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Background/Objectives: The primary method used to pharmacologically arrest cancer development and its metastasis is to disrupt the cell division process. There are a few approaches that may be used to meet this objective, mainly through inhibiting DNA replication or mitosis. Despite intensive studies on new chemotherapeutics, the biggest problem remains the side effects associated with the inhibition of cell division in non-tumoural host cells. Methods: The efficacy and selectivity of the kojic acid derivative (L1) was studied in vitro with the use of tumoural (Caco2, SW480, HT29, T98G) and non- tumoural (HEK293T, RAW) cell lines. Light and electron microscopy observations were supported by the next generation sequencing (NGS), cytoflow, and spectroscopy analysis of mRNA and biomolecules, respectively. Results: The light and electron microscopy observations showed that L1 treatment leads to significant morphological changes in Caco2 cells, which are characteristic of mitosis arrest. Moreover, the fluorescent tubulin staining revealed the formation of tubulin ring structure associated with the apoptotic stage. Mitotic exit into apoptosis was further conformed by the cytoflow of early/late apoptosis stages and caspase-3 analysis. NGS investigation showed differentiated expressions of genes involved in mitosis and apoptosis processes. The observed IC50 in tumoural cell lines were as follows: Caco2 (IC50 = 68.2 mM), SW480 (IC50 = 15.5 mM), and HT29 (IC50 = 4.7 mM). Conclusions: The findings presented here suggest that L1 could be a valid candidate for oral prevention and/or chemotherapy in colorectal cancer. Considering high selectivity of L1 versus tumoural cell lines, more in-depth mechanistic studies could reveal unknown stages in carcinogenesis. [ABSTRACT FROM AUTHOR]

Environmental stressors such as micro‐ and nanosized plastic particles (MNPs) or crude oil have a detrimental effect on aquatic animals; however, the impact upon the cardiovascular system of fish remains relatively under‐researched. This study presents a novel approach for investigating the effect of crude oil and MNPs on the cardiac system of fish. We used salmonid larvae and cardiac cell cultures derived from hearts of salmonid fish and exposed them to environmental stressors. Following exposure to plastic particles or crude oil, the larvae exhibited some variation in contraction rate. In contrast, significant alterations in the contraction rate were observed in all cardiac cell cultures. The greatest differences between the control and treatment groups were observed in cardiac cell cultures derived from older brown trout. Following 7 days of exposure to MNPs or crude oil in Atlantic salmon larval hearts or cardiac cell cultures, there were only minor responses noted in mRNA expression of the selected marker genes. These findings show the use of a novel in vitro technique contributing to the existing body of knowledge on the impact of MNPs and crude oil on the cardiovascular system of salmonids and the associated risk. [ABSTRACT FROM AUTHOR]

Alzheimer's disease (AD) is a major contributor to dementia and the most common neurodegenerative disorder. In AD pathophysiology, matrix metalloproteinases (MMPs)—proteolytic enzymes, best known to be responsible for remodeling and degradation of the extracellular matrix—were suggested to play an important role. Due to the diverse nature of the published data and frequent inconsistent results presented in available papers, it was considered essential to analyze all aspects of MMP literature with respect to AD pathophysiology and attempt to outline a unifying concept for understanding their role in AD. Thus, the main contribution of this review article is to summarize the most recent research on the participation of MMP in AD pathophysiology obtained using the cell cultures to understand the molecular principles of their action. Furthermore, an updated comprehensive view regarding this topic based exclusively on papers from human studies is provided as well. It can be concluded that determining the exact role of any particular MMPs in the AD pathophysiology holds promise for establishing their role as potential biomarkers reflecting the severity or progression of this disease or for developing new therapeutic agents targeting the processes that lead to AD. [ABSTRACT FROM AUTHOR]

The nociceptin receptor (NOP) and nociceptin are involved in the pathways of pain and inflammation. The potent role of nuclear factor-κB (NFκB) in the modulation of tumor necrosis factor-α (TNF-α) and interleukin (IL)-1β on the nociceptin system in human THP-1 cells under inflammatory conditions were investigated. Cells were stimulated without/with phorbol-myristate-acetate (PMA), TNF-α, IL-1β, or PMA combined with individual cytokines. To examine NFκB's contribution to the regulation of the nociceptin system, PMA-stimulated cells were treated with NFκB inhibitor BAY 11-7082, JSH-23, or anacardic acid before culturing with TNF-α or IL-1β. NOP and prepronociceptin (ppNOC) mRNA were quantified by RT-qPCR; cell membrane NOP and intracellular nociceptin protein levels were measured by flow cytometry. Phosphorylation and localization of NFκB/p65 were determined using ImageStream. PMA + TNF-α decreased NOP mRNA compared to stimulation with PMA alone, while PMA + IL-1β did not. BAY 11-7082 and JSH-23 reversed the repression of NOP by PMA + TNF-α. TNF-α and IL-1β attenuated PMA's upregulating effects on ppNOC. None of the inhibitors preserved the upregulation of ppNOC in PMA + TNF-α and PMA + IL-1β cultures. TNF-α strongly mediated the nuclear translocation of NFκB/p65 in PMA-treated cells, while IL-1β did not. Proinflammatory cytokines suppressed NOP and ppNOC mRNA in PMA-induced human THP-1 cells. NFκB signaling seems to be an important regulator controlling the transcription of NOP. These findings suggest that the nociceptin system may play an anti-inflammatory role during immune responses. [ABSTRACT FROM AUTHOR]

The rapid progress in nanotechnology has introduced multifunctional iron oxide nanoparticles as promising agents in cancer treatment. This research focused on the synthesis and assessment of citric-acid-coated, folic-acid-conjugated nanoparticles loaded with doxorubicin, evaluating their therapeutic potential in tumor models. An advanced automated continuous technology line (CTL) utilizing a controlled co-precipitation method was employed to produce highly dispersive, multifunctional nanofluids with a narrow size distribution. Various techniques, including dynamic light scattering (DLS), electrophoretic light scattering (ELS), X-ray diffraction (XRD), and transmission electron microscopy (TEM), were employed to examine the particle size, zeta potential, structure, and morphology. Magnetic properties were analyzed through vibrating sample magnetometry (VSM), and surface modifications were confirmed via UV-visible (UV-Vis) and Fourier-Transform Infrared (FTIR) spectroscopy. Cytotoxicity and drug delivery efficiency were evaluated in vitro using RM1 (prostate cancer) and MEC1 (chronic lymphocytic leukemia) cell lines. Fluorescence microscopy demonstrated the successful intracellular delivery of doxorubicin, showcasing the nanoparticles' potential for targeted cancer therapy. However, folic-acid-conjugated nanoparticles exhibited diminished effectiveness over time. This study highlights the importance of nanoparticle optimization for enhancing therapeutic performance. Further research should aim to improve nanoparticle formulations and explore their long-term impacts for the development of safe, targeted cancer treatments. [ABSTRACT FROM AUTHOR]

