MTT Cell Proliferation Assay Research Articles

Investigation of The Effects of Extraction Polarity Change on the Bioactivity of Eruca Vesicaria

Eruca vesicaria (arugula) plant is frequently used today as a daily food source that contains a rich variety of minerals and vitamins, especially vitamin C. It also contains a high percentage of phenolic compounds which are structures that plants develop to protect themselves from harmful organisms. Phenolic compounds found in plants are obtained by different extraction methods and have a high antioxidant effect. In this study, we report, anticancer activity extract from Eruca vesicaria against human prostate cancer cells (PC-3) in vitro. The phenolic substances contained in the plant were obtained in different concentrations by the extraction technique based on the polarity difference. HPLC and total phenol content were determined to perform extraction content analysis. According to in vitro MTT cell proliferation assay, it acted at high concentrations, regardless of polarity differences. The highest cytotoxic effect was observed in extract extracted with 50% ethanol concentration. It has been observed that it has an anticancer effect compared to the determination of total phenol content. Also, as a result of 24-hour MIC analysis, it shows antibacterial properties according to agent concentration. As a result of this study, it adds many new information to the literature, but also provides guidance for future research.

Synthesis, Absolute Configuration, Biological Profile and Antiproliferative Activity of New 3,5-Disubstituted Hydantoins.

Hydantoins, a class of five-membered heterocyclic compounds, exhibit diverse biological activities. The aim of this study was to synthesize and characterize a series of novel 3,5-disubstituted hydantoins and to investigate their antiproliferative activity against human cancer cell lines. The new hydantoin derivatives 5a-i were prepared as racemic mixtures of syn- and anti-isomers via a base-assisted intramolecular amidolysis of C-3 functionalized β-lactams. The enantiomers of syn-5a and anti-hydantoins 5b were separated by preparative high-performance liquid chromatography (HPLC) using n-hexane/2-propanol (90/10, v/v) as the mobile phase. The absolute configuration of the four allyl hydantoin enantiomers 5a was assigned based on a comparison of the experimental electronic circular dichroism (ECD) and vibrational circular dichroism (VCD) spectra with those calculated using density functional theory (DFT). The antiproliferative activity evaluated in vitro against three different human cancer cell lines: HepG2 (liver hepatocellular carcinoma), A2780 (ovarian carcinoma), and MCF7 (breast adenocarcinoma), and on the non-tumor cell line HFF1 (normal human foreskin fibroblasts) using the MTT cell proliferation assay. In silico drug-like properties and ADMET profiles were estimated using the ADMET Predictor ver. 9.5 and the online server admetSAR. Eighteen new 3,5-disubstituted hydantoins were synthesized and characterized. The compound anti-5c showed potent cytotoxic activity against the human tumor cell line MCF7 (IC50 = 4.5 µmol/L) and the non-tumor cell line HFF1 (IC50 = 12.0 µmol/L). In silico analyzes revealed that the compounds exhibited moderate water solubility and membrane permeability and are likely substrates for CYP3A4 and P-glycoprotein and have a high probability of antiarthritic activity.

Polyphenolic extracts from Diospyros kaki and Vitis vinifera by-products stimulate cytoprotective effects in bacteria-cell host interactions by mediation of transcription factor Nrf2

BackgroundThe intestinal and skin epithelium play a strong role against bacterial stimuli which leads to inflammation and oxidative stress when overwhelmed. Polyphenols from fruit-rich diets and by-products show promise against bacterial deleterious effects; however, their antibacterial and health-promoting effects remain understudied. PurposeThis study aimed to assess the impact of polyphenolic extracts of grape (GrPE), persimmon (PePE) and pomegranate (PoPE) by-products on bacterial pathogen-host interactions, focusing beyond growth inhibition to explore their effects on bacterial adhesion, invasion, and modulation of host responses. MethodsThe microdilution method, as well as the tetrazolium based MTT cell proliferation and cytotoxicity assay with crystal violet staining were used to identify extracts sub-inhibitory concentrations that interfere with bacterial adhesion, invasion or lipopolysaccharides (LPS) effect on cell hosts without compromising host viability. The cytoprotective effects of extracts were assessed in a knock-down model of nuclear factor erythroid 2-related factor 2 (Nrf2). ResultsAll extracts demonstrated significant reductions in pathogen adhesion to Caco-2 and HaCaT cells while preserving cellular integrity. Notably, PePE exhibited specific efficacy against Salmonella enterica adhesion, attributed mostly to its gallic acid content, whereas PoPE reduced S. enterica invasion in Caco-2 cells. The extracts supported the prevalence of non-pathogenic and commensal strains of intestinal and skin surfaces, selectively reducing pathogenic adhesion. The extracts mitigated the oxidative stress, enhanced the barrier function, and modulated the pro-inflammatory cytokines in LPS-challenged cells. GrPE, rich in anthocyanins, and PePE were found to mediate their protective effects through Nrf2 activation, while PoPE exerted multifaceted actions independent of Nrf2. ConclusionOur results highlight the therapeutic potential of GrPE, PePE, and PoPE in shaping bacterial-host interactions, endorsing their utility as novel nutraceuticals for both oral and topical applications to prevent potential bacterial infections through innovative mechanisms.

