No full-text available
To read the full-text of this research,
you can request a copy directly from the author.
A Rapid and Sensitive Method for Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye Binding
Abstract
A protein determination method which involves the binding of Coomassie Brilliant Blue G-250 to protein is described. The binding of the dye to protein causes a shift in the absorption maximum of the dye from 465 to 595 nm, and it is the increase in absorption at 595 nm which is monitored. This assay is very reproducible and rapid with the dye binding process virtually complete in approximately 2 min with good color stability for 1 hr. There is little or no interference from cations such as sodium or potassium nor from carbohydrates such as sucrose. A small amount of color is developed in the presence of strongly alkaline buffering agents, but the assay may be run accurately by the use of proper buffer controls. The only components found to give excessive interfering color in the assay are relatively large amounts of detergents such as sodium dodecyl sulfate, Triton X-100, and commercial glassware detergents. Interference by small amounts of detergent may be eliminated by the use of proper controls.
... Total carbohydrate was extracted and measured by phenol-sulfuric acid method (DuBois et al. 1956). Total protein content was examined according to Kaunas blue method developed by Bradford et al. (1976). Amino acid was analyzed using L-8500 amino acid analyzer (Hitachi, Japan). ...
... Protein and carbohydrates measurement: Taking serum albumin and D-glucose as standards, the protein and carbohydrates contents were examined by a modified Kaunas blue method and phenol sulfuric acid method, respectively (Dubois et al. 1956, Bradford et al., 1976. Taking 10 mL of algal sample was subjected to centrifugation (4000 rpm, 10 min, 4 ℃) to collect the cell pellets, and then was re-suspended with 10 mL distilled H 2 O and lysed on ice by an ultrasonic cell disruptor (each 3 s working with a 2 s interval, 300 w, 8 min). ...
Isochrysis galbana is considered a promising feed for aquaculture, but high cultivation cost well as low substance yield and quality limit its application prospects. This study explored the feasibility and regulating mechanism of green tide waste Ulva prolifera for I. galbana substance production. U. prolifera hydrolysate (UPH) was prepared as an alternative substrate from 5 to 50% concentrations to culture I. galbana. Compared to the control f/2 medium, the biomass production of I. galbana added with 40% UPH reached 318.75 mg/L, which was the most effective up to 1.24-fold increase. Meanwhile, the highest pigments, lipid, protein and carbohydrate productions were observed in the 40% UPH substrate, which were around 1.17 to 1.70 times higher than that of the control, respectively. The above parameters were the lowest with the addition of 5% UPH, except for the lipid content, which was slightly higher than control. The photosynthetic activity showed that 5% UPH blocked four electron transfer sites of I. galbana, including from Quinone A (QA) to QB, from plastoquinone (PQ) to photosystem I (PSI), the oxygen evolving complex (OEC) activity on the donor side and the acceptor pool on the PSI. However, 40% UPH enhanced the electron transfer beyond QA, from PQ to PSI and the acceptor pool of PSI. The photosynthetic parameters φPo, φEO, PICS and PIABS, which had strong correlation with the biomass and substance accumulation of I. galbana, were found to be enhanced by 40% UPH. Changes in electron transfer regulated the photosynthetic activity and metabolism of I. galbana. Encouragingly, the production cost of cultivating I. galbana with UPH prepared from green tide waste was almost negligible compared with the f/2 medium (US$ 15.6). Therefore, the study provides a win–win strategy for obtaining cost-effective energy production as well as the resource utilization of green tide waste.
Graphical Abstract
... Merck KGaA) (Bradford, 1976;Rauca et al., 2018). ...
... The culture medium from DOX-treated and control spheroids group was collected and centrifuged at 350 x g for 5 min, to remove cell debris (Mrówczyńska et al., 2022) . Protein concentration in the supernatant was determined by Bradford assay, followed by treatment with non-reducing buffer (4% SDS, 20% glycerol, 0.01% bromophenol blue, 125 mM Tris-HCl, H2O) (Bradford, 1976) . Thereafter, 50 µg proteins/group were separated via SDS-PAGE, under nonreducing conditions, using a polyacrylamide 7.5% separation gel containing 0.1% gelatin (Licarete et al., 2020;Toth & Fridman, 2001) . ...
Chemoresistance continues to remain a significant barrier to the effective treatment of various cancers, including melanoma. Previous studies, including our own, have demonstrated the limited efficacy of doxorubicin (DOX) against melanoma cells and tumors, both in vitro and in vivo, primarily due to acquired chemoresistance. The lack of favorable outcomes in metastatic melanoma patients treated with liposomal DOX highlights the need to identify early DOX-induced resistance biomarkers and develop combination strategies to improve efficacy. This study explores early adaptive responses in B16.F10 melanoma spheroids after two sub-inhibitory DOX exposures (≈IC30), focusing on chemoresistance mechanisms in cell cycle regulation, DNA repair, apoptosis, angiogenesis, and invasiveness. Melanoma spheroids were cultured in ultra-low attachment plates and treated with DOX in two 48-hour phases separated by 48-hour recovery periods, followed by viability, morphology, gene/protein expression, and enzyme activity assessments using RNA-seq, RT-qPCR, Western blot, and gelatinase assay, respectively. Functional analysis confirmed downregulation of cell cycle pathways, suppression of stress response and survival pathways (e.g., TNFA_SIGNALING_VIA_NFKB and MTORC1_SIGNALING). Despite mRNA data showing cell cycle arrest (downregulated CDK1, Ticrr, Cdc20) and major DNA damage and impaired repair (downregulated BRIP1, BARD1, Rad51, Rmi2), after first exposure to DOX, cells probably bypassed cell cycle checkpoints by upregulating transcripts and proteins linked to pro-survival (Tnfrsf19, cIAP-2) and angiogenesis (VEGF, eotaxin-1), and by upregulating transcripts related to cell adhesion (Pcdh beta family members), and ECM remodeling (Ecm1, Jam2), suggesting early adaptation mechanisms to DOX. With a second DOX exposure, melanoma spheroids showed stronger chemoresistance, evidenced by enhanced angiogenesis (upregulated Aqp1, VEGF, and Ackr3), increased invasiveness (upregulated MMP-2, favored MMP-9 activity), and a 40% increase in pNF-kB levels, reinforcing drug tolerance and aggressiveness. This study provides valuable insights into the molecular drivers of chemoresistance, revealing that early DOX resistance in melanoma arises from adaptive mechanisms enhancing survival, angiogenesis, and invasiveness. Repeated exposure reinforces these traits, sustaining drug tolerance.
... Protein concentrations were adjusted to 0.6 -0.8 mg/ml for the cerebral cortex and 0.8 mg/ml for the hippocampus. Protein measurement was performed using the Bradford method with bovine serum albumin as the standard [32]. AChE activity was determined according to the method previously described [33]. ...
... The cell-containing pellet was resuspended in 500 μL of saline and used for analyses. Protein concentration was measured using the Bradford method with bovine serum albumin as the standard [32]. ...
Alzheimer’s Disease (AD) is characterized by memory decline, dysregulation in cholinergic and purinergic signaling, neuroinflammation, and oxidative stress. Current treatments are limited, highlighting the need for new compounds to prevent or delay AD progression. Thiazolidinones have emerged as promising candidates due to their antioxidant, anti-inflammatory, and anticholinesterase properties. The aim of this study was to evaluate the effects of 3-(3-(diethylamino)propyl)-2-(4-(methylthio)phenyl)thiazolidin-4-one (DS27) in a rat model of scopolamine-induced memory deficits. Male rats were divided into groups: I - Control, II - Scopolamine (SCO) (1 mg/kg), III - SCO and DS27 (5 mg/kg), IV - SCO and DS27 (10 mg/kg), V - SCO and donepezil (5 mg/kg). The animals were treated orally with DS27 or donepezil for seven days. On the 8th day, they underwent the open field test and inhibitory avoidance training, followed by intraperitoneal administration SCO. Twenty-four hours later, an inhibitory avoidance test was conducted. Acetylcholinesterase (AChE) activity, oxidative stress, and inflammatory and purinergic parameters were analyzed in the cerebral cortex, hippocampus, cerebrospinal fluid, serum, lymphocytes, and liver. DS27 prevented memory deficits, alterations in AChE activity, and oxidative damage induced by SCO in brain structures. Additionally, DS27 prevented SCO-induced decrease in IL-10 levels, and increase in IL-6, and TNF-α expression in the cerebral cortex, and normalized ATP and ADP hydrolysis in cerebrospinal fluid and lymphocytes. DS27 did not induce oxidative stress in the liver or alter serum biochemical parameters. These findings suggest that DS27 has significant neuroprotective properties and could be a promising alternative for treating neurodegenerative diseases like AD.
