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The study investigated the efficacy of Campomanesia xanthocarpa infusions on human melanoma cells (SK-MEL-28). The phytochemical profile revealed 18 phenolic compounds in the leaf infusion and 9 in the fruit infusion. After 24 h of treatment, the infusions demonstrated antineoplastic effects, reducing cell viability at all tested concentrations for the leaf infusion. For the fruit infusion, a significant reduction in cell viability was observed specifically at the 800 μg/mL concentration. Fluorescence microscopy and mitochondrial membrane potential results indicated that the leaf infusion was more effective in reducing cell viability and mitochondrial function in SK-MEL-28 cells, possibly due to its greater variety of phenolic compounds compared to the fruit infusion. The leaf infusion also induced higher production of intracellular reactive oxygen species compared to the fruit infusion. Protein sulfhydryl levels were reduced for the leaf infusion. Epigallocatechin gallate, Isoquercitrin, Rutin, Kaempferol-3-O-rutinoside, Chlorogenic acid, and Ellagic acid were identified as the main compounds with activity against SK-MEL-28 cells. Molecular docking analysis underscored factors such as affinity, cavity size, binding mode, and contact residues with specific compounds chosen for their favorable properties in targeting BRAF, CDK4, CDK6, MEK1, and MEK2. The variability in binding affinities may directly influence the compounds' ability to inhibit different signaling pathways related to cancer cell growth and proliferation. The results suggest that phenolic compounds from C. xanthocarpa extracts have therapeutic potential and could contribute to melanoma therapies. [ABSTRACT FROM AUTHOR]

Ischemic stroke consists of rapid neural death as a consequence of brain vessel obstruction, followed by damage to the neighboring tissue known as ischemic penumbra. The cerebral tissue in the core of the lesions becomes irreversibly damaged, however, the ischemic penumbra is potentially recoverable during the initial phases after the stroke. Therefore, there is real need for emerging therapeutic strategies to reduce ischemic damage and its spread to the penumbral region. For this reason, we tested the effect of Extreme Low Frequency Electromagnetic Stimulation (ELF-EMS) on in vitro primary neuronal and microglial cultures under oxygen-glucose deprivation (OGD) conditions. ELF-EMS under basal non-OGD conditions did not induce any effect in cell survival. However, ELF-EMS significantly reduced neuronal cell death in OGD conditions and reduced ischemic induced Ca2+ overload. Likewise, ELF-EMS modulated microglia activation and OGD-induced microglia cell death. Hence, this study suggests potential benefits in the application of ELF-EMS to limit ischemic irreversible damages under in vitro stroke conditions, encouraging in vivo preclinical validations of ELF-EMS as a potential therapeutic strategy for ischemic stroke. [ABSTRACT FROM AUTHOR]

ABSTRACT This study investigated the survival dynamics of Lactobacillus plantarum BG24, a probiotic strain, within reconstituted skim milk (RSM) and yeast extract (YE) matrices during the spray–drying (SD) process, encompassing of inlet/outlet air temperatures. Notably, optimum SD parameters were found to be an inlet air temperature of 150°C and outlet air temperature of 83°C, that achieving high viability (92.23%), and reducing both moisture content (MC) (3.57%) and water activity (aw) (0.266). The use of soy protein isolate (SPI), gum Arabic (GA), RSM, maltodextrin (MD), sucrose (SUC), and lactose in binary mixtures or alone was investigated in terms of the best survival rate of probiotic bacteria, and RSM alone and RSM + GA and SPI alone were found to be the best drying carriers giving higher viability during SD. SD at optimum process temperatures and freeze drying (FD) were compared in the survival rate of probiotic bacteria in the carrier of RSM with YE, and FD samples showed a higher survival rate (97.69%) than SD samples. It was determined that the storage temperature (4°C and 20°C) had an impact on the glass transition temperature, MC, aw, and cell viability. Increased storage temperature led to a greater decrease in cell viability, especially for SD probiotic powders. These findings furnish critical insights into the intricate interplay among process parameters, carrier agents, drying techniques, and storage conditions, thereby elucidating avenues for refining probiotic preservation strategies within the ambit of SD, and by extension, in the domains of food and pharmaceutical sciences. [ABSTRACT FROM AUTHOR]

Epidermal growth factor receptor (EGFR)-activating mutations are critical factors in the development of EGFR-driven non-small-cell lung cancer (NSCLC), and molecular targeted therapies have focused on inhibiting these mutations. EGFR tyrosine kinase inhibitors (TKIs) have remarkable inhibitory effects on NSCLC with EGFR mutations. However, acquired resistance limits the clinical application of EGFR-TKIs, highlighting the need for discovery of novel therapeutic strategies. 1,4,5,6-tetrahydroxy-7,8-diprenylxanthone is a natural product derived from Garcinia xanthochymus, has shown potential anti-tumor activity, but the underlying mechanism needs further elucidation. In this study, we developed Ba/F3 and NIH/3T3 cells harboring EGFR L858R/T790M/C797S mutation, and then found that 1,4,5,6-tetrahydroxy-7,8-diprenylxanthone exerted inhibitory effects against cells with EGFR L858R/T790M/C797S mutation. Additionally, 1,4,5,6-tetrahydroxy-7,8-diprenylxanthone exhibited potent anti-tumor activity against cells harboring the triple-mutant EGFR by promoting apoptosis and inducing changes in cell cycle distribution. Furthermore, 1,4,5,6-tetrahydroxy-7,8-diprenylxanthone was found to significantly reduce tumor growth via suppressing the phosphorylation of EGFR in tumor tissues. These effects are associated with binding of 1,4,5,6-tetrahydroxy-7,8-diprenylxanthone to EGFR resulting in the suppression of extracellular signal-regulated kinase (Erk) phosphorylation. In conclusion, our results suggest that 1,4,5,6-tetrahydroxy-7,8-diprenylxanthone may be a potential novel candidate for further investigation and treatment of NSCLC with the triple-mutant EGFR. [ABSTRACT FROM AUTHOR]

Cancer is a type of non-communicable disease that is responsible for numerous deaths worldwide. Cancer incidence and mortality rates are on the rise due to a combination of factors, such as a growing population, aging, and poor dietary habits. The Allium turcicum Özhatay & Cowley plant is an endemic plant in the area where it grows and is consumed by the public due to its various benefits. This endemic plant, which generally grows in high-altitude regions, is sold in bunches because it is costly, mixed with rock salt, crushed into powder, and consumed as a spice. The cytotoxic and growth-inhibitory effects of A. turcicum Özhatay & Cowley herb extract on human glioblastoma U373 cells, human colorectal carcinoma cell HCT-116, and healthy HUVEC cell lines were determined by the MTT method. After 24 and 48 h of application, logIC50 values in HUVEC, HCT-116, and U373 cells were defined as 3.737, 3.765; 3.513, 3.696, 4.476, and 4.104 μg/mL, respectively. We conducted a cell migration experiment to study the A. turcicum Özhatay & Cowley Extract (ATÖCE) impact on cancer cells' metastatic behavior. Our findings indicate that ATÖCE has an inhibitory effect on the migration potential of the cells used in the study. We conducted experiments using DPPH, ABTS, CUPRAC, and total phenolic content to assess the antioxidant properties of ATÖCE. The findings from the antioxidant activity experiments revealed an activity level of 0.20 ± 0.046 at IC50. Additionally, the total phenolic content was measured to be 0.26 ± 0.044 mg GAE/g. [ABSTRACT FROM AUTHOR]

Oxidative phosphorylation is becoming increasingly important in the induction and development of endometriosis. Recently, it has been reported that ring finger protein 43 (RNF43) is involved in the process of oxidative phosphorylation, but the mechanism remains unclear. Our investigation is to delve into the roles of RNF43 in endometriosis and elucidate the related mechanisms. We found RNF43 was downregulated in ectopic endometrial tissue and primary ectopic endometrial stromal cells (ECESCs). Knockdown of RNF43 enhanced cell viability and migration by activating oxidative phosphorylation in eutopic endometrial stromal cells (EUESCs), while overexpression of RNF43 led to the opposite results. Moreover, RNF43 reinforced the ubiquitination and degradation of NADH dehydrogenase Fe‐S protein 1 (NDUFS1) by interacting with it. Likewise to RNF43 overexpression, NDUFS1 silencing inhibited cell viability, migration, and oxidative phosphorylation in ECESCs. NDUFS1 was a downstream target of RNF43, mediating its biological role in endometriosis. Interestingly, the expression and stability of RNF43 mRNA were regulated by the Methyltransferase‐like 3 (METTL3)/IGF2BP2 m6A modification axis. The results of rat experiments showed decreased RNF43 expression and increased NDUFS1 expression in endometriosis rats, which was enhanced by METTL3 inhibition. Those observations indicated that m6A methylation‐mediated RNF43 negatively affects viability and migration of endometrial stromal cells through regulating oxidative phosphorylation via NDUFS1. The discovery of METTL3/RNF43/NDUFS1 axis suggested promising therapeutic targets for endometriosis. [ABSTRACT FROM AUTHOR]

