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Protein measurement with the Folin phenol reagent

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... Brain tissue was homogenised in 0.1 M PBS (pH 7.4). The homogenate (10% w/v) was subjected to cold centrifuge (10 000 g for 15 min). The supernatant thus obtained was used for further estimations. The protein estimation was done by the method prescribed by Lowry et. al. 1951(Lowry et al. 1951). ...
... The protein estimation was done by the method prescribed by Lowry et. al. 1951(Lowry et al. 1951). ...
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D-galactose (D-gal) induced senescence in rodents is a widely used model for assessment of molecules affecting brain ageing. Chronic administration of D-gal causes neuroinflammation leading to cognitive deficit and memory impairment which represent Alzheimer’s dementia. In present study, we investigated the neuroprotective effects of the natural phenol, p-Coumaric acid (PCA) and its underlying mechanism in the chronic D-gal treated mice. Subcutaneous administration of D-gal (150 mg/kg) to Swiss albino mice for 42 consecutive days resulted in cognitive impairment as observed in Morris water maize (MWM) and Y maze test, which was ameliorated by concurrent treatment with PCA (80 mg/kg, and 100 mg/kg, p.o.). Importantly, PCA treatment attenuated the D-gal induced oxidative stress and significantly inhibited acetylcholinesterase (AChE) activity in mice brain. Furthermore, PCA treatment significantly lowered levels of inflammatory marker nuclear factor kappa B (NFκB) and reduced levels of proapoptotic enzyme caspase3. We also observed that PCA treatment exhibited β-secretase enzyme (BACE1) inhibitory effect. However, our results revealed that PCA treatment failed to decrease the level of advanced glycation end products both in vitro and in vivo. Taken together, current study demonstrated the significant neuroprotective effect of PCA against D-gal induced oxidative stress, neuroinflammation, cognitive impairment and apoptosis.
... The mixture was heated in a water bath at 85 • C for 45 min and centrifuged at 9690 g for 10 min at 4 • C (Gao et al., 2021a). The protein concentration of the supernatant was determined by Lowry et al. (1951) method and adjusted to 2 mg mL − 1 . Then, the protein solution (2 mg mL − 1 ) was mixed with the loading buffer at 4:1 (protein: buffer). ...
... The homogenate was kept at 4 • C for 60 min before centrifuging at 9690 g for 10 min. The content of peptides in the supernatant was determined according to Lowry et al. (1951) method. ...
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The effects of transglutaminase (TGase) addition (0.4–1.2 g/100g), ultrasound (120–720 W, 20 min), and their combination on the gel properties and in vitro digestion characteristics of bay scallop adductor muscle were studied. The gel strength of the gel sample with TGase content of 0.8 g/100g (TG-0.8) was 58.2% higher than that of the control sample (CON). The gel sample treated with ultrasound at 480 W (UT-480) had the highest gel strength. The strength of the gel prepared by combination of 0.8 g/100g TGase and 360 W ultrasound (UT-TG) was 82.3% higher than that of CON. The whiteness and water holding capacity of the gel increased regardless of the addition of TGase or ultrasound treatment. SDS-PAGE patterns showed that the myosin heavy chain of the treated samples became thinner, and the changes of actin and tropomyosin were not significant. The scanning electron microscopy results of gel samples prepared by ultrasound combined with TGase showed a denser structure, which was related to the lowest total sulfhydryl content and TCA-soluble peptide content. The results of dynamic rheology show that the UT-TG sample had the highest G′ value, followed by TG-0.8. The in vitro digestion characteristics of the selected gel samples were also discussed. The degree of protein hydrolysis and the content of free amino acids in TG-0.8 samples were the lowest, which improved after ultrasound treatment. Overall, the combination of appropriate ultrasound treatment and TGase addition provides an effective means for improving gel properties and digestibility of scallop surimi product.
... Soil moisture content was monitored using moisture meter (Delta-T Soil moisture kit -Model: SM150, Delta-T Devices, Cambridge) periodically and re-watering was done when the soil moisture reached below 20 per cent and leaves were completely rolled and started drying at tips and margins. Soluble protein content of the leaf was estimated by following the procedure described by Lowry et al. (1950) [4] and expressed as mg g -1 of fresh weight. Catalase was estimated according to Teranishi et al. (1974) [7] and expressed as μg of H 2 O 2 reduced min -1 g -1 fresh weight. ...
