Extracts Of Cultured Cells Research Articles

Investigation of miniature goat cheese produced from Cynara scolymus L. pistil cell suspension culture extracts elicited with melatonin and salicylic acid.

The present study aimed to evaluate the potential of enzymes produced by artichoke (Cynara scolymus L.) flower cell suspension cultures elicited by melatonin (5 μm) and salicylic acid (50 μm) on the production and characteristics of miniature goat cheeses. In this study, five types of fresh miniature goat cheese were produced using whole artichoke flower extract, salicylic acid (50 μm) or melatonin (5 μm) treated artichoke suspension cell culture extracts, a culture extract without elicitor treatment (control) and rennin enzyme. The milk clotting activity values of the enzymes were measured in the range 0.52-0.74. The effect of the enzymes on the titratable acidity, water-soluble nitrogen, ripening index, αs-caseins and β-caseins of goat cheese was significant (P < 0.05). The highest level of casein degradation was observed in the cheese produced with the enzyme containing melatonin, followed by the cheese produced with the enzyme containing salicylic acid. For the enzymes produced by the suspension cell culture method, the addition of melatonin and salicylic acid had a slightly positive effect on the proteolytic activity of the extracts. It was also found that the enzymes obtained from artichokes by the suspension cell culture method did not achieve successful cheese production in terms of chemical, textural and biochemical aspects compared to those obtained from animal enzymes. © 2024 Society of Chemical Industry.

Piggybacking functionalized DNA nanostructures into live-cell nuclei.

DNA origami nanostructures (DOs) are promising tools for applications including drug delivery, biosensing, detecting biomolecules, and probing chromatin substructures. Targeting these nanodevices to mammalian cell nuclei could provide impactful approaches for probing, visualizing, and controlling biomolecular processes within live cells. We present an approach to deliver DOs into live-cell nuclei. We show that these DOs do not undergo detectable structural degradation in cell culture media or cell extracts for 24 hours. To deliver DOs into the nuclei of human U2OS cells, we conjugated 30-nanometer DO nanorods with an antibody raised against a nuclear factor, specifically the largest subunit of RNA polymerase II (Pol II). We find that DOs remain structurally intact in cells for 24 hours, including inside the nucleus. We demonstrate that electroporated anti-Pol II antibody-conjugated DOs are piggybacked into nuclei and exhibit subdiffusive motion inside the nucleus. Our results establish interfacing DOs with a nuclear factor as an effective method to deliver nanodevices into live-cell nuclei.

Characterization of Extracellular Chitinase from Bacillus cereus SAHA 12.13 and Its Potency as a Biocontrol of Curvularia affinis

Bacillus cereus SAHA 12.13 can produce chitinase, an enzyme that digests chitin in the main compounds of cell walls, mycelia, and spores in pathogenic fungi that cause leaf spots on oil palm plants such as Curvularia affinis. This study aims to determine the properties of the chitinase enzyme B. cereus SAHA 12.13 that can inhibit the growth of C. affinis. Chitinase enzyme production and characterization were measured using the Spindler method. Antagonism test against pathogenic fungi using dual culture method by testing cell culture and enzyme crude extract. This result showed that the isolate produced a high level of specific chitinase activity at 37 °C for 45 h of incubation with 8.45 U mg-1 proteins with a growth rate (k) of 0.25 generation/h, and the generation time was 3.96 h/generation. The optimum chitinase activity was achieved at pH 7.0 and 45 °C and was stable for 3 h with a half-life (t1/2) of 770 min. The crude enzyme and cell culture of strain can inhibit the growth of C. affinis by 36.27±0.043% and 34.25±0.041%, respectively. These characteristics indicate that B. cereus strain SAHA12.13 can be used to inhibit C. affinis, which causes leaf blight of oil palm, under varying pH and temperature conditions.

In Vivo Phosphorylation of the Cytosolic Glucose-6-Phosphate Dehydrogenase Isozyme G6PD6 in Phosphate-Resupplied Arabidopsis thaliana Suspension Cells and Seedlings.