Graphical Abstract
... Proteins were recovered within the supernatant. Protein assays were performed using Bradford method (Bradford 1976). The standard curve was done by using BgG in MQ-water as follows: 0 µg.µL −1 , 1 µg.µL −1 , 2 µg.µL −1 , 5 µg.µL −1 , 8 µg.µL −1 , 10 µg.µL −1 . ...
The increasing demand for sustainable agriculture has highlighted the need for innovative strategies to enhance crop productivity while reducing environmental impact. This study evaluates the potential of Limnospira indica-based biostimulants to promote plant growth in Solanum lycopersicum (tomato) and modulate rhizosphere and endospheric microbiota. Two culture derived fraction of L. indica –culture media (CM) and biomass extracts (BM)– were prepared from nitrate-sufficient (N+) and nitrate-deficient (N-) cultures and applied through amendment or foliar treatments. Phenotypic analyses revealed that biostimulant extracts produced from L. indica CM N- significantly enhanced flower and fruit production, as well as microbial diversity compared to water controls and N+ treatments. 16S sequencing analyses demonstrated that biostimulant extracts enriched plant growth-promoting genera known for their roles in nitrogen fixation, phytohormone production, stress resilience and bioremediation. While numerous metabolites impacting plants are known, polysaccharides excreted under nitrate-deficient conditions were suggested as key drivers of microbial dynamics, promoting biofilm formation and enhancing plant-microbe interactions. Temporal analyses highlighted shifts in microbial communities further correlated with phenotypic improvements. These findings underscore the potential of nitrate-deficient L. indica-based biostimulant extracts to sustainably optimise plant growth by targeting specific microbiota interactions. This study provides a foundation for advancing biostimulant formulations and promoting environmentally friendly agricultural practices.
... For each enzyme assay, five replicates per treatment were used. The protein content was determined using the Coomassie brilliant blue assay (Bradford, 1976). ...
The interest in botanical insecticides has surged due to environmental apprehensions and insect resistance to synthetic insecticides. In this study, we explored the insecticidal, biochemical, and physiological effects of Mentha pulegium essential oil (EO) and Artemisia camphorata methanol extract (ME) on the fourth-instar larvae of Spodoptera littoralis. By GC–MS analysis, the main compounds identified in M. pulegium EO were menthone (22.45%), 1,8-cineole or eucalyptol (19.37%), and pulegone (8.67%). The abundant flavonoid compounds were apigenin, luteolin, quercetin, and naringin, whereas the abundant phenolic compounds were 4-hydroxybenzoic, ferulic, and chlorogenic acids in A. camphorata ME after analysis by HPLC. The LC50 values after 24 h of S. littoralis treatment with M. pulegium EO, A. camphorata, and novaluron were 4458.3, 7349.9, and 9.8 mg/L, whereas the LC50 values after 48 h of treatment were 2495.3, 4024.5, and 7.7 mg/L, respectively. The activities of mixed-function oxidase, α and β-esterases, and glutathione S-transferase were significantly increased in S. littoralis larvae after 24 and 48 h of treatment with LC25 and LC50 concentrations of novaluron, M. pulegium EO, and A. camphorata ME. The mean number of prohemocytes, granulocytes, and plasmatocyte cells was significantly decreased at 24, 48 and 72 h of treatment with LC25 of M. pulegium EO and A. camphorata ME. In contrast, the number of oenocytes and spherulocytes was increased compared to the control. On the other hand, novaluron caused a significant decrease in all hemocyte numbers. The hemocyte surface areas increased dramatically in all hemocyte types after treatment with M. pulegium EO and A. camphorata ME. In contrast, hemocyte surface areas significantly decreased after novaluron treatment compared to the control. These findings suggest that M. pulegium and A. camphorata may be eco-friendly alternatives for synthetic insecticides in S. littoralis control.
... Samples were then centrifuged at 15,000 x g for 15 min at 4°C, and the supernatants were collected. Protein concentrations were determined by the Bradford method (Bradford, 1976). ...
Introduction
Metabolic dysfunction-associated steatotic liver (MASL), the initial, asymptomatic stage of the metabolic dysfunction-associated steatotic liver disease, is directly involved in the progression to steatohepatitis. Healthy lifestyle and dietary measures are currently the only treatments for MASL. Given the high prevalence of MASL in the human population, candidate drugs for its prevention or treatment should have an acceptable safety profile. Repurposing drugs already in clinical use could help to identify effective and safe drug treatments for MASL. We have characterized a high-fat, high-fructose rat dietary model of simple hepatic steatosis to evaluate the potential anti-steatotic effect of mirabegron, which is already in clinical use for the treatment of overactive bladder. We have previously reported that mirabegron administration was unable to reduce liver triglyceride content in our rat model.
Methods
In the present work, we analyse stored liver, adipose tissue (perigonadal and brown), serum and faecal samples from our previous study and present new biochemical, faecal metabolomic and microbiome data.
Results and discussion
We show that oral administration of mirabegron significantly increases the expression of uncoupling protein 1 in brown adipose tissue and β3-Adrenergic receptor protein in perigonadal white adipose and liver tissues. Furthermore, mirabegron treatment changes the relative abundance of several genus and families of rat faecal microbiota, albeit without restoring the global biodiversity and evenness indexes observed in control rats, as well as faecal bile acids composition. These changes are probably due to a direct effect of mirabegron on the gut microbiome, rather than being mediated by changes in bile acid induced by drug treatment.
... Total proteins were determined according to a specific method, using bovine albumin as standard. Aliquots of the hemolyzed solution were added to the Bradford reagent and kept in the dark at room temperature for 5 min, and the absorbance reading was performed at 595 nm [39] with results expressed as μg/μl protein. ...
Natural antioxidants present in fruits have attracted considerable interest due to their presumed safety and potential nutritional value. Even though antioxidant activities of many fruits have been reported, the effects of phytochemicals of goji berry (GB) in patients with metabolic syndrome have not been investigated. In this study, we examined anthropometric and biochemical parameters in patients with metabolic syndrome after the consumption of GB. The patients were divided into two groups, control (C) and supplemented (S), and followed up for 45 days. Participants were individually instructed to carry out a healthy diet, but additionally, an inclusion of 14 g of the natural form of goji berry in the diet during 45 days for the S group was proposed. After 45 days of study, a significant reduction in transaminases as well as an improvement in lipid profile in the S group was observed. Likewise, a significant reduction in the waist circumference of the S group was observed when compared with that of the C group, and increased glutathione and catalase levels associated with a reduction of lipid peroxidation. These results suggest that this is an effective dietary supplement for the prevention of cardiovascular diseases in individuals with metabolic syndrome.
... Gpx was measured at 470 nm in a mixture with the assay buffer, 2 nm h 2 o 2 and 20 mM guaiacol. the total protein content in the enzyme extracts was determined according to Bradford (1976). ...
Currently, the use of biostimulants is considered an environmentally friendly alternative to synthetic fertilizers. In this work, we investigated the use of a biostimulant obtained from winemaking sludge by enzymatic hydrolysis processes in peat enrichment. The aim was to observe how this biostimulant affects the growth and development of green pepper (Capsicum annuum L. cv. Italian) seedlings. The biostimulant was applied in three doses (1, 2, and 3 g/L) for a total of four applications. Seedlings were harvested at a height of 8–16 cm and with 5–7 leaves. Plant height, weight of aboveground and belowground parts, macro- and micronutrient contents, photosynthetic pigments in leaves, and enzymes related to plant stress were determined. The results obtained indicated that plant height, weight of the aboveground and belowground parts, and macro- and micronutrient content in the aboveground and root parts were higher in plants treated with the highest dose of biostimulant. Compared with the control treatment (without amendments), the contents of chlorophyll a, chlorophyll b, and total carotenoids increased by 57%, 51.9%, and 53.1%, respectively, in plants treated with the highest dose of biostimulant. These results suggest that peat enrichment with this biostimulant could be useful for improving bell pepper quality.
... Quantitation of total protein. The protein concentration in the samples was estimated by the Bradford method with bovine serum albumin as a standard (Bradford, 1976). ...
Two lines of the parasitoid Habrobracon hebetor were analyzed: one infected with the endosymbiotic bacterium Wolbachia and one not infected. Differences in morphological characteristics between the lines were observed, in particular in the degree of cuticle melanization and in body size of adult parasitoids.
Sizes of both male and female parasitoids were significantly larger in the infected line. Wolbachia was found to affect levels of dopamine and tyrosine, phenoloxidase activity, and proline concentration in whole-body homogenates from different stages of parasitoid development.
... (total protein) Bradford. (1976) , pepsin amylase Worthington (1991) , trypsin chymotrypsin Erlanger et al. (1961) , lipase Borlongan (1990) . ...
... The ash content was determined by incineration at 600 ± 15 °C. Proteins were determined by the principle of protein-dye binding according to Bradford [24]. Acidity was determined by titration with NaOH 0.01 mol/l solution up to pH 8.2. ...
