Effect Of Different Concentrations Research Articles

Effects of Different Concentrations of Weak Acid Fracturing Fluid on the Microstructure of Coal

Effects of Different Concentrations of Weak Acid Fracturing Fluid on the Microstructure of Coal

Impact of Methyl Jasmonate on Terpenoid Biosynthesis and Functional Analysis of Sesquiterpene Synthesis Genes in Schizonepeta tenuifolia.

This study investigates the impact of methyl jasmonate (MeJA) on the volatile oil composition of Schizonepeta tenuifolia and elucidates the function of the StTPS45 gene, a key player in terpenoid biosynthesis. The effect of different concentrations of MeJA (0, 50, 100, 200, and 300 μmol/L) on the growth of S. tenuifolia adventitious bud clusters was analyzed over a 20 d period. Using gas chromatography-mass spectrometry (GC-MS), 17 compounds were identified from the adventitious bud clusters of S. tenuifolia. Significant changes in the levels of major monoterpenes, including increased contents of (+)-limonene and (+)-menthone, were observed, particularly at higher concentrations of MeJA. Analysis of transcriptome data from three groups treated with 0, 100, and 300 μmol/L MeJA revealed significant changes in the gene expression profiles following MeJA treatment. At 100 μmol/L MeJA, most terpene synthase (TPS) genes were overexpressed. Additionally, gene expression and functional predictions suggested that StTPS45 acts as germacrene D synthase. Therefore, StTPS45 was cloned and expressed in Escherichia coli, and enzyme activity assays confirmed its function as a germacrene D synthase. Molecular docking and structural prediction of StTPS45 further suggested specific interactions with farnesyl diphosphate (FPP), aligning with its role in the terpenoid synthesis pathway. These findings provide valuable insights into the modulation of secondary metabolite pathways by jasmonate signaling and underscore the potential of genetic engineering approaches to enhance the production of specific terpenoids in medicinal plants.

Negative effects of cigarette butt leachate on freshwater phytoplankton communities.

We experimentally tested the effects of different concentrations of cigarette butt leachate on freshwater phytoplankton chlorophyll-a, species richness, cell density, and community composition. For this, we sampled the phytoplankton from a eutrophic lake and acclimated it for 24 h in microcosms. We then conducted the experiment in microcosms maintained for 96 h. The experiment consisted of four treatments: control and leachate from 1 butt L-1 (T1), 5 butts L-1 (T5), and 10 butts L-1 (T10), which were prepared by diluting a stock solution of leachate from 50 butts L-1. We found that algal chlorophyll-a content was not affected by different leachate concentrations. In contrast, phytoplankton cell density decreased in a dose-dependent manner as concentrations of the leachate increased. Similarly, the number of species was highest in the control group relative to all other treatments, with T1 and T5 showing higher species richness than T10. Additionally, the exposition to different concentrations of the leachate impacted community composition across all treatments in comparison to the control group. Our results suggest that cigarette butt leachate alters the number of cells and species, as well as the distribution of abundance, without necessarily reducing chlorophyll-a concentrations. Our findings indicate that to gain a comprehensive understanding of the effects of cigarette butt leachate on freshwater ecosystems, it is essential to evaluate more realistic scenarios that incorporate aquatic communities, rather than isolated species.

Evaluation of the lethal effect of hydrogen peroxide on the root-knot nematode, Meloidogyne javanica, and the disruption in the activity of enzymes related to its biosynthesis

Summary In this study, the accumulation of hydrogen peroxide (H2O2) in different parts (healthy root, gall, root attached to gall, aerial parts) of healthy tomato plants and plants infected by the root-knot nematode, Meloidogyne javanica, was investigated after treating the roots with β-aminobutyric acid (BABA). In addition, the effects of different concentrations of H2O2 (0-2 mM) on mortality, body structures and the digestive system of second-stage juveniles (J2) were also investigated in in vitro experiments. Results showed that exposure to concentrations of more than 1 mM H2O2 is lethal for J2, while the digestive system is not able to tolerate concentrations of more than 0.2 mM. Evaluating the activity level of enzymes related to biosynthesis and removing H2O2 in treated plants also showed that the nematode has significant suppressive effects on the biosynthesis enzymes of this compound, and this effect is greater in the site of nematode activity than in other parts of the plant. By contrast, the activity of catalase enzyme, as one of the H2O2-removing enzymes, shows a significant increase in all parts of nematode infected plants (especially in galls).