Meldonium, as a potential neuroprotective agent, promotes neuronal survival by protecting mitochondria in cerebral ischemia–reperfusion injury

BackgroundStroke is a globally dangerous disease capable of causing irreversible neuronal damage with limited therapeutic options. Meldonium, an inhibitor of carnitine-dependent metabolism, is considered an anti-ischemic drug. However, the mechanisms through which meldonium improves ischemic injury and its potential to protect neurons remain largely unknown.MethodsA rat model with middle cerebral artery occlusion (MCAO) was used to investigate meldonium’s neuroprotective efficacy in vivo. Infarct volume, neurological deficit score, histopathology, neuronal apoptosis, motor function, morphological alteration and antioxidant capacity were explored via 2,3,5-Triphenyltetrazolium chloride staining, Longa scoring method, hematoxylin and eosin staining, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay, rotarod test, transmission electron microscopy and Oxidative stress index related kit. A primary rat hippocampal neuron model subjected to oxygen–glucose deprivation reperfusion was used to study meldonium’s protective ability in vitro. Neuronal viability, mitochondrial membrane potential, mitochondrial morphology, respiratory function, ATP production, and its potential mechanism were assayed by MTT cell proliferation and cytotoxicity assay kit, cell-permeant MitoTracker® probes, mitochondrial stress, real-time ATP rate and western blotting.ResultsMeldonium markedly reduced the infarct size, improved neurological function and motor ability, and inhibited neuronal apoptosis in vivo. Meldonium enhanced the morphology, antioxidant capacity, and ATP production of mitochondria and inhibited the opening of the mitochondrial permeability transition pore in the cerebral cortex and hippocampus during cerebral ischemia–reperfusion injury (CIRI) in rats. Additionally, meldonium improved the damaged fusion process and respiratory function of neuronal mitochondria in vitro. Further investigation revealed that meldonium activated the Akt/GSK-3β signaling pathway to inhibit mitochondria-dependent neuronal apoptosis.ConclusionOur study demonstrated that meldonium shows a neuroprotective function during CIRI by preserving the mitochondrial function, thus prevented neurons from apoptosis.

In silico and in vitro analyses to investigate the effects of vitamin C on VEGF protein

ObjectivesThis study was conducted to evaluate the effects of vitamin C on apoptotic and proliferative genes in injured HepG2 cells. MethodsIn silico analysis was performed using molecular docking of chemical compounds with vascular endothelial growth factor (VEGF). The different computational tools used were AutoDock Vina, BIOVIA DISCOVERY studio, and PyMOL. Drug likeness and toxicity were analyzed by SWISS ADMET. Cells that were 60–70% confluent were treated with different concentrations of hydrogen peroxide (H2O2) (100–2000 μM) and ascorbic acid (30, 60, 90 μg/mL). The MTT cell proliferation assay was performed to compare the proliferative potential of HepG2 cells treated with H2O2 or ascorbic acid with untreated HepG2 cells using 96-well plates. ResultsThe lowest binding energy of VEGF with vitamin C –5.2 kcal/mol and L-ascorbic acid-2 glycoside −4.7 kcal/mol was observed by in silico analysis. Vitamin C was selected because it exhibited a high interaction with VEGF and fulfilled Lipinski's rule, and had better oral viability and pharmacokinetics compared to L-ascorbic acid-2 glycoside. Cell viability assays showed that vitamin C had significant apoptotic effects (P < 0.0001). After treating HepG2 cells with ascorbic acid, reduced VEGF (angiogenesis) was observed as determined by apoptotic and proliferative gene expression. Ascorbic acid treatment of HepG2 cells led to downregulation of the proliferation markers, proliferating cell nuclear antigen, Ki67, and DNA topoisomerase II alpha. Increased apoptosis after treatment with vitamin C was observed due to upregulation of p53 and annexin V. ConclusionThe results of this study showed that vitamin C inhibited the growth of cancer cells, thus protecting HepG2 cells from oxidative stress. Vitamin C exhibited antiproliferative activity as observed in silico and in vitro, as well as by the inhibited expression of genes involved in protein synthesis.