The formulation of a new functional beverage from Opuntia stricta juice enriched with whey cheese was investigated. Five beverages were developed with different percentages of juice. The storage stability was evaluated at 4 °C during 60 days in terms of physicochemical analysis (pH, acidity, total, and reducing sugar). The prepared beverages have a red color and a highly acceptable taste, the overall acceptability varies from 6.12 to 6.80 and from 7.16 to 8 respectively for the two categories of consumers 10-25 and over 25 years old. The relation between acceptability of beverages (for two categories of consumers) and sensory parameters (color, juice percentage, taste, and texture) was modelized using linear regression model with a correlation coefficient (R 2) of 72.9% and 70.5% respectively for consumers 10-25 and over 25 years old. In this vein, the beverages were appreciated differently according to the age range of consumers (10-25 years or over 25 years old). A mathematical relationship between the responses (pH, acidity, reducing, and total sugars) and covariates (storage period and juice concentration) was obtained, described by first-order polynomial equations with an R 2 of 87%, 83%, 89.5%, and 88.7% respectively for pH, acidity, reducing, and total sugar. The study showed that the drinkable beverage based on whey and Opuntia stricta juice with a proportion of juice varying from 35 to 25% is the best regarding sensory properties and physicochemical stability.
... SOTI06 was grown in 100 mL of the DB liquid medium containing calcite, and incubated in a shaker at 120 rpm for 5 days at 30 °C. The cultures were harvested by centrifugation at 10,000 rpm for 10 min, and the cell-free culture supernatants obtained at periodic intervals (0 to 5 days) were subjected to the analysis of calcium [27], calcium carbonate [23], carbonates, bicarbonates [28], phosphatase [25], protein concentration [29], pH, TA, and bacterial growth. ...
Soil degradation due to sodicity is a major constraint to agricultural development in arid and semi-arid regions. The accumulation of exchangeable Na⁺ ions affects soil physicochemical properties, which subsequently increases pH, thus reducing crop yield and nutrient availability. Several practices have been followed for revitalizing salt-affected soils, such as the addition of inorganic or organic amendments. Since most of these soils are calcareous (CaCO3) in nature, they can serve as a cationic source to release Ca²⁺ to replace the Na⁺ from the clay-complex. Though CaCO3 is poorly soluble, dissolution can be easily achieved by calcite dissolving bacteria. The present study aimed to evaluate the efficiency of Brevibacterium sp. SOTI06, in reclaiming calcareous sodic soil, along with organic and inorganic inputs through a soil incubation study. The reduction in pH, Na⁺, ESP (Exchangeable sodium percent), and free CaCO3 during the incubation period confirms the efficiency of amendments. The pH was drastically reduced from 9.10 to 8.20 in gypsum-applied soils. The combined effect of bacterium and press mud reduced higher rates of CaCO3 in soil (16.6 to 14.5%), with a rise of Ca²⁺ ions (23.5 to 28.7 meq 100 g soil–1). The mean calcite dissolution was higher in bioinoculant and gypsum-applied soils (9.04%). The SEM (Scanning Electron Microscopy) images confirmed the colonization and calcite dissolution potential of Brevibacterium sp. SOTI06 by pit formation in calcite stones. Hence, this study revealed that the combined application of bioinoculants with organic and inorganic amendments can effectively reclaim calcareous sodic soils.
... The leaves were crushed and homogenized before the soluble sugar content was measured using the anthrone method (Clegg 1956). The soluble protein content was determined using the Coomassie Brilliant Blue method (Bradford 1976). The vitamin C content was quantified via iodometric titration (Ballentine 1941). ...
Red and blue lights are the most effective spectral components absorbed by plants and are typically applied in a constant spectrum in plant factory agricultural practices. Research and application of non‐constant light modes are relatively rare. In this study, we examined the impacts of varying blue light photon intensity and durations at 5‐, 15‐, 30‐, and 45‐min intervals of intermittent exposure on the growth and development of lettuce (Lactuca sativa) in plant factories while maintaining a constant red light photon flux and daily light integral (DLI). Compared to the constant light condition, intermittent blue light irradiation treatments accelerated the emergence of the leafy head trait in lettuce without compromising photosynthetic capacity and biomass. Specifically, intermittent blue light treatment with 15‐min intervals led to a reduction of 8 days in the time needed for half of the lettuce plants to reach the heading stage. Furthermore, the petiole bending angle in treated lettuce was just 70.2% of that observed under constant light conditions, with strong correlations between multiple hormone levels and bending angle in petioles. Transcriptome sequencing analyses revealed significant differential expression of signaling‐related genes between constant and intermittent blue light treatments. The transient and dynamic expression of light‐responsive and hormone metabolism‐related genes indicated that 15‐min intermittent blue light exposure better maintained the rhythmic differential expression of response genes, leading to different hormone accumulations and consequently accelerating the development of leafy heads.
... Protein concentration in urine and plasma were determined using a Milton Roy Spectronic 1201 spectrophotometer with the Bradford technique [19]. ...
Standardized diets for laboratory rodents become increasingly important. Dietary imbalances can negatively impact experimental outcomes. Commercially available diets often include ingredients that cannot be experimentally manipulated and may not meet all nutritional and experimental requirements. Current growth and maintenance diets for experimental animals are regulated by the American Institute for Nutrition (AIN). In our laboratory, we developed a diet based on egg-white protein (EWP) using endogenous ingredients that are safe for experimental rats and can be experimentally manipulated according to AIN standards. Therefore, this study aims to test and validate the nutritional quality of the EWP diet for the Dahl-SS rat strain, which is known to be nutrient-sensitive and genetically predisposed to develop hypertension. 14 pairs of Dahl-SS rats (14 females and 14 males) were used for mating. The rats were randomly divided into two groups: 7 pairs were fed a commercial standard diet, and the other 7 pairs were fed an EWP diet. From here, 10 pregnant rats (5 from each group), 14 weeks of age, and 1 week of gestation, were selected for the study and were fed their respective diets. The offspring were weighed at birth and weaned at 4 weeks of age. Five pups were randomly selected from each group (standard diet group and EWP diet group) and continued with their mother’s respective diets for an additional 6 weeks. During the study they were subjected to Biological (clinical-behavioral status, metabolic analysis, relative organ-weight), Biochemical (mineral analysis, hemogram, protein analysis) and Physiological parameters analysis (Blood Pressure determination, enzymatic activity of Na+ and Na/K+ pump in renal cortex). To complete analysis of the different parameters, at 10 weeks, the rats were sacrificed by exsanguination via cardiac puncture under intraperitoneal anesthesia with thiopental sodium (20-25 mg/kg). Blood was collected in heparinized and centrifuged tubes at 3500 g for 10 minutes at 4°C. Plasma was separated and stored at -20°C until analysis. The kidneys, heart, spleen, and liver were collected and weighed. Dahl-SS rats fed the EWP diet showed normal behavioral, motor and metabolic values, faster and greater body weight gain, blood and urine biochemistry values within normal range, stable blood pressure, normal enzyme activity, and adequate kidney function in comparison to the rats fed the standard diet. According to the parameters assessed, it was possible to conclude that the EWP diet has proven to be a suitable option for supporting maintenance, growth, and blood pressure regulation in Dahl-SS rats.
... The concentration of MDA was measured by liquid chromatography according to a modification of the method of Richard et al. [81] as previously described [82]. The protein content of the samples was determined using the Bradford method [83]. ...
The rd2 mouse model, characterized by a mutation in the Prph2 gene, exhibits abnormal development of photoreceptor outer segments, resulting in progressive retinal degeneration. While the correlation between poly-ADP-ribose polymerase (PARP) activity and the degeneration of rod photoreceptors is established in the rd2 model, the specific mechanism driving cone degeneration in this model remains unclear. Furthermore, it is yet to be determined whether inhibiting PARP activity can effectively impede the degeneration of cone photoreceptors in this context. We demonstrated that PARP inhibitors Olaparib, BMN-673, and 3-aminobenzamide (3AB), effectively reduced photoreceptor cell loss in the rd2 retina. Notably, rd2 retinas exhibited decreased cone density, but treatment with PARP inhibitors significantly protected cone photoreceptors. The PARP inhibitors, particularly BMN-673, demonstrated a significant protective effect as evidenced by increased rhodopsin expression within the outer segment and a concurrent decrease in Müller cell activity indicated by GFAP expression. The treatment also resulted in significant changes for markers of oxidative stress, such as glutathione (GSH), and oxidized glutathione (GSSG). Notably, the administration of PARP inhibitors also reduced CD9 expression (extracellular vesicle marker), which were significantly increased within the outer nuclear layer (ONL) in the rd2 retinas. Among PARP inhibitors, BMN-673 demonstrated the highest efficacy in preserving photoreceptors, particularly benefiting cone cells.