Diabetic retinopathy is a major eye complication in patients with diabetes mellitus, and it is the leading cause of blindness and visual impairment in the world. Chronic hyperglycemia induces endothelial damage with consequent vascular lesions, resulting in global vasculitis, which affects the small vessels of the retina. These vascular lesions cause ischemic conditions in certain areas of the retina, with a consequent increase in the release of pro-angiogenic mediators. In addition to pharmacological interventions for controlling the blood glycaemic level, the main strategies for treating diabetic retinopathy are the intravitreal injections of drugs, surgical treatments, and vitrectomies. The complexity of diabetic retinopathy is due to its close interactions with different cell types (endothelial cells, astrocytes, and Müller cells). The evaluation of the efficacy of novel pharmacological strategies is mainly performed through in vivo models. However, the use of different animal species leads to heterogenic results and ethical concerns. For these reasons, the development of new and reliable in vitro models, such as cell co-cultures and eye organoids, represents an urgent need in this area of research. This review features an overview of the in vitro models used to date and highlights the advances in technology used to study this pathology. [ABSTRACT FROM AUTHOR]

Skeletal muscle formation involves tight interactions between muscle cells and associated connective tissue fibroblasts. Every muscle displays the same type of organisation, they are innervated in the middle and attached at both extremities to tendons. Myonuclei are heterogeneous along myotubes and regionalised according to these middle and tip domains. During development, as soon as myotubes are formed, myonuclei at muscle tips facing developing tendons display their own molecular program. In addition to molecular heterogeneity, a subset of tip myonuclei has a fibroblastic origin different to the classical somitic origin, highlighting a cellular heterogeneity of myonuclei in foetal myotubes. To gain insights on the functional relevance of myonucleus heterogeneity during limb development, we used 2D culture and co-culture systems to dissociate autonomous processes (occurring in 2D-cultures) from 3D-environment of tissue development. We also assessed the role of muscle contraction in myonucleus heterogeneity in paralysed limb muscles. The regionalisation of cellular heterogeneity was not observed in 2D cell culture systems and paralyzed muscles. The molecular signature of MTJ myonuclei was lost in a dish and paralysed muscles indicating a requirement of 3D-enviroment and muscle contraction for MTJ formation. Tip genes that maintain a regionalized expression at myotube tips in cultures are linked to sarcomeres. The behaviour of regionalized markers in cultured myotubes and paralyzed muscles allows us to speculate whether the genes intervene in myogenesis, myotube attachment or MTJ formation. Highlights: • The molecular signature of MTJ myonuclei is lost in cultured myotubes and paralysed muscles • Genes expressed in muscle tips that maintain their regionalised expression in cultured myotubes are linked to sarcomeric proteins • Cellular heterogeneity of myonuclei is observed in cultured myotubes but with no regionalisation • BMP signalling regulates fibroblast nucleus incorporation into cultured myotubes [ABSTRACT FROM AUTHOR]

Iron oxide nanoparticles (FeO-NPs) have gained global attention as bio safe elicitors for incrementing medicinal metabolites in plants. This study, for the first time evaluated the effects of FeO-NPs under varying light intensities to enhance growth, biomass and production of anti-leishmanial natural products in Artemisia scoparia cell cultures. Results showed that optimal levels of FeO-NPs (10–15 mg/L) significantly enhanced callus growth attributes and metabolites production on solid MS media. Highest callus induction frequency (92%) and biomass accumulation (33 g/L), were observed in explants treated with 15 mg/L FeO-NPs combined with 1.5 mg/L 2,4-D. However, higher levels of Total phenolic content (TPC:37 mg GAE/g DW), Total flavonoid content (TFC:9.2 mg QE/g DW) and DPPH antioxidant activity (88%) respectively, were recorded in callus cultures grown at 15 mg/L FeO-NPs alone. In shake flasks, maximum callus biomass accumulation (191 g/L) was observed on day 24 of the growth curve, when cell cultures were treated with 15 mg/L FeO-NPs combined with 1.5 mg/L 2,4-D and maintained in darkness for ten days followed by exposure to normal light conditions. Antioxidant potential of cell suspension cultures was significantly enhanced compared to callus grown on solid media with the highest values of TPC (57 mg GAE/g DW), TFC (8.9 mg QE/g DW) and antioxidant activity (91%) were observed. Phenylalanine ammonia-lyase activity (PAL) was found to inversely correlate with biomass and antioxidant metabolites production. Significant variations in antioxidant enzymes activities were detected in cell cultures subjected to different light regimens, with dark pre-treatment followed by light exposure induced the highest superoxide dismutase (SOD; 4.1 U/mg protein) and peroxidase (POD; 4.5 U/mg protein) activities. Cultures grown in complete darkness also resulted in substantial expression of SOD, POD, catalase (CAT) and ascorbate peroxidase (APx) enzymes. Nonetheless, the antileishmanial potential of the cell cultures was evaluated and the putative metabolites responsible for this activity were identified and quantified through the robust Gas chromatography- mass spectrometry (GC–MS) analysis.Key message : Iron oxide nanoparticles and varying light intensity significantly influence the biomass production, antioxidant capacity and antileishmanial activity in callus cultures of Artemisia scoparia. [ABSTRACT FROM AUTHOR]

Abstract Spontaneous forward–reverse mutations were reported by us earlier in clinical samples from various types of cancers and in HeLa cells under normal culture conditions. To investigate the effects of chemical stimulations on such mutation cycles, the present study examined single nucleotide variations (SNVs) and copy number variations (CNVs) in HeLa and A549 cells exposed to wogonin-containing or acidic medium. In wogonin, both cell lines showed a mutation cycle during days 16–18. In acidic medium, both cell lines displayed multiple mutation cycles of different magnitudes. Genomic feature colocalization analysis suggests that CNVs tend to occur in expanded and unstable regions, and near promoters, histones, and non-coding transcription sites. Moreover, phenotypic variations in cell morphology occurred during the forward–reverse mutation cycles under both types of chemical treatments. In conclusion, chemical stresses imposed by wogonin or acidity promoted cyclic forward–reverse mutations in both HeLa and A549 cells to different extents.

Spontaneous forward–reverse mutations were reported by us earlier in clinical samples from various types of cancers and in HeLa cells under normal culture conditions. To investigate the effects of chemical stimulations on such mutation cycles, the present study examined single nucleotide variations (SNVs) and copy number variations (CNVs) in HeLa and A549 cells exposed to wogonin-containing or acidic medium. In wogonin, both cell lines showed a mutation cycle during days 16–18. In acidic medium, both cell lines displayed multiple mutation cycles of different magnitudes. Genomic feature colocalization analysis suggests that CNVs tend to occur in expanded and unstable regions, and near promoters, histones, and non-coding transcription sites. Moreover, phenotypic variations in cell morphology occurred during the forward–reverse mutation cycles under both types of chemical treatments. In conclusion, chemical stresses imposed by wogonin or acidity promoted cyclic forward–reverse mutations in both HeLa and A549 cells to different extents. [ABSTRACT FROM AUTHOR]

