Microalgae species encounter oxidative stress in their natural environments, prompting the development of species-specific adaptation mechanisms. Understanding these mechanisms can offer valuable insights for biotechnological applications in microalgal metabolic manipulation. In this study, we investigated the response of Tetraselmis chuii , an industrially important microalga, to H 2 O 2 -induced oxidative stress. Exposure to 0.5-mM H 2 O 2 resulted in reduced cell viability, and higher concentrations led to a drastic decline. After 1 h of exposure to H 2 O 2 , photosynthetic capacity (Qy) was negatively impacted, and this reduction intensified after 6 h of continuous stress. Global multi-omics analysis revealed that T. chuii rapidly responded to H 2 O 2 -induced oxidative stress within the first hour, causing significant changes in both transcriptomic and metabolomic profiles. Among the cellular functions negatively affected were carbon and energy flow, with photosynthesis-related PSBQ having a 2.4-fold downregulation, pyruvate kinase decreased by 1.5-fold, and urea content reduced by threefold. Prolonged exposure to H 2 O 2 incurred a high energy cost, leading to unsuccessful attempts to enhance carbon metabolism, as depicted, for example, by the upregulation of photosystems-related PETC and PETJ by more than twofold. These findings indicate that T. chuii quickly responds to oxidative stress, but extended exposure can have detrimental effects on its cellular functions. Key points • 0.5-mM H 2 O 2 –induced oxidative stress strongly affects T. chuii • Distinct short- and long-term adaptation mechanisms are induced • Major metabolic adaptations occur within the first hour of exposure
Liver cancer ranks among the most prevalent malignancies globally and stands as a leading cause of cancer-related mortality. Numerous isothiazolone derivatives and analogues have been synthesized and investigated for their potential as anticancer agents; however, limited data exist regarding their efficacy against liver cancer. In the present study, two nitrophenyl-isothiazolones, the 5-benzoyl-2-(4-nitrophenyl)isothiazol-3(2H)-one (IsoA) and the 2-(4-nitrophenyl)isothiazol-3(2H)-one (IsoB), were preliminarily investigated for their cytotoxicity against hepatoma human (Huh7) cells as a liver cancer model and Immortalized Human Hepatocytes (IHHs) as a model of non-cancerous hepatocytes. IsoB, derived from IsoA after removal of the benzoyl moiety, demonstrated the highest cytotoxic effect against Huh7 cells with CC50 values of 19.3 μΜ at 24 h, 16.4 μΜ at 48 h, and 16.2 μΜ at 72 h of incubation, respectively. IsoB also exhibited selective toxicity against the liver cancerous Huh7 cells compared to IHH cells, reinforcing its role as a potent and selective anticancer agent. Remarkably, the cytotoxicity of IsoB was higher when compared with the standard chemotherapeutical agent 5-fluorouracil (5-FU), which also failed to exhibit higher toxicity against the liver cancerous cell lines. Moreover, IsoB-treated Huh7 cells presented a noteworthy reduction in mitochondrial membrane potential (ΔΨm) after 48 and 72 h, while mitochondrial superoxide levels showed an increase after 24 h of incubation. The molecular mechanism of the IsoB cytotoxic effect was also investigated using RT-qPCR, revealing an apoptosis-mediated cell death along with tumor suppressor TP53 overexpression and key-oncogene MYCN downregulation.
The high genetic heterogeneity of small ruminant lentiviruses (SRLV) renders the genetic characterization of the circulating strains crucial for the epidemiological investigation and the designation of effective diagnostic tools. In Greece, research data regarding the genetic diversity of the circulating SRLV strains is scarce, hindering the implementation of efficient surveillance and control programs. The objective of the study was to genetically characterize SRLV strains isolated from intensive dairy sheep farms in Greece and evaluate the variability of the immunodominant regions of the capsid protein. For this reason, a total of 12 SRLV-infected animals from four intensive dairy sheep farms with purebred Chios and Lacaune ewes were used for the amplification and sequencing of an 800 bp gag-pol fragment. The phylogenetic analyses revealed a breed-related circulation of strains; Chios ewes were infected with strains belonging exclusively to a separate group of genotype A, whereas strains belonging to subtype B2 were isolated from Lacaune ewes. Immunodominant epitopes of capsid protein were quite conserved among the strains of the same genotype, except for the Major Homology Region which showed some unique mutations with potential effects on viral evolution. The present study contributes to the extension of the current knowledge regarding the genetic diversity of SRLV strains circulating in sheep in Greece. However, broader genetic characterization studies are warranted for the exploration of possible recombinant events and the more comprehensive classification of the circulating strains.