... Soil moisture content was monitored using moisture meter (Delta-T Soil moisture kit -Model: SM150, Delta-T Devices, Cambridge) periodically and re-watering was done when the soil moisture reached below 20 per cent and leaves were completely rolled and started drying at tips and margins. Soluble protein content of the leaf was estimated by following the procedure described by Lowry et al. (1950) [4] and expressed as mg g -1 of fresh weight. Catalase was estimated according to Teranishi et al. (1974) [7] and expressed as μg of H 2 O 2 reduced min -1 g -1 fresh weight. ...
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A field experiment was conducted to determine the physiological performance of traditional rice landraces under water stress given at reproductive stage. Drought is one of the serious threats on rice productivity. Traditional rice landraces shows substantial adaptability to drought so Six popular rice land races, namely Rascadam, Kothamalli samba, Kaattu samba, Kallundai, Kuliyadichan and Milagu samba from Tamilnadu, India and two popular rice check varieties, namely IR64 (susceptible check), N22 (tolerant check) were used to study the physiological performance for drought tolerance. These genotypes were subjected to drought under field condition at Agricultural Research Station, Bhavanisagar, TNAU, Coimbatore. The crops were imposed with drought during reproductive stage by withholding irrigation till the soil moisture reaches below 20 per cent. The experimental design for the trial was factorial randomized block design (FRBD). Chlorophyll Index (CSI), Soluble protein, Catalase activity and Nitrate reductase activity were measured.
... Protein estimation of enzyme extract was determined by the method of Lowry et al. [20]. 1 ml NaOH was added to 1 ml of enzyme extract (crude and purified), placed in boiling water for 10 min and cooled under tap water. The mixture was incubated with reagent A (1 ml of 1% Na-K tartarate solution containing 0.5% CuSO 4 into 50 ml 2% Na 2 CO 3 solution). ...
... During determination of the effect of l-phe (20,40, 60, 80, 100, 120, 140, 160 and 180 mM) substrate on Spirulina CPCC-695 PAL activity at 30 °C and pH 8.0. Maximum PAL activity was noted at 100 mM (significant at P < 0.05) (Fig. 3c). ...
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Phenylalanine ammonia lyase (PAL) catalyzes the deamination of phenylalanine to cinnamic acid and ammonia. It plays a crucial role in the formation of secondary metabolites through the phenylpropanoid pathway. Recently there has been growing interest in exploring the biochemical properties of PAL for its clinical and commercial applications. PAL as a key component has been used in metabolic engineering and synthetic biology. Due to its high substrate specificity and catalytic efficacy, PAL has opened a new area of interest in the biomedical field. PAL has been frequently used in the enzyme replacement therapy of phenylketonuria, cancer treatment and microbial production of l-phe the precursor of noncalorific sweetener aspartame (Methyl l-α-aspartyl-l-phenylalaninate), antimicrobial and health supplements. PAL occurs in few plants, fungi, bacteria, and cyanobacteria. The present investigation is a preliminary study in which an attempt has been made for the isolation, partial purification, and biochemical characterization of PAL (crude and partially purified) from Spirulina CPCC-695. Partially purified PAL exhibited higher enzymatic activity and protein content than the crude enzyme. Molecular weight of the crude and partially purified PAL was ~ 66 kDa. The optimum temperature and pH for PAL activity was observed as 30 ℃ and 8.0 respectively. l-Phe was the most preferred substrate (100 mM) whereas gallic acid showed maximum inhibition of PAL activity. Enzyme kinetics suggested good catalytic efficacy of the PAL enzyme and affinity towards substrate. Both the enzyme (crude and partially purified) showed less than 5% haemolysis suggesting the biocompatible nature of PAL.
... The precipitated proteins were recovered by centrifugation at 7000 × g for 20 min at 4 °C and dissolved in a minimal volume of 50 mM Tris-HCl buffer (pH 7.5) and dialyzed against the same buffer for 24 h at 4 °C [16]. The dialyzed fractions were concentrated, assayed for keratinase activity, and the protein content was determined using the Lowry et al. method, using bovine serum albumin (BSA) as standard [17]. ...