The Short-term Exposure Effect of Chlorpyrifos (20% EC) on Haematological, Biochemical and Histopathological Response of Striped Catfish Pangasianodon hypophthalmus

An experiment of acute toxicity was conducted to find out the lethal concentration 50 (LC50) of chloropyrifos (CPF) for Pangasianodon hypophthalmus. Our findings indicated that the 96-h LC50 value of CPF for P. hypophthalmus was 0.106 mg L-1. Effects of different concentrations of CPF (0.09, 0.1, 0.11, 0.12, and 0.13 mg L-1) for 96-h on hemato-biochemical, enzyme and histological effects were evaluated in this study. Acute exposure induced significant differences in enzymes acid phosphatase, alkaline phosphatase, aspartate aminotransferase, alanine aminotransferase and acetylcholine of blood serum of the treated group as compared to control group. However, significant decreases in serum total protein, globulin, albumin, and triglycerides levels while serum glucose enhanced with consequent increasing doses of CPF exposure. Also, CPF induced significant declines in RBC count, haemoglobin and hematocrit level whereas the exposure groups showed significant rises in WBC and blood indices levels as compared to control group. Furthermore, CPF exposure causes gill curling, epithelium lifting, necrosis, and lamellae degeneration. Hemosiderin, sinusoid dilations, necrosis, malanomacrophage, hepatocyte hypertrophy were observed in liver histology. Reduction of lumen tubule, hyperplasia, increased bowman space and degenerations were seen in kidney. Overall findings indicates that the acute CPF exposure to P. hypophthalmus resulted in significant adverse changes in biochemical, hematological, and histopathological response.

Development of Hybrid Electrospun Nanofibers: Improving Effects of Cellulose Nanofibers (CNFs) on Electrospinnability of Gelatin.

Cellulose nanofibers (CNFs) were used to improve the electrospinnability of the gelatin protein in a water/ethanol/acetic acid (3:2:3, v/v) solution. The effects of different concentrations of CNFs (0.5-4%) on the important physical properties of the gelatin solution (15%), including rheology, conductivity, and surface tension, were investigated. The apparent viscosity and shear-thinning behavior were increased by increasing the CNF concentration from 0 to 4% at a low shear rate (<10 s-1). CNFs also increased the electrical conductivity and surface tension of the gelatin solution. Scanning electron microscopy (SEM) images revealed uniformly ordered structures with good continuity without fracture or bead formation in all hybrid nanofibers. They also showed that the average diameters of fibers decreased from 216 nm in the pure gelatin nanofibers to 175.39 nm in the hybrid gelatin/CNF (4%) ones. Differential scanning calorimetry (DSC) results showed that CNFs increased Tg, and X-ray diffraction (XRD) analysis showed that the electrospinning process caused the formation of more amorphous structures in the gelatin/CNF hybrid nanofibers. The tensile test indicated that by adding 2% CNFs, the ultimate tensile strength (UTS) and strain at break (SB) of nanofiber mats increased from 4.26 to 10.5 MPa and 3.3% to 6.25%, respectively. The current study indicated that incorporating CNFs at the optimal concentration into a gelatin solution can improve the resulting hybrid nanofibers' morphology, average diameter, and mechanical properties.

Regulation of winter wheat-originated antifreeze glycoprotein on rooster spermatozoa freezability

The freezability of chicken spermatozoa is low, therefore, effective cryoprotectants is desiderated. Antifreeze proteins (AFPs) are widely found in cold-tolerant species and help them to survive in freezing environments. This study was the first to evaluate the effects of different concentrations of plant-originated antifreeze glycoprotein (AFGP) (0, 0.1, 1, and 5 μg/mL) on post-thawed sperm motion characteristics, morphology, mitochondrial function, antioxidant activity, and fertilizing potential in chickens. Results showed that the total motility of 0.1 to 1 μg/mL AFGP groups were significantly higher than those of the 5 μg/mL AFGP group (P < 0.05). The post-thawed sperm viability of 0.1 μg/mL AFGP group was significantly higher than any of test groups (P < 0.05). Higher abnormal morphology rate of post-thawed sperm was observed in the control group (0 μg/mL AFGP) than in the 0.1, 1, and 5 μg/mL AFGP groups (P < 0.05). The concentrations of malondialdehyde (MDA) decreased gradually with the increase of AFGP concentration. ATP was significantly higher in the 0.1 and 1 μg/mL AFGP groups than those of control and any of test groups (P < 0.05). The 0.1 to 1 μg/mL AFGP groups had increased mitochondrial membrane potential (MMP) level (P > 0.05). The 0.1 μg/mL AFGP group had the highest average fertility (61.36%) compared with control group (57.02%) and any of test groups of chickens at 31 wk of age, and the 1 μg/mL AFGP group had the highest average fertility (37.72%) compared with control group (21.73%) and any of test groups of chickens at 65 wk of age. In conclusion, the results from this study suggest lower concentration of AFGP (0.1-1 μg/mL) showed positive effect for sperm function. This study inspires the continuous evaluation and seeking right way of adopting different kinds of AFPs in rooster semen cryopreservation.

