Cultures Of Normal Cells Research Articles

Evaluation of Spliceosome Protein SmD2 as a Potential Target for Cancer Therapy

The core spliceosome Sm proteins are gaining attention as potential targets for cancer treatment. Here, we evaluate this, with focus on SmD2. A pan-cancer analysis including 26 solid tumor types revealed that the SmD2-encoding SNRPD2 gene was overexpressed in almost all cancers. In several cancers, high SNRPD2 expression was associated with a poor prognosis. To investigate the vulnerability of human cells to the loss of SmD2 expression, we silenced SNRPD2 using a short hairpin-expressing lentiviral vector in established cancer cell lines; in short-term cultured melanoma cells; and in several normal cell cultures, including cancer-associated fibroblasts cultured from non-small cell lung cancer resections. Additionally, we analyzed publicly available cell viability datasets for the dependency of cancer cell lines to SmD2 expression. Together, these studies clearly established SmD2 as a cancer-selective lethal target. Delving into genes with similar essentiality profiles to SNRPD2, we uncovered the intersected lethal stress between the loss of SmD2 and the loss of gene products participating in not only different mRNA processing steps including mRNA splicing, but also processes for coordinated protein production, as well as mitosis. Furthermore, we could correlate SNRPD2 expression to the responses of cancer cells to several FDA-approved anti-tumor drugs, especially to drugs inhibiting the cell cycle. Overall, our study confirms the anticipated role for targeting SmD2 in cancer treatment and reveals non-canonical SmD2 functions beyond mRNA splicing that could contribute to the dependency of cancer cells to high SNRPD2 expression.

Novel Pyrroloquinoline Quinone-Modified Cerium Oxide Nanoparticles and Their Selective Cytotoxicity Under X-Ray Irradiation

Ionizing radiation leads to the development of oxidative stress and damage to biologically important macromolecules (DNA, mitochondria, etc.), which in turn lead to cell death. In the case of radiotherapy, both cancer cells and normal cells are damaged. In this regard, the development of new selective antioxidants is relevant. In this study, we first investigated the redox activity of cerium oxide-pyrroloquinoline quinone nanoparticles (CeO2@PQQ NPs) and their cytotoxic effects on normal (mouse fibroblasts, L929) and cancer (mouse adenocarcinoma, EMT6/P) cell cultures. Furthermore, the biological activity of CeO2@PQQ NPs was evaluated in comparison with that of CeO2 NPs and PQQ. The nanoparticles demonstrated pH-dependent reductions in the content of hydrogen peroxide after X-ray exposure. Our findings indicate that viability of EMT6/P cells was more adversely affected by CeO2@PQQ NPs at lower concentrations (0.1 μM) compared to L929. Following X-ray irradiation at a dose of 5 Gy, significant changes in mitochondrial potential (by 29%) and decreased glutathione levels (by 32%) were also observed in EMT6/P culture following irradiation and incubation with CeO2@PQQ NPs. Furthermore, EMT6/P exhibited a 2.5-fold increase in micronuclei and a 2-fold reduction in survival fraction compared to L929. It is hypothesized that CeO2@PQQ NPs may exhibit selective cytotoxicity and radiosensitizing properties against EMT6/P cancer cells. The findings suggest that CeO2@PQQ NPs may have potential as a selective redox-active antioxidant/pro-oxidant in response to X-ray radiation.

Assessment of the Role of miR-30a-5p on the Proliferation and Apoptosis of Hair Follicle Stem Cells.