Study on the Lupine-hulls Based Cellular Scaffolds for Human Cardiac Tissue Regeneration

It's becoming more popular to develop new biomaterials enabling in vitro cell cultivation with microbial environments of biological structures. The novel strategies for creating native cellular environments that facilitate the growth of human cardiac progenitor cells (hCPCs) (Sca-1pos) and scaffolding more closely resemble authentic cell function microenvironments with offer a wide range of appealing in vivo and in vitro uses. Both the biologically active substances and the mechanical properties of the hull scaffold were studied. SEM analysis of the hull's microstructure reveals that the inside surface shows its large and naturalistic activity, while the outside surface displays the celular-structures. Tensile properties, high-performance liquid chromatography (HPLC), and fourier transform infrared spectroscopy (FTIR) spectra are further characteristics of the lupine-hulls. Scaffolds made from lupine-hulls are cheap, simple to make, and from a renewable material. The MTT assay for cell proliferation served to evaluate the impact of the hulls that are solubility substance on the Sca-1pos. The cell lines utilized in this study were able to multiply and maintain high viability even after being cultured for 7 straight days. Results also demonstrate that the nuclei of the cells were visible under fluorescence microscopy and remained viable within the cellular structures of the hulls in vitro. This research showed that scaffold made from lupine hulls are being used as biological scaffolds for in vitro cell growth, which may play a role in the tissue engineering of hCPCs.

Synchronized Chemotherapy/Photothermal Therapy/Sonodynamic Therapy of Human Triple-Negative and Estrogen Receptor-Positive Breast Cancer Cells Using a Doxorubicin–Gold Nanoclusters–Albumin Nanobioconjugate

ObjectiveNovel strategies for treating triple-negative breast cancer (TNBC) are ongoing because of the lack of standard-of-care treatment. Nanoframed materials with a protein pillar are considered a valuable tool for designing multigoals of energy-absorbing/medication cargo and are a bridge to cross-conventional treatment strategies. MethodsNanobioconjugates of gold nanoclusters–bovine serum albumin (AuNCs–BSA) and doxorubicin–AuNCs–BSA (Dox–AuNCs–BSA) were prepared and employed as a simultaneous double photosensitizer/sonosensitizer and triple chemotherapeutic/photosensitizer/sonosensitizer, respectively. ResultsThe highly stable AuNCs–BSA and Dox–AuNCs–BSA have ζ potentials of –29 and –18 mV, respectively, and represent valuable photothermal and sonodynamic activities for the combination of photothermal therapy and sonodynamic therapy (PTT/SDT) and synchronized chemotherapy/photothermal therapy/sonodynamic therapy (CTX/PTT/SDT) of human TNBC cells, respectively. The efficiency of photothermal conversion of AuNCs–BSA was calculated to be a promising value of 32.9%. AuNCs–BSA and Dox–AuNCs–BSA were activated on either laser light irradiation or ultrasound exposure with the highest efficiency on the combination of both types of radiation. CTX/PTT/SDT of MCF-7 and MDA-MB-231 breast cancer cell lines by Dox–AuNCs–BSA were evaluated with the MTT cell proliferation assay and found to progress synergistically. ConclusionResults of the MTT assay, detection of the generation of intracellular reactive oxygen species and occurrence of apoptosis in the cells confirmed that CTX/PTT/SDT by Dox–AuNCs–BSA was attained with lower needed doses of the drug and improved tumor cell ablation, which would result in the enhancement of therapeutic efficacy and overcoming of therapeutic resistance.

Abstract 5945: Exploring the antiproliferative potential of KHS101 on triple negative breast cancer cell lines

Abstract Breast cancer is the second leading cause of cancer deaths in the world. Among its subtypes, triple-negative breast cancer (TNBC) is highly aggressive, particularly affecting African American (AA) women, with limited treatment choices. HSP60 protein promotes tumorigenesis and is being studied as a possible biomarker to treat cancers such as TNBC. The small molecule KHS101 specifically binds and inhibits the mitochondrial chaperone HSP60. In this study, we hypothesized that KHS101 would inhibit the proliferation of TNBC cells and therefore may have a therapeutic potential.We studied the antiproliferative effects of KHS101 on TNBC cell lines (MDA-MB-231, MDA-MB-468, HCC1806) through MTT cell proliferation and colony forming assays at varying doses and time points (24, 48, 72, and 96 hours). We performed cell cycle analysis to understand the cell cycle response and apoptotic events in treated or control cells. Additionally, the gene expression and frequently altered pathways resulting from KHS101 treatment were analyzed by RNA sequencing of cells treated with vehicle (DMSO) or with various concentrations of KHS101(1, 2.5, 5, and 7.5 uM) for 72 hours. We analyzed the data using GraphPad Prism and Microsoft Excel, with statistical significance set at p ≤ 0.05. TNBC cell lines exhibited significant dose-response and antiproliferative effects with increased KHS101 doses and treatment durations. Notably, AA breast cancer cells (MDA-MB-468) displayed heightened cytotoxicity and reduced proliferation compared to Caucasian American (CA) breast cancer cells (MDA-MB-231 and HCC1806). Cell cycle analysis revealed G2/M phase arrest, consistent with RNA sequencing data showing downregulation of the activation of ATM/ATR response in KHS101-treated cell lines compared to the controls. Gene set enrichment analysis indicated downregulation of the cell cycle control genes in MDA-MB-231 cells after 72 hours of KHS101 treatment (7.5 µM). Reactome pathway analysis in KHS101-treated MDA-MB-231 cells showed upregulation of IFN-gamma signaling and degradation of extracellular matrix. Similarly, Kegg pathway analysis showed upregulation in cell adhesion molecules, cytokine-cytokine receptor interaction, antigen processing and presentation, and Natural Killer cell-mediated cytotoxicity in KHS101-treated cancer cells compared to the controls. KHS101 demonstrates varying cytotoxic effects on TNBC cells, with potential implications for antitumor immune responses. Our findings highlight the distinct susceptibility of AA breast cancer cells and suggest KHS101 as a promising avenue for further research in understanding its mechanism of action and validating therapeutic potential in TNBC. Citation Format: Reshma Gurung, Alehegne W. Yirsaw, Muhammad G. Khodary, Temesgen Samuel, Maninder Sandey, Clayton C. Yates, Deepa Bedi. Exploring the antiproliferative potential of KHS101 on triple negative breast cancer cell lines [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5945.