Glucose-6-phosphate dehydrogenase (G6PD) catalyzes the first committed step of the oxidative pentose phosphate pathway (OPPP). Our recent phosphoproteomics study revealed that the cytosolic G6PD6 isozyme became hyperphosphorylated at Ser12, Thr13 and Ser18, 48 h following phosphate (Pi) resupply to Pi-starved (-Pi) Arabidopsis thaliana cell cultures. The aim of the present study was to assess whether G6PD6 phosphorylation also occurs in shoots or roots following Pi resupply to -Pi Arabidopsis seedlings, and to investigate its relationship with G6PD activity. Interrogation of phosphoproteomic databases indicated that N-terminal, multi-site phosphorylation of G6PD6 and its orthologs is quite prevalent. However, the functions of these phosphorylation events remain unknown. Immunoblotting with an anti-(pSer18 phosphosite-specific G6PD6) antibody confirmed that G6PD6 from Pi-resupplied, but not -Pi, Arabidopsis cell cultures or seedlings (i.e., roots) was phosphorylated at Ser18; this correlated with a significant increase in extractable G6PD activity, and biomass accumulation. Peptide kinase assays of Pi-resupplied cell culture extracts indicated that G6PD6 phosphorylation at Ser18 is catalyzed by a Ca2+-dependent protein kinase (CDPK), which correlates with the 'CDPK-like' targeting motif that flanks Ser18. Our results support the hypothesis that N-terminal phosphorylation activates G6PD6 to enhance OPPP flux and thus the production of reducing power (i.e., NADPH) and C-skeletons needed to establish the rapid resumption of growth that ensures Pi-resupply to -Pi Arabidopsis.

Influence of Dioscorea Extracts on Physical Load Tolerance in the Setting of Hypoxic and Temperature Factors

A comparative evaluation of the effectiveness of dry alcoholic extracts of Dioscorea from pharmacopoeial raw materials and cell cultures was carried out on a model of combined exposure to hypoxic and temperature factors and swimming of laboratory animals (mice) with a load of 5% of body weight. Dioscorea preparations had a pronounced thermoprotective effect and increased physical performance; however, the aftereffect was confirmed only for the cell culture extract.

Antimicrobial Activity and Biodiversity Study of a Homemade Vegetable Puree Treated with Antimicrobials from Paenibacillus dendritiformis

Paenibacillus dendritiformis UJA2219 isolated from carrot produces broad-spectrum antibacterial activity. The aim of the present study was to determine the impact of partially-purified cell-culture extracts of strain UJA2219 on the microbial load and bacterial diversity of a homemade vegetable puree. The puree was challenged with an overnight culture of strain UJA2219 or with cultured broth extracts partially purified by cation exchange (CE) chromatography or reversed-phase (RP) chromatography and incubated for 7 days at temperatures of 4 °C or 25 °C. The best results were obtained at 25 °C with the RP extract, decreasing counts of the presumptive Enterobacteriaceae below detectable levels. The bacterial diversity of control and treated puree was studied by Illumina paired-end sequencing, using DNA extracted from the puree samples incubated at 25 °C for 24 h. The controls and the puree inoculated with the UJA2219 strain showed an almost-identical bacterial diversity profile, with Pseudomonadota (mainly Fam. Pseudomonadaceae -gen. Pseudomonas- and Enterobacteriace as the most abundant groups). The greatest differences in bacterial diversity were obtained in the puree treated with RP extract, showing a decrease in the relative abundance of Pseudomonadota (especially gen. Pseudomonas) and an increase of Bacillota (mainly of the genera Bacillus, Enterococcus and Lactococcus). Results from the study suggest that the antimicrobial preparations from strain UJA2219 have a potential for application in food biopreservation.

Changes in the Contents of Podophyllotoxin and other Phenolic Compounds in Acetone Extracts of Suspension Cell Cultures of Podophyllum peltatum L. at Different Stages of Cultivation

Changes in the Contents of Podophyllotoxin and other Phenolic Compounds in Acetone Extracts of Suspension Cell Cultures of Podophyllum peltatum L. at Different Stages of Cultivation

Safety Assessment of Centella asiatica-Derived Ingredients as Used in Cosmetics.