Graphical Abstract
... The cell lysates were centrifuged at 15,000 × g for 1 min at 4 °C and the supernatant was collected. Total protein content was determined using Bradford assay [33]. The assay was performed in a total volume of 100 μl in 96-well plates. ...
Stemona tuberosa is widely recognized for its traditional applications as an anti-cancer agent. This study aimed to assess the anti-cancer properties of S. tuberosa in human lung adenocarcinoma A549 cells. Among the various solvent extracts of S. tuberosa, the methanolic extract showed the highest toxicity against A549 cells. The S. tuberosa extract elicited cytotoxic effects and suppressed colony formation in A549 cells in a dose-dependent manner. S. tuberosa activity was further supported by AO/EtBr staining, increased caspase 3/6 activity, upregulation of pro-apoptotic genes, DNA damage , and elevated lipid peroxidation, with decreasing antioxidant levels. LC-MS analysis identified 80 predominant secondary metabolites in the methanolic extracts of S. tuberosa. A network pharmacology study identified SRC as the primary target of compounds identified from S. tuberosa. SRC protein is crucial for advancing lung cancer because of its function in cell proliferation, survival, and metastasis. Among the various compounds identified from S. tuberosa extract, 4-Azatricyclo [4.3.1.13,8] undecan-5-one (ADE) (− 10.88 kcal/mol) and Dihydro-normorphine, 3-desoxy-(DNY) (− 10.83 kcal/mol) exhibited notable binding affinities for SRC. Further analysis using molecular dynamics simulations (100 ns) validated the stability of SRC-ligand complexes, with RMSD of 1.8 and 2.2 Å for ADE and DNY, respectively, alongside the establishment of essential hydrogen bonds with pivotal residues, including ASP408, ALA403, and THR438. Finally, gmx._MMPBSA showed favourable ΔGbind values for ADE (− 15.06 ± 0.11 kcal/mol) and DNY (− 15.66 ± 0.25 kcal/ mol), which highlights the significant potential of ADE and DNY as effective SRC inhibitors, suggesting S. tuberosa as a novel candidate for cancer therapy.
... Total soluble proteins were measured in accordance with the Bradford (1976) method. Bovine serum albumin (BSA) solution was used as a protein standard, and its absorbance was computed at 595 nm. ...
Melatonin, a multifunctional, non‐toxic regulatory molecule, plays a crucial role in enhancing tolerance to abiotic stress, which is tightly linked to S metabolism. Despite the proven efficacy of sulfur (S) in enhancing abiotic stress tolerance, the combined effect of S and melatonin in stress mitigation remains to be elucidated. This is particularly relevant in the context of climate change, where the increased occurrence of waterlogging stress increases the risk of reduced S availability, leading to reduced yield and quality in rapeseed. The objective of this study is to examine the impact of a combination of foliar melatonin and sulfur, when administered to soil or leaves, on the response of plants to waterlogging stress. The experimental design involved the supplementation of rapeseed ( Brassica napus L.) plants with sulfur (S) to either the soil (0.2 g kg ⁻¹ ) or the leaves (300 ppm) 5 days prior to stress induction. The plants were subjected to waterlogging at BBCH–31 for a period of 7 days, preceded by a pretreatment 2 days prior to the stress with melatonin (200 μM). In comparison, untreated plants subjected to waterlogging showed a significant reduction in growth, nutrient uptake, photosynthetic activity, and sugar content but an increase in the antioxidant defense system. However, the application of melatonin significantly mitigated the adverse effects of waterlogging stress. In comparison with the control, soil‐S application exhibited higher efficacy than foliar S application in increasing plant resistance, as reflected by improved dry weight (+50%), photosynthesis (+12%), stomatal conductance (+40%), sulfur (+40%), magnesium (+59%), and reduced hydrogen peroxide (−22%) and lipid peroxidase (−26%). This combination also increased antioxidant defense by increasing catalase (+43%), glutathione reductase (+17%), ascorbate peroxidase (+47%), ascorbate (+39%), and glutathione (+40%) contents, in contrast to untreated waterlogged plants. The study underlines the potential of melatonin and sulfur as effective agents to alleviate waterlogging stress.
... Supernatants were collected from homogenized samples by centrifugation at 1000 g for 15 min at 4°C. Protein levels were quantified using the Bradford method [42]. Antioxidative enzyme activities were assessed by measuring superoxide dismutase (SOD) activity via the pyrogallol autoxidation method and catalase (CAT) activity using hydrogen peroxide (H₂O₂) [43,44]. ...
Cancer remains a significant global health challenge with limited therapeutic success, prompting the need for innovative treatment strategies. This study investigates the anticancer potential of nano‐encapsulated metal derivatives (CS‐QM2) using a zebrafish model with chemically induced cellular neoplasia. Characterization of CS‐QM2 nanoparticles revealed successful synthesis with a high entrapment efficiency and enhanced drug release under acidic conditions. Zebrafish embryos exposed to 7,12‐Dimethylbenz[a]anthracene (DMBA) exhibited significant malformations, macrophage accumulation, and abnormal tissue growth, which were markedly reduced by CS‐QM2 treatment. CS‐QM2 significantly increases intracellular ROS, resulting in higher LPO and induces apoptosis in neoplasm tissues. Furthermore, CS‐QM2 treatment alters the tumor microenvironment, reducing macrophage accumulation by decreasing neutral lipid droplets, disrupting TAM metabolic support and limiting their protumorigenic activities. Biochemical assays demonstrated restored activities of antioxidant enzymes SOD, CAT, and GSH. Gene expression analysis showed upregulation of apoptosis and tumor suppressor genes ( cas3 , p53 ) and downregulation of inflammatory genes ( cox‐2 , nf‐kb ). Histological assessment and SEM analysis confirmed reduced neoplasm occurrence and tissue abnormalities. These findings suggest that CS‐QM2 nanoparticles effectively inhibit neoplasm growth and modulate the tumor microenvironment through oxidative stress induction and gene expression regulation.
... The homogenates were then centrifuged at 13,000× g for 30 min at 4 • C, and the resulting supernatant was collected for further assays. The protein content in the extracts was measured following the method described by [27]. ...
In this study, plasma-activated water (PAW) was generated using a large-volume (5 L) plasma reactor with a quasi-stationary, water-cathode glow-type discharge in atmospheric pressure air. Tap water was activated up to 75 min. PAW exhibited high concentrations of long-lived reactive nitrogen species (RNSs), reaching 8 mM, which is between 4 and 26 times higher than those reported in previous studies. The reactor reached an RNS synthesis efficiency of 61 nmol/J and an RNS production rate of 526 μmol/min, both among the highest reported. PAW was evaluated on tomato and bell pepper. Seedling emergence was determined in a nutrient-free substrate. To assess plant growth, seedlings were transplanted into pots filled with either nitrogen-free or nutrient-rich substrate. PAW-irrigation significantly promoted seedling emergence and leaf expansion, especially in tomato plants. The plant growth-stimulating effects of PAW were more pronounced in nitrogen-free substrate: fresh weight of tomato and bell pepper increased up to 13.1-fold and 2.6-fold, respectively. In contrast, the effect on the nutrient-rich substrate was negligible. Tomato plants grown in the nitrogen-free substrate and irrigated with 75-min PAW reached a dry weight comparable to those grown in nutrient-rich substrate. PAW irrigation did not induce oxidative stress, as confirmed by malondialdehyde (MDA) levels and antioxidant enzyme activity.
The aim of the study was purification and biochemical characterization of a keratinase enzyme from indigenous microorganism and its promising potential biotechnological applications. The increasing demand of keratinases in different industrial sectors calls for the need of more robust and stable keratinases with potential industrial applications. Keratinase produced by Bacillus D2 strain was purified by gel filtration and Q-Sepharose chromatograph. The enzyme showed a specific activity of 525 U/mg with overall recovery of 23.8%. Molecular size of purified enzyme after SDS-PAGE and zymogram was found 40 kDa. Purified keratinase had an optimal pH 8.5 and optimal temperature 50oC. Enzyme displayed pH stability in pH range of 7.5–9.0 and thermal stability up to 60oC. Amongst reducing agents, sodium sulfite and dithiothreitol (DTT) reduced relative activity however; SDS increased activity at 5 mM concentration. Keratinase enzyme was found solvent stable at 0.5% and 1% concentration. MgCl2 enhanced activity up to 108% and 104% at 5 and 10 mM concentrations, respectively. Enzymatic treatment of goat skin and cow hide resulted in dehairing and removal of scud and keratin skin layer resultantly smooth intact skin surface was observed. However, conventionally treated hides produced dark and hard skin surface due to partial elimination of keratin layer. Potential of Bacillus strain for conversion of feather keratin waste in to valuable feather meal suggests its usefulness in poultry industry and eventually reducing environmental pollution hazards. Key words: Characterization, Feather waste, Keratinase, Leather dehairing, Protein purification
The treatment of lung cancer continues to be a significant challenge for many oncologists and their patients. Treatment using epidermal growth factor receptor inhibitors is connected to a positive outcome. Cetuximab, a chimeric monoclonal antibody targeting the epidermal growth factor receptor (EGFR), in conjunction with monoterpene phenol thymol, is recommended for the treatment of lung cancer. While a mild acne-like skin rash is quite frequent in patients using cetuximab, a severe rash is rare. The goal of the current study was to assess whether thymol could enhance the anticancer effectiveness of cetuximab in A-549, non-small cell lung cancer (NSCLC) cell line. We found that the combination of cetuximab and thymol synergistically suppressed cell proliferation by inducing membrane damaging, oxidative stress, and apoptosis in A-549 cells. Taken together, our results indicate that the combination of thymol and cetuximab could improve anticancer responses and may notably enhance treatment outcomes in NSCLC.