Medicinal signaling cells (MSC) hold promise for regenerative medicine due to their ability to repair damaged tissues. However, their effectiveness can be affected by how long they are cultured in the lab. This study investigated how passage number influences key properties for regenerative medicine of pig bone marrow MSC. The medicinal signiling cells derived from pig bone marrow (BM-MSC) were cultured in D-MEM High Glucose supplemented with 15% foetal bovine serum until the 25th passage and assessed their growth, viability, ability to differentiate into different cell types (plasticity), and cell cycle activity. Our findings showed that while the cells remained viable until the 25th passage, their ability to grow and differentiate declined after the 5th passage. Additionally, cells in later passages spent more time in a resting phase, suggesting reduced activity. In conclusion, the number of passages is a critical factor for maintaining ideal MSC characteristics. From the 9th passage BM-MSC exhibit decline in proliferation, differentiation potential, and cell cycle activity. Given this, it is possible to suggest that the use of 5th passage cells is the most suitable for therapeutic applications. [ABSTRACT FROM AUTHOR]

In order to meet growing consumer demands in terms of naturalness, the pharmaceutical, food, and cosmetic industries are looking for active molecules of plant origin. In this context, hairy roots are considered a promising biotechnological system for the sustainable production of compounds of interest. Poplars (genus Populus, family Salicaceae) are trees of ecological interest in temperate alluvial forests and are also cultivated for their industrial timber. Poplar trees also produce specialized metabolites with a wide range of bioactive properties. The present study aimed to assess the hybrid poplar hairy root extracts for antimicrobial and antibiofilm activities against four main life-threatening strains of Gram-positive (Staphylococcus aureus, Bacillus subtilis) and Gram-negative (Escherichia coli, Pseudomonas aeruginosa) bacteria. Ethyl acetate extracts from two hairy root lines (HP15-3 and HP A4-12) showed significant antibacterial properties as confirmed by disc diffusion assay. Antibiofilm activities were found to be dose dependent with significant biofilm inhibition (75–95%) recorded at 1000 µg.mL−1 in all the bacterial strains tested. Dose-dependent enhancement in the release of exopolysaccharides was observed in response to treatment with extracts, possibly because of stress and bacterial cell death. Fluorescence microscopy confirmed loss of cell viability of treated bacterial cells concomitant with increased production of reactive oxygen species compared to the untreated control. Overall, this study demonstrates for the first time a high potential of poplar hairy root extracts as a natural and safe platform to produce antimicrobial agents in pharmaceutical, food, industrial water management, or cosmetic industries. [ABSTRACT FROM AUTHOR]

Cell suspension culture has the potential to be a valuable source for the bioactive compound productions. In this study, an optimized procedure was established for callus and cell suspension culture of Physalis alkekengi for the first time, and the impact of static magnetic field (SMF, 6 mT) was studied on the high-value metabolic compounds through investigation of signaling molecules and gene expressions at the late log-to-stationary phase. Results showed that the growth regulators of 6-benzyl amino purine (BAP, 1.5 mg−1 L) and 1-naphthaleneacetic acid (NAA, 0.4 mg−1 L) induced the highest fresh weight, callus rate, callus index, and total withanolides. Cell suspension culture was established in the liquid MS medium supplied with BAP (1.5 mg−1 L) and NAA (0.1 mg−1 L). SMF application decreased slightly the cell growth and viability and enhanced the number of round-shaped cells. The hydrogen peroxide (H2O2) and nitric oxide (NO) levels increased at an all-time series after SMF exposure, and their maximum contents were observed after 12 h. A significant alteration of malondialdehyde content was also identified after 12 h of SMF exposure. The expression of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR), 1-deoxyD-xylulose 5-phosphate synthase (DXS), squalene synthase (SQS), sterol Δ7-reductase (DWF5), and C-7,8 sterol isomerase (HYD1) genes was upregulated significantly after 24 and 48 h. An increase in the total withanolides was related to more activity of HMGR and DXS enzymes in SMF-exposed cells and the maximum physalin A (12.8 mg g−1 DW) and physalin B (1.92 mg g−1 DW) obtained after 24 h compared to controls. Findings suggest that SMF can play a supportive factor in inducing steroidal compounds in P. alkekengi through modulating H2O2 and NO levels and the related-gene expressions. [ABSTRACT FROM AUTHOR]

This research assessed the mutagenicity and DNA damage of a novel type of nano-hydroxyapatite-silica glass ionomer cement (nano-HA-SiO 2 -GIC) and a conventional GIC (cGIC) using Ames and Comet assays. Cell viability was tested on human periodontal ligament fibroblasts (HPLFs) using 3.125 mg/ml, 6.25, 12.5, 25, 50, 100 and 200 mg/ml, on both types of GICs employing MTT assay. For the Comet assay, HPLFs were treated with IC 50 , IC 25 and IC 10 of test materials and the tail moments were measured. In the Ames test, four genotypic variants of strains of Salmonella typhimurium (TA100, TA98, TA1537 and TA1535) and a strain of Escherichia coli (WP2 uvrA) were employed. The material tested was extracted using sterile distilled water (0.2 g per ml) at 37 °C for 72 h. This was considered as 100 %, which was diluted to 50, 25, 12.5 and 6.25 % utilizing sterile distilled water. These five concentrations were incubated with the bacterial strains with and without metabolic activation (S9), along with appropriate positive controls. The number of revertant colonies was used to evaluate the outcome. The highest cell viability (159.4 %) for nano-HA-SiO 2 -GIC was noticed at 3.125 mg/ml, while the lowest (24.26 %) was observed at 200 mg per ml. IC 50 , IC 25 and IC 10 values were 95.27, 51.4 and 20.1 mg/ml for cGIC, 106.9, 55.8 and 22.9 mg/ml for nano-HA-SiO 2 -GIC, respectively. The IC 10 of both test materials showed no significant DNA damage compared to that of the negative control based on the Comet assay. The plate treated with nano-HA-SiO 2 -GIC showed less than double the average number of revertant colonies compared to that of negative control with regard to the Ames test. It can be concluded that nano-HA-SiO 2 -GIC is non-mutagenic based on the Ames test and did not cause DNA damage at the lowest concentration of IC 10 based on the Comet assay. [ABSTRACT FROM AUTHOR]

Melanoma is a highly malignant cancer with a high differentiation potential and metastatic capacity. Sodium butyrate, known for its anti-cancer activity, is used in various types of solid tumors. This study aimed to investigate the effects of sodium butyrate on B16 melanoma cells using in vitro and in vivo mouse models. The study utilized MTT assay, flow cytometry, and immunoblot analysis. Mice were treated with normal saline (control) or 1 mM, 2 mM, 3 mM, or 5 mM sodium butyrate. Results showed that cell viabilities were significantly reduced in 2 mM, 3 mM, and 5 mM sodium butyrate groups after 24 to 48 hours (p < 0.01 for all). Moreover, sodium butyrate exhibited a tumor suppression effect that was time-dependent and lasted for 30 days (p < 0.01 for all). A significant tumor suppression effect was observed in the case of 5 mM sodium butyrate after 30 days (p < 0.001 for all). As compared to control (no sodium butyrate), tumor-associated macrophages were decreased in 2 mM, 3 mM, and 5 mM sodium butyrate groups (<0.01 for all). The maximum reduction was observed in 5 mM sodium butyrate groups. Sodium borate decreased the release of interleukin-10, vascular endothelial growth factor, transforming growth factor beta, and β-actin (<0.01 for all). A significant reduction was observed in the case of 5 mM concentration. Overall, these findings suggest that sodium butyrate is effective in the treatment of melanoma and may offer a promising new avenue for melanoma therapy. [ABSTRACT FROM AUTHOR]

Cinobufagin has inhibitory effects on various tumors, but there are few studies on gynecological tumors. This study explored the function and molecular mechanism of cinobufagin in endometrial cancer (EC). Different concentrations of cinobufagin treated EC cells (Ishikawa and HEC-1). Clone formation, methyl thiazolyl tetrazolium (MTT), flow cytometry, and transwell assays were used to detect malignant behaviors. A Western blot assay was performed to detect protein expression. Cinobufacini was sensitive to the inhibition of EC cell proliferation in a time- and concentration-dependent manner. Meanwhile, EC cell apoptosis was induced by cinobufacini. In addition, cinobufacini impaired the invasive and migratory abilities of EC cells. More importantly, cinobufacini blocked the nuclear factor kappa beta (NF-κB) pathway in EC by inhibiting p-IkBα and p-p65 expression. Cinobufacini suppresses malignant behaviors of EC by blocking the NF-κB pathway. [ABSTRACT FROM AUTHOR]

B-cell lymphoma 2 protein (Bcl-2) is an important regulator of cell apoptosis. Inhibitors that mirror the structural domain 3 (BH3) of Bcl-2 can activate apoptosis in cancer cells, making them a promising target for anticancer treatment. Hence, the present study aimed to investigate potential BH3-mimetic peptides from two vicilin-derived legume proteins from soybean and cowpea bean. The proteins were isolated and sequentially hydrolyzed with pepsin/pancreatin. Peptides < 3 kDa from vicilin-derived proteins from soybean and cowpea beans experimentally inhibited the growth of cultivated breast and prostate cancer cells. In silico analysis allowed the identification of six potential candidates, all predicted to be able to interact with the BH3 domain. The VIPAAY peptide from the soybean β-conglycinin β subunit showed the highest potential to interact with Bcl-2, comparable to Venetoclax, a well-known anticancer drug. Further experiments are needed to confirm this study's findings. [ABSTRACT FROM AUTHOR]