Effect of Tea Tree Essential Oil on the Quality, Antioxidant Activity, and Microbiological Safety of Lightly Processed Lily (Lilium brownii var. viridulum) during Storage.

The Lanzhou lily is a regionally distinctive vegetable; the emergence of lightly processed lilies has addressed the inconvenience of consuming fresh lilies. However, the cleaning and impurity removal during the processing of lightly processed lily may strip off its original protective barrier and affect the edible quality. As one of the preservation methods, tea tree essential oil (TTEO) has the characteristics of being green, safe, and efficient preservative properties. This study focused on investigating the effects of different concentrations (25 μL/L, 50 μL/L, and 100 μL/L) of TTEO on the quality and microbiological safety of lightly processed lily. The results showed that compared with the control, appropriate concentrations of TTEO treatment could delay weight loss, improve appearance, firmness, and sensory quality, and maintain microbiological safety with the best effect observed at 50 μL/L. Meanwhile, TTEO treatment induced phenylalanine ammonia-lyase activity, thereby increasing the total phenolic content. Furthermore, TTEO enhanced the superoxide dismutase (SOD) and ascorbate peroxidase (APX) activity, which reduced O2-· production rate and H2O2 content. TTEO inhibited lipoxygenase (LOX) activity, reducing the relative conductivity and malondialdehyde content, thereby delaying lipid peroxidation and quality deterioration. This indicates that TTEO could enhance antioxidant capacity by regulating reactive oxygen species (ROS) metabolism and delay the quality deterioration of lightly processed lily by inhibiting lipid peroxidation.

Ginsenoside Rb1 Inhibits the Proliferation of Lung Cancer Cells by Inducing the Mitochondrial-mediated Apoptosis Pathway.

Lung cancer is one of the more common malignant tumors posing a great threat to human life, and it is very urgent to find safe and effective therapeutic drugs. The antitumor effect of ginsenosides has been reported to be a treatment with a strong effect and a high safety profile. This paper aimed to investigate the inhibitory effect of ginsenoside Rb1 on 95D and NCI-H460 lung cancer cells and its pathway to promote apoptosis. We performed the CCK-8 assay, fluorescence staining assay, flow cytometry, scratch healing assay, and Transwell assay to detect the effects of different concentrations of ginsenoside Rb1 on the antitumor activity of 95D and NCI-H460 cells and Western Blot detected the mechanism of antitumor effect. Ginsenoside Rb1 treatment significantly increased the inhibition and apoptosis rates of 95D and NCIH460 cells and inhibited the cell cycle transition from S phase to G2/M. Rb1 induces apoptosis by altering the levels of P53, Bax, Cyto-c, Caspase-8, Caspase-3, Cleaved Caspase-3, Bcl-2, MMP-2, and MMP-9 proteins and activating the external apoptotic pathway. Ginsenoside Rb1 inhibits proliferation and migration and induces apoptosis of 95D and NCI-H460 lung cancer cells by regulating the mitochondrial apoptotic pathway to achieve antitumor activity.

Cadmium exposure induces changes in gut microbial composition and metabolic function in long-tailed dwarf hamsters, Cricetulus longicaudatus.

