Search Results (60)

Full-field optical coherence tomography (FF-OCT) is a high-resolution interferometric imaging technique that enables label-free visualization of cellular structural changes. In this study, we employed a custom-built time-domain FF-OCT system to monitor morphological alterations in HeLa cells undergoing doxorubicin-induced apoptosis and ethanol-induced necrosis at the single-cell level. Apoptotic cells showed characteristic features such as echinoid spine formation, cell contraction, membrane blebbing, and filopodia reorganization. In contrast, necrotic cells exhibited rapid membrane rupture, intracellular content leakage, and abrupt loss of adhesion structure. These dynamic events were visualized using high-resolution tomography and three-dimensional surface topography mapping. Furthermore, FF-OCT-based interference reflection microscopy (IRM)-like imaging effectively highlighted changes in cell–substrate adhesion and cell boundary integrity during the cell death process. Our findings suggest that FF-OCT is a powerful imaging platform for distinguishing cell death pathways and assessing dynamic cellular states, with potential applications in drug toxicity testing, anticancer therapy evaluation, and regenerative medicine. Full article

Background: This paper deals with the detection of amino acid composition of Iraqi Ocimum basilicum (basil) leaves and evaluation of the cytotoxic effects of the plant leaf extract on human colorectal cancer cells. Methods: Leaves of Ocimum basilicum were collected from Iraq in November 2024. After drying and powdering, the plant material went through cold methanol extraction. Initial phytochemical screening was conducted to identify the presence of alkaloids, flavonoids, coumarins, and terpenoids. Amino acid analysis was completed by an amino acid analyzer with fluorescence detection. The cytotoxic effect was evaluated via the MTT assay on HRT-18 cell lines. Morphological changes were further tested using dual Propidium Iodide/Acridine Orange assay fluorescent staining. Results: Seventeen amino acids were detected in the plant extract. The extract showed dose-dependent cytotoxic effects on HRT-18 cells, with significant reduction in cell viability at concentrations of more than 25 µg/mL. Morphological alterations of membrane blebbing and cell shrinkage were observed, suggesting apoptotic activity. The IC₅₀ value confirmed strong cytotoxic potential. Conclusions: The extract of Ocimum basilicum leaf cultivated in Iraq shows a rich amino acid profile and significant cytotoxic activity against colorectal cancer cells that highlights its potential effect as a natural source of anticancer compounds. Full article

Trabeculectomy with mitomycin C is a key surgical intervention for managing glaucoma when conservative treatments fail. The success of trabeculectomy is influenced by various factors, including preoperative ocular characteristics like conjunctival vascularity. This study aims to explore the relationship between the preoperative conjunctival vascular area and post-trabeculectomy outcomes in glaucoma patients. By analyzing the conjunctival vascular density, intraocular pressure (IOP), bleb morphology, laser suture lysis (LSL) frequency, and postoperative eye drops, this research sheds light on the impact of preoperative vascularity on surgical success. Results show that lower preoperative conjunctival vessel density is associated with favorable outcomes, such as better bleb formation and reduced need for postoperative interventions, while higher conjunctival vessel density correlates with complications like hyphema. These findings emphasize the importance of assessing preoperative conjunctival vascularity to optimize trabeculectomy outcomes and personalize treatment strategies for glaucoma patients. Full article

Pancreatic cancer cells exhibit a remarkable ability to tolerate nutrient deprivation, a phenomenon termed “austerity,” which enables their survival within the hypovascular tumor microenvironment. Conventional anticancer therapies frequently fail to effectively target these resilient neoplastic cells, posing a significant challenge to the therapeutic management of pancreatic cancer. Consequently, targeting austerity, the ability of cancer cells to tolerate nutrient starvation, represents a promising anti-austerity strategy for developing novel pancreatic cancer therapeutics. In this study, we investigated calliviminone A (CVM-A), a phloroglucinol–meroterpenoid isolated from Callistemon citrinus leaves, for its anti-austerity activity against PANC-1 human pancreatic cancer cells. Calliviminone A exhibited potent preferential cytotoxicity in nutrient-deprived medium (NDM) with a PC₅₀ of 0.57 µM, while showing minimal toxicity in nutrient-rich Dulbecco’s Modified Eagle’s medium (IC₅₀ = 45.2 µM), indicating a favorable therapeutic index. Real-time live-cell imaging revealed that CVM-A induced significant morphological changes, including cell shrinkage and membrane blebbing, leading to cell death within 24 h of NDM. Furthermore, under normal nutrient conditions in Dulbecco’s Modified Eagle’s Medium (DMEM), CVM-A significantly inhibited PANC-1 cell migration (up to 47% reduction at 20 µM) and colony formation (over 80% suppression at 25 µM), suggesting its antimetastatic potential. Western blot studies demonstrated that CVM-A downregulated key survival components of the PI3K/Akt/mTOR signaling pathway, completely inhibiting Akt and p-Akt at 2.5 µM in NDM, and suppressing insulin-induced Akt activation. These findings highlight CVM-A as a promising lead compound for developing novel anticancer therapies that target the adaptive survival mechanisms and metastatic potential of pancreatic cancer in nutrient-deprived microenvironments. Full article