The Expert Panel for Cosmetic Ingredient Safety (Panel) assessed the safety of 9 Centella asiatica-derived ingredients, which reportedly function primarily as skin conditioning agents in cosmetic products. The Panel reviewed relevant data relating to the safety of these ingredients. The Panel concluded that Centella Asiatica Extract, Centella Asiatica Callus Culture, Centella Asiatica Flower/Leaf/Stem Extract, Centella Asiatica Leaf Cell Culture Extract, Centella Asiatica Leaf Extract, Centella Asiatica Leaf Water, Centella Asiatica Meristem Cell Culture, Centella Asiatica Meristem Cell Culture Extract, and Centella Asiatica Root Extract are safe in the present practices of use and concentration in cosmetics, as described in this safety assessment, when formulated to be non-sensitizing.

Myrtus communis and Celastrol enriched plant cell culture extracts control together the pivotal role of Cutibacterium acnes and inflammatory pathways in acne.

Acne is a multifactorial inflammatory disease of the pilosebaceous unit in which Cutibacterium acnes is one of the main triggers. A strong predominance of C.acnes phylotype IA1 is present in acne skin with higher biofilm organization and virulence, promoting local immuno-inflammation, especially the Th17 pathway. We evaluated the single and combined pharmacological properties of the plant extracts, Myrtus communis (Myrtacine®) and Celastrol enriched plant cell culture (CEE) extracts on the C.acnes/Th17 pathway. The effect of Myrtacine® on the virulence of C.acnes phylotype IA1 was quantified according to the expression of several related genes. The activity of Myrtacine® and CEE on the inflammatory cascade was assessed using monocytes-derived dendritic cells (Mo-DC) stimulated with membranes or biofilms of the C.acnes phylotype IA1. Finally, the effect of CEE on the Th17 pathway was studied using C.acnes stimulated sebocyte 2D cultures and 3D skin tissue models containing preactivated Th17 cells. Myrtacine® had an anti-virulence effect, evident as a significant and strong inhibition of the expression of several virulence factor genes by 60%-95% compared to untreated controls. Myrtacine® and CEE significantly inhibited proinflammatory cytokine (IL-6, IL-8, IL-12p40 and TNF-α) production by Mo-DC in response to C.acnes phylotype IA1. Interestingly, these two ingredients resulted in synergistic inhibition of most cytokines when used in combination. Finally, we demonstrated an inhibitory effect of CEE, in solution or formulated at 0.3%, specifically on IL-17 release by Th17 lymphocytes in a C.acnes-stimulated sebocyte 2D cultures and by Th17-lymphocytes integrated in a 3D skin models. 2D and 3D models were developed to represent relevant and specific pathways involved in acne. Myrtacine® and CEE were shown to alter one or more of these pathways, indicating their potential beneficial effects on this disease.

Change in Cutibacterium acnes phylotype abundance and improvement of clinical parameters using a new dermocosmetic product containing Myrtus communis and Celastrol enriched plant cell culture extracts in patients with acne vulgaris.

Acne is a multifactorial chronic inflammatory disease of the pilosebaceous unit, where Cutibacterium acnes plays a main role. Recent papers demonstrated that specific C.acnes phylotypes were correlated with the severity of inflammatory acne and reported a specific loss of C.acnes phylotype diversity in this context. The aim of this exploratory study was to evaluate the efficacy of a new dermocosmetic product containing Myrtus communis and Celastrol-enriched plant cell culture extracts on C.acnes phylotype abundance and clinical parameters in subjects with mild to moderate acne vulgaris. Cutibacterium acnes phylotype diversity was evaluated by single-locus sequence typing sequencing on the nonlesional areas of the forehead, that is, areas excluding inflammatory lesions (papules and pustules) on day 1 (D1) and after 56 days (D57) of twice daily application of the dermocosmetic product on the whole face. Clinical efficacy on acne was also assessed by acne lesion counting and Global Evaluation Acne (GEA) score on D1 and D57. Our study confirmed the link between the presence of some C.acnes phylotypes and acne severity. The dermocosmetic cream was linked to a positive impact on C.acnes phylotypes: a significant decrease in pro-pathogen phylotype IC and increase in nonpathogen phylotype IB were observed in the nonlesional areas of acne on D57 compared to D1. In parallel, the clinical results showed a significant decrease in inflammatory and comedonal acne lesions and a significant improvement in the acne severity according to the GEA score. This study showed that the application of a new dermocosmetic product containing M.communis and Celastrol-enriched plant cell culture extracts was linked to a change in the C.acnes phylotype abundance and an improvement in acne severity.