To investigate the role of miR-30a-5p on the proliferation and apoptosis of hair follicle stem cells (HFSCs) and whether the Wnt/β-catenin signaling pathway is involved. HFSCs derived from the vibrissa of mammary rats were obtained by enzymatic digestion, and subsequently the obtained HFSCs were treated with Lipofectamine 2000 cell transfection and divided into normal cell culture group (control), miR-30a-5p overexpression group (miR-30a-5p mimic), miR-30a-5p empty vector group (miR-NC), miR-30a-5p inhibitor group (in-miR-30a-5p), and in-miR-30a-5p empty vector group (in-miR-NC). After transfection, the cell proliferation and apoptosis rates were examined separately. In addition, the mRNA expression of β-catenin, proliferating cell nuclear antigen (PCNA) and apoptosis-related genes (Bax and Bcl-2) were examined. The results of cell proliferation ability showed that in-miR-30a-5p group promoted cell proliferation of HFSCs relative to other groups, along with significant upregulation of gene levels of PCNA. Apoptosis analysis indicated that apoptosis rate was reduced in the in-miR-30a-5p group, and the expression of Bax was suppressed, while that of Bcl-2 was promoted. Wnt/β-catenin signaling pathway investigation revealed a significant increase in the levels of β-catenin in HFSCs in the in-miR-30a-5p group. Downregulation of miR-30a-5p levels inhibited HFSCs apoptosis and simultaneously promoted proliferation, furthermore, the increased expression of β-catenin indirectly confirmed the activation of the Wnt/β-catenin signaling pathway.

5178 TNFα-dependent altered miRNA expression regulates endometrial stromal cell proliferation

Abstract Disclosure: I. Chowdhury: None. S. Banerjee: None. W. Xu: None. A. Driss: None. C. Nezhat: None. N. Sidell: None. R.N. Taylor: None. W.E. Thompson: None. Abstract: Aberrant upregulation of cytokines, including tumor necrosis factor α (TNFα) and activation of nuclear factor κB (NF-κB) with disbalance of microRNAs (miRNAs) are often found in endometrium of endometriosis subjects, which causes directly or indirectly pelvic inflammation, pain, and infertility, Although, the pathophysiology of endometriosis is not clearly established yet, inflammation is an essential feature. MicroRNAs are emerging as potential regulatory factors in cellular processes such as cell growth, survival, migration, and invasion. In the current study, we examined the direct effects of exogenous recombinant TNFα dependent regulation of miRNAs involved in proliferation in primary cultures of normal endometrial stromal cells (NESC) and compared with the untreated cells derived from endometriosis-ESC (EESC). NanoString nCounter-based assays and quantitative real-time polymerase chain reaction (RT-PCR) were used to identify and validate differentially expressed miRNAs linked to cell proliferation in EESC compared to NESC and TNFα treated NESCs. The expression level of miRNAs linked to cell cycle was downregulated significantly in EESCs compared to NESCs. Interestingly, a dose and time-dependent exogenous TNFα treatment in NESCs showed a similar cell cycle regulator miRNAs expression pattern with EESCs. In further studies, protein analysis by Western Blots of selective cell cycle regulators such as PCNA, Ki67, CDK2, CDK4, CDK6, p21, and p27, and cell proliferation assay demonstrated an inverse correlation between cell cycle regulators and miRNA expression pattern both in EESC and TNFα treated NESCs. These findings suggest that TNFα dependent dysregulation of miRNAs in EESCs may promote proliferation of endometriosis. Further research is needed to investigate the detailed mechanisms by which TNFα regulates miRNA expression and determine these findings' potential therapeutic implications. Acknowledgments: This study was supported in part by National Institutes of Health Grants 1SC3 GM113751, 1SC1 GM130544, U01 HD66439, 1R01HD057235, U54 CA118948, HD41749, S21MD000101, and G12-RR03034. This investigation was conducted in a facility constructed with support from Research Facilities Improvement Grant #C06 RR18386 from NIH/NCRR. Presentation: 6/2/2024

In Vitro Evaluation of Anti-Hemolytic and Cytotoxic Effects of Traditional Mexican Medicinal Plant Extracts on Human Erythrocytes and Cell Cultures.