Environmental stresses, particularly drought and salinity, significantly impair plant growth and productivity. This study explores the novel synergistic interaction between biochar and arbuscular mycorrhizal fungi (AMF) in enhancing the resilience of sweet pepper plants subjected to the individual or combined stresses of drought and salinity. The impact of these biostimulants on growth parameters, photosynthetic efficiency, and biochemical traits was assessed. Sweet pepper plants were subjected to drought stress (35 and 75% of field capacity (FC)), salinity (0 and 150 mM NaCl), and their combined effects (150 mM NaCl +35% of FC), with treatments including biochar (2.5 g/kg soil), AMF, and their combination. Under drought stress, the dual application of biochar and AMF notably improved plant growth indicators such as shoot fresh weight, shoot height, and number of leaves by 50, 14, and 3%, respectively compared to the control plants. Under drought and salinity combined, this combination also enhanced photosynthetic pigments content by 144% for Chl a, 316% for Chl b, 212% for Chl T and 302% for carotenoids content respectively compared to the control plants. Additionally, AMF and Biochar combined reduced the oxidative effect of malondialdehyde (MDA) by 37% and hydrogen peroxide (H2O2) by 43%, indicating a reduction in oxidative damage. Furthermore, a significant increase in antioxidant enzyme activities was observed, with peroxidase activity (POX) rising by 33% and polyphenol oxidase activity (PPO) increasing by 212%, indicating enhanced stress tolerance. This study underscores the efficacy of using biochar and AMF together to bolster sweet pepper plant resilience, offering a viable strategy for improving plant performance under challenging environmental conditions.
The knowledge gaps in the nutritional composition and quality of traditionally textured plant-based products eaten as meat is affecting the global acceptance despite the acclaimed health, environmental, ethical, religious, and social benefits. This paper aimed to prepare and evaluate the nutritional quality of Nyam ngub for potential valorization and vulgarization. Standard methods were used to determine the chemical composition and to evaluate the quality of protein. Protein fractions were used to estimate the solubility and individual amino acids were analysed with rapid amino acid analyser. Nutrient bio accessibility was determined firstly by calculation through the phytate: mineral ratio for iron and while the simulated in-vitro gastrointestinal test evaluated the protein digestibility and mineral accessibility. Results indicated that nyam ngub had an ash content of 13.021.14g/g at a moisture content of 89.562.43% and dry matter of 12.860.30%. The reducing and total sugar content were 0.80.02 g/1000mL and 51.424.26 g/1000mL respectively yielding a moderate energy supply (67.260.72 Kcal/mol) compared to other tubers. The crude fibre, fat and protein were respectively 6.70.3 (g/100g), 3.070.42 (g/100g) and 6.030.15 (g/100g). The Calcium, iron, Zinc and Copper contents were 0.010.00 mg/100g, 1.60 g/100g, 0.250.04 mg/100g and 2.870.00 g/g respectively while vitamin A after conversion from -carotene was 1.650.77gRE/g and vitamin C was 5.0430.54 mg/100g. The albumin, globulin, prolamin, and glutelin fractions were 70.512.48, 65.931.44, 16.413.21 and 18.461.35 mgBSA/100g respectively. Iron and zinc were 57.320.58% and 51.730.23% accessible while protein had the greatest digestibility in the gastric phase (74.63%) compared to 70.15% in the intestines. The essential amino acids quantified in mg/ 100g were Arg (1.39), His (0.61), Leu (2.04), Lys (1.52) Met (0.59), Phe (1.40), and Thr (1.11). Despite the limited protein content and lack of some essential amino acids, the protein of nyam ngub was relatively soluble and available and the micronutrients are accessible.
Eucalypt wastes (EW), comprising bark, leaf and branches, when retained and distributed on-site are a valuable source of nutrients for soil and microbial biomass. This study investigates the acceleration of EW biodegradation through the introduction of the edible fungus Lentinula edodes in-vitro cultures. Production of enzymes, ergosterol and organic matter consumption (OMC) were assessed in the spent mushroom compost over 90-days incubation. Concurrently, the residual solid decreased to 71.9%, with 35% reduction in lignin and hemicellulose and 28.5% loss of cellulose. EW substrate was supplemented with rice bran (RB) and soybean bran (SB) in different ratios to explore L. edodes enzyme production during substrate decomposition. Composition of EW supplemented with cereal brans (CB) provided high production of endoglucanase (2,300 U / kg) within 45 days, β-glucosidase (375 U / kg) in 60 days. Productions of 80,000 U / kg of xylanase and 195 U / kg of β-xylosidase occurred in 45 days of cultivation. Laccase (3,750 U / kg) was determined in 60 days, along with manganese peroxidase (MnP) (6,000 U / kg) in 75 days in EW: SB (90:10). Ergosterol extraction (45 mg / kg) and L. edodes mycelial biomass (11 g / kg) was obtained in all combinations of EW and CB. The maximum biological efficiency (BE) of 67% was achieved in a single cycle with EW: RB 90:10. Overall, L. edodes emerges as a promising resource for EW biodegradation, and this work sheds light on applications for fungus-based degradation in forest eucalypt plantations. For fungus-based degradation in forest eucalyptus plantations. Parte superior do formulário.
Nectar and pollen attractants as well as visitors in Helleborus viridis ssp. viridis and H. purpurascens (Ranunculaceae), i.e. European species, were studied during two flowering seasons. The long-lived flowers of these species are characterized by long nectar availability lasting 15–17 days, i.e. until the end of pollen presentation. On average, approx. 2-fold more nectar and sugar mass was available in the flowers of H. viridis ssp. viridis compared to H. purpurascens. Both species are protogynous, having an initial female phase lasting for 3 days. The nectar and sugar mass was consistently male-biased. At the end of the flower’s life span, the nectar mass and the sugar concentration decreased considerably. In both species, sucrose predominated in the nectar, which also contained small quantities of other disaccharides (maltose and trehalose) and monosaccharides (glucose and fructose). The chemical composition of the nectar sugar varied between the sexual phases; sucrose in the female-phase nectar was replaced mainly by fructose. In addition to nectar, the flowers offered pollen (between 2 and 4 and 10–17 days of anthesis). In the study area, both Helleborus species were foraged by queens of bumble bees and honey bees; however, their distribution differed between the species. Bumble bees dominated in H. viridis subsp. viridis, whereas both pollinator groups were noted with a similar frequency in H. purpurascens.
To understand better the differences related to manufacturers’ koji making methods, we evaluated the environment during koji making using trays or boxes, and assessed the qualities of the resulting koji at the measured locations. During koji making using boxes, spatial temperature in the box was lower than the koji temperature, whereas during koji making using trays, spatial temperature in the tray was similar to the koji temperature. Moreover, the spatial humidity in the tray was higher than that in the box. These differences were considered to affect the koji qualities. The water content in koji made using trays was higher than that in koji made using boxes. Enzyme activities, mycelium content, and mycelial growth level on rice（haze） in koji made using boxes were higher than those in koji made using trays. However, the glucoamylase to α-amylase activity ratio in koji made using trays was higher than that in koji made using boxes. This study showed the differences in the environment during koji making with trays and boxes.
Bacillus zhangzhouensis OBB Liu et al. (Caryophanales: Bacillaceae) isolate was obtained from Polyphylla fullo (L., 1758) (Coleoptera: Scarabaeidae) larvae in the Microbiology Laboratory of the Biology Department of the Faculty of Science of Atatürk University in 2024. Additionally, the presence of the cry1 gene was identified. After endospore staining of the isolate, the presence of crystal protein was detected by phase contrast microscopy and SEM analysis. As a result of PCR, only the presence of the cry1 gene was detected and confirmed. The total protein contents were compared with those of Bacillus thuringiensis (Berliner, 1915) (Bacteria: Bacillaceae) by performing SDS-PAGE analysis using a crystal protein and spore mixture. B. zhangzhouensis OBB showed bands ̴ 250 kDa and ̴ 80 kDa, while B. thuringiensis showed bands corresponding to ̴ 70 kDa and ̴ 45 kDa. Probit analysis was used to determine the LC50 value of the isolates, and the Abbott method was used to determine the mortality percentages of the larvae. Spore-crystal mixtures of B. thuringiensis and B. zhangzhouensis OBB isolates were tested against P. fullo larvae at doses of 1000, 2000, and 4000 ppm. The highest mortality rate was determined in the spore-crystal mixture of B. zhangzhouensis OBB isolate at 4000 ppm dose.