Background/Objectives: This study aims to evaluate the long-term efficacy and safety of ab interno techniques using minimally invasive glaucoma surgery (MIGS), specifically XEN gel stent implantation, by evaluating its 2-year outcomes in patients with primary open-angle glaucoma (POAG) and pseudoexfoliation glaucoma (PXG). Methods: This retrospective single-center study consecutively included 31 eyes of 31 patients with POAG or PXG who underwent XEN gel stent implantation. Patients were followed for 24 months, with assessments at multiple time points. Success was defined as achieving an IOP of less than 14 mmHg and a reduction of more than 20% from preoperative IOP without additional glaucoma surgery. Bleb morphology was evaluated using anterior segment optical coherence tomography (AS-OCT) and slit-lamp photographs. Postoperative interventions and complications were also recorded. Results: At 24 months, complete success and qualified success rates were 35.5% (11/31) and 51.6% (16/31), respectively. There was no difference in surgical success rates at 2 years based on the tip location (intraconjunctiva, intratenon, and uviform) on the 1st postoperative day. Patients with high sparse wall on AS-OCT imaging or avascular bleb morphology via slit-lamp photography at 6 months postoperatively had higher complete success rates at 2 years than those without (p = 0.007, p = 0.009, respectively). Patients with avascular bleb types at 6 months postoperatively had higher qualified success rates at 2 years compared with the vascular types (p = 0.038). Needling was performed in 32.3% of eyes, with secondary surgical procedures required in 16.1% of eyes. The most common adverse event was hypotony, occurring in 67.7% of eyes on the 1st postoperative day but resolving within 6 months. Conclusions: The ab interno XEN gel stent is an effective and minimally invasive option for managing POAG and PXG, with long-term success predicted by the AS-OCT assessment of bleb morphology at 6 months. Proactive postoperative management, emphasizing early intervention and monitoring, is crucial for maintaining optimal outcomes. Full article

This work examines how multiphase flow behavior during CO₂ and N₂ displacement in a microfluidic chip under capillary-dominated circumstances is affected by interfacial tension (IFT) and the viscosity ratio. In order to simulate real pore-scale displacement operations, microfluidic tests were performed on a 2D rock chip at flow rates of 1, 10, and 100 μL/min (displacement of water by N₂/supercritical CO₂). Moreover, core flooding experiments were performed on various sandstone samples collected from three different geological basins in Australia. Although CO₂ is notably denser and more viscous than N₂, the findings show that its displacement efficiency is more influenced by the IFT values. Low water recovery in CO₂ is the result of non-uniform displacement that results from a high mobility ratio and low IFT; this traps remaining water in smaller pores via snap-off mechanisms. However, due to the blebbing effect, N₂ injection enhances the dissociation of water clots, resulting in a greater swept area and fewer remaining water clusters. The morphological investigation of the residual water indicates various displacement patterns; CO₂ leaves more retained water in irregular shapes, while N₂ enables more uniform displacement. These results confirm earlier studies and suggest that IFT has a crucial role in fluid displacement proficiency in capillary-dominated flows, particularly at low flow rates. This study emphasizes the crucial role of IFT in improving water recovery through optimizing the CO₂ flooding process. Full article