Plant extracts of fifteen plants of ethnomedicinal use in Mexico were analyzed to provide scientific knowledge of their medicinal properties through the evaluation of different biological activities such as anti-hemolytic, antioxidant, and cytotoxic effects in normal cells. Therefore, methanolic extracts were obtained from each of the plants by the Soxhlet extraction. The hemolytic activity in human erythrocytes was evaluated, as was their potential to protect the erythrocyte membrane against the 2,2'-azobis (2-methylpropionamidine) dihydrochloride (AAPH) and 1,1-diphenyl-2-picryl hydrazyl (DPPH) radicals. Finally, the toxicity of the extracts in normal cell cultures of African green monkey kidney cells (Vero) and peripheral blood mononuclear cells (PBMC) was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction method. Most of the extracts showed low hemolytic activity and high anti-hemolytic activity as well as high selectivity indices (SI) and antioxidant effects. Extracts of H. inuloides, J. dioica, and J. spicigera induced cell proliferation of the Vero cells. K. daigremontiana, A. adstringens, S. mexicanum, J. spicigera, L. tridentata, and M. tenuiflora extracts showed PBMC cell proliferation. In the present study, it was observed that the evaluated extracts did not present hemolytic activity, and some presented low toxicity when Vero and PBMC cell cultures were exposed. In conclusion, traditionally used plants possess beneficial health properties, and it is hoped that this study will serve as a basis for understanding the biological effects of traditionally used plants and may complement future studies.

Effects of Rubus idaeus extracts on some Cancerous Cell Lines in vitro Study Using MTT Assay

Our study investigated the cytotoxicity of Rubus idaeus alcoholic and aqueous extracts prepared from ripe blackberries were collected from the farmlands north of Baghdad, the extracts prepared at the same starting concentration, and tested on two cell line, the result show high toxic effects increase with the increase of concentration. The three extracted components from Rubus idaeus (purified flavonoid, polysaccharides, and carotenes) showed cytotoxic effect towards both: the primary cell culture of normal hepatic cells (HepG2 and L20B cell), and cancer hepatic cell lines (HepG2 and L20B cell) at 100μg/ml concentration for 24 hours treatment. Purified flavonoid exerted the potent effect on both cell lines among the other two extracted components. The cytotoxicity assay was held only for the purified flavonoid to investigate the mechanism by which the purified flavonoid affected living cells toward apoptosis. The most significant reduction (p≤0.05) in cell viable count was at the concentration 100μg/ml which appear to cause the induction of cell death via mitochondrial pathway for HepG2 and L20B cell line after 24 hours exposure. The purified flavonoid had no effect on the HepG-2 cell cycle. The Immunomodulation effect for the purified flavonoid and the extracted polysaccharides on normal human peripheral blood lymphocytes showed that the purified flavonoid suppress lymphocytes proliferation, while the later increased lymphocytes proliferation significantly. The immune stimulating effect of the polysaccharides caused alteration in IL-17 and level estimated by ELISA technique towards IL-2 elevation for the normal human blood lymphocytes against IL-17 level after 4 hours exposure at concentrations (250 and 500 μg/ml), while toxicity results were shown after 4 hours.

Pulmonary surfactant and prostaglandin E2 in airway smooth muscle relaxation of human and male guinea pigs.

Pulmonary surfactant serves as a barrier to respiratory epithelium but can also regulate airway smooth muscle (ASM) tone. Surfactant (SF) relaxes contracted ASM, similar to β2-agonists, anticholinergics, nitric oxide, and prostanoids. The exact mechanism of surfactant relaxation and whether surfactant relaxes hyperresponsive ASM remains unknown. Based on previous research, relaxation requires an intact epithelium and prostanoid synthesis. We sought to examine the mechanisms by which surfactant causes ASM relaxation. Organ bath measurements of isometric tension of ASM of guinea pigs in response to exogenous surfactant revealed that surfactant reduces tension of healthy and hyperresponsive tracheal tissue. The relaxant effect of surfactant was reduced if prostanoid synthesis was inhibited and/or if prostaglandin E2-related EP2 receptors were antagonized. Atomic force microscopy revealed that human ASM cells stiffen during contraction and soften during relaxation. Surfactant softened ASM cells, similarly to the known bronchodilator prostaglandin E2 (PGE2) and the cell softening was abolished when EP4 receptors for PGE2 were antagonized. Elevated levels of PGE2 were found in cultures of normal human bronchial epithelial cells exposed to pulmonary surfactant. We conclude that prostaglandin E2 and its EP2 and EP4 receptors are likely involved in the relaxant effect of pulmonary surfactant in airways.