Numerous studies have demonstrated that exposure to cadmium disrupts the diversity and composition of the gut microbiota, resulting in damage to organ tissue. However, there remains a lack of comprehensive understanding regarding the broader ecological reality associated with this phenomenon. In this study, we conducted a thorough evaluation of the effects of different concentrations of Cd (6, 12, 24, and 48 mg/L) over a period of 35 consecutive days on the organ viscera and the gut microbiota of long-tailed dwarf hamsters, Cricetulus longicaudatus (Rodentia: Cricetidae), using histopathological analysis, 16S rDNA, and metagenome sequencing. Our findings revealed that the results suggest that Cd exposure induced liver, spleen, and kidney damage, potentially leading to increased intestinal permeability and inflammation. These alterations were accompanied by significant perturbations in the gut microbiota composition, particularly affecting potentially pathogenic bacteria such as Prevotella and Treponema within the gut ecosystem. Consequently, host susceptibility to underlying diseases was heightened due to these changes. Notably though, Cd exposure did not significantly impact the overall structure of the gut microbiota itself. Additionally, Cd exposure induced significant changes in the metabolic functions, with the pathways related to disease and environmental information processing notably enhanced, possibly indicating stronger innate defense mechanisms against external injuries among wild mammals exposed to Cd. This study offers a novel approach to comprehensively evaluate the significant impact of Cd pollution on ecosystems by investigating both structural and functional alterations in the digestive system, as well as disruptions in intestinal flora among wild mammals.

Toxic effect of mussel Mytilus galloprovincialis exposed to Ag–TiO2 and ZnTi2O4–TiO2 bicomponent nanoparticles

Nanoparticles (NPs) are widely used in various fields, including antifouling paints for ships and industrial structures submerged in water. The potential impact of NPs on aquatic organisms, particularly their potential toxicity, is a significant concern, as their negative impact has been relatively poorly studied. In this study, we evaluated the effect of different concentrations of bimetallic Ag–TiO₂ and ZnTi₂O₄-TiO₂ NPs, which could potentially be used in antifouling coatings, on the hemocytes of the Mediterranean mussel Mytilus galloprovincialis. Hemocytes were exposed to NPs at concentrations of 0.1–1 mg/L for 1 and 2 h, and the production of reactive oxygen species (ROS), levels of DNA damage, and number of dead cells were measured. Exposure to Ag–TiO₂ NPs at 1 mg/L concentration for 1 h suppressed ROS production in hemocytes and reduced the relative number of agranulocytes in cell suspensions, without inducing DNA damage or cell death. Exposure to ZnTi2O4–TiO2 NPs did not cause changes in the ratio of granulocytes to agranulocytes in suspensions, nor did it affect other functional parameters of hemocytes. However, after a 2 h exposure period, ZnTi2O4–TiO2 NPs (1 mg/L) significantly reduced the production of ROS by hemocytes. These findings suggest that Ag–TiO2 and ZnTi2O4–TiO2 NPs have low acute toxicity for marine bivalves.

Effect and mechanism of Compound Shougong Powder in attenuating triple-negative breast cancer progression by reversing epithelial-to-mesenchymal transition

The study investigated the effect of Compound Shougong Powder(CSGP) on the biological functions of triple-negative breast cancer(TNBC) cells and whether its mechanism of action was related to the epithelial-mesenchymal transition(EMT) signaling pathway. TNBC cells(MDA-MB-231 and BT-549) were treated with different concentrations of CSGP-containing serum. MTS assay was used to detect the effect of CSGP on the proliferation of TNBC cells. The EdU staining was used to detect the effect of CSGP on the proliferation of TNBC cells. Flow cytometry was used to examine the impact of CSGP on apoptosis of TNBC cells. Wound-healing and Transwell assays were used to evaluate the effects of different concentrations of CSGP on the migration and invasion capabilities of TNBC cells. RNA sequencing technology was utilized to elucidate its mechanism. Subsequently, qRT-PCR was performed to measure the mRNA expression levels of E-cadherin, N-cadherin, Slug, Snail, Vimentin, Twist, Zinc finger E-box-Binding homeobox 1(Zeb1), and Zinc finger E-box-Binding homeobox 2(Zeb2). Western blot was used to assess the protein expression levels of Slug, Vimentin, and E-cadherin. After intervention with CSGP, the proliferation of MDA-MB-231 and BT-549 cells significantly decreased, while the apoptosis rate markedly increased. The expression levels of the epithelial marker protein E-cadherin significantly increased, while the expression levels of the EMT-related transcription factors Slug and Vimentin showed a decrease. In conclusion, CSGP inhibits the EMT, thereby suppressing the malignant progression of TNBC.