Curcuminoids are the major specialized metabolites in Curcuma species that are used as natural dyes and therapeutic agents with multifold biological activities. Curcuminoids are produced by field cultivation of plants; however, this method suffers from long growth periods and genotypic and environmental conditions. Therefore, research efforts have been made to procure curcuminoids from plant tissue cultures of Curcuma species, especially Curcuma longa, C. aromatica, and C. zedoaria. Cell, hairy root, shoot, and microrhizome cultures have been established to produce curcuminoids in vitro. Elicitation processes have been implemented to improve curcuminoid accumulation in plant tissue cultures. In this review, we present the research conducted on curcuminoid production in plant tissue cultures. In addition, efforts have been made to summarize the research accomplishments in the production of curcuminoids in heterologous systems. Curcuminoids produced from cell and organ cultures are used for food coloring, flavoring, preservation, and therapeutic purposes. [ABSTRACT FROM AUTHOR]

Ginkgo biloba is an ancient plant that has survived up until the present day. Gingko biloba is a rich source of valuable secondary metabolites, particularly terpene trilactones (TTLs) such as ginkgolides and bilobalides, which are obtained from the leaves and seeds of the plant. TTLs have pharmacological properties, including anticancer, anti-dementia, antidepressant, antidiabetic, anti-inflammatory, anti-hypertensive, antiplatelet, immunomodulatory, and neuroprotective effects. However, ginkgo is a very-slow-growing tree that takes approximately 30 years to reach maturity. In addition, the accumulation of TTLs in these plants is affected by age, sex, and seasonal and geographical variations. Therefore, plant cell cultures have been established in ginkgo to produce TTLs. Extensive investigations have been conducted to optimize the culture media, growth regulators, nutrients, immobilization, elicitation, and precursor-feeding strategies for the production of TTLs in vitro. In addition, metabolic engineering and synthetic biology methods have been used for the heterologous production of TTLs. In this review, we present the research strategies applied to cell cultures for the production of TTLs. [ABSTRACT FROM AUTHOR]

Three-dimensional cancer cell cultures have been a valuable research model for developing new drug targets in the preclinical stage. However, there are still limitations to these in vitro models. Scaffold-based systems offer a promising approach to overcoming these challenges in cancer research. In this study, we show that two-photon polymerization (TPP)-assisted printing of scaffolds enhances 3D tumor cell culture formation without additional modifications. TPP is a perfect fit for this task, as it is an advanced 3D-printing technique combining a μm-level resolution with complete freedom in the design of the final structure. Additionally, it can use a wide array of materials, including biocompatible ones. We exploit these capabilities to fabricate scaffolds from two different biocompatible materials—PEGDA and OrmoClear. Cubic spheroid scaffolds with a more complex architecture were produced and tested. The biological evaluation showed that the human ovarian cancer cell lines SKOV3 and A2780 formed 3D cultures on printed scaffolds without a preference for the material. The gene expression evaluation showed that the A2780 cell line exhibited substantial changes in CDH1, CDH2, TWIST, COL1A1, and SMAD3 gene expression, while the SKOV3 cell line had slight changes in said gene expression. Our findings show how the scaffold architecture design impacts tumor cell culture 3D spheroid formation, especially for the A2780 cancer cell line. [ABSTRACT FROM AUTHOR]

In vitro plant cell and organ cultures are appealing alternatives to traditional methods of producing valuable specialized metabolites for use as: pharmaceuticals, food additives, cosmetics, perfumes, and agricultural chemicals. Cell cultures have been adopted for the production of specialized metabolites in certain plants. However, in certain other systems, adventitious roots are superior to cell suspension cultures as they are organized structures that accumulate high levels of specialized metabolites. The cultivation of adventitious roots has been investigated in various bioreactor systems, including: mechanically agitated, pneumatically agitated, and modified bioreactors. The main relevance and importance of this work are to develop a long-lasting industrial biotechnological technology as well as to improve the synthesis of these metabolites from the plant in vitro systems. These challenges are exacerbated by: the peculiarities of plant cell metabolism, the complexity of specialized metabolite pathways, the proper selection of bioreactor systems, and bioprocess optimization. This review's major objective is to analyze several bioreactor types for the development of adventitious roots, as well as the advantages and disadvantages of each type of bioreactor, and to describe the strategies used to increase the synthesis of specialized metabolites. This review also emphasizes current advancements in the field, and successful instances of scaled-up cultures and the generation of specialized metabolites for commercial purposes are also covered. [ABSTRACT FROM AUTHOR]

Betalains are nitrogen-containing natural pigments that are water soluble and they comprise of the red-violet betacyanin and the yellow betaxanthin which are abundant in plants such as red/yellow beet, amaranth, prickly pear, pitaya, and others. They are widely used as food coloring agents for many centuries. Betalains are used in pharmaceuticals, functional foods, and cosmeceuticals, since they have tremendous potential to scavenge free radicals and prevent diseases, such as hypertension, dyslipidemia, cancer, neurological disorders, and vascular stenosis. Betalains are proven to be toxicologically safe and have health benefits, they have been approved as food additives in the United States of America, and European countries. Although betalains can be found in natural resources, there are differences in their composition, amounts, and seasonality. For this reason, researchers have developed alternative methods of producing these valuable compounds using cell and organ culture techniques. In several plants, cell and organ cultures are established, and bioreactor technologies have been used to produce betalains on a wide scale. In this review, we discuss the varied biotechnological methods and approaches applied for the biosynthesis of betalains including metabolic engineering approaches.Key message: The primary objective of the present research was to present the progress that has been made in the production of betalains in plant cell and organ cultures. [ABSTRACT FROM AUTHOR]

The rising market interest for Nigella sativa (Black seeds) necessitates the development of cultivation strategies to enhance metabolites production. Zinc oxide nanoparticles (ZnO-NPs) have drawn global attention as efficient and bio safe elicitors for in vitro cultures, to enhance secondary metabolites production in medicinal plants. In this study, ZnO-NPs were utilized for establishment of callus and cell cultures in black seeds for the first time. Hypocotyl explants were cultured on Murashige and Skoog (MS) media with varying levels of ZnO-NPs, resulted in callus induction and biomass formation. Optimal response in callus growth parameters were observed when explants were grown on MS media supplemented with 60 mg/L ZnO-NPs, resulting in 71.2 % callus induction frequency, 28.2 g/L fresh biomass, 9.7 g/L dry biomass, and 63 % water content. A substantial increase in callus growth was observed when ZnO-NPs were combined with 6-Benzylaminopurine (BA) at ratio of 45:1.5 mg/L, resulting in 91.2 % callus induction frequency and 42.2 g/L fresh biomass. In cell suspension cultures, ZnO-NPs alone at 45 mg/L produced optimum callus biomass (60.9 g/L). However, in combination with BA, callus biomass did not increase significantly in cell cultures. Maximum accumulation of total phenolic content (TPC: 26.8 mg GAE/g DW; Gallic acid equivalent dry weight) and total flavonoid content (TFC: 19.5 mg QE/g DW; Quercetin equivalent dry weight) was observed in cell cultures treated with higher concentration (70 mg/L) of ZnO-NPs in the 5th week of the growth curve. Moreover, ZnO-NPs incremented substantially the Phenylalanine lyase (PAL), Superoxide dismutase (SOD) and Peroxidase (POD) enzyme activities in cell cultures. Nonetheless, Reverse Phase High Performance Liquid Chromatography (RP-HPLC) analysis indicated peak production of thymoquinone (168.5 mg/g FW) in cell cultures treated with 45 mg/L ZnO-NPs alone. This study offers a promising approach for commercial production of Nigella sativa biomass and bioactive metabolites.