α-Tocopheryl Succinate Induces ER Stress, Disruption of Lipid Metabolism, and Apoptosis in a Culture of Normal and Tumor Cells of Epidermal Origin

α-Tocopheryl Succinate Induces ER Stress, Disruption of Lipid Metabolism, and Apoptosis in a Culture of Normal and Tumor Cells of Epidermal Origin

Investigating the Anticancer Potential of Zinc and Magnesium Alloys: From Base Materials to Nanocoated Titanium Implants.

In this work, we show the in vitro anticancer potential of surgical wires, obtained from zinc (ZnMg0.004) or magnesium (MgCa0.7) alloys by spatial technology comprising casting, extrusion, and final drawing processes. We also present the selective anticancer effects of applied soluble multilayer nanocoatings of zinc and magnesium onto titanium surfaces using the pulse laser deposition method. In the latter, the titanium samples were produced via 3D printing using the selective laser melting method and coated with various combinations of zinc and magnesium layers. For cytotoxicity studies, human dental pulp-derived stem cells (hDPSCs) and human osteosarcoma SaOS-2 cell line were used as representatives of healthy and cancer cells. Cells were examined against the 0.3-3.0 cm2/mL material extract ratios obtained from experimental and steel surgical wires, the latter being the current clinical industry standard. The MgCa0.7 alloy wires were approx. 1.5 times more toxic to cancer cells at all examined extract ratios vs. the extracts from steel surgical wires that exhibited comparable toxicity towards healthy and cancer cells. The ZnMg0.004 alloy wires displayed increased toxicity towards cancer cells with decreasing extract ratios. This was also reflected in the increased anticancer effectiveness, calculated based on the viability ratio of healthy cells to cancer cells, from 1.1 to 4.0 times. Healthy cell viability remained at 80-100%, whereas cancer cell survival fluctuated at 20-75%, depending on the extract ratio. Furthermore, the culture of normal or cancer cells on the surface of Zn/Mg-coated titanium allowed us to select combinations of specific coating layers that yielded a comparable anticancer effectiveness to that observed with the experimental wires that ranged between 2 and 3. Overall, this work not only demonstrates the substantial anticancer properties of the studied wires but also indicates that similar anticancer effects can be replicated with appropriate nanocoatings on titanium samples. We believe that this work lays the groundwork for the future potential development of the category of new implants endowed with anticancer properties.

Overcoming the inconsistent cell culture performance caused by single-use bioreactors during scaling-up the manufacturing process for a therapeutic bispecific antibody

Single use technology has been widely used in modern biopharmaceutical process development and manufacturing. However, there are concerns about the adverse effects of disposable materials on bioprocess, product quality and patient safety. Recently, we observed slow cell growth, increased lactate accumulation, and decreased production titer during scaling up a bispecific antibody (BsAb) upstream process from 3 L to 500 L bioreactor. To investigate the phenomenon, we conducted medium incubation and leachable spike-in experiments. We found that cell culture medium incubated in CX5–14 bag caused poor cell culture performance, which was correlated with the concentration of bis(2,4-di-tert-butylphenyl)-phosphate (bDtBPP), a breakdown product of the antioxidant Irgafos®168 in polyethylene film. In addition, when the media were spiked with 0.15 μg/ml or a greater concentration of bDtBPP, slow cell growth and reduced BsAb expression level were observed. In contrast, Aegis5–14 film produced bDtBPP at a level of below detection during medium incubation. Thus, the bispecific-expressing CHO cell line restores normal cell culture performance and production titer when Aegis5–14 single-use bioreactor was employed in large-scale manufacturing. Our results reveal the importance of evaluating leachable profiles from disposable materials during process development of bispecifics and other novel therapeutic modalities.