CYP1-Activation and Anticancer Properties of Synthetic Methoxylated Resveratrol Analogues.

Naturally occurring stilbenoids, such as the (E)-stilbenoid resveratrol and the (Z)-stilbenoid combretastatin A4, have been considered as promising lead compounds for the development of anticancer drugs. The antitumour properties of stilbenoids are known to be modulated by cytochrome P450 enzymes CYP1A1 and CYP1B1, which contribute to extrahepatic phase I xenobiotic and drug metabolism. Thirty-four methyl ether analogues of resveratrol were synthesised, and their anticancer properties were assessed, using the MTT cell proliferation assay on a panel of human breast cell lines. Breast tumour cell lines that express CYP1 were significantly more strongly affected by the resveratrol analogues than the cell lines that did not have CYP1 activity. Metabolism studies using isolated CYP1 enzymes provided further evidence that (E)-stilbenoids can be substrates for these enzymes. Structures of metabolic products were confirmed by comparison with synthetic standards and LC-MS co-elution studies. The most promising stilbenoid was (E)-4,3',4',5'-tetramethoxystilbene (DMU212). The compound itself showed low to moderate cytotoxicity, but upon CYP1-catalysed dealkylation, some highly cytotoxic metabolites were formed. Thus, DMU212 selectively affects proliferation of cells that express CYP1 enzymes.

Organic Extract of Streptomyces Induces Apoptosis, Anti-Proliferation and Cell-Cycle Arrest in Colon Cancer Cells: Caco-2 and HCT116 Cell Line as Surrogate Model

Background: Colorectal cancer (CRC) is a prevalent and very lethal malignancy, resulting in more than one million deaths and nearly two million newly diagnosed cases in the year 2018. Streptomyces bacteria are a potential source of conventional compounds, as their extract contains bioactive chemicals with antiproliferative effects in cell culture settings. The aim of this research was to investigate the antiproliferative effects of the Streptomyces ethyl acetate fraction against the Caco-2 and HCT116 cell line in addition to its fundamental mechanisms. Methods: We examined the effects of Streptomyces extract on colon cancer cell lines Caco-2 and HT-116 by incubating the cells with and without the extract. Antiproliferative effects are analyzed using the MTT cell proliferation assay, cell cycle analysis and flow cytometric detection of Annexin V/PI apoptosis. The formation of intracellular ROS was measured, in addition to Western Blot for Bax, Bcl2 and caspase-3 activation. Result: Our findings showed the dose-dependent inhibition of cell growth by streptomyces extract. Both Caco-2 and HT-116 cells treated with streptomyces extract displayed the beginning of apoptotic events and G1 cell-cycle arrest. Additionally, treat with the streptomyces extract significantly increased (P less than 0.05) ROS production, Bax and cleaved-caspase-3 were increased, while Bcl-2 was downregulated, as determined by western blotting.