ABSTRACT Vine tea extract (VTE), from the traditional Chinese herbal tea, was added to reconstituted skimmed milk; the mixture was fermented with Lacticaseibacillus casei, and fermentation characteristics, flavonoid content, antioxidant capacity (AOC), and viability of L. casei were measured. 2 mg/mL VTE promoted L. casei growth and 8 mg/mL VTE inhibited growth, an effect consistent with observed pH changes. Total flavonoid content and AOC increased with increasing VTE dosage. Dihydromyricetin was partially metabolized during fermentation and accounted for most of the antioxidant function of VTE. 2 mg/mL VTE was optimal for maintenance of probiotic culture and pH stability during cold storage and improved AOC during product shelf life. VTE has the potential to increase the health benefits of probiotic dairy products, and the resulting mixture may be suitable to use as a daily milk‐based health drink. [ABSTRACT FROM AUTHOR]

Some potentially probiotic strains of lactic acid bacteria (LAB) and yeast that inhabit the digestive tract of humans are known to detoxify xenobiotics, including acrylamide (AA). The objective of the subsequent research was to evaluate the AA-detoxification capability of LAB and yeast isolated from various sources. Namely, the effect of AA was tested on the growth of LAB and yeast strains, as well in the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Subsequently, the AA-binding ability of LAB and yeast was investigated in various environments, including the pH, incubation temperature, cell density, and with inanimate cells. The ability of selected LAB and yeast to reduce the genotoxicity of AA was tested on Caco-2 and Hep-G2 cell lines. The results showed that all tested strains exhibited strong resistance to AA at concentrations of 5, 10, and 50 µg/mL. Also, AA was detected in the intracellular and membrane extracts of tested strains. The most effective binding strain was Pediococcus acidilactici 16 at pH = 5, cell density = 109 CFU/mL, and incubation temperature = 37 °C (87.6% of AA removed). Additionally, all tested strains reduced the genotoxicity of AA, with the greatest reduction observed at the highest concentration of 50 µg/mL. The phenomena of detoxification by potentially probiotic strains could reduce the toxic and harmful effects of AA exposure to humans every day. [ABSTRACT FROM AUTHOR]

Isoniazid (INH) is amongst the first-line antibiotics that have been employed for the treatment of Tuberculosis (TB). Despite its potent anti-tubercular action, susceptibility to rapid hepatic first-pass metabolism and elimination largely limits its oral bioavailability, and has been associated with the induction of drug resistance and adverse effects. This presents study aims at the development and evaluation of unique mannosylated INH loaded solid lipid nanoparticles (Mn-INH-SLNs) for the treatment of TB. The Mn-INH-SLNs demonstrated a particle size of 466 ± 11 nm, which was acceptable for macrophage targeting and had % entrapment efficiency of 80.41 ± 1.37% (n = 6). The dissolution studies depicted a dual-phase drug release profile, i.e., burst release followed by a sustained release, revealing the best fit with the Korsmeyer-Peppas model. The MTT assay (cytotoxicity study) performed utilizing J774A.1 cells with optimized Mn-INH-SLNs deemed them to be safe and nontoxic. Mannosylated SLNs tagged with coumarin-6 (C6 – an established fluorescent marker) showed a substantially high intracellular internalization (1.11-folds) when analyzed through flow cytometric analysis (FACS). The in vivo pharmacokinetic evaluation following per-oral administration in rats demonstrated a significant rise in relative bioavailability (∼5.5-folds) with Mn-INH-SLNs compared to pure drug solution. The bio-distribution (drug disposition) studies exhibited greater lung accumulation of Mn-INH-SLNs (2.13-folds) in comparison to the unconjugated INH-SLNs (Un-INH-SLNs). The promising results of the study depict that the targeted-optimized Mn-INH-SLNs may be a propitious and potent tool to fight TB. [ABSTRACT FROM AUTHOR]

Ovarian cancer is the fifth most prevalent cancer in women. Chemotherapy is a major treatment option for patients with advanced ovarian cancer (OC). Quinoline-2-thione and its derivatives are potential candidates for tumor therapy. In this study, we investigated the anticancer activity of the quinoline-2-thione derivative KA3D against ovarian cancer. The effect of KA3D on the viability of ovarian cancer cells was evaluated using MTT assay, and its effects on apoptosis and the cell cycle were detected using flow cytometry. Western blotting was performed to identify apoptosis-and cell cycle-related proteins altered by KA3D treatment. A xenograft model was used to verify the inhibitory effect of KA3D in vivo. H&E staining, biochemical indicator detection, and blood cell counts were used to observe the toxicity and side effects of KA3D. KA3D treatment impeded cell viability, induced apoptosis, and impeded the G2 phase of the cell cycle in ovarian cancer cells. Mechanistically, we found that KA3D enhanced the expression of proapoptotic molecules such as BAX and Caspase 3, while antiapoptotic proteins such as BCL2 were inhibited. The G0/G1 phase-related protein cyclin D1 was reduced and the G2 phase-related protein cyclin B1 was upregulated. In vivo, KA3D displayed potent anticancer activity, with no apparent toxicity in BABLC/c nude mice bearing SKOV3 cells. KA3D demonstrated remarkable chemotherapeutic drug efficacy in terms of significant cancer suppression in vitro and in vivo with low toxicity. [ABSTRACT FROM AUTHOR]

Background: An ideal aesthetic restorative material should be attached to the tooth tissues by adhesion, have a smooth surface as possible, should not cause toxic reactions in the pulp and discoloration and microleakage. This study aims at comparatively assess the cytotoxicity of current adhesive systems on human dental pulp cells. Materials and methods: The adequate density of human pulp cells was observed from the ready cell line. The passaging was performed and the 3rd passage cells were selected. Adhesive systems and MTA were used on the cultures. Trypan blue staining was conducted on the cells at the 1st, 2nd, 3rd days and a count of live and dead cells using a light microscope. The dead cells whose membrane integrity was impaired by staining with trypan blue and the viability rate was determined using live and dead cell numbers. Data analysis was performed using IBM SPSS Statistics 22. Results: A significant difference in vialibity rates between adhesive systems was observed on the first day. No significant statistical differences were observed on the 2nd and 3rd days (p < 0.05). Conclusion: Futurabond M showed similar biocompatibility with MTA on human pulp cells and it can be applied in cavities with 1–1.5 mm hard tissue between pulp and dentine. [ABSTRACT FROM AUTHOR]

Introduction: The purpose of this study was to investigate the effects and potential mechanisms of ferroptosis‐related gene heat shock protein beta‐1 (HSPB1) on acute myeloid leukemia (AML). Methods: The RNA‐seq and clinical data of AML samples were obtained from the Genomic Data Commons database, and the FerrDb database was used to screen the marker, drive and suppressor of ferroptosis. Besides, DESeq2 was applied for differential expression analysis on AML samples and screening for differentially expressed genes (DEGs). The screened DEGs were subjected to the intersection analysis with ferroptosis‐related genes to identify the ferroptosis‐related DEGs. Next, the functional pathways of ferroptosis‐related DEGs were further be discussed by Gene Ontology as well as Kyoto Encyclopedia of Genes and Genomes enrichment analysis of DEGs. Additionally, lasso regression analysis was employed to determine the differential genes related to prognosis in patients with AML and the survival analysis was performed. Subsequently, quantitative real‐time polymerase chain reaction and western blot assay were applied to detect the mRNA and protein expression levels of HSPB1 in normal/AML bone marrow tissues and human normal (HS‐5)/AML (HL‐60) bone marrow cells, respectively. Furthermore, HSPB1 was knocked down to assess the expression changes of glutathione peroxidase 4 and acyl‐CoA synthetase long‐chain family member 4. Ultimately, the viability and oxidative stress levels of HL‐60 were analyzed by Cell Counting Kit‐8 and biochemical detection. Results: A total of 4986 DEGs were identified in AML samples, with 3324 up‐regulated and 1662 down‐regulated. The enrichment analysis illustrated that ferroptosis‐related DEGs were significantly enriched in response to metal irons, oxidative stress, and other pathways. After lasso regression analysis, 17 feature genes related to the prognosis of patients with AML were obtained, with HSPB1 exhibiting a significant correlation. The reliability of our models was verified by Cox regression analysis and survival analysis of the hazard model. Furthermore, the outcomes of quantitative real‐time polymerase chain reaction and western blot showed that mRNA and protein expression levels of HSPB1 were significantly increased in the AML Group and HL‐60 cells. The knockdown of HSPB1 in HL‐60 cells reduced the protein level of glutathione peroxidase 4, increased the protein level of acyl‐CoA synthetase long‐chain family member 4, decreased the cell viability, and aggravated oxidative stress. Conclusion: Ferroptosis‐related gene HSPB1 is highly expressed in patients with AML. In addition, HSPB1 may be involved in the occurrence and development of AML by regulating oxidative stress and ferroptosis‐related pathways. This study provides new clues for further understanding of AML molecular mechanisms. Also, HSPB1 is expected to be a potential therapeutic target for AML in the future. [ABSTRACT FROM AUTHOR]