Biocompatibility characterisation of CMOS-based Lab-on-Chip electrochemical sensors for in vitro cancer cell culture applications

Lab-on-Chip electrochemical sensors, such as Ion-Sensitive Field-Effect Transistors (ISFETs), are being developed for use in point-of-care diagnostics, such as pH detection of tumour microenvironments, due to their integration with standard Complementary Metal Oxide Semiconductor (CMOS) technology. With this approach, the passivation of the CMOS process is used as a sensing layer to minimise post-processing, and Silicon Nitride (Si3N4) is the most common material at the microchip surface. ISFETs have the potential to be used for cell-based assays however, there is a poor understanding of the biocompatibility of microchip surfaces. Here, we quantitatively evaluated cell adhesion, morphogenesis, proliferation and mechano-responsiveness of both normal and cancer cells cultured on a Si3N4, sensor surface. We demonstrate that both normal and cancer cell adhesion decreased on Si3N4. Activation of the mechano-responsive transcription regulators, YAP/TAZ, are significantly decreased in cancer cells on Si3N4 in comparison to standard cell culture plastic, whilst proliferation marker, Ki67, expression markedly increased. Non-tumorigenic cells on chip showed less sensitivity to culture on Si3N4 than cancer cells. Treatment with extracellular matrix components increased cell adhesion in normal and cancer cell cultures, surpassing the adhesiveness of plastic alone. Moreover, poly-l-ornithine and laminin treatment restored YAP/TAZ levels in both non-tumorigenic and cancer cells to levels comparable to those observed on plastic. Thus, engineering the electrochemical sensor surface with treatments will provide a more physiologically relevant environment for future cell-based assay development on chip.

Effects of EZH2 inhibitor, trichostatin A, and 5-azacytidine combinatorial treatment on osteogenic differentiation of dental pulp stem cells

ObjectiveEpigenetic mechanisms play regulatory roles in dental pulp stem cell (DPSC) differentiation. The molecules that modulate these mechanisms can be used to enhance DPSC differentiation in experimental studies and clinical applications. We investigated the combined effects of an epigenetic modulator enhancer of zeste homologue 2 inhibitor (EZH2i), trichostatin A (TSA), and 5-azacytidine (5-AZA) on the osteogenic differentiation of DPSCs. ResultsTo assess osteogenic differentiation, we measured alkaline phosphatase activity, calcium deposition, and expression of osteogenic differentiation marker genes (RUNX2, BMP2, and ALPL) after 7 or 21 days of combinatorial drug treatment in normal cell culture medium or osteo-inductive medium (OIM). No synergistic effects were observed for any possible combination of EZH2i, TSA, or 5-AZA. However, the effects of these drugs and their combinations depend on the time and culture conditions. DiscussionWe confirmed that EZH2i and TSA have positive effects on the osteogenic differentiation of DPSCs. EZH2i activates the expression of key regulatory genes (RUNX2, BMP2, and ALPL) directly, whereas TSA interacts with signalling pathways induced by supplements in OIM to activate these genes.

Anti-anemic properties of Pterocarpus marsupium latex and its cytotoxic and embryotoxic effects.