Mung bean is a promising crop in Egypt, but the small seed size (≤ 4 g) of high-yielding varieties like Kawmy-1 limits its widespread adoption. Larger-seeded varieties with higher yield potential, such as VC1973 A (100-seeds > 4 g), are preferred by farmers. However, the impact of plant population densities on growth and yield of these contrasting varieties has not been well studied. This research was conducted during the 2023 and 2024 in summer seasons, aimed to evaluate the effects of varying plant population densities (75, 150, 225, and 300 thousand plants per fed, equivalent to 4200 m ² ) on the growth, yield, and physiological responses of two mung bean varieties, Kawmy-1 and VC1973 A, under biological stress. The results revealed that Kawmy-1 exhibited tolerance to high-density stress (300,000 plants per fed), maintaining favorable growth and yield, while VC1973 A showed superior vegetative growth across parameters such as dry matter accumulation, leaf area, and leaf weight ratio. In contrast, Kawmy-1 excelled in attributes like leaf area ratio, specific leaf area, relative growth rate, and net assimilation rate. Increased plant density significantly reduced several growth parameters, but some traits like leaf area index, leaf area ratio, and leaf weight ratio showed reversible trends. In terms of yield, Kawmy-1 outperformed VC1973 A in pod and seed yield plant －1 , while VC1973 A achieved better plant height and 100-seed weight. Interestingly, higher plant densities enhanced protein content but decreased overall yield and carbohydrate levels. This study underscores the importance of optimizing plant population density to balance yield and quality in mung bean cultivation. Future research should explore the genetic potential of larger-seeded varieties like VC1973 A, as well as strategies to enhance their performance under varying agronomic conditions.
Light quality is a critical factor influencing plant morphogenesis in tissue culture. In-vitro culture environments benefit from using light-emitting diodes (LEDs), which offer customizable light spectra tailored to specific plant needs and are more cost-effective than fluorescent lamps. However, the impact of the light spectrum on the morphogenesis of Colletia paradoxa remains largely unexplored. This study aimed to evaluate the effects of different LED lighting systems on the photosynthetic and antioxidant metabolism of C. paradoxa cultivated in-vitro. We compared the morphogenic responses, photosynthetic pigment content, chlorophyll a fluorescence, and antioxidant enzyme activities in explants grown under monochromatic white, red, and blue LEDs and a combination of red and blue LEDs (1:1 ratio). The red and blue LED mixture (1:1) was found to be the most favorable for micropropagation, as it resulted in the highest number (5.44) and average length of shoots (4.13 cm), levels of chlorophyll pigments, optimal efficiency in light energy absorption, conservation, and transfer within the photosynthetic complexes, and the lowest levels of antioxidant enzyme activity. These findings suggest reduced reactive oxygen species production and minimal light-induced stress in C. paradoxa shoots under this lighting condition. Therefore, the red and blue LED combination is recommended to develop an efficient micropropagation protocol for C. paradoxa.
This study examined the effects of plant growth-promoting rhizobacteria (R), arbuscular mycorrhizal fungi (M), and compost (C) on wheat growth, yield, and drought tolerance.
Methods
Conducted with a randomized complete block design, the experiment involved two different irrigation water regimes: well-watered (WW) at 75% field capacity (FC) and drought-stressed (DS) at 25% FC.
Results
DS reduced plant growth, yield, and antioxidant enzyme activity but these effects were alleviated through the application of biostimulants. Shoot dry weight increased by 187% with AMF and 159% with C, while seed number and yield improved most significantly with C, C + M, C + R, and C + M + R treatments compared to the control. The application of C, R, and M either individually or in combination effectively mitigated DS by increasing root dry matter and improving the uptake of key minerals such as phosphorus through MR and CMR treatments, by 93% and 524%, respectively. Notably, C alone or combined with M improved the auxin content by 121% and 111%, which contributed to enhanced plant development. This combination strengthened the antioxidant system (including CAT, POX, PPO, and APX), promoted the synthesis of compatible solutes, and increased soil glomalin content. These effects reduced lipid peroxidation and H₂O₂ levels, thereby improving plant resilience under drought conditions.
Conclusions
Biostimulants significantly improved soil physico-chemical properties and fertility, and enhanced the antioxidant system, including key enzymes like alkaline phosphatase, boosting overall soil health and resilience.
Rhizospheric microorganisms, individually or in consortia, interact with root exudates and influence plant growth. This study investigated the effect of three Streptomyces isolates (CLV89, CLV104, and CLV179) individually or in co-inoculation, as plant growth-promoting rhizobacteria on maize (Zea mays L.). This study examined their impact on maize growth and the metabolic profile of root exudates. Maize seeds were treated with Streptomyces at single, dual, or triple co-inoculation, and vegetative growth was assessed. Root exudates were collected hydroponically and analyzed for concentration of reducing sugars, amino acids, proteins, and phenolic compounds. Co-inoculation with two or three isolates positively influenced plant growth, with the triple inoculation promoting greater shoot growth compared to combinations involving CLV104. However, root length reduced with the triple inoculation compared to single and dual inoculations of CLV104. Biomass accumulation was highest in plants co-inoculated with CLV104 + CLV179, which resulted in greatest root biomass despite shorter root length. These findings highlight that co-inoculation can be more effective than triple inoculation, emphasizing the importance of carefully selecting consistent strains to optimize plant growth. Additionally, root-Streptomyces interactions modulated the exudation of phenolic acids, with pathways varying depending on the specific strains involved. Ferulic acid was detected in all treatments assessed, whereas hydroxybenzoic acid was found only in the exudates from the roots of the triple-inoculated plants.
Euglena is one of the alternative natural resources for medicine, food, and energy, and it is important to develop its metabolic contents to fulfill human demands. Improvement of metabolic content in Euglena was conducted in several ways, such as by adding the phytohormone. Brassinolide is one of the phytohormones and is well-known for its ability to stimulate and protect the plant from stressful environments. The application of brassinolide is still lacking. In addition, previous studies have never applied this phytohormone to Euglena sp. cultures. This research aimed to analyze the effect of brassinolide on the growth, metabolic content and wax fermentation in Euglena sp. The growth rate was measured during cultivation, and the metabolic content was analyzed at the late exponential phase before entering the fermentation process. Gas Chromatography-Mass Spectrometry (GC-MS) was carried out to reveal the wax ester content after the fermentation process. The result showed that brassinolide significantly increased the growth rate and metabolic content at lower concentrations, while high concentration tends to inhibit the effect. The high metabolite content, including carbohydrate, lipid, protein, and paramylon, was 0.47 ± 0.02 g/L, 0.20 ± 0.01 g/L, 15.91 ± 1.21 × 10-3 g/L, and 145 ± 0.10 × 10-3 g/L, respectively. Interestingly, wax esters at lower brassinolide concentrations showed contrasting results compared to the control treatment. These findings provide information about the effect of brassinolide in Euglena sp., and advanced research is needed to reveal the mechanism of brassinolide in Euglena sp.
Objective: The main objective of this study was to enhance the functional food properties of Swiss chard (Beta vulgaris var. cicla L.) through controlled elicitation using hydrogen peroxide (H₂O₂). Theoretical Framework: Chard is a highly demanded leafy vegetable due to its low cost, ease of production, and high nutritional value, containing minerals, soluble sugars, fatty acids, vitamins, and phenolic compounds. Plants respond to these stressors leading to the production of antioxidant (AOX) compounds that mitigate stress-induced damage. Hydrogen peroxide (H₂O₂) exogenous application can stimulate specialized metabolite production and enhance antioxidant capacity in plant tissues. Method: Swiss chard plants were subjected to seven H₂O₂ concentrations. It was evaluated DPPH• and ABTS•+, Catalase (CAT), Superoxide dismutase (SOD), Phenylalanine ammonia-lyase (PAL) enzymatic activity, total phenolic content and concentration, and Nitrate concentration. Results and Discussion: The 75 mM H₂O₂ treatment acted as a bio-stimulant, significantly increasing antioxidant activity and the synthesis of phenolic compounds without negatively affecting nitrate concentration. Enzymatic activity of PAL was enhanced, indicating an upregulation of secondary metabolism, leading to higher levels of phenolic compounds and antioxidants in the plant. Research Implications: This study provides an alternative approach to improving leafy vegetable quality using a non-toxic, cost-effective elicitor (H₂O₂). The results suggest that controlled H₂O₂ application could be implemented in agriculture to enhance plant metabolic responses, leading to higher antioxidant content and better functional properties in crops. Originality/Value: This study demonstrates a novel bio-stimulation approach that does not compromise nitrate concentrations, addressing concerns related to excess nitrate accumulation in leafy greens.