КОРРЕКЦИЯ ЦИТОСТАТИЧЕСКОЙ МИЕЛОТОКСИЧНОСТИ ЭКСТРАКТОМ КУЛЬТУРЫ КЛЕТОК БОЛИГОЛОВА ПЯТНИСТОГО

The study aims to analyze furocoumarin extracts isolated from poison hemlock cell culture influencing the correction of cisplatin-caused cytostatic myelosuppression. The experiments were carried out on 160 CD1 female mice weighing 18–25 g. Cytostatic myelosuppression was simulated by administering cisplatin intraperitoneally once at a maximum tolerated dose of 10 mg/kg. The control group received physiological saline injections. Poison hemlock (Conium maculatum L.) cell culture extract was administered at a dose of 30 mg/kg to prevent the development of disorders. 4 mg/kg of Warfarin Nycomed was used for comparison examination. The correctors were administered intragastrically on the sixth day after cisplatin injected for four days. The indicators were examined on the 1st, 2nd, 5th, 7th, 10th, 15th, 20th and 30th days following the administration of cisplatin. The bone marrow and peripheral blood values were examined using the standard method. Statistical analysis was carried out using Stat Plus Pro (build 7.3.0.0). Under conditions of cytostatic myelosuppression caused by the administration of 10 mg/kg (maximum tolerated dose) of cisplatin, the use of the poison hemlock cell culture extract contributes to the restoration of myeloid and erythroid hematopoietic germs, as well as the normalization of bone marrow and peripheral blood values. With increase in the number of mature neutrophils and lymphocytes, erythroblasts, and normoblasts, the total number of myelokaryocytes increased. The content of erythrocytes and leukocytes increased in the peripheral blood as the number of segmented neutrophils and lymphocytes increased. The dynamics of hematopoietic sprout recovery with poison hemlock cell culture extract are similar to those of Warfarin.

Noncanonical NLRP3 Inflammasome Activation: Standard Protocols.

The canonical activation of multimeric inflammasomes usually occurs through caspase-1 activation, and it is characterized by the presence of extracellular IL-1β and IL-18 or measuring danger signal proteins, such as HMGB1 using enzyme-linked immunosorbent assay (ELISA) or Western blots; these assays differentiate non-cleaved and cleaved forms of these two cytokines (the cleaved form is the mature and active form). Similar techniques can be used to assess noncanonical inflammasome activation. Real-time PCR can measure the relative mRNA expression for a specific gene, whereas Western blots or immunocytochemistry can detect the presence of proteins by binding of specific antibodies to their antigens in biological samples. Moreover, noncanonical inflammasome activation can be evaluated through the cleavage of the amino and the carboxy terminals of one important component, gasdermin D (GSDMD), whose cleavage induces its pyroptotic activity. Thus, the analysis of cleaved GSDMD is an ideal pathway to study the noncanonical inflammasome. ELISA and immunoblot can be performed on cell culture supernatants or cell extracts.

The Elicitor Effect of Yeast Extract on the Accumulation of Phenolic Compounds in Linum grandiflorum Cells Cultured In Vitro and Their Antiradical Activity

This paper examines the elicitor effect of yeast extract (YE) in various concentrations (200–1000 mg/L) on the accumulation of phenolic compounds (PC) in flowering flax (Linum grandiflorum Desf.) cells cultured in vitro and their antiradical activity. It is established that the total PС content and the content of phenylpropanoids increase in the cell culture, especially at high YE concentrations in the medium (500 and 1000 mg/L). The antiradical activity of flax culture extracts remains in most cases at the control level. Therefore, the elicitation of flowering flax in vitro cultures by YE activates the PC biosynthesis resulting in the accumulation of these secondary metabolites, while the antiradical activity of cell culture extracts does not decrease compared to the control level.

Green Synthesis of Silver Nanoparticles Using Randia aculeata L. Cell Culture Extracts, Characterization, and Evaluation of Antibacterial and Antiproliferative Activity.

