived from non-small cell lung cancer (NSCLC) cell lines to enable greater opportunity for successful cultivation of patient-derived samples.MethodsFour NSCLC cell lines, two adenocarcinomas (A549, NCI-H1975) and two squamous cell carcinomas (HCC-95, HCC-1588), were first cultured in traditional 2D settings. Their expected expression profiles concerning TTF-1, CK7, CK5, and p40 status were confirmed by immunohistochemistry (IHC) before the generation of 3D cultures. Tumoroids were established in the hydrogel GrowDex®-T, Nunclon™ Sphera™ flasks, BIOFLOAT™ plates, and Corning® Elplasia® plates. Western blot was used to verify antigen protein expression. Hematoxylin-eosin staining was used to evaluate the cell morphology in the 2D and 3D cultures. Mutational analysis of KRAS and EGFR by PCR on extracted DNA from 3D tumoroids generated from cells with known mutations (A549; KRAS G12S mutation, NCI-H1975; EGFR L858R/T790M mutations).ResultsWe successfully established 3D cultures from A549, NCI-H1975, HCC-95, and HCC-1588 with all four used cultivation methods. The adenocarcinomas (A549, NCI-H1975) maintained their original IHC features in the tumoroids, while the squamous cell carcinomas (HCC-95, HCC-1588) lost their unique markers in the cultures. PCR analysis confirmed persistent genetic changes where expected.ConclusionThe establishment of tumoroids from lung cancer cell lines is feasible with various methodologies, which is promising for future tumoroid growth from clinical lung cancer samples. However, analysis of relevant markers is a prerequisite and may need to be validated for each model and cell type.