Introduction. It has been established that the presence of homologous recombination deficiency in a breast tumor is associated with the effectiveness of treatment. But despite the high chemosensitivity of the tumor to DNA-damaging agents, complete pathological responses to treatment are very rare. And this process may be based on a change in the somatic status of BRCA1, that is, a reversion and return of the wild-type allele occurs and the DNA repair function is restored.Aim. To evaluate changes in the presence of chromosomal aberrations and the expression profile of the main genes of homologous recombination in cell models of breast cancer under the influence of cisplatin and docetaxel.Materials and methods. The study was conducted on breast cancer tumor cell cultures: MCF-7, MDA-MB-231 and MDA-MB-468. A cell model of drug resistance was obtained for two drugs: cisplatin and docetaxel. RNA and DNA were isolated from cell suspension using the RNeasy Plus Mini Kit and QIAamp DNA Mini Kit (Qiagen, Germany), respectively. The expression level of homologous recombination genes was assessed using reverse transcription polymerase chain reaction. To assess the presence of chromosomal aberrations, microarray analysis was performed on DNA chips.Results. Restoration of normal copy number for the BRCA1, CDK12, CHEK1 and RAD51D genes in MCF-7 under the influence of cisplatin was shown. For BRCA2 and PALB2, amplifications were detected. A statistically significant increase in the expression of the BRCA1 (p = 0.04), BRCA2 (p = 0.02), PALB2 (p = 0.01) and RAD51D (p = 0.05) genes was also shown. MDAMB-231 shows that all identified loci with deletions, where the BRCA2, BARD1, CHEK2, PALB2 and RAD54L genes are localized, are restored to normal copy number by cisplatin. The appearance of amplifications was registered for BRCA1, BRIP1, FANCL, RAD51B, PARP1. A similar result was shown for docetaxel. An increase in the expression level is typical for the genes BRCA1 (p = 0.02), BRCA2 (p = 0.02), CHEK2 (p = 0.05), FANCL (p = 0.04), PALB2 (p = 0.05), RAD51C (p = 0.02), PARP1 (p = 0.02), which corresponds to the appearance of amplifications. In the MDA-MB-468 cell culture, an increase in the copy number of only the BRCA1 gene is observed. The effect of docetaxel has no effect on this cell culture. The level of BRCA1 expression increases in direct proportion to the duration of drug action.Conclusion. Thus, the study showed that under the influence of cisplatin, reversion of not only homologous recombination gene mutations, but also other disorders can occur.

Xanthohumol (Xn) is an antioxidant flavonoid mainly extracted from hops (Humulus lupulus), one of the main ingredients of beer. As with other bioactive compounds, their therapeutic potential against different diseases has been tested, one of which is Alzheimer's disease (AD). Adenosine is a neuromodulatory nucleoside that acts through four different G protein-coupled receptors: A1 and A3, which inhibit the adenylyl cyclases (AC) pathway, and A2A and A2B, which stimulate this activity, causing either a decrease or an increase, respectively, in the release of excitatory neurotransmitters such as glutamate. This adenosinergic pathway, which is altered in AD, could be involved in the excitotoxicity process. Therefore, the aim of this work is to describe the effect of Xn on the adenosinergic pathway using cell lines. For this purpose, two different cellular models, rat glioma C6 and human neuroblastoma SH-SY5Y, were exposed to a non-cytotoxic 10 µM Xn concentration. Adenosine A1 and A2A, receptor levels, and activities related to the adenosine pathway, such as adenylate cyclase, protein kinase A, and 5′-nucleotidase, were analyzed. The adenosine A1 receptor was significantly increased after Xn exposure, while no changes in A2A receptor membrane levels or AC activity were reported. Regarding 5′-nucleotidases, modulation of their activity by Xn was noted since CD73, the extracellular membrane attached to 5′-nucleotidase, was significantly decreased in the C6 cell line. In conclusion, here we describe a novel pathway in which the bioactive flavonoid Xn could have potentially beneficial effects on AD as it increases membrane A1 receptors while modulating enzymes related to the adenosine pathway in cell cultures. [ABSTRACT FROM AUTHOR]

[Display omitted] Tissue-specific homing peptides have been shown to improve chemotherapeutic efficacy due to their trophism for tumor cells. Other sequences that selectively home to the placenta are providing new and safer therapeutics to treat complications in pregnancy. Our hypothesis is that the placental homing peptide RSGVAKS (RSG) may have binding affinity to cancer cells, and that insight can be gained into the binding mechanisms of RSG and the tumor homing peptide CGKRK to model membranes that mimic the primary lipid compositions of the respective cells. Following cell culture studies on the binding efficacy of the peptides on a breast cancer cell line, a systematic translational characterization is delivered using ellipsometry, Brewster angle microscopy and neutron reflectometry of the extents, structures, and dynamics of the interactions of the peptides with the model membranes on a Langmuir trough. We start by revealing that RSG does indeed have binding affinity to breast cancer cells. The peptide is then shown to exhibit stronger interactions and greater penetration than CGKRK into both model membranes, combined with greater disruption to the lipid component. RSG also forms aggregates bound to the model membranes, yet both peptides bind to a greater extent to the placental than cancer model membranes. The results demonstrate the potential for varying local reservoirs of peptide within cell membranes that may influence receptor binding. The innovative nature of our findings motivates the urgent need for more studies involving multifaceted experimental platforms to explore the use of specific peptide sequences to home to different cellular targets. [ABSTRACT FROM AUTHOR]

Breast cancer, a formidable global health challenge, needs continuous translational research to understand the complexity of mechanisms and improve therapeutic and diagnostic strategies. Breast cancer cell lines are of paramount importance as they significantly contribute to the initial stage of research to understand cancer biology. This review provides insights into targeted therapies and immunotherapies that have emerged using in vitro models and microbiome analysis. It focuses on therapeutic development using cell lines and the limitations of tumor heterogeneity and microenvironment. We explore the evolving landscape of breast cancer cell lines from two‐dimensional (2‐D) cultures to patient‐derived xenograft (PDX) models advancing both fundamental and translational research. Patient‐derived xenografts, cell line‐derived xenografts (CDX), three‐dimensional (3‐D) cultures, organoids, and circulating tumor cells (CTC) models provide promising alternatives that capture the intricacies of the tumor microenvironment. This review bridges the gap between traditional cell lines and newer developments exploring the therapeutic and diagnostic advancements and needs for cell lines to expedite the progress in breast cancer research and treatment. [ABSTRACT FROM AUTHOR]