AIM: To investigate the impact of platycodin D （PD） on the viability, migration, invasion, apoptosis and cell cycle of osteosarcoma cells in vitro, along with its underlying mechanisms. METHODS: Human osteosarcoma cells MG63 and U2OS were divided into control group （0 μmol/L） and PD treatment group （6. 25, 12. 5, 25, 50 and 100 μmol/L, respectively）. Human osteosarcoma cells MG63 and U2OS were categorized into control groups （0 μmol/L PD） and PD treatment groups （6. 25, 12. 5, 25, 50 and 100 μmol/L）. The CCK-8 assay determined cell viability and identified effective treatment concentrations. MG63 （15 μmol/L PD） and U2OS （25 μmol/L PD） were specifically analyzed. Cell scratch and Transwell assays assessed migration and invasion. Hoechst 33342 staining examined nuclear morphological changes. Flow cytometry analyzed cell cycle distribution and apoptosis rate. Western blot measured protein expression levels: cleaved caspase-3, cleaved PARP, c-Jun N-terminal kinase （JNK）, p-JNK, B-cell lymphoma-2 （Bcl2）, Bcl-2-ssociated X protein （BAX）, matrix metalloproteinase 2 （MMP-2）, MMP-9, cyclin-dependent kinase 4 （CDK4）, cyclin D1, CDK1, cyclin B1, extracellular signal-regulated kinase （ERK） and p-ERK. Proteome sequencing of MG63 cells was performed. RESULTS: PD treatment significantly decreased cell survival, scratch healing rate, and invasive cell numbers, while increasing apoptosis rates （P<0. 05）. Morphological changes such as nuclear hyperchromatism and fragmentation were observed in PD-treated cells. PD induced G2/M phase arrest in MG63 and G0/G1 phase arrest in U2OS cells. PD treatment upregulated BAX, cleaved caspase-3, cleaved PARP, and p-JNK/JNK, while downregulating Bcl-2, MMP-2, MMP-9, CDK4, cyclin D1, CDK1, cyclin B1, and p-ERK/ERK（ P<0. 05）. Proteome sequencing revealed PD's involvement in cell division, cell cycle regulation, focal adhesion, apoptosis, and the MAPK signaling pathway. CONCLUSION: PD inhibits cell viability, migration, and invasion of osteosarcoma cells in vitro, while promoting apoptosis and inducing cell cycle arrest. These effects are likely mediated through modulation of the MAPK signaling pathway. [ABSTRACT FROM AUTHOR]

In this study, we aim to synthesize self-assembled amphiphilic diblock poly(PEGA-b-HEA-PCL) copolymers through RAFT living polymerization, targeting the delivery of hydrophobic anticancer drugs. The synthesized self-assembled diblock copolymers polymeric micelles (PMs) comprising poly(ethylene glycol) methyl ether acrylate (PEGA), as a hydrophilic segment and 2-hydroxyethyl acrylate-polyhexanoate monomer (HEA-PCL) with different block lengths, as a hydrophobic segment. The chemical structures, compositions, and self-assembled behavior were identified through 1H NMR spectroscopy. The thermal stability was assessed through TGA and DSC. Furthermore, DOX was encapsulated into all PMs. The drug-loaded PMs exhibited enhanced drug release profiles in acidic medium. Particle diameter was measured through DLS and TEM techniques. The cell viability of diblock polymers and selected DOX-loaded PMs were evaluated against non-cancerous (L929) and cancerous cells (SK-N-AS), respectively, through well-known MTT assay. Micellar aggregates with mean diameters of approximately 127.2–145.3 nm formed in aqueous solution. The diameters of PMs increased to 141.5–173.1 nm upon the incorporation of DOX. The drug loading content and encapsulation efficiency of PMs were approximately 8.09–18.84% and 30.43–54.07%, respectively. The MTT assay results indicated that all synthesized materials had minimal effects on the viability of L929 cells, while DOX-loaded materials inhibited the viability of neuroblastoma cells by 68.7%. The highest drug release was 89.20% at pH 7.4, while 83.45% at pH 5.0 for 40 h. These findings suggest that the synthesized amphiphilic PMs are promising candidates for drug delivery systems. [ABSTRACT FROM AUTHOR]

BACKGROUND: Breast cancer is one of the main Cancers affecting patients all over the world; however, till now, there has been no successful treatment without any side effects on patient health, but there are groups of medications similar to MDM2 inhibitors that have promising effects. The aim of this study was to find out whether the use of mdm2 inhibitors can help treat breast cancer using three cell lines. The use of treatments belonging to the MDM2 inhibitor alone or with the use of doxorubicin together with a combination of Nutlin-3, Miladometan, Yh239-EE, and doxorubicin in breast cancer cell lines HP100, MCF7. MATERIALS AND METHODS: Cell lines were treated with different concentrations of MDM2 inhibitors 1,5,10, and 20 micromolar alone or in combinations with doxorubicin. After that, cell viability was estimated by the MTT assay method. Then, we have assessed the expression of caspase as an indicator of cell apoptosis after treatment, and the expression of P53 and MDM2 after and before treatment. RESULTS: The IC50 of Doxorubicin in the MCf7 cell line was about 1.12 µM. The best IC50% in MDM2 inhibitors was for Nutlin, about 5.9 µM, then for Yh239-EE about 8.45 µM, and Milademetan about 11.07 µM the high IC50% values in normal epithelial cell HBl100. Also, the MDM2 inhibitor increased P53 levels, as did doxorubicin. CONCLUSION: Our research concluded that Nutlin 3 has a superior effect over Yh239-EE and Miladometan in treating Breast cancer; moreover, the combination group has shown to be more effective than treatment with Doxorubicin or MDM2 inhibitors alone. Interesting information is that Doxorubicin also causes an increase in P53 levels. This result provided us with a promising therapeutic strategy for the treatment of breast cancer. However, more research is required to be conducted on more types of cell lines and in human or animal models [ABSTRACT FROM AUTHOR]

Aflatoxin B1 (AFB1), the most potent toxic and carcinogenic secondary fungal metabolite, has frequently been reported in food/feed. Nowadays, herbal extracts are considered safe dietary additives to reduce the toxicity of such compounds. The protective capability of various combinations of hydro‐alcoholic extracts (HAEs) of ginger, turmeric, and Shirazi thyme, against the toxicity of AFB1 on the RAW264.7 cell line was investigated. The RAW264.7 cells were exposed to six different concentrations of AFB1 (0.09, 0.18, 0.37, 0.75, 1.5, and 3 μg mL−1) for 48 h to determine the IC50 of AFB1. AFB1 was estimated to have an IC50 of 1.5 μg mL−1 for RAW264.7 cells. Then, the cells were simultaneously incubated with 1.5 μg mL−1 AFB1 and the HAEs for 24 h. The HAEs significantly reduced the toxicity of AFB1 in RAW264.7 cells. HAE of Shirazi thyme showed the highest amount of total phenol content (TPC) and the highest DPPH• activity. In addition, a combination of ginger, turmeric, and Shirazi thyme extract showed the highest antioxidant activity. Rutin, quercetin, and apigenin were the main phenolic components of ginger HAE. A significantly positive correlation was observed between TPC of hydro‐alcoholic extract with ferric reducing antioxidant power (FRAP) and 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) values. Consequently, the simultaneous consumption of such extracts is recommended to protect the cells against dietary toxins. [ABSTRACT FROM AUTHOR]

Purpose: Corneal fibroblasts are involved in the wound healing of the cornea with proliferation, migration, and differentiation processes. Coenzyme Q10 (CoQ10) and vitamin E can enhance corneal wound healing when applied after a corneal lesion as an eye drop. Thus, this study was performed to determine the potential efficiency of a CoQ10 ophthalmical solution containing a CoQ10 and vitamin E D-α-tocopherol polyethylene glycol 1000 succinate (TPGS)-derived formulation in human corneal fibroblasts (HCFs) in vitro. Methods: Primary HCFs were obtained from cadaveric corneal tissue, and cell viability was determined using MTT assay at 24 and 72 h. Cell migration was evaluated using an in vitro wound healing assay, and mRNA expressions of collagen type I (COL-I), collagen type III (COL-III), lumican, hyaluronan, matrix metalloproteinase (MMP)-1, MMP-2, MMP-9, tissue inhibitors of MMP (TIMP)-1, TIMP-2, interleukin (IL)-1β, IL-6, IL-8, and IL-10 were assessed using reverse transcription polymerase chain reaction at 24 and 72 h. Results: At various concentrations of CoQ10 ophthalmical solution (CoQ10-os), cell viability and wound healing rates of HCFs increased compared with the control group. The expressions of COL-I, COL-III, lumican, and hyaluronan were increased by CoQ10-os, whereas those of MMP-1, MMP-2, MMP-9, TIMP-1, TIMP-2, and TIMP-3 were not affected by CoQ10-os at 24 and 72 h. In treating HCFs with a CoQ10-os medium, IL-1β, IL-6, and IL-8 decreased, whereas IL-10 was significantly increased in a time- and dose-dependent manner. Conclusions: The findings indicate that CoQ10 and vitamin E-TPGS are potent regulators of the bioactivity of HCFs, thus supporting their potential application as ophthalmical solutions in therapies aimed at the fast regeneration of damaged cornea tissues. [ABSTRACT FROM AUTHOR]