Mangifera indica L. kernel ethanol extract inhibits cell viability and proliferation with induction of cell cycle arrest and apoptosis in lung cancer cells

ABSTRACT In this study, we investigated the effects of an ethanolic extract of Mangifera indica L. kernel on the viability and proliferation of human lung cancer cells. We utilized MTT and BrdU cell proliferation assays, morphological assessments, cell cycle analyses, and apoptosis assays to investigate the extract’s effects on lung cancer (A549 and NCI-H292) and normal lung (MRC-5) cells. The extract demonstrated a toxicity toward cancer cells compared to normal cells with dose-dependent anti-proliferative effect on lung cancer cells. The extract also caused differential effects on the cell cycle, inducing G0/G1 arrest and increasing the Sub-G1 population in both lung cancer and normal lung cells. Notably, the extract induced loss of membrane integrity, shrinkage, membrane blebbing, and apoptosis in lung cancer cells, while normal cells exhibited only early apoptosis. Furthermore, the extract exhibited higher toxicity towards NCI-H292 cells, followed by A549 and normal MRC-5 cells in decreasing order of potency. Our results suggest that the ethanolic extract of M. indica L. kernel has significant potential as a novel therapeutic agent for treating lung cancer cells, given its ability to induce apoptosis in cancer cell lines while causing minimal harm to normal cells.

Synthesis and evaluation of tetrahydropyrrolo[1,2-a]quinolin-1(2H)-ones as new tubulin polymerization inhibitors.

Here we explored new 1,5-disubstituted pyrrolidin-2-ones 1, 2 and 5-aryl-3,3a,4,5-tetrahydropyrrolo[1,2-a]quinoline-1(2H)-ones 3 as inhibitors of tubulin polymerization. We evaluated their effects on microtubule dynamics in vitro and on the proliferation of A549 cells, using flow cytometry-based cell cycle analysis. The results were verified with phase-contrast microscopy in three cancer cell lines: A549, HeLa and MCF-7. Guided by molecular modeling of the interactions between tubulin and the most active of the identified compounds, we designed, synthesized, and tested the 3-hydroxyphenyl-substituted compound 3c. This compound was further shown to bind to the colchicine site of tubulin and reduce microtubule growth rates in vitro. Moreover, compound 3c arrested division of the A549 cells in the low micromolar range (IC50 = 5.9 μM) and exhibited cytotoxicity against four different cell lines in the MTT assay for cell proliferation. Our findings demonstrate that 5-aryltetrahydropyrrolo[1,2-a]quinoline-1(2H)-one is a promising scaffold for the development of novel tubulin polymerization inhibitors.

Development of phenyllactic acid ionic liquids and evaluation of cytotoxicity to human cervical epithelial cells.

Phenyllactic acid (PLA), is a naturally produced, broad-spectrum antimicrobial compound with activity against bacteria and fungi. PLA can be produced by a variety of lactic acid bacteria, including vaginal Lactobacillus species, which are healthy constituents of the vaginal microbiome with a protective role against invading pathogenic bacteria and/or fungi. Additionally, PLA has been shown to exhibit anti-inflammatory and immunomodulatory properties, overall indicating its therapeutic potential as an intravaginally delivered compound for modulation of the vaginal microbiome. However, PLA has low kinetic solubility in water. Hence, strategies to improve the solubility of PLA are necessary to facilitate its intravaginal delivery. Using biocompatible cations, choline and carnitine, we successfully transformed both d- and l-enantiomers of crystalline PLA into amorphous low-melting ionic liquids (ILs) with high water solubility. We further evaluated the in vitro cytotoxicity of PLA ILs to human cervical epithelial cells. Microscopic visualisation of cellular morphology using crystal violet staining and MTT cell proliferation assay revealed that PLA ILs result in minimal morphological changes and low cytotoxicity to human cervical epithelial cells. Overall, we successfully demonstrated that transforming PLA into ILs efficiently enhances its solubility in water and these formulations are not toxic to human epithelial cells. This investigation lays the groundwork for future testing of PLA ILs for their antimicrobial properties and metabolic activity within the cervicovaginal microenvironment.

Abstract C052: Effect of ginger extracts on hepatocellular carcinoma cell lines-derived from Caucasian, Asian, and African American patients