Nilaparvata lugens (Stål) is a major pest in rice, causing significant yield losses. This study evaluated 21 rice landraces and resistant varieties, including ASD7, Babawee, Chinasaba, IR36, ARC10550, IR64, IR-71033-121-15, Milyang, MUTNS1, OM4498, Pokkali, Ratu Heenati, RP 2068-18-3-5, Swarna, T12, CR-3006-8-2, CR-317, CR-805, and Salkathi along with resistant check PTB33 and susceptible check TN1, to investigate resistance mechanisms. Damage was assessed on a 1–9 scale: PTB33 and Salkathi were highly resistant (score 1, <8.5% damage), while TN1 and Swarna were fully susceptible (score 9, 100% damage).Resistant genotypes exhibited lower honeydew excretion (PTB33: 45.33 mm²; Salkathi: 52 mm²) and reduced nymphal survival (PTB33: 21%; Salkathi: 24.67%), compared to susceptible varieties (TN1: 197.33 mm²; 91.33%). Biochemical analysis revealed that BPH infestation reduced sugar and protein levels, with minimal losses in resistant genotypes (PTB33: 27.44% sugars, 8.95 % proteins) and significant reduction in TN1 (67.82% sugars, 53.82% proteins). Phenol levels increased post-infestation, with the highest increment in susceptible genotypes (TN1:52.19%; Swarna: 50.55%) and the lowest in resistant genotype (PTB33: 4.34%; Salkathi: 4.43%).These findings suggest that reduced sugar and protein levels and high phenol contentin plants contribute to antibiosis-based resistance in resistant genotypes. This study highlights the importance of resistant landraces in sustainable pest management for rice cultivation.
Polyethylene terephthalate (PET) is a polyester used in the beverage bottling industry that generates a serious pollution problem. Films were obtained to reuse discarded PET bottles, and their surface was modified to determine their effectiveness in supporting the immobilization of the laccase enzyme applied to a catechol biodegradation assay. Radiofrequency (RF) plasma was used to modify the PET surface using different precursors: (a) with the use of air as precursor, the effect of the etching caused by the air on the greater or lesser immobilization was observed; (b) with the use of ethylenediamine, a mixture of N2/H2, or aniline as precursors, it was observed which of these three precursors presented the greater or lesser number of amino groups deposited on the PET surface. After plasma modification, the films were cross-linked with glutaraldehyde to immobilize the laccase enzyme. Finally, the catechol test was performed. It was found that the best etching time using air as a precursor was 90 min, and the precursor that caused a higher insertion of amino groups on the surface was ethylenediamine, which reached a density of amino groups of 3.98 ± 0.10 g·mm−2. The highest percentage of laccase immobilization achieved on the surface of ethylenediamine-modified PET was 97.30%. In the catechol assay, the highest retention was 86.11%. This research reveals how the effect of plasma increases the surface area on a PET surface and, in conjunction with ethylenediamine as the best precursor of the three precursors evaluated, can immobilize a greater amount of enzyme and oxidize more catechol. There is no scientific evidence from previous studies that used air plasma technology to erode and then used three different precursors to modify a surface to immobilize the laccase enzyme and remove a water contaminant.
Cadmium (Cd), an unnecessary trace element in plants, destructively affects ecosystems. This pollutant exists everywhere in the environment, enters the plant through the soil, and causes extensive changes from the macro to the cellular level. This study investigated the effect of cadmium chloride (CdCl2) at three concentrations of 0, 1, and 1.5 mM on morphological, biochemical, and molecular indicators in the Nicotiana tabacum plant as a significant agricultural plant and a common model organism. The results showed that an increase in CdCl2 concentration significantly decreased plant growth and the concentration of photosynthetic pigments compared to the control plant. In contrast, the amounts of soluble sugar, free amino acids, and total phenol increased in plants under stress. In addition, the amount of nicotine and the expression of genes related to the pathway of nicotine biosynthesis increased in plants under Cd stress. The expression of NtA622, NtQPT, NtODC, and NtPMT genes at 1 mM CdCl2 increased by 2, 1.7, 2.4, and 2.9 times, respectively, and at 1.5 mM, CdCl2 increased by 2.9, 2.2, 2.7, and 3.8 times, respectively, compared to the control plant. Therefore, among the four studied genes, the relative expression of the NtPMT gene (the significant gene of the nicotine biosynthesis pathway) was higher. The nicotine content increased by 17.9% and 25.4% with 1- and 1.5-mM CdCl2 compared to the control plant. Tobacco cultivation in Cd-contaminated environments should be managed and it seems that using Cd can increase the biosynthesis of nicotine for agricultural and medicinal purposes.
In 2020, the World Health Organization declared that COVID-19, caused by the SARS-CoV-2 virus, is a pandemic. This led to severe respiratory syndromes and overwhelmed hospital capacities alongside the widespread, yet unproven, use of drugs like ivermectin. Amidst growing concerns over the consequences of frequent ivermectin use, this study aims to examine its toxicological effects following repeated dosage in rats. Female Wistar rats received a daily dose of 12 mg/kg of ivermectin intragastrically for 5 days. Two groups were studied: one euthanized 24 h post the final dose (early protocol) and the other 14 days later (late protocol). The rats underwent tests for locomotion and anxiety- and depression-like behaviors. Additionally, blood and cortex samples were analyzed for acetylcholinesterase and Na+/K+-ATPase activities, oxidative stress levels, and liver and kidney function markers. The early protocol results showed decreased locomotion and increased signs of anxiety and depression in the rats, along with Na+/K+-ATPase inhibition and oxidative stress. In the late protocol, signs of persistent depression-like behavior and hyperlocomotion were observed, coupled with heightened oxidative stress, as indicated by increased reactive oxygen species and disrupted catalase activity. Moreover, the dual inhibition of acetylcholinesterase and Na+/K+-ATPase activities seems to underlie the behavioral alterations seen in the late protocol. The study also noted ivermectin’s potential hepatotoxic effects, corroborating previous findings of elevated liver enzyme levels and severe drug-induced liver injury cases, as well as delayed neuropsychiatric and behavioral changes.
The determination that compounds used for chemotherapeutic purposes in cancer treatment cause cytotoxic effects on healthy cells as well as target cells has led researchers to work on reducing treatment-related toxicity, and accordingly, research on the effectiveness of herbal resources in cancer treatment has gained importance. In our study, ethanol, methanol and chloroform extracts were obtained from the aboveground parts of Origanum majorana L. (O. majorana) and their cytotoxic effects on A-549 cells (non-small cell lung cancer (NSCLC)) were investigated using MTT test. Additionally, the free radical scavenging activities of extracts were demonstrated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) test. It has been shown that cytotoxic effects of extracts increased depending on concentrations on A-549 cells. It was determined that methanol, ethanol and chloroform extracts of O. majorana caused an increase in the activity of lactate dehydrogenase (LDH), and apoptotic enzyme, caspase-3 activity, compared to the control. It has been demonstrated that methanol, ethanol, and chloroform extracts of O. majorana had membrane damaging and apoptotic effects on A-549.
Antibiotics have the potential to affect the health of humans and other living organisms even at slight concentrations. Therefore, there has been a growing global awareness of the environmental impacts associated with antibiotics as emerging pollutants. Cephalexin (CPX) is classified as a first-generation cephalosporin and exhibits a significant efficacy in combating bacterial infections. The current work was conducted to examine the capability of the microalga Chlorella vulgaris to mitigate CPX contamination in aquatic environments. The results indicated that the growth of microalgae diminished in a dose-dependent manner after a 6-day exposure to concentrations of 200–800 mg L− 1 CPX. The analysis conducted through scanning electron microscopy revealed alterations in cell morphology, specifically shrinkage and wrinkling, following the application of CPX. These effects became more significant as the concentration of CPX increased. The results from flow cytometry revealed a notable decrease in cell viability for all concentrations of CPX used, with the highest concentration yielding a viability rate of less than 30%. In addition, CPX caused a decrease in levels of photosynthetic pigments and non-enzymatic antioxidants, including phenols and flavonoids. However, the activity levels of the main antioxidant enzymes considerably increased, achieving their peak at 800 mg L⁻¹. Moreover, the algal cells demonstrated the capability to decrease the concentration of CPX present in the contaminated media, with the most effective reduction observed at 400 mg L− 1. The data obtained confirmed the significant toxicity of CPX on Chlorella vulgaris, while also emphasizing the ability of microalgal cells to withstand antibiotic contamination.