Many clinical procedures and biomedical research workflows rely on microscopy, including diagnosis of cancer, genetic disorders, autoimmune diseases, infections, and quantification of cell culture. Despite its widespread use, traditional image acquisition and review by trained microscopists is often lengthy and expensive, limited to large hospitals or laboratories, precluding use in point‐of‐care settings. In contrast, lensless or lensfree holographic microscopy (LHM) is inexpensive and widely deployable because it can achieve performance comparable to expensive and bulky objective‐based benchtop microscopes while relying on components that cost only a few hundred dollars or less. Lab‐on‐a‐chip integration is practical and enables LHM to be combined with single‐cell isolation, sample mixing, and in‐incubator imaging. Additionally, many manual tasks in conventional microscopy are instead computational in LHM, including image focusing, stitching, and classification. Furthermore, LHM offers a field of view hundreds of times greater than that of conventional microscopy without sacrificing resolution. Here, the basic LHM principles are summarized, as well as recent advances in artificial intelligence integration and enhanced resolution. How LHM is applied to the above clinical and biomedical applications is discussed in detail. Finally, emerging clinical applications, high‐impact areas for future research, and some current challenges facing widespread adoption are identified. [ABSTRACT FROM AUTHOR]

Objective: To evaluate the effect of the deterioration of computer aided design/computer aided manufacturing (CAD/CAM) burs during zirconia milling, on surface roughness, contact angle, and fibroblast viability. Materials and Methods: Ceramic blocks were milled and 75 ceramic disks (8 × 1.5 mm) made and allocated into three groups (n = 25): G1‐brand new 2L and 1L burs, G2‐2L bur at the end of lifetime and brand new 1L bur and G3‐both burs at the end of their lifetimes. Roughness (Ra, Rq, and Rz) was evaluated using a 3D optical profilometer, the contact angle by the sessile drop method and the cell viability of the mouse NIH/3T3 fibroblast, using the Alamar Blue assay at intervals of 24, 48, and 72 h (ISO 10993‐5). Data were analyzed by one‐way ANOVA and Kruskal–Wallis tests (p ≤ 0.05). Results: Roughness increased as the burs deteriorated and G3 (0.27 ± 0.04) presented a higher value for Ra (p < 0.001). The highest contact angle was observed in G3 (86.2 ± 2.66) when compared with G1 (63.7 ± 12.49) and G2 (75.3 ± 6.36) (p < 0.001). Alamar Blue indicated an increase in cell proliferation, with no significant differences among the groups at 24 and 72 h (p > 0.05). Conclusions: The deterioration of the burs increased the surface roughness and decreased the wettability, but did not interfere in cell viability and proliferation. Clinical Significance: The use of custom zirconia abutments represents an effective strategy for single crowns restorations. Our findings suggest that these abutments can be efficiently milled using CAD/CAM burs within their recommended lifetime. [ABSTRACT FROM AUTHOR]

Microalgae are unicellular photosynthetic microorganisms typically found in aquatic environments and they play a vital role in the global carbon and energy cycles. Discrimination of dead and live cells is an important factor in microalgae research and environmental monitoring. Numerous research on the effects of various microalgae has been conducted concerning cell viability. Recently, dyes such as Trypan Blue (TB), Evans Blue (EB), and Neutral Red (NR) have been employed to assess the viability of microalgae. Existing approaches for identifying dead and living microalgal cells all have flaws, such as the requirement for staining and pre-treatment. A machine learning method was created to distinguish the living and dead microalgal cells by using of a digital holography microscopy, and the accuracy of this technique was greater. The machine learning method offers a new way of studying both freshwater and marine microalgal cell cultures. This review focuses on the existing methods and emerging technology for determining dead and living microalgae cells. This review work will enlighten the new research for the detection of live or dead microalgae. [ABSTRACT FROM AUTHOR]

Aim: This study aimed to compare the effects of the molecular targeted drugs, sorafenib and lenvatinib, on the survival, invasion, and angiogenesis of hepatocellular carcinoma cells. Additionally, we investigated the involvement of transforming growth factor beta (TGF‐β) signaling in their molecular mechanisms. Methods: To investigate the effects of sorafenib and lenvatinib, we conducted cell viability, invasion, and angiogenesis assays, as well as western blotting analyses. Results: In human hepatocellular carcinoma cells (HepG2), sorafenib demonstrated potent inhibitory effects on cell proliferation, but induced cell invasion similar to TGF‐β. In contrast, lenvatinib showed weaker cytotoxicity compared with sorafenib, but suppressed cell invasion induced by TGF‐β. The actions of these two molecular targeted drugs were suggested to involve the regulation of the TGFβR2/ERK pathway. Moreover, in human umbilical vein endothelial cells, Sorafenib showed weaker cytotoxicity and enhanced the effects of TGF‐β on angiogenesis. Conversely, lenvatinib showed potent cytotoxic abilities and suppressed angiogenesis induced by TGF‐β. The actions of these two molecular targeted drugs were suggested to involve the regulation of the crosstalk between TGF‐β signaling and vascular endothelial growth factor signaling. Conclusions: Our findings indicate that both sorafenib and lenvatinib possess anticancer abilities by inducing the cytotoxicity of hepatocellular carcinoma cells. Furthermore, they show opposing effects on TGF‐β‐induced cell invasion and angiogenesis, thereby enhancing the understanding of the multifaceted functions of molecular targeted drugs in treating hepatocellular carcinoma. [ABSTRACT FROM AUTHOR]

The high consumption of dietary supplements was a fundamental driver for the creation of the regulatory framework by the Brazilian governmental authorities. However, the regulatory agencies lack official low-cost methodologies to evaluate the quality of food supplements. A preliminary screening method by HPLC-DAD was proposed and validated for screening and quantification of adulterants in dietary supplements. The limits of detection and quantification were <0.11 and 0.37 µg.g−1, respectively. The method was applied for the investigation of ten unauthorized substances (spironolactone, hydrochlorothiazide, furosemide, clenbuterol, testosterone, testosterone propionate, yohimbine, vardenafil, tadalafil, and sildenafil) with a time of analysis of <5 min. Sixteen percent of the 44 samples analyzed had at least one adulterant at or above therapeutic concentrations. Subsequently, in vitro evaluations were performed of the potential cytotoxicity to evaluate the cell viability, DNA damage, determination of nitric oxide levels, and quantification of reactive oxygen species. Despite the necessity of further studies, the results indicate a relationship between the presence of adulterants in food supplements and a potential cytotoxic effect. [ABSTRACT FROM AUTHOR]

Polymers derived from renewable sources, particularly those based on furan, have attracted significant attention within the scientific community due to their potential as an ecologically viable alternative, aiming to minimize the negative impacts arising from the accumulation of plastics derived from fossil hydrocarbons. However, few toxicogenetic studies have been conducted. Therefore, the objective of this study was to assess the cytotoxicity and mutagenicity of starting materials (hexamethylenediamine, maleic anhydride, furan and furan maleic anhydride adduct), as well as a new polymer derived from the furan maleic anhydride adduct. Hexamethylenediamine, maleic anhydride, and furan maleic anhydride adduct exhibited comparable cytotoxic profiles, whereas furan demonstrated lower cytotoxic potential (IC50 of 72.94 ± 2.49 mg/mL), yet it was the only reagent capable of inducing mutagenicity. Furan's mutagenic activity by Ames test was observed in the TA100 strain, both with and without metabolization. The polymer, in turn, exhibited low cytotoxic effect (cell viability exceeding 70%) and demonstrated non-mutagenicity under the experimental conditions, emphasizing its safety concerning toxicological risks. In conclusion, this study contributes to the genotoxicological safety assessment of starting materials, and a new polymer sourced from renewable origins. The absence of mutagenicity and the low cytotoxic potential of the assessed samples contribute to the battery of tests required to indicate safe applications, thereby fostering acceptance, and minimizing associated risks. Among the evaluated reagents, special caution is warranted in handling furan due to its potential to induce gene mutations. [ABSTRACT FROM AUTHOR]

Herein, silver ions modified α-Fe2O3 (Ag: α-Fe2O3) nanoparticles were synthesized using a sol-gel auto-combustion method comprising of dual phase i.e. trigonal structure of α-Fe2O3 and FCC structure of Ag. The refinement results verified the existence of both the phase with R-3c: R s and Fm-3 m space group. Transmission Electron Microscopy (TEM) showed the crystallite size around 34.24 ± 2.00 nm. The percentage of Ag0 and Ag+ verified through X-ray Photoelectron Spectroscopy (XPS) clearly indicate that around 74% of the Ag present is in the metallic form. The cyclic voltammetry showed that the Csp of the Ag: α-Fe2O3 NPs modified electrode is 132 Fg − 1 and 100 Fg − 1 when recorded at a scan rate of 50 mVs− 1 and 75 mVs− 1, respectively. The Zones of Inhibition (ZOI) against bacterial strains generated by utilizing Ag: α-Fe2O3 NPs are clearly showed the ZOI of 19 ± 1 mm against Bacillus subtilis. The synthesized nanoparticles exhibited higher inhibition against the bacterial strains in comparison to the standard antibiotic ampicillin. Further, cell viability analysis suggested to use a concentration of 9 to 12 µg/ml for Ag: α-Fe2O3in invitro experiments. [ABSTRACT FROM AUTHOR]