Abstract Anemia is a hematological disorder, and a large number in the developing world are affected with significant morbidity and mortality. Among them, most depend on traditional medicine to strengthen the hematopoietic action. The availability, low cost, and minor side effects are thought to be associated with this high selection rate of traditional medicine. The Pterocarpus marsupium is a medicinal plant native to Sri Lanka, India, and Nepal. In which the different components are used by traditional medical practitioners in treating different disease conditions, specifically, latex in enhancing hemoglobin production in anemic patients. However, the scientific evidence for using the Pterocarpus marsupium latex as an anti-anemic agent is minimal. Therefore, this study is planned to evaluate its anti-anemic effect, erythropoietin (EPO)-like effect, toxicity, phytochemical composition, and bioactive ingredients. Consequently, we aim to apply in vitro and in vivo assays to investigate the toxicity and favorable anti-anemic and other hematopoietic potentials of the plant latex using normal human cell cultures, rat-derived hematopoietic stem cell cultures, zebrafish embryos, and phenyl hydrazine-induced anemic rats with identification of bioactive ingredients that might mediate such potentials. If we discover a similar action of EPO in this plant latex, it would be beneficial in the way of treating anemia-associated disease conditions such as chronic renal failure, myelodysplasia, rheumatoid arthritis, HIV, and cancer as a replacement for recombinant human EPO (rhEPO). This will be beneficial in the way of reducing the high cost of rhEPO treatment as well. Finally, an attempt to discover a therapeutic agent for anemia from a safe, natural source will considerably contribute to traditional medicine/herbalism due to the crucial role of traditional medicine in the prophylaxis or therapy of hematological diseases in Sri Lanka. Keywords: Pterocarpus marsupium; Indian kino; Anemia; Toxicity; Latex; Erythropoietin

Selective elimination of tumorigenic cells from mixed culture of normal and tumorigenic cells using hybrid liposomes aimed at realizing of cell therapy

While induced pluripotent stem (iPS) cells are expected to be a cell source for regenerative medicine, they also have tumorigenic properties owing to their proliferative potential. During the manufacturing of regenerative medicine products, undifferentiated iPS cells and malignant transformed cells may be mixed in the cell culture population. Therefore, it is essential to eliminate tumorigenic cells selectively. In this study, a mixed culture of normal human fetal hepatocytes (Hc cells) and human hepatocellular carcinoma cells (HuH-7 cells) was used as a cell population model to be used as regenerative medicine products, and the selective elimination of HuH-7 cells by hybrid liposomes (HL) was analyzed. HL tended to fuse and accumulate more in HuH-7 cells due to larger fluidity of plasma membrane for HuH-7 cells than that for Hc cells. In a mixed culture of Hc and HuH-7 cells, HL selectively eliminated HuH-7 cells while allowing Hc cells to remain viable. In addition, HL treatment for the mixed culture of Hc and HuH-7 cells suppressed the tumorigenicity of HuH-7 cells. Therefore, HL selectively fused and accumulated in tumorigenic cells in a mixed cell culture of normal and tumorigenic cells, and eliminated tumorigenic cells while allowing normal cells to remain viable. The results of this study suggest the potential of HL in eliminating tumorigenic cells during the manufacturing of regenerative medicine products. Thus, HL could be expected to contribute to the development of safe regenerative medical products.

Cytotoxic activity of Fomitopsis betulina against normal and cancer cells - a comprehensive literature review.

In recent years, there has been increasing interest in research on fungi, including Fomitopsis betulina, known for its potential therapeutic properties. The aim of this paper is to systematically review the literature on the cytotoxic effects of Fomitopsis betulina on normal and cancer cell lines. The review was conducted by searching PubMed, Google Scholar, and Web of Science databases up to July 2024, using specific MeSH terms and keywords related to cytotoxicity, cancer cells, and normal cells. Articles were included if they investigated both cancer and normal cell cultures, reported IC50 values, and used validated cytotoxicity assays (e.g. MTT, LDH). Out of 450 articles screened, 5 met the inclusion criteria. The analysed studies demonstrated that Fomitopsis betulina extracts, particularly methanolic and ethanolic, exhibited selective cytotoxicity against cancer cells while having minimal impact on healthy cells. The strongest effects were observed in prostate cancer and melanoma cell lines. The cytotoxic effects were largely attributed to bioactive compounds such as triterpenes and glucans. Fomitopsis betulina extracts, particularly those derived from fruiting bodies and mycelial cultures, show promising cytotoxic properties against cancer cells, with limited impact on healthy cells. However, further research is needed to fully understand the mechanisms of action and to optimise extraction methods.