Abstract Hepatocellular Carcinoma (HCC) is a highly fatal disease with mortality running parallel to its incidence. For HCC, race/ethnicity plays a vital role in determining incidence, mortality, and survival rates. The incidence of HCC is highest in Asia and Africa. Furthermore, there is a statistically significant increase in incidence and mortality and a decrease in 5-year survival rates in African American (AA)/Black patients compared to non-Hispanic white patients. There is a knowledge gap in our understanding of the molecular mechanism underlying the HCC racial disparity between AA/Black, White, and Asian patients. To understand the underlying cause, we performed bioinformatics on existing gene expression data. We found that type I Interferon (IFN-I) signaling pathway showed statistically significant activation in AA/Black patients compared to white patients. Over 60% of Gen Xers/Millennials are taking a more holistic approach to their diet to prevent chronic diseases, including cancer. We hypothesized that dietary compounds exert anticancer effects on HCC, and because of their anti-inflammatory property, they might modulate the IFN-I signaling pathway. We tested ginger extracts on HCC derived from white (HepG2), Black (Hep3B and O/20), and Asian (Hu7) cancer patients. A dose-response of these extracts was used to determine IC50s on these cell lines using an MTT cell proliferation assay. Activation of INF-1-mediated downstream signaling proteins, including JAK1, TYK2, STAT1, and STAT2 phosphorylation status, was determined on a Western blot analysis using phospho-specific primary antibodies. The expression of Interferon Signaling Genes (ISGs), including Myxovirus resistance gene 1 (MX1), 2′,5′-oligoadenylate synthetase (OAS1), Interferon-alpha inducible protein 6 (IFI6), and Interferon stimulated gene 15 (ISG15) was assayed using a quantitative Real-Time Polymerase Chain Reaction (RT-PCR). Ginger has a significantly (P&amp;lt;0.05) lower IC50s (mg/ml) on cell lines from Black patients (Hep3B=156 ± 3, and O/20=162 ± 3) than cell lines from white (HepG2=176±5) or Asian (Hu7=174 ± 5) patients. Our data show that untreated cells exhibited phosphorylation/activation of IFN-1 downstream mediators, including JAK1, TYK2, STAT1, and STAT2 proteins. Ginger treatment reduced the phosphorylation of these IFN downstream mediators in all HCC cell lines. Furthermore, expression of MX1, ISG15, IF16, and OAS1 was also reduced in all HCC cells lines in a dose-dependent manner; however, the effects on gene expression were more sensitive to a lower concentration of ginger extract in Black patients derived HCC cell lines than other HCC cell lines. Currently, research is underway to identify the active chemical agent in ginger and test it on other patient-derived HCC lines and in an in vivo model. In conclusion, our data suggest that ginger can potentially attenuate IFN-mediated signaling pathways in HCC, and cells from Black HCC patients may be more sensitive to ginger (Funded: P20 CA264068-01). Citation Format: Sadia Kanwal, Eva Davis, Seung Lee, Milton Omar Faison, Devanand Sarkar, Rafat Ali Siddiqui. Effect of ginger extracts on hepatocellular carcinoma cell lines-derived from Caucasian, Asian, and African American patients [abstract]. In: Proceedings of the 16th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2023 Sep 29-Oct 2;Orlando, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2023;32(12 Suppl):Abstract nr C052.

Antimalarial evaluation of alkyl-linked bis-thiadiazine derivatives in murine model infected with two Plasmodium strains.

Plasmodium falciparum and P. vivax are responsible for most malaria cases in humans in the African Region and the Americas; these parasites have developed resistance to classic antimalarial drugs. On the other hand, previous investigations of the alkyl-linked bis tetrahydro-(2H)-1,3,5-thiadiazine-2-thione (bis-THTT) derivatives compounds show satisfactory results against protozoan parasites such as Trypanosoma cruzi, Trypanosoma vaginalis, Trypanosoma brucei rhodesiense and Leishmania donovani. Therefore, it is possible to see some effect of bis-THTT derivatives on other protozoan parasites, such as Plasmodium. This study aimed to perform an in vivo biological evaluation of bis-THTT (JH1 to JH6) derivatives compounds as possible anti-malaria drugs in BALB/c mice infected with Plasmodium berghei ANKA and Plasmodium yoelii 17XL strains. In this work, we evaluated the compounds as potential antimalarial drugs in BALB/c mice infected with Plasmodium strains. For each compound, we assess the percentages of parasitemia by smears from tail blood and the humoral response by indirect ELISA test using each compound as an antigen. We also evaluated the B lymphocyte response and the cytotoxicity of the bis-THTT derivatives compounds with MTT cell proliferation assays. Our results show that the bis-THTT derivatives JH2 and JH4 presented effective parasitemia control in mice infected with P. berghei; JH5 and JH6 compounds have similar infection control results as chloroquine in mice infected P. yoelii strain. The evaluation of bis-THTT derivatives compounds in a model of BALB/c mice infected with P. berghei and P. yoelii allowed us to conclude that some of them have an antimalarial effect; however, none of the tested compounds exceeded the efficiency of chloroquine.

Role of Ibea Gene in Hbmecs Invasion and Biofilm Formation Induced By Neonatal Meningitis Escherichia coli