Cryopreservation of germ cells would facilitate the availability of cells at any time allowing the selection of donors and maintaining quality control for further applications such as transplantation and germline recovery. In the present study, we analyzed the efficiency of four cryopreservation protocols applied either to isolated cell suspensions or to testes fragments from Senegalese sole. In testes fragments, the quality of cryopreserved germ cells was analyzed in vitro in terms of cell recovery, integrity and viability, DNA integrity (fragmentation and apoptosis), and lipid peroxidation (malondialdehyde levels). Transplantation of cryopreserved germ cells was performed to check the capacity of cells to in vivo incorporate into the gonadal primordium of Senegalese sole early larval stages (6 days after hatching (dah), pelagic live), during metamorphosis (10 dah) and at post-metamorphic stages (16 dah and 20 dah, benthonic life). Protocols incorporating dimethyl sulfoxide (DMSO) as a cryoprotectant showed higher number of recovered spermatogonia, especially in samples cryopreserved with L-15 + DMSO (0.39 ± 0.18 × 106 cells). Lipid peroxidation and DNA fragmentation were also significantly lower in this treatment compared with other treatments. An important increase in oxidation (MDA levels) was detected in samples containing glycerol as a cryoprotectant, reflected also in terms of DNA damage. Transplantation of L-15 + DMSO cryopreserved germ cells into larvae during early metamorphosis (10 dah, 5.2 mm) showed higher incorporation of cells (27.30 ± 5.27%) than other larval stages (lower than 11%). Cryopreservation of germ cells using testes fragments frozen with L-15 + DMSO was demonstrated to be a useful technique to store Senegalese sole germline. [ABSTRACT FROM AUTHOR]

The aim of this work was to evaluate the toxicological action of AH Plus (AHP), Bio-C Sealer (BCS), and EndoSequence BC Sealer (ESB), using Drosophila melanogaster as the model organism performing in vivo and ex vivo analysis. D. melanogaster were exposed for 10 days to three concentrations (5 mg/ml, 10 mg/ml, and 20 mg/ml) of AHP, BCS, and ESB sealers mixed with 10 ml of standard diet. During this period, the mortality of flies was evaluated. On the 11th day, the locomotor activity test was performed and the flies were euthanized for oxidative damage analysis (reactive species and lipid peroxidation) and cell viability (resazurin reduction). For the mortality curves evaluation, the log-rank test (Mantel–Cox) was used. For the analysis of other data, a one-way analysis of variance (ANOVA) was applied, followed by Tukey's post hoc test (α = 0.05). Regarding mortality, there were no significant differences. The locomotor activity was reduced, mainly in the two highest concentrations of AHP and BCS. Besides, reactive species generation was bigger in the AHP 20 mg/ml group. AHP induced a lipid peroxidation increase in all three concentrations tested, when compared to other sealers. Considering cell viability, the two highest concentrations of AHP reduced this parameter; while in other sealers, viability was reduced only in the highest concentration. AHP showed changes in oxidative markers that led to greater damage to the flies. [ABSTRACT FROM AUTHOR]

Alginate-based materials have gained significant recognition in the medical industry due to their favorable biochemical properties. As a continuation of our previous studies, we have introduced a new composite consisting of cellulose nonwoven fabric charged with a metallic copper core (CNW-Cu0) covered with a calcium alginate (ALG−Ca2+) layer. The preparation process for these materials involved three main steps: coating the cellulose nonwoven fabric with copper via magnetron sputtering (CNW → CNW-Cu0), subsequent deposition with sodium alginate (CNW-Cu0 → CNW-Cu0/ALG−Na+), followed by cross-linking the alginate chains with calcium ions (CNW-Cu0/ALG−Na+ → CNW-Cu0/ALG−Ca2+). The primary objective of the work was to supply these composites with such biological attributes as antibacterial and hemostatic activity. Namely, equipping the antibacterial materials (copper action on representative Gram-positive and Gram-negative bacteria and fungal strains) with induction of blood plasma clotting processes (activated partial thromboplastin time (aPTT) and prothrombin time (PT)). We determined the effect of CNW-Cu0/ALG−Ca2+ materials on the viability of Peripheral blood mononuclear (PBM) cells. Moreover, we studied the interactions of CNW-Cu0/ALG−Ca2+ materials with DNA using the relaxation plasmid assay. However, results showed CNW-Cu0/ALG−Ca2+'s cytotoxic properties against PBM cells in a time-dependent manner. Furthermore, the CNW-Cu0/ALG−Ca2+ composite exhibited the potential to interact directly with DNA. The results demonstrated that the CNW-Cu0/ALG−Ca2+ composites synthesized show promising potential for wound dressing applications. [ABSTRACT FROM AUTHOR]

The substance principal used to synthesize these materials is fibroin, obtained from the silkworm's cocoon (Bombyx mori) in the form of a filament, but can be converted into nanometric particles. After the synthesis, the obtained samples were characterized by size and Zeta potential (via nanosizer), X-Ray diffraction (XRD), specific area determination by the BET method, Infrared Spectroscopy with Fourier Transform with attenuated diffuse reflectance (FTIR), transmission electron microscopy (TEM). The silk fibroin nanoparticles (SFNp) were immobilized on polylactic acid (PLA)-knitted fabric and functionalized with chitosan for FTIR and XRD analyses, in addition to cytotoxicity analysis with colorimetric test (MTT) and cell adhesion. The DLS, FTIR and MET results confirm the formation of spherical silk fibroin nanoparticles with average sizes between 48 and 156nm, and zeta potential of − 19.95mV which characterizes a good dispersibility. Cell-viability assessment showed that SFNp is a non-cytotoxic material at low concentrations up to 48h, confirming that the nanoparticle synthesis method using the ternary system is an effective and low-cost alternative. This technique has great potential for use in the development of biomaterials, due to the production of knitted tissues with PLA functionalized with chitosan and immobilized with SFNp, exhibiting mechanical strength and biocompatibility. [ABSTRACT FROM AUTHOR]

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We present a single-step nonthermal plasma method for the synthesis of near-infrared (NIR)-emitting and water-soluble Si quantum dots (QDs) for bioimaging applications. Oxygen gas and water vapor were introduced together with acrylic acid (AA) into the afterglow region of the synthesis plasma leading to the surface functionalization of the upstream synthesized Si QDs. The simultaneous surface oxidation and ligand grafting enabled solubility and colloidal stability of the Si QDs in water, as evidenced by strongly reduced hydrodynamic diameters. Aged Si QDs in water emitted NIR photoluminescence (PL) at around 830 nm. The PL quantum yield of the Si QDs in water increased over time from initially undetectable to ∼30% after 8 days. Cell viability tests showed that >70% of 3T3 cells survived for 24 h at a concentration of 200 μg/mL of oxidized AA grafted Si QDs. The water solubility, NIR emission with a high quantum yield, and cell viability make the Si QDs promising for bioimaging applications. [ABSTRACT FROM AUTHOR]

Breast cancer remains a significant global health challenge, spurring ongoing investigations into innovative treatment approaches. Low-level laser therapy (LLLT) has emerged as a promising non-invasive therapeutic avenue of interest. This research delves into the impact of LLLT on the cytotoxicity of the MCF-7 breast cancer cell line, employing lasers emitting various wavelengths. The objective is to assess whether diverse LLLT wavelengths elicit disparate cytotoxic responses, shedding light on LLLT's potential as a targeted breast cancer treatment. MCF-7 cell cultures were subjected to lasers of varying wavelengths, including blue (473 nm), red (660 nm), and near-infrared (780 nm). Each wavelength was delivered at four different power levels: 10, 25, 45, and 65 mW, with exposure durations of 60, 300, 600, and 900 s. Cellular responses, encompassing factors such as cell viability, and cytotoxicity were assessed using WST-1 assays technique. Statistical analysis was performed to discern the wavelength-specific impacts of low-level laser therapy (LLLT) on MCF-7 cells. The study revealed that the blue laser had the least noticeable adverse impact on MCF-7 breast cancer cell lines, leading to the highest cell survival rate of 107.62% after 24 h. The most severe toxicity occurred when the laser was used at 45 mW for 900 s, resulting in cell viability ranging from 81.85% to 107.62%. As for cell viability after exposure to the red laser, the mildest harmful effect was observed at 45 mW power for 60 s, resulting in a cell survival rate of 147.62%. Conversely, the most significant toxic response occurred at 10 mW power for 60 s, resulting in a cell viability of 91.56%. In contrast, when employing infrared laser irradiation, the least substantial cytotoxic effect on MCF-7 cells was observed at 10 mW power for 600 s, resulting in the highest cell viability of 109.37% after 24 h. The most pronounced cytotoxic effect was observed by infrared laser (780 nm) at 25 mW power for 900 s, leading to the lowest viability of 32.53%. [ABSTRACT FROM AUTHOR]