The development of cell cultures to produce monoclonal antibodies is a multi-step, time-consuming, and labor-intensive procedure which usually lasts several years and requires heavy investment by biopharmaceutical companies. One key aspect of process optimization is improving the feeding strategy. This step is typically performed though design of experiments (DoE) during process development, in such a way as to identify the optimal combinations of factors which maximize the productivity of the cell cultures. However, DoE is not suitable for time-varying factor profiles because it requires a large number of experimental runs which can last several weeks and cost tens of thousands of dollars. We here suggest a methodology to optimize the feeding schedule of mammalian cell cultures by virtualizing part of the experimental campaign on a hybrid digital model of the process to accelerate experimentation and reduce experimental burden. The proposed methodology couples design of dynamic experiments (DoDE) with a hybrid semi-parametric digital model. In particular, DoDE is used to design optimal experiments with time-varying factor profiles, whose experimental data are then utilized to train the hybrid model. This will identify the optimal time profiles of glucose and glutamine for maximizing the antibody titer in the culture despite the limited number of experiments performed on the process. As a proof-of-concept, the proposed methodology is applied on a simulated process to produce monoclonal antibodies at a 1-L shake flask scale, and the results are compared with an experimental campaign based on DoDE and response surface modeling. The hybrid digital model requires an extremely limited number of experiments (nine) to be accurately trained, resulting in a promising solution for performing in silico experimental campaigns. The proposed optimization strategy provides a 34.9% increase in the antibody titer with respect to the training data and a 2.8% higher antibody titer than the optimal results of two DoDE-based experimental campaigns comprising different numbers of experiments (i.e., 9 and 31), achieving a high antibody titer (3,222.8 mg/L) —very close to the real process optimum (3,228.8 mg/L).

SCIENTIFIC RELEVANCE. The Tox21 (Toxicology in the 21st Century) programme was developed by the US Tox21 Consortium with the aim to replace animal-based toxicity assessments of chemicals with a wide range of in vitro and in silico testing approaches and has since been successfully applied in practice.AIM. The study aimed to review information on alternative in vitro models developed as part of the Tox21 programme for testing the toxicity of chemical compounds.DISCUSSION. According to the information provided by the National Toxicology Program, Environmental Protection Agency, National Center for Advancing Translational Sciences, and other Tox21 Consortium members on their official websites and in the literature, the Tox21 Consortium has developed a quantitative high-throughput screening technology for testing the safety of chemicals and created the Tox21 10K library of chemical compounds using this screening technology. The library has been successfully used to create models that predict the toxicity of chemicals prior to preclinical studies. Researchers have proposed new approaches to studying the safety of chemical compounds in human cell lines to replace in vivo studies. Innovative organ-on-chip, multi-organ-on-chip, and organoid models are free from the drawbacks and limitations of cell-line models and offer more accurate representations of complex cell–matrix and organ–organ interactions. Developed under the Tox21 programme to search for new chemical toxicity biomarkers and gene signatures, novel transcriptomics (toxicogenomics) technologies can be used to classify toxicants according to their health risks and to identify potential side effects long before discovering any pathological changes in the body. The Interagency Coordinating Committee on the Validation of Alternative Methods conducts technical evaluation of alternative testing methods and promotes their implementation into regulatory practice.CONCLUSIONS. Thus, new tools and technologies provide an opportunity for switching from in vivo toxicity testing of candidate medicinal products to in silico and in vitro methods.

The aim of the review is to give a brief characteristic of cell cultures obtained from mammalian tissues and to consider the current possibilities and prospects for their use in virology. The analysis of the literature data presented in the main databases, such as Web of Science, PubMed, Scopus, Elsevier, Google Scholar and RSCI (as of July 2023), indicates that various types of cell cultures are currently used in virological studies. The use of cell culture has a number of advantages over other in vitro and in vivo methods of virological research. The review provides numerous examples on the development of new methods of obtaining cell cultures for the cultivation of viruses. Among them are sensitive and reporter cell systems, the design of which can be a promising tool for diagnostics of existing and new unknown viral infections. Cell cultures are characterized as potential in vitro models in virology for developing new diagnostic test-systems and antiviral drugs. An important area of cell cultures application is their use as a substrate for the production of culture-derived vaccines. Another aspect of the cell cultures application is also highlighted, such as the study of the effect of the viruses on the host immune system or the mechanisms of immunopathogenesis of viral infections. It is concluded that the use of cell cultures remains currently and in the near future one of the most important methods in practical virology and in scientific research.

The rapid progress in nanotechnology has introduced multifunctional iron oxide nanoparticles as promising agents in cancer treatment. This research focused on the synthesis and assessment of citric-acid-coated, folic-acid-conjugated nanoparticles loaded with doxorubicin, evaluating their therapeutic potential in tumor models. An advanced automated continuous technology line (CTL) utilizing a controlled co-precipitation method was employed to produce highly dispersive, multifunctional nanofluids with a narrow size distribution. Various techniques, including dynamic light scattering (DLS), electrophoretic light scattering (ELS), X-ray diffraction (XRD), and transmission electron microscopy (TEM), were employed to examine the particle size, zeta potential, structure, and morphology. Magnetic properties were analyzed through vibrating sample magnetometry (VSM), and surface modifications were confirmed via UV-visible (UV-Vis) and Fourier-Transform Infrared (FTIR) spectroscopy. Cytotoxicity and drug delivery efficiency were evaluated in vitro using RM1 (prostate cancer) and MEC1 (chronic lymphocytic leukemia) cell lines. Fluorescence microscopy demonstrated the successful intracellular delivery of doxorubicin, showcasing the nanoparticles’ potential for targeted cancer therapy. However, folic-acid-conjugated nanoparticles exhibited diminished effectiveness over time. This study highlights the importance of nanoparticle optimization for enhancing therapeutic performance. Further research should aim to improve nanoparticle formulations and explore their long-term impacts for the development of safe, targeted cancer treatments.

Astrocytes and ependymal cells have been reported to be able to switch from a mature cell identity towards that of a neural stem/progenitor cell. Astrocytes are widely scattered in the brain where they exert multiple functions and are routinely targeted for in vitro and in vivo reprogramming. Ependymal cells serve more specialized functions, lining the ventricles and the central canal, and are multiciliated, epithelial-like cells that, in the spinal cord, act as bi-potent progenitors in response to injury. Here, we isolate or generate ependymal cells and post-mitotic astrocytes, respectively, from the lateral ventricles of the mouse brain and we investigate their capacity to reverse towards a progenitor-like identity in culture. Inhibition of the GSK3 and TGFβ pathways facilitates the switch of mature astrocytes to Sox2-expressing, mitotic cells that generate oligodendrocytes. Although this medium allows for the expansion of quiescent NSCs, isolated from live rats by "milking of the brain", it does not fully reverse astrocytes towards the bona fide NSC identity; this is a failure correlated with a concomitant lack of neurogenic activity. Ependymal cells could be induced to enter mitosis either via exposure to neuraminidase-dependent stress or by culturing them in the presence of FGF2 and EGF. Overall, our data confirm that astrocytes and ependymal cells retain a high capacity to reverse to a progenitor identity and set up a simple and highly controlled platform for the elucidation of the molecular mechanisms that regulate this reversal. [ABSTRACT FROM AUTHOR]