The Impact of the Addition of Vitamins on a Silicone Lining Material to the Oral Mucosa Tissue-Evaluation of the Biocompatibility, Hydrolytic Stability and Histopathological Effect.

Background and Objectives: One's quality of life depends on overall health, and in particular, oral health, which has been and continues to become a public health issue through frequent manifestations in various forms, from simple oral stomatitis (inflammations of the oral cavity) to the complicated oral health pathologies requiring medical interventions and treatments (caries, pulp necrosis and periodontitis). The aim of this study focused on the preparation and evaluation of vitamins (vitamin A, B1 and B6) incorporated into several silicone-based lining materials as a new alternative to therapeutically loaded materials designed as oral cavity lining materials in prosthodontics. Materials and Methods: Silicone-based liners containing vitamins were prepared by mixing them in solution and becoming crosslinked, and then they were characterized using Fourier-transform infrared (FT-IR) spectroscopy to confirm the incorporation of the vitamins into the silicone network; scanning electron microscopy (SEM) to evidence the morphology of the liner materials; dynamic vapor sorption (DVS) to evaluate their internal hydrophobicity, swelling in environments similar to biological fluids and mechanical test to demonstrate tensile strength; MTT to confirm their biocompatibility on normal cell cultures (fibroblast) and mucoadhesivity; and histopathological tests on porcine oral mucosa to highlight their potential utility as soft lining materials with improved efficiency. Results: FT-IR analysis confirmed the structural peculiarities of the prepared lining materials and the successful incorporation of vitamins into the silicone matrix. The surface roughness of the materials was lower than 0.2 μm, while in cross-section, the lining materials showed a compact morphology. It was found that the presence of vitamins induced a decrease in the main mechanical parameters (strength and elongation at break, Young's modulus) and hydrophobicity, which varied from one vitamin to another. A swelling degree higher than 8% was found in PBS 6.8 (artificial saliva) and water. Hydrolytic stability studies in an artificial saliva medium showed the release of low concentrations of silicone and vitamin fragments in the first 24 h, which increased the swelling behavior of the materials, diffusion and solubility of the vitamins. The microscopic images of fibroblast cells incubated with vitamin liners revealed very good biocompatibility. Also, the silicone liners incorporating the vitamins showed good mucoadhesive properties. The appearance of some pathological disorders with autolysis processes was more pronounced in the case of vitamin A liners. Conclusions: The addition of the vitamins was shown to have a beneficial effect that was mainly manifested as increased biocompatibility, hydrolytic stability and mucoadhesiveness with the mucosa of the oral cavity and less of an effect on the mechanical strength. The obtained lining materials showed good resistance in simulated biological media but caused a pronounced autolysis phenomenon, as revealed by histopathological examination, showing that these materials may have broad implications in the treatment of oral diseases.

Evaluation of the Biological Effect of Non-UV-Activated Bergapten on Selected Human Tumor Cells and the Insight into the Molecular Mechanism of Its Action.