ABSTRACT Introduction Methods Results Conclusions The standard flow cytometry method for viability testing using 7‐aminoactinomycin D (7‐AAD) determines cells in necrosis and late apoptosis. The colony‐forming unit (CFU) assay, which evaluates the proliferation ability of HSCs, is also used in graft quality assessment despite known deficiencies that make this assay impractical in routine clinical settings. The aim was to compare the effectiveness of the flow cytometry 7‐AAD/annexin V method with the 7‐AAD method in assessing the quality of HSCs in autologous and allogeneic peripheral blood stem cell (PBSC) products.Thirty autologous and 30 allogeneic fresh and thawed cryopreserved PBSC products were included in this study. The viability of HSCs was determined using the 7‐AAD method and 7‐AAD/annexin V method on a flow cytometer, while their clonogenic capacity was assessed by CFU assay.There was an excellent correlation for CD34+ cell viability between the 7‐AAD and the 7‐AAD/annexin V method for fresh samples (Rs = 0.930, p < 0.001) and a good correlation for thawed PBSC samples (Rs = 0.739, p < 0.001). Excellent correlation was observed for post‐thaw CD34+ cell recovery between the two methods for viability (Rs = 0.980, p < 0.001). Statistical analysis showed a weak correlation between CFU‐GM recovery and CD34+ cell recovery, regardless of which viability testing method was used (7‐AAD method p = 0.021, Rs = 0.298; 7‐AAD/annexin V method p = 0.029, Rs = 0.282).Results of this study showed that in the quality assessment of cryopreserved PBSC product viability, the 7‐AAD/annexin V method had no added value compared to the 7‐AAD method, which was suitable enough for routine quality control of cryopreserved autologous and allogeneic PBSC samples. [ABSTRACT FROM AUTHOR]

In the pharmaceutical sector, solid lipid nanoparticles (SLN) are vital for drug delivery incorporating a lipid core. Chondroitin sulfate (CHON) is crucial for cartilage health. It is often used in osteoarthritis (OA) treatment. Due to conflicting results from clinical trials on CHON's efficacy in OA treatment, there has been a shift toward exploring effective topical systems utilizing nanotechnology. This study aimed to optimize a solid lipid nanoparticle formulation aiming to enhance CHON permeation for OA therapy. A 3 × 3 × 2 Design of these experiments determined the ideal parameters: a CHON concentration of 0.4 mg/mL, operating at 20,000 rpm speed, and processing for 10 min for SLN production. Transmission electron microscopy analysis confirmed the nanoparticles' spherical morphology, ensuring crucial uniformity for efficient drug delivery. Cell viability assessments showed no significant cytotoxicity within the tested parameters, indicating a safe profile for potential clinical application. The cell internalization assay indicates successful internalization at 1.5 h and 24 h post-treatment. Biopharmaceutical studies supported SLNs, indicating them to be effective CHON carriers through the skin, showcasing improved skin permeation and CHON retention compared to conventional methods. In summary, this study successfully optimized SLN formulation for efficient CHON transport through pig ear skin with no cellular toxicity, highlighting SLNs' potential as promising carriers to enhance CHON delivery in OA treatment and advance nanotechnology-based therapeutic strategies in pharmaceutical formulations. [ABSTRACT FROM AUTHOR]

Cryopreservation is essential for the broad clinical application of mesenchymal stem cells (MSCs), yet its impact on their cellular characteristics and cardiomyogenic differentiation potential remains a critical concern in translational medicine. This study aimed to evaluate the effects of cryopreservation on the biological properties and cardiomyogenic capacity of rat adipose-derived MSCs (AD-MSCs). We examined their cellular morphology, surface marker expression (CD29, CD90, CD45), trilineage differentiation potential (adipogenic, osteogenic, chondrogenic), and gene expression profiles for the pluripotency marker REX1 and immunomodulatory markers TGFβ1 and IL-6. After inducing cardiomyocyte differentiation, we assessed cardiac-specific gene expressions (Troponin I, MEF2c, GSK-3β) using quantitative RT-qPCR, along with live/dead cell staining and immunofluorescence for cardiac-specific proteins (Troponin T, α-actinin, Myosin Heavy Chain). Cryopreserved AD-MSCs preserved their morphology, surface markers, and differentiation potential, but exhibited a reduced expression of REX1, TGFβ1, and IL-6. Additionally, cryopreservation diminished cardiomyogenic differentiation, as indicated by the lower levels of Troponin I, MEF2c, and GSK-3β seen compared to non-cryopreserved cells. Despite this, high cell viability (>90%) and maintained cardiac protein expression were observed post-cryopreservation. These findings highlight the necessity of optimizing cryopreservation protocols to ensure the full therapeutic potential of AD-MSCs, particularly in applications related to cardiac regenerative medicine. [ABSTRACT FROM AUTHOR]

Background: Fetal calf serum (FCS), an existing cell culture supplement, is effective but has several drawbacks, including being expensive, requiring a lengthy process of production, and requiring a hard currency. With this in mind, we planned to evaluate chick embryo extract and egg yolk extracts in cell culture as alternatives to fetal calf serum (FCS). Methods: Specific pathogen-free eggs were purchased from the National Veterinary Institute, Bishoftu, Ethiopia, and incubated in a humidified incubator at 37 °C for 11 days. Egg yolk extract (EYE) and chick embryo extract (CEE) were collected after the egg was opened with caution not to destroy the yolk sack or the chick embryo itself. Chick fibroblasts and Vero cells were cultured in minimum essential medium (MEM) supplemented with egg yolk extract or chick embryo extract at ratios of 0:10, 1:9, 2.5:7.5, and 5:5% fetal calf serum. Results: Fibroblast cell attachment was better in media supplemented with 5% CEE and 5% FCS. The confluency was also greater than 50% at this concentration. Vero cells cultured with 5% CEE and 5% FCS also exhibited very good cell attachment and a confluency of up to 70%. Viability and confluency were also observed at 5%:5% ratios of 50 and 70%, respectively. Conclusion: This investigation evaluated these two extracts as cell culture supplements and revealed promising results as alternatives to fetal calf serum. The limitation of this study is that it only used two cell types and additional cell lines, and different ratios should be tested. With the above findings, further research using different cell lines, ratios and conditions is warranted. [ABSTRACT FROM AUTHOR]

Delivering foreign molecules and genetic material into cells is a crucial process in life sciences and biotechnology, resulting in great interest in effective cell transfection methods. Importantly, physical transfection methods allow delivery of molecules of different chemical composition and are, thus, very flexible. Here, we investigated the influence of microwave radiation on the transfection and survival of mammalian cells. We made use of an optimized microwave-poration device and analyzed its performance (frequency and electric field strength) in comparison with simulations. We, then, tested the effect of microwave irradiation on cells and found that 18 GHz had the least impact on cell survival, viability, cell division and genotoxicity while 10 GHz drastically impacted cell physiology. Using live-cell fluorescence microscopy and image analysis, we tested the uptake of small chemical substances, which was most efficient at 18 GHz and correlated with electric field strength and frequency. Finally, we were able to obtain cellular uptake of molecules of very different chemical composition and sizes up to whole immunoglobulin antibodies. In conclusion, microwave-induced poration enables the uptake of widely different substances directly into mammalian cells growing as adherent cultures and with low physiological impact. [ABSTRACT FROM AUTHOR]

Hyperplastic scar (HS) is an overreaction of tissue to skin injury caused by local fibroblast proliferation and excessive collagen production. Histone posttranslational modification patterns are important epigenetic processes that control various biological activities. This study was designed to investigate the effects of histone lactylation on HS and the underlying mechanism. Western blot was used to analyse the lactylation level in HS patients and fibroblasts (HSFs). In vitro experiments, western blot, cell counting kit‐8, and immunofluorescence staining were performed to detect the collagen level, cell viability, and autophagy, respectively. The relationship between snai2 (SLUG) and phosphatase and tensin homologue (PTEN) was assessed by RNA immunoprecipitation and dual‐luciferase reporter assays. The results showed that the histone lactylation level was upregulated in HS tissues and HSFs. HSFs showed increased collagen production and cell viability, and decreased autophagy. Silencing of lactate dehydrogenase A (LDHA) promoted the transcription of PTEN by inhibiting SLUG, thus promoting autophagy. Knockdown of LDHA inhibited collagen deposition and cell viability, and increased autophagy in HSFs, and the results were reversed after PTEN inhibition. In summary, histone lactylation inhibited the transcription activity of PTEN by promoting SLUG, thereby suppressing autophagy and promoting collagen deposition and cell viability of HSFs, which might provide effective therapeutic strategies in HS. [ABSTRACT FROM AUTHOR]