Autophagy, apoptosis, and inflammation response of Sebastiscus marmoratus to different concentrations of hydroxychloroquine

Hydroxychloroquine (HCQ) is an important public health therapeutic agent widely used in the prevention and treatment of malaria and autoimmune diseases, with some antiviral effects, as well as a common autophagy inhibitor. Its autophagy-inhibiting effect attracts great research interest in mammals but is still little studied in fish. We even have no idea about the effects of HCQ on different tissues of fish and what concentrations should be used for safety studies. This study investigated the effect of different concentration of HCQ treatments on the survival, tissue structure, and expressions of genes related to autophagy, apoptosis, and inflammation in the gill, spleen, testis, and ovary of Sebastiscus marmoratus. The results showed that the higher HCQ concentration (77.40 and 154.80 mg/mL) led to mass mortality within an hour. The half-lethal concentration (LC50, 24 h) of HCQ for S. marmoratus was approximately 48.95 mg/mL. When exposed to 30.96 mg/mL HCQ for 24 hours, autophagy was blocked as revealed by electron microscopy from gill, spleen and testis of fish. The expressions of autophagy-related genes (LC3/Cx43), apoptosis-related genes (Cas3/p53), and inflammation-related genes (TNF-α/IL8) exhibited tissue-specific and dose-dependent responses. The gonads showed preferential expression of all these genes and were found to be sensitive and regular after HCQ treatment. For example, at a concentration of 30.96 mg/mL, the testis demonstrated a regularity that suggests it is an ideal candidate tissue for studying the role of HCQ or autophagy. This study systematically revealed the response of S. marmoratus to different concentrations of HCQ and provided optional assay concentrations for key tissues, serving as an important reference for the future studies on HCQ and autophagy in S. marmoratus. Furthermore, the potential crosstalk between autophagy, apoptosis, and inflammatory pathways initially identified in this study could be helpful for the future research on autophagy regulation in marine fish.

Effects on fatty acids, biochemical composition and growth of rotifer (Brachionus plicatilis) fed with different concentrations of Nannochloropsis sp.

AbstractThe density of microalgae and nutrient content have impact on the population growth rate, egg rate and nutritional values of rotifers. The aim of this study was to investigate the effects of different concentrations of the alga Nannochloropsis sp. (N10: 10x106, N20: 20x106, N30: 30x106 cells mL-1) on the growth, egg rate, biochemical composition and fatty acid profile of the rotifer Brachionus plicatilis. To maintain a consistent algae density, an equivalent quantity was supplemented to compensate for any reduction. The study was started with three replicates, each with an initial inoculation density of 200 Brachionus individuals mL-1 . The investigation spanned 5 days, during which population parameters (total number of individuals, proportion of individuals with eggs and growth rate) were calculated. After the study period, rotifers were harvested for subsequent biochemical and fatty acid analyses. Notably, the N30 group showed the highest proportions of crude protein (54.41%) and crude lipid (34.5%). The N20 group displayed the most substantial content of total fatty acids, recording a value of 70.69%. Consequently, the N30 group, with a concentration of 30x106 cells mL-1, emerged as the most proficient group, demonstrating superior performance in both population growth and biochemical composition.

Pengaruh Penambahan Variasi Konsentrasi Karbon Aktif pada Media Kultur In Vitro untuk Pertumbuhan Anggrek Dendrobium welirang

This research aims to determine the effect of different concentrations of added activated carbon and determine the optimal concentration in the in vitro culture growing medium for Dendrobium welirang orchids to increase plant growth. The type of research used is true experimentation. This research used a Completely Randomized Design (CRD), and the sample used in this research was the orchid species Dendrobium welirang. The instrument used is in the form of a table. The type of data used is quantitative data. The data were analyzed statistically using the ANOVA test. Data collection was carried out in the third month after planting. The data taken includes leaf length, number of leaves, root length, and number of roots. The results of the research showed that adding variations in activated carbon concentration to the in vitro culture medium affected the parameters of root number, root length, leaf length, and number of leaves of the Dendrobium welirang orchid. In conclusion, the addition of activated carbon has a real influence on the growth of Dendrobium welirang orchids. Keywords: Dendrobium welirang, Activated Carbon, In vitro Culture.