Borosilicate glass surpasses polystyrene in optical quality; however, it is less frequently used for cell culture due to poor protein and cell adhesion. To overcome this impasse, the surface of glass coverslips requires functionalization to enable facile covalent attachment of proteins to promote cell attachment and differentiation. Herein, a novel approach is presented to covalently attach proteins to glass by depositing a thin layer of radical‐rich carbon film using a plasma polymerization process. The surface chemistry of these plasma‐activated coatings can be controlled by varying the gas composition used during the deposition. Mass spectrometry reveals different protein profiles attached to functionalized glass coverslips when they are exposed to cell culture media. Mouse embryonic stem cell adhesion and subsequent differentiation into neural lineage on plasma‐treated coverslips are significantly enhanced compared to bare coverslips. Importantly, the coatings are in the nanometer range, preserve the optical properties of the glass coverslips for imaging, and remain stable for at least 4 weeks in simulated body fluid. These results demonstrate the utility of covalently attaching proteins to glass for enhanced cell attachment and stem cell differentiation and provide a promising technique to achieve better outcomes in cell culture in a range of biomedical applications. [ABSTRACT FROM AUTHOR]

Antiviral therapy based on neuraminidase (oseltamivir) or polymerase (baloxavir marboxil) inhibitors plays an important role in the management of influenza infections. However, the emergence of drug resistance and the uncontrolled inflammatory response are major limitations in the treatment of severe influenza disease. Protectins D1 (PD1) and DX (PDX), part of a family of pro‐resolving mediators, have previously demonstrated anti‐influenza activity as well as anti‐inflammatory properties in various clinical contexts. Herein, we synthetized a series of simplified PDX analogs and assessed their in vitro antiviral activity against influenza A(H1N1) viruses, including oseltamivir‐ and baloxavir‐resistant variants. In ST6GalI‐MDCK cells, the PDX analog AN‐137B reduced viral replication in a dose‐dependent manner with IC50 values of 23.8 for A/Puerto Rico/8/1934 (H1N1) and between 32.6 and 36.7 µM for susceptible and resistant A(H1N1)pdm09 viruses. In MTS‐based cell viability experiments, AN‐137B showed a 50% cellular cytotoxicity (CC50) of 638.7 µM with a resulting selectivity index of 26.8. Of greater importance, the combination of AN‐137B with oseltamivir or baloxavir resulted in synergistic and additive in vitro effects, respectively. Treatment of lipopolysaccharide (LPS)‐stimulated macrophages with AN‐137B resulted in a decrease of iNOS activity as shown by the reduction of nitrite production, suggesting an anti‐inflammatory effect. In conclusion, our results indicate that the protectin analog AN‐137B constitutes an interesting therapeutic modality against influenza A virus, warranting further evaluation in animal models. [ABSTRACT FROM AUTHOR]

Porcine parvovirus 6 (PPV6) was first identified in aborted swine fetuses in China in 2014. Since its identification, an increased number of PPV6 cases have been reported in many countries with developed pig breeding. In this study, the first identification of porcine parvovirus 6 in Russia, its phylogenetic analysis, and its characterization in vitro are reported. During the investigation, 521 serum samples collected from pigs of different ages from seven regions of the Russian Federation were tested. In four regions, the DNA of the virus was detected. The overall prevalence of porcine parvovirus 6 in Russia was 9.4%. Fattening pigs were the group with the most frequent detection of the virus genome. Phylogenetic analysis of the Russian isolate detected in a domestic boar indicated high homology with strains from Spain. In vitro studies revealed that the most promising cell cultures for PPV6 isolation are SPEV and SK. Our results demonstrated that PPV6 induced typical apoptotic features in cells, including DNA fragmentation, chromatin margination, nuclear condensation, pyknosis of nuclei, symplast formation, and various pathological mitoses. [ABSTRACT FROM AUTHOR]

Preclinical and clinical studies indicate that psychostimulants, in addition to having abuse potential, may elicit brain dysfunctions and/or neurotoxic effects. Central toxicity induced by psychostimulants may pose serious health risks since the recreational use of these substances is on the rise among young people and adults. The present review provides an overview of recent research, conducted between 2018 and 2023, focusing on brain dysfunctions and neurotoxic effects elicited in experimental models and humans by amphetamine, cocaine, methamphetamine, 3,4-methylenedioxymethamphetamine, methylphenidate, caffeine, and nicotine. Detailed elucidation of factors and mechanisms that underlie psychostimulant-induced brain dysfunction and neurotoxicity is crucial for understanding the acute and enduring noxious brain effects that may occur in individuals who use psychostimulants for recreational and/or therapeutic purposes.

Ginkgo biloba is an ancient plant that has survived up until the present day. Gingko biloba is a rich source of valuable secondary metabolites, particularly terpene trilactones (TTLs) such as ginkgolides and bilobalides, which are obtained from the leaves and seeds of the plant. TTLs have pharmacological properties, including anticancer, anti-dementia, antidepressant, antidiabetic, anti-inflammatory, anti-hypertensive, antiplatelet, immunomodulatory, and neuroprotective effects. However, ginkgo is a very-slow-growing tree that takes approximately 30 years to reach maturity. In addition, the accumulation of TTLs in these plants is affected by age, sex, and seasonal and geographical variations. Therefore, plant cell cultures have been established in ginkgo to produce TTLs. Extensive investigations have been conducted to optimize the culture media, growth regulators, nutrients, immobilization, elicitation, and precursor-feeding strategies for the production of TTLs in vitro. In addition, metabolic engineering and synthetic biology methods have been used for the heterologous production of TTLs. In this review, we present the research strategies applied to cell cultures for the production of TTLs.

Polymers are of great interest for medical and cosmeceutical applications. The current trend is to combine materials of natural and synthetic origin in order to obtain products with appropriate mechanical strength and good biocompatibility, additionally biodegradable and bioresorbable. Citric acid, being an important metabolite, is an interesting substance for the synthesis of materials for biomedical applications. Due to the high functionality of the molecule, it is commonly used in biomaterials chemistry as a crosslinking agent. Among citric acid‐based biopolyesters, poly(1,8‐octanediol citrate) is the best known. It shows application potential in soft tissue engineering. This work focuses on a much less studied polyester, poly(1,3‐propanediol citrate). Porous and non‐porous materials based on the synthesized polyesters are prepared and characterized, including mechanical, thermal, and surface properties, morphology, and degradation. The main focus is on assessing the biocompatibility and antimicrobial properties of the materials. [ABSTRACT FROM AUTHOR]

With the remarkable development of DNA nanotechnology, interest in DNA molecules has expanded beyond its biological role to building blocks in materials science. As a unique branch of DNA‐based materials, DNA hydrogels have exhibited many fascinating characteristics, including broad biocompatibility, precise programmability, convenient modification, and tunable mechanical properties, which make DNA hydrogels ideal biomaterials. Moreover, by combining with functional nucleic acids, such as aptamers, i‐motif nanostructures, CpG oligodeoxynucleotides, and DNAzymes, DNA hydrogels can be further tailored to provide additional target recognition, therapeutic potential, and catalytic activities, allowing them to play important roles in biosensing and medical applications. This review, aims to provide readers with an up‐to‐date overview of the important developments of biomedical DNA hydrogels. First, it introduces different synthetic strategies of hydrogels that utilize DNA as building materials and functional units within the hydrogel networks and discuss their advantages in biomedical applications. Subsequently, new approaches and applications of biomedical DNA hydrogels in the recent years are highlighted, such as therapeutic systems, cell culture platforms, tissue engineering materials, and biosensors. Finally, future perspectives and remaining challenges of DNA hydrogels in biomedicine are presented. [ABSTRACT FROM AUTHOR]