Cellular therapy development and manufacturing has focused on providing novel therapeutic cell-based products for various diseases. The International Organization for Standardization (ISO) has provided guidance on critical quality attributes (CQAs) that shall be considered when testing and releasing cellular therapeutic products. Cell count and viability measurements are two of the CQAs that are determined during development, manufacturing, testing, and product release. The ISO Cell Counting Standard Part 1 and 2 addressed the needs for improving the quality of cell counting results. However, there is currently no guidance on the qualification and selection of a fit-for-purpose cell viability detection method. In this work, we present strategies for the characterization and comparison of AO/PI and AO/DAPI staining methods using the heat-killed (HK) and low temperature/nutrient-deprived (LT/ND) cell death models to evaluate the comparability of cell viability measurements and identify potential causes of differences. We compared the AO/PI and AO/DAPI staining methods using HK and LT/ND-generated dead cells, investigated the staining time effects on cell viability measurements, and determined their viability linearity with different mixtures of live and dead cells. Furthermore, we validated AO/PI and AO/DAPI cell viability measurement with a long-term cell proliferation assay. Finally, we demonstrate a practical example of cell viability measurement comparison using AO/PI and AO/DAPI on antibiotic-selected transduced Jurkat and THP-1 cells to select a fit-for-purpose method for functional genomics screening. The proposed strategies may potentially enable scientists to properly characterize, compare, and select cell viability detection methods that are critical for cellular therapeutic product development and manufacturing. [ABSTRACT FROM AUTHOR]

This paper reports an experimental study on the compatibility of human bronchial epithelial (HBE) cells in a collagen–alginate bioink. The compatibility was assessed using the culture well method with three bioink compositions prepared from a 10% alginate solution and neutralized TeloCol-10 mg/mL collagen stock solution. Cell viability, quantified by (live cell count—dead cell count)/live cell count within the HBE cell-laden hydrogel, was evaluated using the live/dead assay method from Day 0 to Day 6. Experimental results demonstrated that the collagen–alginate 4:1 bioink composition exhibited the highest cell viability on Day 6 (85%), outperforming the collagen–alginate 1:4 bioink composition and the alginate bioink composition, which showed cell viability of 75% and 45%, respectively. Additionally, the live cell count was highest for the collagen–alginate 4:1 bioink composition on Day 0, a trend that persisted through Days 1 to 6, underscoring its superior performance in maintaining cell viability and promoting cell proliferation. These findings show that the compatibility of HBE cells with the collagen–alginate 4:1 bioink composition was higher compared with the other two bioink compositions. [ABSTRACT FROM AUTHOR]

Imidazole derivatives are considered potential chemical compounds that could be therapeutically effective against several harmful pathogenic microbes. The chemical structure of imidazole, with a five-membered heterocycle, three carbon atoms, and two double bonds, tends to show antibacterial activities. In the present study, novel imidazole derivatives were designed and synthesized to be evaluated as antimicrobial agents owing to the low number of attempts to discover new antimicrobial agents and the emerging cases of antimicrobial resistance. Two imidazole compounds were prepared and evaluated as promising candidates regarding in vitro cytotoxicity against human skin fibroblast cells and antimicrobial activity against several bacterial strains. The synthesized imidazole derivatives were chemically identified using nuclear magnetic resonance (NMR) and Fourier-transform infrared spectroscopy (FTIR). The results demonstrated a relatively high cell viability of one of the imidazole derivatives, i.e., HL2, upon 24 and 48 h cell exposure. Both derivatives were able to inhibit the growth of the tested bacterial strains. This study provides valuable insight into the potential application of imidazole derivatives for treating microbial infections; however, further in vitro and in vivo studies are required to confirm their safety and effectiveness. [ABSTRACT FROM AUTHOR]

Simple Summary: Our study identified specific conditions for optimal cell viability in enzymatic dissociation protocols across various endocrine tumors. For adrenal medullary tumors, the critical factor was the dissociation time, with 20 min of incubation proving most effective. In the case of adrenocortical tumors, using Collagenase IV or the MTDK enzyme kit, along with a post-dissociation procedure involving a debris removal kit, resulted in higher cell viability. No significant patterns were observed for thyroid carcinomas and pituitary neuroendocrine tumors, indicating that the effectiveness of dissociation protocols may vary significantly depending on the tissue type and its unique morphological characteristics. These findings underscore the importance of tailored dissociation protocols for different tumor types to ensure optimal outcomes in cell isolation processes. The enzymatic dissociation of human solid tissues is a critical process for disaggregating extracellular matrix and the isolation of individual cells for various applications, including the immortalizing primary cells, creating novel cell lines, and performing flow cytometry and its specialized type, FACS, as well as conducting scRNA-seq studies. Tissue dissociation procedures should yield intact, highly viable single cells that preserve morphology and cell surface markers. However, endocrine tissues, such as adrenal gland tumors, thyroid carcinomas, and pituitary neuroendocrine tumors, present unique challenges due to their complex tissue organization and morphological features. Our study conducted a morphological examination of these tissues, highlighting the intricate structures and secondary degenerative changes that complicate the dissociation process. We investigated the effects of various dissociation parameters, including the types of enzymes, incubation duration, and post-dissociation purification procedures, such as debris removal and nontarget blood cell lysis, on the viability of cells derived from different tumor types. The findings emphasize the importance of optimizing tissue digestion protocols to preserve cell viability and integrity, ensuring reliable outcomes for downstream analyses. [ABSTRACT FROM AUTHOR]

Introduction: Vitex negundo has a myriad of medicinal uses, such as antioxidant, analgesic and anthelminthic properties, and it is used for dysmenorrhoea. The plant contains various phytochemicals, such as flavonoids, vitamins and casticin. All components of the plant are utilised medicinally due to the minimal occurrence of adverse drug reactions, making it a potential drug target for various chronic diseases. However, the role of this plant in neurodegenerative disorders, such as Alzheimer's Disease (AD), has not yet been extensively studied. Aim: To evaluate the in-silico and in-vitro activity of the anti-Alzheimer properties of the Methanolic Extract of Vitex negundo (MEVN) leaves. Materials and Methods: The present in-silico docking study and in-vitro study were conducted in the Department of Pharmacology at Sri Ramachandra Institute of Higher Education and Research, Porur, Chennai, Tamil Nadu, India, over a period of two months. The in-silico analysis was performed to determine the intermolecular interactions between the ligand and protein of the top five scored molecules using PyMol software. To assess the Acetylcholinesterase (AChE) inhibitory property, the findings were presented as docking scores. Cell viability and cytotoxicity were evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, with results expressed as percentages for various concentrations: 6.25 µg/mL, 12.5 µg/mL, 25 µg/mL, 50 µg/mL, 75 µg/mL, and 100 µg/mL of MEVN. Results: The MEVN exhibited competitive inhibition of the AChE enzyme. There were active interactions between Agnuside, Isochlorogenic Acid B, Isochlorogenic Acid C, Kaempferol-3-O-rutinoside, and Quercetin found in the MEVN with AChE. The percentage of cell viability for the concentrations of 6.25 µg/mL, 12.5 µg/mL, 25 µg/mL, 50 µg/mL, 75 µg/mL, and 100 µg/mL, as determined by the MTT assay, was 98.62%, 98.86%, 97.19%, 96.66%, 91.86%, and 78.82%, respectively. The results indicated that MEVN does not exhibit any toxicity. Conclusion: The findings demonstrated that MEVN possesses AChE inhibitory properties and maintains cell viability, both of which are indicative of anti-Alzheimer activity. Therefore, MEVN can be further evaluated for its potential in preventing cell cytotoxicity and treating AD. [ABSTRACT FROM AUTHOR]

When humans consume seafood contaminated by lipophilic polyether phycotoxins, such as azaspiracids (AZAs), the toxins are mainly leached and absorbed in the small intestine, potentially causing intestinal damage. In this study, human intestinal epithelial Caco-2 cells were used to investigate the adverse effects of azaspiracid-2 (AZA-2) on human intestinal epithelial cells. Cell viability, apoptosis, oxidative damage and mitochondrial ultrastructure were investigated, and ribonucleic acid sequence (RNA-seq) analysis was applied to explore the potential mechanisms of AZA-2 toxicity to Caco-2 cells. Results showed that AZA-2 significantly reduced the proliferation of Caco-2 cells in a concentration-dependent response, and the 48 h EC50 of AZA-2 was 12.65 nmol L−1. AZA-2 can induce apoptosis in Caco-2 cells in a dose-dependent manner. Visible mitochondrial swelling, cristae disintegration, membrane rupture and autophagy were observed in Caco-2 cells exposed to AZA-2. Reactive oxygen species (ROS) and malondialdehyde (MDA) content were significantly increased in Caco-2 cells after 48 h of exposure to 1 and 10 nmol L−1 of AZA-2. Transcriptome analysis showed that KEGG pathways related to cellular oxidative damage and lipid metabolism were affected, mainly including mitophagy, oxidative phosphorylation, cholesterol metabolism, vitamin digestion and absorption, bile secretion and the peroxisome proliferator-activated receptor signaling pathway. The cytotoxic effects of AZA-2 on Caco-2 cells may be associated with ROS-mediated autophagy and apoptosis in mitochondrial cells. Results of this study improve understanding of the cytotoxicity and molecular mechanisms of AZA-2 on Caco-2 cells, which is significant for protecting human health. [ABSTRACT FROM AUTHOR]