A significant number of research papers regarding biosensor-related assays for key food safety pathogens based on the use of mammalian cells has been reported. In this study, the Salmonella typhimurium infection progression was monitored in the human colon cancer cell line Caco-2 and the mucus-secreting HT29-MTX-E12, after treatment with five different bacterial MOI for 30 min by comparing the alterations of frequencies recordings with impedance spectroscopy measurements. For this purpose, bacterial adhesion and invasion assays were initially performed. Then, the data obtained from impedance spectroscopy recordings were compared to cell viability data derived from the MTT uptake cell proliferation assay as well as from live cell analysis assays of mitochondrial membrane potential alterations. From our findings a concentration-dependent increase in bacterial colonies occurring from invaded cells was observed upon a higher multiplicity of infection (MOI) bacterial infection at both cell lines. On the contrary, the bacteria infection did not have any impact on the viability of the cells after 1 h of treatment. Differential results were obtained from the measurement of mitochondrial potential at both cell lines. Finally, the impedance values recorded from the 2D, and 3D cultures were concentration-dependent for both cell lines whereas a characteristic pattern specific to each cell line was revealed. Our results indicate that human cell-based bio-electric assays can be a valuable tool for obtaining a unique fingerprint for each bacterial infection in the near future.
The aim of the current study wat to comparatively assess the impact of different nitrogen (N) fertilization schemes on fresh pod yield and yield quality in either organically or conventionally grown common beans (Phaseolus vulgaris L.). Prior to common bean crop establishment, the experimental field site was cultivated following either organic (a) or conventional (b) farming practices with a winter non-legume crop (Brassica oleracea var. italica) (BR), or (c) with field bean (Vicia faba sp.) destined to serve as a green manure (GM) crop. At the end of the winter cultivation period the broccoli crop residues (BR) and green manure biomass (GM) were incorporated into the soil and the plots that accommodated the treatments (a) and (c) were followed by an organically cultivated common bean crop, while the conventional broccoli crop was followed by a conventionally cropped common bean crop. Additional to the plant residues (BR), sheep manure (SM) at a rate of 40 kg N ha(-1) was also applied to the organically treated common beans, while the plots with a conventionally cropped common bean received 75 kg N ha(-1). Organic common bean treated with SM + BR produced smaller pods of higher dry matter and bioactive compound content, responses that are correlated with limited soil N availability. No significant variations were observed on yield components and N levels of pods cultivated under organic (SM + GM) and conventional cropping systems. Pod sugar and starch content was not influenced by the different fertilization practices. In conclusion, we have demonstrated that the combined application of SM + GM can be considered as an efficient N-fertilisation strategy for organic crops of common bean, benefiting their nutritional value without compromising yield.
In the pursuit of sustainable sources for food, energy, and health products, microalgae have gained attention. In the present study, the lagoonal system of the Nestos River delta was selected as a sampling point in order to search for opportunistic and robust species. Two new strains of Tetraselmis are described with regards to their taxonomic features (as observed using light and transmission electron microscopy and molecular phylogenetics) and their biochemical properties (total lipid, total protein, and total carbohydrate content, photosynthetic pigments, and antioxidant capacity). The studied strains were identified as representatives of Tetraselmis verrucosa f. rubens. Furthermore, both strains exhibited an interesting biochemical profile coupled with high growth rates and promising antioxidant activity, without the use of enhancement and induction culture methods, warranting further investigation and showing potential for biotechnological use.