The demand for metallic nanoparticles synthesized using green methods has increased due to their various therapeutic and clinical applications, and plant biotechnology may be a potential resource facilitating sustainable methods of AgNPs synthesis. In this study, we evaluate the capacity of extracts from Randia aculeata cell suspension culture (CSC) in the synthesis of AgNPs at different pH values, and their activity against pathogenic bacteria and cancer cells was evaluated. Using aqueous CSC extracts, AgNPs were synthesized with 10% (w/v) of fresh biomass and AgNO3 (1 mM) at a ratio of 1:1 for 24 h of incubation and constant agitation. UV-vis analysis showed a high concentration of AgNPs as the pH increased, and TEM analysis showed polydisperse nanoparticles with sizes from 10 to 90 nm. Moreover, CSC extracts produce reducing agents such as phenolic compounds (162.2 ± 27.9 mg gallic acid equivalent/100 g biomass) and flavonoids (122.07 ± 8.2 mg quercetin equivalent/100 g biomass). Notably, AgNPs had strong activity against E. coli, S. pyogenes, P. aeruginosa, S. aureus, and S. typhimurium, mainly with AgNPs at pH 6 (MIC: 1.6 to 3.9 µg/mL). AgNPs at pH 6 and 10 had a high antiproliferative effect on cancer cells (IC50 &lt; 5.7 µg/mL). Therefore, the use of cell suspension cultures may be a sustainable option for the green synthesis of AgNPs.

Mass Spectrometry-Based Untargeted Metabolomics and Lipidomics Platforms to Analyze Cell Culture Extracts.

Metabolites represent the most downstream level of the cellular organization. Hence, an in vitro untargeted metabolomics approach is extremely valuable to deepen the understanding of how endogenous metabolites in cells are altered under a given biological condition. This chapter describes a robust liquid chromatography-high-resolution mass spectrometry-based metabolomics and lipidomics platform applied to cell culture extracts. The analytical workflow includes an optimized sample preparation procedure to cover a wide range of metabolites using liquid-liquid extraction and validated instrumental operation procedures with the implementation of comprehensive quality assurance and quality control measures to ensure high reproducibility. The lipidomics platform is based on reversed-phase liquid chromatography for the separation of slightly polar to apolar metabolites and covers a broad range of lipid classes, while the metabolomics platform makes use of two hydrophilic interaction liquid chromatography methods for the separation of polar metabolites, such as organic acids, amino acids, and sugars. The chapter focuses on the analysis of cultured HepaRG cells that are derived from a human hepatocellular carcinoma; however, the sample preparation and analytical platforms can easily be adapted for other types of cells.

33556 Clinical efficacy of a new sunscreen containing Myrtus communis and Celastrol enriched plant cell culture extracts on reducing acne and limiting lesions’ recurrence post-sun exposure in acneic patients

Introduction: This clinical study aimed to assess the efficacy and tolerance of a facial SPF50+ sunscreen containing Myrtus communis and Celastrol extracts after 8 weeks of use and its benefits on the autumnal recurrence of acne “Post-Sun Exposure” (P-S.E.).

33563 Change in Cutibacterium acnes phylotype diversity and improvement of clinical parameters using a new dermocosmetic product containing Myrtus communis and Celastrol-enriched plant cell culture extracts in patients with acne vulgaris

Introduction: Cutibacterium acnes, the main bacterial species of the pilosebaceous unit, plays a major role in acne. Recent papers demonstrated that specific C acnes phylotypes were correlated with the severity of inflammatory acne and reported a specific loss of C acnes phylotype diversity in this context. Objectives: The aim of the study was to evaluate the efficacy of a new dermocosmetic product containing Myrtus communis and Celastrol-enriched plant cell culture extracts on C acnes phylotype abundance and clinical parameters in subjects with mild to moderate acne vulgaris. Materials/methods: Efficacy on C. acnes phylotype diversity was evaluated by single-locus sequence typing sequencing after 57 days of twice daily application of the product. Clinical parameters of acne and product tolerance were also assessed. Results: Our study confirmed the link between the presence of some C. acnes phylotypes and acne. At day 57, the product had a positive impact on C. acnes phylotypes: a significant decrease of propathogen phylotype IC and increased of nonpathogen phylotype IB were observed. In parallel, the clinicals results showed a significant decrease of inflammatory and total acne lesions, and a significant improvement of global acne evaluation score. The tolerance of the product was assessed very good. Conclusion: This study showed the topic modulation of C. acnes phylotype diversity by this new dermocosmetic product containing Myrtus communis and Celastrol enriched plant cell culture extracts associated with a significant clinical efficacy.