Filtration bleb (FB) fibrosis represents the primary risk factor for glaucoma filtration surgery (GFS) failure. We reviewed the most recent literature on post-GFS fibrosis in humans, focusing on novel molecular pathways and antifibrotic treatments. Three main literature searches were conducted. First, we performed a narrative review of two models of extra-ocular fibrosis, idiopathic pulmonary fibrosis and skin fibrosis, to improve the comprehension of ocular fibrosis. Second, we conducted a systematic review of failed FB features in the PubMed, Embase, and Cochrane Library databases. Selected studies were screened based on the functional state and morphological features of FB. Third, we carried out a narrative review of novel potential antifibrotic molecules. In the systematic review, 11 studies met the criteria for analysis. Immunohistochemistry and genomics deemed SPARC and transglutaminases to be important for tissue remodeling and attributed pivotal roles to TGFβ and M2c macrophages in promoting FB fibrosis. Four major mechanisms were identified in the FB failure process: inflammation, fibroblast proliferation and myofibroblast conversion, vascularization, and tissue remodeling. On this basis, an updated model of FB fibrosis was described. Among the pharmacological options, particular attention was given to nintedanib, pirfenidone, and rapamycin, which are used in skin and pulmonary fibrosis, since their promising effects are demonstrated in experimental models of FB fibrosis. Based on the most recent literature, modern patho-physiological models of FB fibrosis should consider TGFβ and M2c macrophages as pivotal players and favorite targets for therapy, while research on antifibrotic strategies should clinically investigate medications utilized in the management of extra-ocular fibrosis. Full article

Unruptured intracranial aneurysms, affecting 2–5% of the population, are characterized by localized wall weakening and irregular morphologies, including features such as blebs, lobulations, or asymmetries, which are significant predictors of rupture risk. Although up to 57% of ruptured intracranial aneurysms exhibit irregular dome geometry, its influence on aneurysm stability remains underexplored. Irregular geometries are associated with adverse hemodynamic forces, such as increased wall shear stress (WSS), amplifying wall stress at specific regions, and promoting flow disturbances, which may increase aneurysm vulnerability. This study investigates the influence of aneurysm dome morphology, particularly in IAs with irregular domes that may include daughter blebs, using Computational Fluid Dynamics (CFD). Unlike prior CFD studies that modeled blood as Newtonian or non-Newtonian, this work employs a three-phase blood flow model, representing plasma and red blood cells (RBCs) as distinct phases. Numerical simulations, conducted via the Finite Volume Method, solve the Navier–Stokes equations to capture complex flow dynamics within cerebral vasculature. Key hemodynamic metrics, such as Wall Shear Stress (WSS), Wall Shear Stress Gradient (WSSG), and Viscous Dissipation Rate, are analyzed to assess the interplay between dome morphology and hemodynamic stressors. Full article

Focal swellings of dendrites (“dendritic blebbing”) together with structural damage of mitochondria and the endoplasmic reticulum (ER) are morphological hallmarks of glutamate neurotoxicity, also known as excitotoxicity. These pathological alterations are generally thought to be caused by the so-called “overactivation” of N-methyl-D-aspartate receptors (NMDARs). Here, we demonstrate that the activation of extrasynaptic NMDARs, specifically when forming a protein–protein complex with TRPM4, drives these pathological traits. In contrast, strong activation of synaptic NMDARs fails to induce cell damage despite evoking plateau-type calcium signals that are comparable to those generated by activation of the NMDAR/TRPM4 complex, indicating that high intracellular calcium levels per se are not toxic to neurons. Using confocal laser scanning microscopy and transmission electron microscopy, we show that disrupting the NMDAR/TRPM4 complex using the recently discovered small-molecule TwinF interface inhibitor FP802 inhibits the NMDA-induced neurotoxicity-associated dendritic blebbing and structural damage to mitochondria and the ER. It also prevents, at least in part, the disruption of ER–mitochondria contact sites. These findings establish the NMDAR/TRPM4 complex as the trigger for the structural damage of dendrites and intracellular organelles associated with excitotoxicity. They also suggest that activation of the NMDAR/TRPM4 complex, in addition to inducing high-amplitude, plateau-type calcium signals, generates a second signal required for glutamate neurotoxicity (“two-hit hypothesis”). As structural damage to organelles, particularly mitochondria, is a common feature of many human neurodegenerative diseases, including Alzheimer’s disease and amyotrophic lateral sclerosis (ALS), TwinF interface inhibitors have the potential to provide neuroprotection across a broad spectrum of these diseases. Full article