The search for and testing of drugs with senolytic activity is a new direction in gerontology. The increasing number of "senescent" cells with age contributes to the development of age-related diseases and chronic non-infectious inflammation. Removing "senescent" cells or suppressing their influence on surrounding tissues seems like a logical step to improve quality of life and possibly prolong it. However, drugs with senolytic and senomorphic activity in model systems induce the development of a number of side effects in clinical trials. In this review, we discuss the main advances in senotherapy, the prospects for the use of senotherapeutics, and the limitations that researchers and clinicians may encounter. [ABSTRACT FROM AUTHOR]

Drug-induced liver injury (DILI) is the reason for 15–18% of medicinal product recalls from the market. Since interspecies differences often limit the relevance of standard non-clinical tests in vivo, a promising alternative is to develop cell-based in vitro methods.The aim of the study was to review current advances in cell modelling for the in vitro identification of DILI.In vitro mechanistic studies of DILI require cells that exhibit activity specific to hepatic metabolising enzymes and transporters. This article reviews the main cell cultures (primary human hepatocytes, immortal cell lines, stem cell-derived hepatocyte-like cells, co-cultures of hepatocytes and non-parenchymal liver cells) and their configurations. The optimisation of cell systems is directed towards enhancing their viability, functionality, compositional and configurational complexity, thus bringing them closer to in vivo models. Potential DILI causes include chemically reactive metabolites, oxidative stress, mitochondrial damage, intracellular accumulation of toxic bile acids resulting from transporter inhibition, and adaptive immune system activation. Accordingly, DILI studies rely on various methods, including innovative technologies for acquisition, storage, and analysis of large datasets (e.g. high-content screening, transcriptomics, proteomics, and metabolomics). Cell models are applicable to both DILI identification and mechanistic studies. Currently, the most promising technologies are omics, complex co-culture models, and organ-on-a-chip systems.

Traditional methods for determining superoxide dismutase (SOD) content and catalase (CAT) activity rely on measuring the absorbance of individual tissue (biological) samples using a cuvette and spectrophotometer, rather than cell cultures. Although there are kits available for SOD and CAT assays, these allow for high-throughput analysis of samples and might be too expensive for research laboratories in countries from the Global South, such as South Africa. This paper describes a simple and cost-effective method to determine SOD content and CAT activity in mammalian cell cultures following exposure to environmental chemical mixtures by measuring absorbance in 96-well microplates. Moreover, the equipment used for this method is considered standard for cell culture laboratories, while the reagents and consumables are easily obtainable. • Antioxidant enzyme levels can be measured in vitro in cell cultures. • The supernatant obtained can be used to determine protein concentration, SOD content, and CAT activity. • This method is simple and affordable, allowing for the analysis of multiple samples (up to 32 samples per microplate).

Bioresource collection of the N. N. Blokhin National Medical Research Center of Oncology is a unique structured biobank that combines different types of primary and subsidiary samples and ensures its comprehensive characterization, including both generally accepted indicators and specific types of research. Samples of paraffin blocks, frozen material, blood and its derivatives are deposited in the clinical department of the center. The research institute of the center has collected commercial and unique human cell lines and transplantable strains obtained from clinical material, as well as cells from laboratory animals, mainly of mouse origin. The provided cell lines undergo multi- stage quality control, including confirmation of authenticity, assessment of viability and determination of optimal cultivation conditions, analysis of interspecific and intraspecific contamination, study of tumorigenicity and reproducibility in athymic Balb/ c-nude mice, etc. In addition, the Bioresource Collection has hybridoma clones that produce antibodies to a wide range of targets. The Center provides the opportunity to conduct preclinical research using samples from the Bioresource Collection to obtain scientific and regulatory data on the antiproliferative activity of new agents or methods for cancer treatment.

Cell cultures are an important part of the research and treatment of autoimmune connective tissue diseases. By culturing the various cell types involved in ACTDs, researchers are able to broaden the knowledge about these diseases that, in the near future, may lead to finding cures. Fibroblast cultures and chondrocyte cultures allow scientists to study the behavior, physiology and intracellular interactions of these cells. This helps in understanding the underlying mechanisms of ACTDs, including inflammation, immune dysregulation and tissue damage. Through the analysis of gene expression patterns, surface proteins and cytokine profiles in peripheral blood mononuclear cell cultures and endothelial cell cultures researchers can identify potential biomarkers that can help in diagnosing, monitoring disease activity and predicting patient's response to treatment. Moreover, cell culturing of mesenchymal stem cells and skin modelling in ACTD research and treatment help to evaluate the effects of potential drugs or therapeutics on specific cell types relevant to the disease. Culturing cells in 3D allows us to assess safety, efficacy and the mechanisms of action, thereby aiding in the screening of potential drug candidates and the development of novel therapies. Nowadays, personalized medicine is increasingly mentioned as a future way of dealing with complex diseases such as ACTD. By culturing cells from individual patients and studying patient-specific cells, researchers can gain insights into the unique characteristics of the patient's disease, identify personalized treatment targets, and develop tailored therapeutic strategies for better outcomes. Cell culturing can help in the evaluation of the effects of these therapies on patient-specific cell populations, as well as in predicting overall treatment response. By analyzing changes in response or behavior of patient-derived cells to a treatment, researchers can assess the response effectiveness to specific therapies, thus enabling more informed treatment decisions. This literature review was created as a form of guidance for researchers and clinicians, and it was written with the use of the NCBI database. [ABSTRACT FROM AUTHOR]

The delivery of nucleosides represents an interesting research trend in recent years due to their application in various viral infections. The main aims of this study were to develop and to characterize polyurethane particles that are intended to be used for the transport of nucleosides. Three samples have been prepared using aliphatic diisocyanates, a mixture of polyethylene glycol, polycaprolactone, and diols, respectively. The samples were characterized through refractivity measurements, drug loading efficacy, release and penetration rate investigations, FTIR and Raman spectroscopy, thermal analyses, Zetasizer, SEM, HDFa cells viability, and irritation tests on mice skin. The results indicate the obtaining of particles with sizes between 132 and 190 nm, positive Zeta potential values (28.3–31.5 mV), and a refractivity index around 1.60. A good thermal stability was found, and SEM images show a medium tendency to agglomerate. The samples' color, pH, and electrical conductivity have changed only to a small extent over time, and the evaluations indicate an almost 70% encapsulation efficacy, a prolonged release, and that around 70% of particles have penetrated an artificial membrane in the first 24 h. The synthesized products should be tested in further clinical trials, and the current tests on cell cultures and mice skin revealed no side effects. [ABSTRACT FROM AUTHOR]