Patched-Related Is Required for Proper Development of Embryonic Drosophila Nervous System.

Patched-related (Ptr), classified primarily as a neuroectodermal gene, encodes a protein with predicted topology and domain organization closely related to those of Patched (Ptc), the canonical receptor of the Hedgehog (Hh) pathway. To investigate the physiological function of Ptr in the developing nervous system, Ptr null mutant embryos were immunolabeled and imaged under confocal microscopy. These embryos displayed severe alterations in the morphology of the primary axonal tracts, reduced number, and altered distribution of the Repo-positive glia as well as peripheral nervous system defects. Most of these alterations were recapitulated by downregulating Ptr expression, specifically in embryonic nerve cells. Because similar nervous system phenotypes have been observed in hh and ptc mutant embryos, we evaluated the Ptr participation in the Hh pathway by performing cell-based reporter assays. Clone-8 cells were transfected with Ptr-specific dsRNA or a Ptr DNA construct and assayed for changes in Hh-mediated induction of a luciferase reporter. The results obtained suggest that Ptr could act as a negative regulator of Hh signaling. Furthermore, co-immunoprecipitation assays from cell culture extracts premixed with a conditioned medium revealed a direct interaction between Ptr and Hh. Moreover, in vivo Ptr overexpression in the domain of the imaginal wing disc where Engrailed and Ptc coexist produced wing phenotypes at the A/P border. Thus, these results strongly suggest that Ptr plays a crucial role in nervous system development and appears to be a negative regulator of the Hh pathway.

553 Change in c. acnes phylotype abundance and improvement of clinical parameters using a new dermocosmetic product containing myrtus communis and celastrol enriched plant cell culture extracts in patients with acne vulgaris

Introduction: Cutibacterium acnes (C. acnes), the main bacterial species of the pilosebaceous unit, plays a major role in acne. Recent papers demonstrated that specific C. acnes phylotypes were correlated with the severity of inflammatory acne and reported a specific loss of C. acnes phylotype diversity in this context. Objectives: The aim of the study was to evaluate the efficacy of a new dermocosmetic product containing Myrtus communis and Celastrol enriched plant cell culture extracts on C. acnes phylotype abundance and clinical parameters in subjects with mild to moderate acne vulgaris.

Inhibitory activity of soybean (Glycine max L. Merr.) Cell Culture Extract on tyrosinase activity and melanin formation in alpha-melanocyte stimulating Hormone-Induced B16-F10 melanoma cells.

Hyperpigmentation, which causes excessive melanin synthesis and accumulation, is an important issue in the cosmetic industry. Since compounds developed against hyperpigmentation often come with side effects such as skin irritation and contact dermatitis, new studies focus on the use of natural agents that have no side effects. In this study, it was found that the effects of soybean cell culture extract (SCE) on alpha-melanocyte-stimulating hormone (α-MSH) induced melanogenesis in B16-F10 murine melanoma cells. The cells were incubated with SCE for 48h after treatment with α‑MSH for 24h to analysis the melanin content, cellular tyrosinase activity, and gene and protein expression. SCE at 1mg/mL decreased melanin content and tyrosinase activity by 34% and 24%, respectively, compared to the α-MSH-treated group, which did not decrease cell viability. In addition, SCE (1mg/mL) downregulated the expression of tyrosinase (TYR), tyrosinase-related protein (TRP)-1, tyrosinase-related protein (TRP)-2, and microphthalmia-associated transcription factor (MITF) genes 1.5-, 1.5-, 2-, and 2-fold, respectively. Furthermore, SCE inhibited the expression of TYR, TRP1, and TRP2 by decreasing the expression of MITF, as shown in a western blot assay. This study suggests that SCE reveals dose-dependent inhibition of melanin synthesis through the suppression of tyrosinase activity as well as molecular levels of TYR, TRP1, TRP2, and MITF in B16-F10 murine melanoma cells. Therefore, SCE has the potential to be an effective and natural skin-whitening agent for application in the cosmetic industry.