The red blood cell (RBC) membrane is unique and crucial for maintaining structural–functional relationships. Maternal smoking induces significant changes in the morphological, rheological, and functional parameters of both maternal and foetal RBCs, mainly due to the continuous generation of the free radicals. The major aim of this study was to follow the consequences of a secondary stressor, like fungal infection, on the already compromised RBC populations. The impact of Candida infection, a growing health concern, was investigated on four blood sample groups: mothers and their neonates originating from non-smoking versus smoking populations. Here, we searched for phenotypical and molecular markers that precisely reflected the effect of Candida infection on the RBC membrane; this included the level of hemolysis, appearance of morphological variants, formation of the lipid peroxidation marker 4-hydroxyl-nonenal, arrangement of the Band 3 molecules and activation of the Caspase 3. In most of the examined cases, the fungal infection increased the adverse symptoms induced by smoking, indicating a general stress response, likely due to an altered redox state of the cells. However, we were able to identify an atypical phenotype (clustered populations with shrinkage and membrane blebbing) in both the non-smoking and smoking populations, which might be a unique marker for Candida spp. infection. Full article

The oviductal fimbria is the first extraovarian anatomical structure that the cumulus–oocyte complex (COC) encounters, and is sensitive to sex hormone changes. The morphology, glycan pattern, expression of heat shock proteins (HSPs), estradiol receptor (ER), and progesterone receptor (PR) were investigated in the oviductal fimbria epithelium of the baboon (Papio hamadryas) during the menstrual cycle. The morphology was investigated by light and scanning electron microscopy; the glycopattern was characterized using conventional and lectin histochemistry; HSPs (60, −70, −90), ER, and PR were localized immunohistochemically. Well-differentiated ciliated and nonciliated cells were present only during the preovulatory phase. The nonciliated cells contained small apical protrusions and thin microvilli. During the preovulatory phase (1) the luminal surface of the fimbria displayed acidic glycans, complex N-glycans containing fucose, and oligolactosamine residues; (2) nonciliated cells expressed HSP60 and HSP90 in the apical blebs, HSP70 in the nucleus and cytoplasm, as well as nuclear ERα and PR; (3) ciliated cells showed HSP70 in the nucleus, cytoplasm, and cilia that also expressed HSP90 and PR. These results are related to the function of the fimbria where the early COC–oviduct crosstalk occurs and may represent a benchmark for translational studies of other primates. Full article

Cannabis sativa is a well-known plant for its psychoactive effects; however, its many derivatives, such as Cannabidiol (CBD), contain several therapeutic applications. Tetrahydrocannabinol (THC) is the main cannabis derivative responsible for psychoactive properties, while CBD is non-psychotropic. For this reason, CBD has been more exploited in the last decade. CBD has been connected to multiple anticancer properties, and when combined with photodynamic therapy (PDT), it is possible to eradicate tumors more effectively. In this study, CBD was utilized to treat MCF-7 breast cancer cells, followed by in vitro PDT combination therapy. Conventional breast cancer treatment modalities such as chemotherapy, radiotherapy, etc. have been reported for inducing a number of undesirable side effects, recurrence of the disease, and low quality of life. In this study, cells were exposed to varying concentrations of CBD (i.e., 1.25, 2.5, 5, 10, and 20 μg/mL) and incubated 12 and 24 h after treatment. The optimal doses were then used in combination therapy. Morphology and biochemical assays, including lactate dehydrogenase (LDH) for membrane integrity, adenosine triphosphate (ATP) for viability, and trypan blue exclusion assay for viability, were used to examine cellular responses after treatments. The optimal concentration was then utilized in Hypericin-Gold nanoparticles mediated PDT combination. The results revealed that, in a dose-dependent manner, conventional morphological characteristics of cell death, such as vacuolization, blebbing, and floating were observed in treated cells. The biochemical responses demonstrated an increase in LDH, a decrease in ATP, and a reduction in viability. This study demonstrated that CBD induces cell death in MCF-7 breast cancer cells cultured in vitro. The immunofluorescence results of combination therapy indicated that cell death occurred via apoptosis. In conclusion, this study proposes that the CBD and PDT combination therapy is effective in killing MCF-7 breast cancer cells in vitro by induction of apoptosis. Full article