Attenuation of the effect of salinity on redwood (Caesalpinia platyloba) by saline compost

The objective of this research is to evaluate the effect of saline compost on the attenuation of salinity stress in palo colorado (Caesalpinia platyloba). A representative sample of saline soil was taken at Ciudad Universitaria Intercultural, sieved (mesh &lt; 5mm) and composted with cattle manure and Cynodon dactylon garden grass pruning, in a 3:1:1 ratio (stubble: manure: saline soil), for 120 days, the characterization consisted of measuring the bulk density (DA), saturation point, pH and EC of the saturated paste extract. The effect of different concentrations of NaCl on the in vitro germination of palo colorado seeds was evaluated in a randomized complete block design with two substrates (blocks), B1) sand-perlite; B2) sand-perlite-salt compost, five treatments 0 M, 0.3 M, 0.4 M, 0.5 M and 0.6 M of NaCl, eight replicates per treatment. The control irrigation was with distilled water. The plants transference to polyethylene cups was fifteen days after sowing; the cups had the same substrate mixture. The variables measured were survival, leaf biomass and dry root biomass. The EC decreased during composting, from 9.45 dS/m low to 2.4 dS/m; the pH of the samples did not show a significant difference. It is concluded that neither the substrate nor the NaCl concentration significantly influenced the germination of palo colorado seeds, and that the saline compost had no effect on the mitigation of salt stress in palo colorado plants.

Effect of Ziziphus Spina-Christi Leaves on the Level of Testosterone Hormone and ‎Semen Parameters of Iraqi Bucks Semen

The study aimed to assess the effects of different concentrations of Ziziphus spina-Christi (ZSC) leaves on the fertility of buck goats. Twelve buck goats aged 12–15 months with an average body weight of 22±2 kg were divided randomly into three dietary treatments: T1, T2, and T3, each given a basal diet daily plus feeds of 20 g/kg (2%) of ZSC for T1 and 40 g/kg (4%) of ZSC for ‎T2; and T3 was used as the control that consumed a basal diet only. The experiment lasted for 3 months. On days 45 and 90, blood samples were collected for plasma testosterone measurement using an immunochromatographic assay. Additionally, semen samples were collected at the same time points for physical (volume, color, and pH) and microscopic (semen concentration, mass and individual motility percentage, and live-dead sperm percentages) evaluations. ‎ The results showed that ZSC at 4% significantly (P&lt;0.05) increased the testosterone hormone concentration and all physical parameters at 45 and 90 days comparing to 2% ZSC and control ‎groups. ‎Additionally, ZSC at 4% significantly improved (P&lt;0.05) the microscopic parameters at both ‎time periods compared to 2% ZSC and control groups. The present findings revealed that ZSC at 4% diet could enhance goat semen processing and ‎goat fertility enhanced significantly (P&lt;0.05) the testosterone hormone that had higher values ‎compared to ‎‎(2%) of the ZSC control group. Similarly, (4%) of ZSC affected the physical ‎assessment of ‎semen, such as the volume, color, and pH values. Thus, adding ZSC with a high ‎concentration ‎‎(4%) could enhance goat semen processing and goat fertility in general.

Thyroxine regulates pig Sertoli cell line proliferation and maturation through the IKK/NFκB and p38 MAPK signaling pathways

The aim of this study was to investigate the signaling pathways involved in the proliferation and differentiation of pig Sertoli cells (SCs) mediated by thyroid hormone (T3) to provide a theoretical and practical basis for enhancing pig semen production. The effects of different concentrations of T3 on the proliferation of pig SCs were evaluated using the CCK8 assay. The impact of T3 on the proliferation and differentiation of pig SCs was further examined using RNA-seq, qPCR, and Western Blotting techniques. Additionally, the involvement of the p38 MAPK and NFκB pathways in mediating the effects of T3 on SCs proliferation and differentiation was investigated. Our findings revealed a strong correlation between the dosage of T3 and the inhibition of pig SCs proliferation and promotion of maturation. T3 regulated the activation state of the NFκB signaling pathway by upregulating IKKα, downregulating IKKβ, and promoting IκB phosphorylation. Furthermore, T3 facilitated SCs maturation by upregulating AR and FSHR expression while downregulating KRT-18. In conclusion, T3 inhibits pig SCs proliferation and promote pig SCs maturation through the IKK/NFκB and p38 MAPK pathways. These findings provide valuable insights into the mechanisms by which T3 influences the proliferation and maturation of pig SCs.