Background: Bacterial meningitis is a severe infectious disease that affects the central nervous system and can result in high mortality rates and neurological complications. In neonates, meningitis is mainly caused by neonatal meningitis Escherichia coli (NMEC), which invades the intestine, leading to hematogenous spreadand potential bacterial meningitis through blood-brain barrier. Previous studies indicated that the ability of NMEC to adhere and invade is associated with biofilm formation and a deficiency in ibeA may result in decreased virulence and biofilm-forming ability of avian pathogenic Escherichia coli (APEC). Furthermore, the ibeA gene is positioned downstream of ibeR. Our previous studies have shown that ibeR and ibeA genes can both regulate the virulence and relevant phenotypes in E. coli K1. However, the effects of ibeR and ibeA on the biofilm formation and invasion of K1 strain remain to be further studied. Recent studies suggest that indole may act as a signaling molecule to regulate the expression of adhesion and biofilm-promoting factors in E. coli. However, the mechanism by which indole affects biofilms is still unclear. This study aimed to investigate the mechanism of ibeA inhibition of indole-induced NMEC invasion and biofilm formation. Methods: The viability and growth of the wild strain E44 and ibeA-deletion mutant strain ZD1 were assessed by MTT cell proliferation assay kit (Coolaber). The absorbance values were measured at 490 nm and 540 nm by ELISA. The invasion activities and the biofilm formation ability of E44 and ZD1 were detected in vitro studies. We conducted a cell invasion test using the Human Brain Microvascular Endothelial Cells (HBMEC) model. The ability to form a biofilm was measured at 600 nm using a specific biofilm formation index. Biofilm phenotypic was observed by an confocal laser-scanning microscope(Olympus FV-1000, Japan). Then, we used indole as a regulatory agency to compare its biofilm inhibitory efficacy between E44 and ibeR-deletion mutant strain BR2. The experimental data were analyzed using the SPSS 19.0 statistical package. A statistically significant difference was indicated by P&amp;lt;0.05. Results: No difference was found in bacterial growth between the E. coli strains E44 and ZD1. The biofilm formation and HBMECs invasion of ZD1 was significantly lower than that of E44. After treatment with 0.5mM indole, the biofilm formation ability and the ibeA protein expressions of E44 were significantly inhibited. However, indole at different concentrations had no obvious inhibition on BR2 biofilm formation and the ibeA protein expressions. Conclusion: In summary, the study found that ibeA had a significant regulatory effect on HBMECs invasion and biofilm formation of NMEC, which may inhibit by indole via regulating ibeA through ibeR. ( Acknowledgements: Grant from School of Public Health of Southern Medical University, Guangzhou, China, No. GW202329; J.Z., Q.L. and L.W. contributed equally to this work; Corresponding author: Hong Cao, gzhcao@smu.edu.cn)

The Effect of Calcium hydroxide, Glass Ionomer and light cured resin modified calcium silicate on viability, proliferation and differentiation of stem cells from human exfoliated deciduous teeth

BackgroundVital pulp therapy, based on the use of stem cells, has promising research and therapeutic applications in dentistry. It is essential to understand the direct effect of capping materials on the dental pulp stem cells of primary teeth, which contribute to the healing powers of the tooth. The aim of this study is to evaluate the effect of different capping materials (Calcium Hydroxide (DyCal®) – Glass Ionomer (Fuji IX®) and light-cured resin modified calcium silicate (TheraCal LC®)) on the viability, proliferation, and differentiation of stem cells from human exfoliated deciduous teeth (SHEDs).MethodsSHEDs were isolated from extracted primary teeth, then divided into four groups and each of the capping materials were applied to the stem cells as follows: group I the controls, group II with Ca(OH)2, group III with the GIC, and group IV with the Theracal LC. For all groups assessment of viability and proliferation rate was done using the MTT cell proliferation assay. Also, Differentiation was evaluated by measuring the gene expression of Alkaline phosphatase enzyme activity (ALP) and Dentin matrix protein-1 (DMP1) through quantitative real-time PCR. Morphological assessment was conducted using Alizarin Red S staining. All evaluations were performed after 7 and 14 days of culture.ResultsTheraCal LC showed the highest values of proliferation, which was significant only compared to the control group after 2 weeks (p = 0.012). After one week, TheraCal LC showed the highest significant values of ALP and DMP1 compared to all other groups (p < 0.001).ConclusionThe three materials under study are biocompatible, maintain viability, and stimulate the proliferation and differentiation of SHEDs. However, TheraCal LC allows better proliferation of SHEDs than Dycal Ca(OH)2 and Fuji IX GIC.

Alteration in the expression of the chemotherapy resistance‑related genes in response to chronic and acute hypoxia in pancreatic cancer.