Although carotenoids generally possess antimicrobial and antioxidant properties, the in vivo synergistic action of carotenoid blends derived from plant-based by-products has not been thoroughly studied. Therefore, the carotenoid characterization and antimicrobial potential of Citrus reticulata extract as well as the impact of this carotenoid-rich extract (CCE) dietary supplementation on the performance, meat quality, and immune-oxidative status of broiler chickens were determined. One hundred and twenty one-day-old hatched chicks (Ross 308) were allocated to two dietary groups, with four replicate pens of 15 birds each. Birds were fed either a basal diet (CON) or the basal diet supplemented with 0.1% CCE (25 mg carotenoid extract included in 1 g of soluble starch) for 42 d. β-Cryptoxanthin, β-Carotene, Zeaxanthin, and Lutein were the prevailing carotenoid compounds in the Citrus reticulata extract. The CCE feed additive exerted inhibitory properties against both Gram-positive (Staphylococcus aureus) and negative (Klebsiella oxytoca, Escherichia coli, and Salmonella typhimurium) bacteria. Both the broiler performance and meat quality did not substantially differ, while the breast muscle malondialdehyde (MDA) concentration tended to decrease (p = 0.070) in the CCE-fed broilers. The inclusion of CCE decreased the alanine aminotransferase and MDA concentration, and the activity of glutathione peroxidase, while the activity of superoxide dismutase was increased in the blood. Catalase and NADPH oxidase 2 relative transcript levels were significantly downregulated in the livers of the CCE-fed broilers. Additionally, Interleukin 1β and tumor necrosis factor (TNF) relative transcript levels were downregulated in the livers of the CCE- fed broilers, while TNF and interferon γ (IFNG) tended to decrease in the spleens and bursa of Fabricius, respectively. The present study provided new insights regarding the beneficial properties of carotenoids contained in Citrus reticulata in broilers' immune-oxidative status. These promising outcomes could be the basis for further research under field conditions.
Crocus sativus L. has various pharmacological properties, known for over 3600 years. These properties are attributed mainly to biologically active substances, which belong to the terpenoid group and include crocins, picrocrocin and safranal. The aim of the current work was to examine the effects of crocins (CRCs) and their methyl ester derivate dimethylcrocetin (DMCRT) on glioblastoma and rhabdomyosarcoma cell lines, in terms of cytotoxicity and gene expression, implicated in proapoptotic and cell survival pathways. Cell cytotoxicity was assessed with Alamar Blue fluorescence assay after treatment with saffron carotenoids for 24, 48 and 72 h and concentrations ranging from 22.85 to 0.18 mg/mL for CRCs and 11.43 to 0.09 mg/mL for DMCRT. In addition, BAX, BID, BCL2, MYCN, SOD1, and GSTM1 gene expression was studied by qRT-PCR analysis. Both compounds demonstrated cytotoxic effects against glioblastoma and rhabdomyosarcoma cell lines, in a dose- and time-dependent manner. They induced apoptosis, via BAX and BID upregulation, MYCN and BCL-2, SOD1, GSTM1 downregulation. The current research denotes the possible anticancer properties of saffron carotenoids, which are considered safe phytochemicals, already tested in clinical trials for their health promoting properties.
Cartilage acidic protein 1A (hCRTAC1-A) is an extracellular matrix protein (ECM) of human hard and soft tissue that is associated with matrix disorders. The central role of fibroblasts in tissue integrity and ECM health made primary human dermal fibroblasts (NHDF) the model for the present study, which aimed to provide new insight into the molecular function of hCRTAC1-A. Specifically, we explored the differential expression patterns of specific genes associated with the presence of hCRTAC1-A by RNA-seq and RT-qPCR analysis. Functional enrichment analysis demonstrated, for the very first time, that hCRTAC1-A is involved in extracellular matrix organization and development, through its regulatory effect on asporin, decorin, and complement activity, in cell proliferation, regeneration, wound healing, and collagen degradation. This work provides a better understanding of putative hCRTAC1-A actions in human fibroblasts and a fundamental insight into its function in tissue biology.