Wheat, a staple crop, faces numerous challenges due to climate change and the increasing demand for sustainable practices. Biostimulants, which enhance plant growth and resilience, have gained attention for their potential to improve wheat productivity in an environment-friendly manner. This study presents a comprehensive bibliometric analysis of field-based research on wheat's response to biostimulants under field conditions from 2000 to 2024. Analyzing 222 studies, the bibliometric analysis reveals a significant rise in research publications on biostimulants, with an annual growth rate of 15.6%. Asia leads with the largest share of publications (59.4%), followed by Europe (18.1%) and Africa (11.6%). North America, South America and Oceania have fewer contributions. Additionally, research institutions in Pakistan, India and Egypt rank as the most productive on this topic. Saudi Arabia stands out with the highest percentage of international collaboration, at 91.7% between countries and 100% among institutions. The findings reveal that biostimulants significantly improve wheat's ability to withstand abiotic stress, optimize nutrient uptake, and enhance overall plant health. Research is transitioning from traditional organic methods and microbial inoculants to advanced biostimulant formulations, improved nutrient management, and reduced environmental impact. However, gaps remain, particularly in understanding the combined effects of multiple biostimulants and their long-term impact on wheat and soil health. This synthesis of research trends lays the groundwork for advancing sustainable wheat production, supporting food security and agricultural resilience amidst environmental challenges.
Amylase has numerous applications in the processing food sector, including brewing, animal feed, baking, fruit juice manufacturing, starch syrups, and starch liquefaction. Practical applications have been the primary focus of recent research on novel properties of bacterial α-amylases. Many amylolytic-active bacterial isolates were obtained from samples of organic-rich, salinity-rich soil. Morphological and 16S rRNA gene sequence studies clearly revealed that the organism belongs to Bacillus sp. and was named Bacillus cereus strain GL2 (PP463909.1 (When pH 6.0, 45°C, and 12 hours of incubation were met the optimal growth conditions for the strain produced the highest amount of α-amylase activity. B. cereus strain GL2 α-amylase isoenzyme was purified to homogeneity using Sephacryl™ S-200 chromatography and ammonium sulfate precipitation. The electrophoretic molecular weight of B. cereus α-amylase was 58 kDa. The optimal pH and temperature for measuring α-amylase activity were 50°C and 6.0, respectively. α-Amylase did not change at 50°C. The purified enzyme improves bread texture by reducing stiffness while improving cohesiveness and flexibility. Purified α-amylase was added to the flour, which improved the rheological properties and overall bread quality. As a result, the α-amylase from B. cereus strain GL2 can be used to promote bread-making.
In Chile, most potato crops are cultivated under rainfed systems, frequently facing
temperature increases and water defcits during the growing season. This feld study
aimed to evaluate the physiological responses and genotypic variations between the
Chilean native potato variety Chona Negra and commercial varieties (Karú INIA and Desiree) when exposed to moderately high temperatures (+5.0 to+5.9 °C) for 40 days during the tuber bulking phase under diferent hydro- (or water availability) conditions. Four treatments were applied: (i) ambient temperature with rainfed conditions (T0H0), (ii) high temperature with rainfed conditions (T1H0), (iii) ambient temperature with irrigation (T0H1), and (iv) high temperature with irrigation (T1H1). The results showed that lower water availability and a moderate temperature increase (+5 °C) afected the leaf pigment content, photosynthetic performance and, chlorophyll fuorescence in both commercial and native potato genotypes. While moderately high temperatures and water defcits generally did not have stronger combined efects than each condition alone, our fndings highlight the critical role of water availability in environments like southern Chile. Notably, the native potato, Chona Negra, exhibited enhanced photosynthetic performance under higher temperatures with irrigation, associated with increases in specifc leaf area, stomatal conductance, photochemical quenching, and chlorophyll content. These findings suggest that Chona Negra may possess adaptive traits that improve its tolerance to heat stress when adequate water is available. In contrast, commercial varieties appeared more susceptible to limitations in photosynthetic activity under higher or prolonged periods of elevated temperatures.
GABA serves as the major metabolite and signaling molecule for plant growth and development under fluctuating environmental conditions. In the present study, the effect of different concentrations of GABA (0, 0.25, 0.5, or 0.75 mM) as foliar spray or seed soaking treatment on K-21 and S-22 varieties of tomato (Solanum lycopersicum L.) under normal and NaCl stress (150 mM) was studied. At 40 days after transplantation (DAT), NaCl stress impaired the photosynthetic apparatus, elevated reactive oxygen species (ROS) and malondialdehyde (MDA) levels, and reduced photosynthetic traits and related enzymes, and consequently compromised plant growth and development. However, among the different concentrations of GABA tested 0.5 mM of GABA applied through foliar mode proved best in improving growth, photosynthetic traits (like PN of K-21 and S-22 varieties by 39.17% and 30.35%, gs by 40.11% and 32.37%; Ci by 39.48% and 33.67%; E by 37.98% and 30.08% respectively, compared to control group) and enzyme activities (Rubisco, succinate dehydrogenase, and fumarase activity) maintained organization of chloroplast in the leaves and also improved lycopene and β-carotene contents in fruits of tomato plants. Moreover, treatment of GABA (0.5 mM) reduced levels of ROS (such as decreased the contents of O2·⁻ by 38.10% and 34.05%; and H2O2 by 35.95% and 30.36% respectively, in K-21 and S-22 varieties, as compared to their control) and cell death compared to non-treated tomato plants. Foliar application of GABA (0.5 mM) also was more effective than seed soaking to overcome the adverse effects of salt stress specifically in K-21 than S-22 variety. This study addresses this gap and uniquely investigates role of GABA in regulating photosynthetic (Rubisco), GABA shunt, and respiratory (fumarase, succinate dehydrogenase) enzyme activities under salt stress, offering new insights into its multifaceted stress-mitigating potential.
1. Our earlier preparations of bovine chymotrypsinogen B isolated by chromatography on O-(carboxymethyl)cellulose have been further purified by O-(carboxymethyl)Sephades chromatography. the product finally obtained (yield, 25%; 400 mg per kg fresh pancreas) is entirely free from deoxyribonuclease and behaves as an homogeneous protein when subjected to free-boundary electrophoresis, ultracetrifugation and equilibrium chromatography. Chromatography on O-(carboxymethyl)-Sephadex also appears to be a good technique for the purifition of bovine deoxyribonuclease.
The Pro-Milk method was compared to the official final action Udy method, 16.037-16.040, for 166 samples of raw milks from individual cows and to the official final action Kjeldahl method, 16.035, for 54 similar samples. Averages ± standard deviations of the differences were 0.019±0.017 and 0.027±0.109% protein for the 2 methods, respectively. Fat test or protein level of the samples did not affect the magnitude of the difference between the Pro-Milk and either of the other 2 methods. The Pro- Milk comparisons have also been made on samples of processed milks with similar results. A number of comparisons of the Pro-Milk method to other methods for protein in milk have been reported in the literature. The principles of the Pro-Milk method differ from the Udy method only in the type of dye used. Periodic calibration and routine control of the Pro-Milk method is required because of variability in assay of the Amido Black 10B dye.
Zusammenfassung Es wird die Interferenz von Natrium Ethylendiaminotetraacetat bei der Eiweissbestimmung nach der Methode vonLowry et al. und deren Verhinderung gezeigt. Die Anwesenheit von Natrium Ethylendiaminotetraacetat interferiert bei der Eiweissbestimmung nach der Methode vonLowry, da sie selbst zur Reduktion des Folin-Reagens führt. Es gelingt, diese Interferenz durch Zusatz von CaCl2 zu verhindern.
Equations for protein in sow's milk by Orange G dye binding procedures were: per cent protein = 9.184-4.526 (absorbance) at Sensitivity 1 for samples above 6.5% protein, and per cent protein = 7.317-6.061 (absorbance) at Sensitivity 2 for samples containing 3.0 to 6.5% protein. Forty-two samples of sow's milk were analyzed with a Beckman Model B spectrophotometer. Sow's milk had more protein than cow's milk, requiring 1 ml milk per 25 ml Orange G dye. Protein analyses by Orange G dye binding were 0.95 correlated with Kjeldahl estimates of protein. These equations differed from those developed for estimating protein of cow's milk by the Orange G procedure.
Glycerol interferes with the estimation of protein by both the Lowry and the biuret procedures. Color development in the presence of protein is lessened by the presence of glycerol in samples tested by either assay. In addition, with the Lowry test significant color development linearly related to glycerol concentration is obtained in the absence of protein. Problems in either assay of protein estimation in the presence of glycerol may be overcome by the use of appropriate blanks and calibration curves.