Improvement of sulforaphane production in hairy root cultures of broccoli (Brassica oleracea L. var. italica) by eliciting Myrosinase gene expression and its effect on breast cancer cells

Plant-derived hairy root cultures have the potential to be a significant biotechnological tool for producing valuable secondary metabolite compounds. This study deals with the production of broccoli’s hairy root cultures via the mediation of Agrobacterium rhizogenes and investigates the effects of different concentrations of copper sulfate (CuSO4) and zinc sulfate (ZnSO4) as two elicitors on the production of sulforaphane in broccoli's hairy root. The expression of the Myrosinase (MY) gene, associated with sulforaphane production, was quantified using real-time PCR. Moreover, the cytotoxic effect of different treatments of sulforaphane on the breast cancer cell line, MDA-MB-231, was estimated and the transcription levels of apoptosis-associated genes were also determined. The highest amount of sulforaphane was produced at 4 µM CuSO4 after 16 h and 16 µM ZnSO4 after 8 h of elicitation. Overexpression of the MY gene was found to be associated with increased production of sulforaphane. All sulforaphane treatments induced a dose-dependent reduction in the viability of cancer cells. The sulforaphane extracted from broccoli's hairy roots treated with CuSO4 exerted a higher cytotoxic effect on the MDA-MB-231 breast cancer cell line than the sulforaphane extracted from broccoli's hairy root treated with ZnSO4. All treatments of sulforaphane up-regulated the Apoptosis Regulator (Bax) gene, Proapoptotic Bcl2 Associated X, Caspase-3, Caspase-8, and Caspase-9, while down-regulating the B-cell lymphoma 2 (Bcl-2) gene (antiapoptotic) transcription. The overall results showed an antiapoptotic effect of sulforaphane extract derived from broccoli's hairy roots on MDAMB-231 breast cancer cells, thereby establishing the potential anticancer activity of the sulforaphane produced in broccoli's hairy root cultures.

Preovulatory follicular fluid secretome added to in vitro maturation medium influences the metabolism of equine cumulus-oocyte complexes

BackgroundIn vitro embryo production is a highly demanded reproductive technology in horses, which requires the recovery (in vivo or post-mortem) and in vitro maturation (IVM) of oocytes. Oocytes subjected to IVM exhibit poor developmental competence compared to their in vivo counterparts, being this related to a suboptimal composition of commercial maturation media. The objective of this work was to study the effect of different concentrations of secretome obtained from equine preovulatory follicular fluid (FF) on cumulus-oocyte complexes (COCs) during IVM. COCs retrieved in vivo by ovum pick up (OPU) or post-mortem from a slaughterhouse (SLA) were subjected to IVM in the presence or absence of secretome (Control: 0 µg/ml, S20: 20 µg/ml or S40: 40 µg/ml). After IVM, the metabolome of the medium used for oocyte maturation prior (Pre-IVM) and after IVM (Post-IVM), COCs mRNA expression, and oocyte meiotic competence were analysed.ResultsIVM leads to lactic acid production and an acetic acid consumption in COCs obtained from OPU and SLA. However, glucose consumption after IVM was higher in COCs from OPU when S40 was added (Control Pre-IVM vs. S40 Post-IVM: 117.24 ± 7.72 vs. 82.69 ± 4.24; Mean µM ± SEM; p < 0.05), while this was not observed in COCs from SLA. Likewise, secretome enhanced uptake of threonine (Control Pre-IVM vs. S20 Post-IVM vs. S40 Post-IVM: 4.93 ± 0.33 vs. 3.04 ± 0.25 vs. 2.84 ± 0.27; Mean µM ± SEM; p < 0.05) in COCs recovered by OPU. Regarding the relative mRNA expression of candidate genes related to metabolism, Lactate dehydrogenase A (LDHA) expression was significantly downregulated when secretome was added during IVM at 20–40 µg/ml in OPU-derived COCs (Control vs. S20 vs. S40: 1.77 ± 0.14 vs. 1 ± 0.25 vs. 1.23 ± 0.14; fold change ± SEM; p < 0.05), but not in SLA COCs.ConclusionsThe addition of secretome during in vitro maturation (IVM) affects the gene expression of LDHA, glucose metabolism, and amino acid turnover in equine cumulus-oocyte complexes (COCs), with diverging outcomes observed between COCs retrieved using ovum pick up (OPU) and slaughterhouse-derived COCs (SLA).