Pancreatic cancer is currently one of the least curable types of human cancer and remains a key health problem. One of the most important characteristics of pancreatic cancer is its ability to grow under hypoxic conditions. Hypoxia is associated with resistance of cancer cells to radiotherapy and chemotherapy. It is a major contributor to pancreatic cancer genetic instability, which local and systemic resistance that may result in poor clinical outcome. Accordingly, identifying gene expression changes in cancer resistance genes that occur under hypoxic conditions may identify a new therapeutic target. The aim of the present study was to explore the association between hypoxia and resistance to chemotherapy and determine the alteration in the expression of cancer resistance-related genes in the presence of hypoxia. Pancreatic cancer cells (PANC-1) were exposed to 8 h hypoxic episodes (<1% oxygen) three times/week for a total of 20 episodes (chronic hypoxia) or 72 h hypoxic episodes twice/week for a total of 10 episodes (acute hypoxia). The alterations in gene expression were examined using reverse transcription-quantitative PCR array compared with normoxic cells. Chemoresistance of hypoxic cells toward doxorubicin was assessed using MTT cell proliferation assay. Both chronic and acute hypoxia induced chemoresistance toward doxorubicin in PANC-1 pancreatic cancer cell line. The greatest changes occurred in estrogen Receptor Alpha Gene (ESR1) and ETS Like-1 protein (ELK1) pathways, in nucleic transcription factor Peroxisome proliferator-activated receptors (PPARs) and in a cell cycle inhibitor cyclin dependent kinase inhibitor 1A (CDKN1A). The present study demonstrated that exposing cells to prolonged hypoxia results in different gene expression changes involving pleotropic pathways that serve a role in inducing resistance in pancreatic cancer.

Development of Liposomal and Liquid Crystalline Lipidic Nanoparticles with Non-Ionic Surfactants for Quercetin Incorporation.

The aim of the present study is the development, physicochemical characterization, and in vitro cytotoxicity evaluation of both empty and quercetin-loaded HSPC (hydrogenated soy phosphatidylcholine) liposomes, GMO (glyceryl monooleate) liquid crystalline nanoparticles, and PHYT (phytantriol) liquid crystalline nanoparticles. Specifically, HSPC phospholipids were mixed with different non-ionic surfactant molecules (Tween 80 and/or Span 80) for liposomal formulations, whereas both GMO and PHYT lipids were mixed with Span 80 and Tween 80 as alternative stabilizers, as well as with Poloxamer P407 in different ratios for liquid crystalline formulations. Subsequently, their physicochemical properties, such as size, size distribution, and ζ-potential were assessed by the dynamic and electrophoretic light scattering (DLS/ELS) techniques in both aqueous and biological medium with serum proteins. The in vitro biological evaluation of the empty nanosystems was performed by using the MTT cell viability and proliferation assay. Finally, the entrapment efficiency of quercetin was calculated and the differences between the two different categories of lipidic nanoparticles were highlighted. According to the results, the incorporation of the non-ionic surfactants yields a successful stabilization and physicochemical stability of both liposomal and liquid crystalline nanoparticles. Moreover, in combination with an appropriate biosafety in vitro profile, increased encapsulation efficiency of quercetin was achieved. Overall, the addition of surfactants improved the nanosystem's stealth properties. In conclusion, the results indicate that the physicochemical properties were strictly affected by the formulation parameters, such as the type of surfactant.

Some Wild Mushrooms with High Antioxidant Capacity Exhibit Potent Anticancer Activity on Cancer Cells using the Apoptotic and Antimigration Cell Death Mechanisms.

This study aims to provide in vitro experimental evidence that wild mushrooms have the potential to be used as a pharmaceutical that could be effective against various types of cancer. Throughout human history, besides food, traditional medicine and natural poisons obtained from mushrooms have been used for the treatment of many diseases. Clearly, edible and medicinal mushroom preparations have beneficial health effects without the known severe adverse side effects. This study was designed to reveal the cell growth inhibitory potential of five different edible mushrooms and the biological activity of Lactarius zonarius was shown here for the first time. The mushrooms fruiting bodies were dried and powdered then extracted with hexane, ethyl acetate, and methanol. The mushroom extracts were screened for possible antioxidant activities by the free radical scavenging activity (DPPH) method. Antiproliferative activity and cytotoxicity of the extracts were investigated in vitro on A549 (human lung carcinoma), HeLa (human cervix carcinoma), HT29 (human colon carcinoma), Hep3B (human hepatoma), MCF7 (human breast cancer), FL (human amnion cells), and Beas2B (normal human cells) cells lines by using MTT cell proliferation assay, lactate dehydrogenase (LDH) assay, DNA degradation, TUNEL, and cell migration assay. Using proliferation, cytotoxicity, DNA degradation, TUNEL, and migration assay, we displayed that hexane, ethyl acetate, and methanol extracts of the Lactarius zonarius, Laetiporus sulphureus, Pholiota adiposa, Polyporus squamosus, and Ramaria flava were effective on the cells even so at low doses (< 45.0 - 99.6 µg/mL) by acting in a way that represses migration, as a negative inducer of apoptosis. It was also demonstrated that mushroom extracts with high antioxidant effect have within the acceptable cytotoxic activity of 20%-30% on the cell membrane at concentrations higher than 60 µg/mL. Overall, all of the mushroom extracts with high antioxidant effects had strong antiproliferative activity and low toxicity for cells. These findings, at least, highlight that these mushroom extracts can be used for the treatment of cancer disease, especially as a supportive therapy against colon, liver, and lung cancer.