The formation of specific cellular protrusions, plasma membrane blebs, underlies the amoeboid mode of cell motility, which is characteristic for free-living amoebae and leukocytes, and can also be adopted by stem and tumor cells to bypass unfavorable migration conditions and thus facilitate their long-distance migration. Not all cells are equally prone to bleb formation. We have previously shown that membrane blebbing can be experimentally induced in a subset of HT1080 fibrosarcoma cells, whereas other cells in the same culture under the same conditions retain non-blebbing mesenchymal morphology. Here we show that this heterogeneity is associated with the distribution of vimentin intermediate filaments (VIFs). Using different approaches to alter the VIF organization, we show that blebbing activity is biased toward cell edges lacking abundant VIFs, whereas the VIF-rich regions of the cell periphery exhibit low blebbing activity. This pattern is observed both in interphase fibroblasts, with and without experimentally induced blebbing, and during mitosis-associated blebbing. Moreover, the downregulation of vimentin expression or displacement of VIFs away from the cell periphery promotes blebbing even in cells resistant to bleb-inducing treatments. Thus, we reveal a new important function of VIFs in cell physiology that involves the regulation of non-apoptotic blebbing essential for amoeboid cell migration and mitosis. Full article

Ferruginol is a promising abietane-type antitumor diterpene able to induce apoptosis in SK-Mel-28 human malignant melanoma. We aim to increase this activity by testing the effect of a small library of ferruginol analogues. After a screening of their antiproliferative activity (SRB staining, 48 h) on SK-Mel-28 cells the analogue 18-aminoferruginol (GI50 ≈ 10 µM) was further selected for mechanistic studies including induction of apoptosis (DAPI staining, p < 0.001), changes in cell morphology associated with the treatment (cell shrinkage and membrane blebbing), induction of caspase-3/7 activity (2.5 at 48 h, 6.5 at 72 h; p < 0.0001), changes in the mitochondrial membrane potential (not significant) and in vitro effects on cell migration and cell invasion (Transwell assays, not significant). The results were compared to those of the parent molecule (ferruginol, GI50 ≈ 50 µM, depolarisation of mitochondrial membrane p < 0.01 at 72 h; no caspases 3/7 activation) and paclitaxel (GI50 ≈ 10 nM; caspases 3/7 activation p < 0.0001) as a reference drug. Computational studies of the antiproliferative activity of 18-aminoferruginol show a consistent improvement in the activity over ferruginol across a vast majority of cancer cells in the NCI60 panel. In conclusion, we demonstrate here that the derivatisation of ferruginol into 18-aminoferruginol increases its antiproliferative activity five times in SK-MEL-28 cells and changes the apoptotic mechanism of its parent molecule, ferruginol. Full article

Trabeculectomy has traditionally been upheld as the gold standard in glaucoma surgery, but recent advancements, including the PRESERFLO^® MicroShunt, have introduced less invasive techniques to mitigate complications and reduce intraocular pressure (IOP). The examination of bleb morphology post-filtering surgery is critical for modulating wound healing and ensuring long-term success. While previous evaluations of PRESERFLO-generated blebs have relied on data from trabeculectomy blebs, the differing surgical techniques used in PRESERFLO and trabeculectomy surgeries suggest potential variations in bleb morphologies. This study conducted a comparative analysis of blebs resulting from PRESERFLO and trabeculectomy procedures. Retrospective descriptive assessments using the Jenaer Bleb Grading System were performed, along with quantitative evaluation using eight-dimensional parameters utilizing anterior segment OCT. We included 93 eyes (57 following PRESERFLO, 36 following trabeculectomy). In the descriptive assessment, PRESERFLO-generated blebs exhibited fewer conjunctival cysts (3.5% vs. 22.2%, p = 0.007) and cavernous changes (10.5% vs. 30.5%, p = 0.021) compared to trabeculectomy. Quantitatively, overall bleb dimensions were comparable (maximal bleb height, width, and length, p > 0.05 for all). However, PRESERFLO blebs displayed a shallower (0.52 ± 0.24 vs. 0.67 ± 0.3 mm, p = 0.017) yet longer (4.12 ± 1.54 vs. 3.23 ± 1.64 mm, p = 0.024) episcleral lake. A thicker bleb wall (1.52 ± 0.46 vs. 1.10 ± 0.37 mm, p < 0.001) along with more posteriorly positioned blebs (distance to limbus = 6.16 ± 1.36 vs. 4.87 ± 1.34 mm, p < 0.001) were observed following PRESERFLO. This study illuminates the nuanced morphological differences between PRESERFLO and trabeculectomy blebs. Understanding these distinctions is vital, empowering clinicians to make informed postoperative decisions and avoid misinterpretation of bleb morphology. Full article