Glycosphingolipids (GSLs) are a glycolipid subtype which plays vital roles in numerous biological processes, cell–cell interactions, as well as oncogenesis and ontogenesis. They are ubiquitous molecules found mostly in cell membranes. Abnormal expression of GSLs as well as altered molecular structure have been linked with progression of cancer and metastasis and are involved in the pathophysiology of neurodegenerative, autoimmune, and infectious diseases as well as inherited enzyme defects—glycosphingolipidoses. Matrix-assisted laser desorption ionization mass spectrometry (MALDI MS) plays a leading role in analyzing and characterizing different GSLs, and thus can help to distinguish altered GSL patterns. This review offers insights into the benefits and limitations when using MALDI MS in this field of lipidomic research, with an emphasis on which are the optimal matrices in analyzing GSLs from different tissues (normal and pathological) as well as highlighting GSLs' particular profiles in various cell cultures, and normal and pathological human tissues obtained by MALDI non-imaging MS (non-IMS). These findings can have implications in further understanding the role of altered GSL expression in various pathological conditions and could be a target for future therapies. [ABSTRACT FROM AUTHOR]

Photodynamic therapy (PDT) represents a therapeutic intervention applied in various pre-malignant and malignant disorders' treatments. The interaction between a photosensitizer (PS), ideal wavelength radiation, and tissue molecular oxygen activates a series of photochemical reactions liable to produce reactive oxygen species. These highly reactive species allow for the decrease cell proliferation and yield cancerous and pre-cancerous cell death. The aim of this work is to carry out a systematic review to investigate the effects of in vitro PDT for oral potential malignant disorders (OPDM) cell lines. This systematic review was conducted according to the PRISMA protocol, and the PROSPERO registration number was CRD42022362349. An electronic search was performed on the following search engines: PubMed, Embase, and Web of Science. The Population, Intervention, Comparison, Outcomes, and Study design (PICOS) has been applied as the method by which to outline our study eligibility criteria. The QUIN tool was employed to interpret the risk of bias of the included studies. Initially, seventy-five records were retrieved through databases, and after the selection steps, seven items finally met our inclusion criteria. The preliminary search resulted in 75 studies, out of which 22 were found to be duplicates. After reviewing the titles and abstracts of the remaining 53 studies, 45 were rejected as they did not meet the inclusion criteria. Further evaluation of the full texts led to the exclusion of only one article, since the full text was not available. As a result, seven studies were ultimately identified and included in the analysis. The main findings confirm the role of in vitro photodynamic therapy using several photosensitizers as a potential treatment for oral potentially malignant disorders. [ABSTRACT FROM AUTHOR]

Now mesenchymal stem cells (MSC) are more widely applied in different areas of medicine. However, despite requirement for considerable quantities of human MSC for clinical practice, there is restricted information concerning culture conditions optimization required for their production. In present article the literature review, concerning various conditions of MSC extraction and culture, is presented. Influence of culture medium, cells passage density, fetal calf serum presence or absence and other conditions on MSC proliferation is surveyed. Data concerning use of three-dimensional structures and various bioreactors for ex vivo expansion are presented. MSC multilinear differentiation features are surveyed.

To date, most endocytosis studies in plant cells have focused on clathrin-dependent endocytosis, while limited evidence is available on clathrin-independent pathways. Since dynamin a is a key protein both in clathrin-mediated endocytosis and in clathrin-independent endocytic processes, this study investigated its role in the uptake of poly-(lactic-co-glycolic) acid (PLGA) nanoparticles (NPs). The experiments were performed on cultured cells and roots of Arabidopsis thaliana. Dynasore was used to inhibit the activity of dynamin-like proteins to investigate whether PLGA NPs enter plant cells through a dynamin-like-dependent or dynamin-like-independent endocytic pathway. Observations were performed by confocal microscopy using a fluorescent probe, coumarin 6, loaded in PLGA NPs. The results showed that both cells and roots of A. thaliana rapidly take up PLGA NPs. Dynasore was administered at different concentrations and exposure times in order to identify the effective ones for inhibitory activity. Treatments with dynasore did not prevent the NPs uptake, as revealed by the presence of fluorescence emission detected in the cytoplasm. At the highest concentration and the longest exposure time to dynasore, the fluorescence of NPs was not visible due to cell death. Thus, the results suggest that, because the NPs' uptake is unaffected by dynasore exposure, NPs can enter cells and roots by following a dynamin-like-independent endocytic pathway. [ABSTRACT FROM AUTHOR]

We established model systems for exploring the roles of symplastic and apoplastic ascorbate in heavy-metal-polluted dicot and monocot cells. Cell-suspension cultures of Arabidopsis and maize were treated with copper, cadmium or nickel; growth and ascorbate metabolism were measured. Growth was halved by ∼80 µM Cu2+, 90 µM Cd2+ or 1200 µM Ni2+ in Arabidopsis, and ∼90 µM Cu2+, 650 µM Cd2+ or 650 µM Ni2+ in maize. Cu2+ (128 µM) and Cd2+ (512 µM) caused partial loss of symplastic ascorbate, especially in Arabidopsis; Ni2+ (512 and 2048 µM) had moderate effects. Added apoplastic l-ascorbate (1 mM) was consumed by the cultures (half-life ∼23 and 44 min in Arabidopsis and maize, respectively), consumption rate being 3–6-fold increased by Cu2+, Cd2+ and Ni2+ in Arabidopsis, and by Cu2+ in maize; Cd2+ and Ni2+ had relatively little effect on apoplastic ascorbate consumption in maize. Radioactivity from exogenous 1 mM l-[1-14C]ascorbate remained extracellular; catabolites formed were dehydroascorbic acid, diketogulonate and oxalyl-threonates. In conclusion, suspension-cultured cells respond to heavy-metal stresses by maintaining symplastic ascorbate concentrations, which may beneficially scavenge symplastic reactive oxygen species (ROS). Apoplastic ascorbate is catabolised in metal-polluted cultures via several oxidative and non-oxidative reactions, the former potentially scavenging stress-related apoplastic ROS. [ABSTRACT FROM AUTHOR]

The aim of the work is to review the professional literature sources from the scientific database PubMed mainly for the last 20 years analyzing the modern view on approaches to experimental modeling of cognitive impairment. Materials and methods. A review of the scientific literature over the past 20 years was performed. The lack of requisite knowledge about the pathogenesis of cognitive impairment and the wide range of risk factors for these conditions continue to be major challenges in the development of guidelines on early diagnosis and treatment. The literary analysis suggests that all modeling approaches to experimental cognitive impairment are currently divided into two groups: cell culture and animal models. Conclusions. Experimental modeling of cognitive impairment remains important in addition to clinical and population-based studies. In recent years, the problem of selecting an adequate model to study cognitive impairment, which is a central clinical manifestation of various neurological diseases (Alzheimer’s and Parkinson’s diseases, traumatic brain injury, vascular, demyelinating, and infectious diseases, metabolic aberrations and hormonal imbalance, neurodegenerative diseases of the central nervous system) is becoming increasingly relevant. The choice of model and experimental material – animals or cultures (invertebrate and mammalian cells) is based on a clear understanding of the study design and depends on the ultimate goal of research.