There is some evidence that non-photoactivated psoralens may be active against breast and colon tumor cells. Therefore, we evaluated the antiproliferative, proapoptotic, and anti-migrative effect of 5-methoxypsoralen (5-MOP) isolated from Peucedanum tauricum MB fruits in human colorectal adenocarcinoma (HT-29 and SW620), osteosarcoma (Saos-2 and HOS), and multiple myeloma (RPMI8226 and U266). Dose- and cell-line-dependent effects of 5-MOP on viability and proliferation were observed, with the strongest inhibitory effect against Saos-2 and a moderate effect against the HOS, HT-29, and SW620 cells. Multiple myeloma showed low sensitivity. The high viability of human normal cell cultures (HSF and hFOB) in a wide range of 5-MOP concentrations tested (6.25-100 µM) was confirmed. Moreover, the migration of treated Saos-2, SW620, and HT-29 cell lines was impaired, as indicated via a wound healing assay. Flow cytometry analysis conducted on Saos-2 cells revealed the ability of 5-MOP to block the cell cycle in the G2 phase and trigger apoptosis, which was accompanied by a loss of mitochondrial membrane potential, caspases (-9 and -3) activation, the altered expression of the Bax and Bcl-2 proteins, and decreased AKT phosphorylation. This is the first report evaluating the antiproliferative and antimigratory impact of non-UV-activated bergapten on the abovementioned (except for HT-29) tumor cells, which provides new data on the potential role of 5-MOP in inhibiting the growth of various types of therapeutic-resistant cancers.

Transverse Myelitis in the Setting of Enterobius vermicularis (Pinworm) Infection: Case Report

Myelitis is a rare inflammatory myelopathy, and known associated etiologies only account for a small number of causes. A significant percentage of cases have an unknown etiology and are considered idiopathic. With 64% to 68% of cases fitting into the idiopathic category, helminth infections, and specifically pinworm parainfections, should be considered in cases that would otherwise be classified as idiopathic. This case report outlines a pediatric patient diagnosed with myelitis given her progressive weakness, fussiness, refusal to bear weight as well as magnetic resonance imaging (MRI) demonstrating T2-hyperintense signal and/or T1 gadolinium enhancement, and/or positive cerebrospinal fluid (CSF) inflammatory markers. This patient had a negative evaluation for typical known etiologies for myelitis including no signs of multiple sclerosis and neuromyelitis optica spectrum disorder on brain MRI, oligoclonal banding and aquaporin-4 autoantibodies, and no evidence of bacterial or viral meningitis given normal cell counts and cultures in CSF. She was found to have a pinworm infection, suggesting a parasitic parainfectious etiology of her myelitis. This case outlines the first case noting the correlation between myelitis and pinworm infection in a pediatric patient.

P14-05: Measuring mucin content and the ratio of goblet cells in three-dimensional cultures of normal human bronchial epithelial cells by confocal microscopy

P14-05: Measuring mucin content and the ratio of goblet cells in three-dimensional cultures of normal human bronchial epithelial cells by confocal microscopy

Topology and adenocarcinoma cell localization dataset on the labyrinthin diapeutic biomarker

ObjectiveThe discovery and characterization of tumor associated antigens is increasingly important to advance the field of immuno-oncology. In this regard, labyrinthin has been implicated as a neoantigen found on the cell surface of adenocarcinomas. Data on the (1) topology, (2) amino acid (a.a.) homology analyses and (3) cell surface localization of labyrinthin by fluorescent activated cell sorter (FACS) are studied in support of labyrinthin as a novel, pan-adenocarcinoma marker.ResultsBioinformatics analyses predict labyrinthin as a type II protein with calcium binding domain(s), N-myristoylation sites, and kinase II phosphorylation sites. Sequence homologies for labyrinthin (255 a.a.) were found vs. the intracellular aspartyl/asparaginyl beta-hydroxylase (ASPH; 758 a.a.) and the ASPH-gene related protein junctate (299 a.a.), which are both type II proteins. Labyrinthin was detected by FACS on only non-permeablized A549 human lung adenocarcinoma cells, but not on normal WI-38 human lung fibroblasts nor primary cultures of normal human glandular-related cells. Microscopic images of immunofluorescent labelled MCA 44-3A6 binding to A549 cells at random cell cycle stages complement the FACS results by further showing that labyrinthin persisted on the cell surfaces along with some cell internalization for greater than 20 min.