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The application of biotechnology through the utilization of enzymes is considered an easy and inexpensive method of producing valuable products from waste resources. The present study describes the production of a keratinolytic enzyme from Pseudomonas aeruginosa S-04, which showed efficiency for feather biodegradation. The production of extracellular keratinase was improved 1.3-fold by optimizing various parameters through the one factor at a time (OFAT) approach. The keratinase was purified to homogeneity by ammonium sulfate precipitation and DEAE-cellulose anion exchange chromatography. The purified keratinase (35.5 kDa) displayed optimal activity at 60 °C and pH 9.5 and was stable between pH 7 and 9.5 and temperatures ranging from 4 to 40 °C for 2 h. The catalytic activity of keratinase was enhanced in the presence of Fe3+ and Mn2+ ions, Triton X-100, Tween 20, DMSO, isopropyl alcohol and ethanol, but reduced activity was recorded in the presence of methanol and acetone. Furthermore, the enzyme activity was deactivated by EDTA, suggesting that this keratinase belongs to a metalloprotease family. The Km and Vmax of the purified keratinase were 7.62 mg/ml and 200 U/mg protein, respectively. The partially purified keratinase displayed remarkable feather degradation capabilities (93% after 24 h), and the nutritional properties of the resulting feather hydrolysate make it a promising candidate for use as a poultry feed ingredient. Thus, thermostable alkaline keratinase from P. aeruginosa S-04 is a promising candidate that converts low-cost chicken feather waste into a value-added product, indicating a novel approach to feather waste treatment and utilization. https://rdcu.be/cTFcg
... The total protein content and total sugar content were determined in this study. Lowry's procedure was used to evaluate the total protein content of purified bioflocculant using Bovine Serum Albumin as a standard 26 . The total sugar content of the bioflocculant was determined using the phenol-sulfuric technique 27 , and glucose was used as the standard solution. ...
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This study was designed to evaluate the potential of bioflocculant producing strains isolated from wastewater sludge. According to the Plackett–Burman design, the response surface revealed glucose, magnesium sulfate, and ammonium sulfate as critical media components of the nutritional source, whereas the central composite design affirmed an optimum concentration of the critical nutritional source as 16.0 g/l (glucose), 3.5 g/l magnesium sulfate heptahydrate (MgSO4.7H2O), and 1.6 g/l ammonium sulfate ( (NH4)2SO4), yielding an optimal flocculation activity of 96.8%. Fourier Transformer Infrared Spectroscopy (FTIR) analysis confirmed the presence of hydroxyl, carboxyl and methoxyl in the structure of the bioflocculant. Additionally, chemical analysis affirmed the presence of mainly a polysaccharide in the main backbone of the purified bioflocculant with no detection of protein. Energy Dispersive X-ray analysis affirmed the presence of chlorine, phosphorous, oxygen and chlorine as representatives of elemental composition. Thermogravimetric (TGA) analysis revealed over 60% weight was retained at a temperature range of 700 °C. The purified bioflocculant remarkably removed chemical oxygen demand, biological oxygen demand and turbidity in brewery wastewater. This study suggested that the bioflocculant might be an alternate candidate for wastewater treatment.
... Thiobarbituric acid-reactive substances (TBARS) shaped up as a byproduct of lipid peroxidation were determined in the homogenized samples by the TBARS assay using thiobarbituric acid as a reagent at 532 nm spectrophotometer (UV-160A Shimadzu), measuring malondialdehyde (MDA) present in the sample with an assay of TBARS [26]. Protein concentration in the homogenized samples was also measured by using the method of Lowry [27]. The levels of estimated parameters are represented as ng/mg of protein for GRP78, CHOP, caspase-3, caspase-12, and nmol/mg of protein for TBARS. ...
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Endoplasmic reticulum (ER) stress and apoptosis are implicated in the pathogenesis of epilepsy. Here we examine the effects of valproic acid (VA) plus 4-phenylbutyric acid (4-PBA) on abnormal electrical brain activity, ER stress and apoptosis in acute seizures induced by pentylenetetrazole (PTZ). Forty male rats were randomly divided into five groups, each consisting of 8 rats as follows: Sham, PTZ, VA+PTZ, 4-PBA+PTZ, and VA plus 4-PBA+PTZ. The treated groups received VA, 4-PBA and VA plus 4-PBA by intraperitoneal application for 7 days prior to PTZ-induced seizure. On the 8th day, acute epileptic seizures were induced by PTZ (50 mg/kg, i.p.) injection, except for the sham group. Then, the seizure stage was observed and ECoG activities were recorded during the 30 min. At 24th post seizures, the hippocampus and blood samples were collected for biochemical and histopathological examinations. Administration of VA plus 4-PBA prior to PTZ-induced seizures significantly decreased seizure stage, the duration of generalized tonic–clonic seizure and the total number of spikes as increased the latency to the first myoclonic jerk when compared to the PTZ group. 4-PBA suppressed the increased levels of ER stress markers GRP78 and CHOP in the hippocampus. VA plus 4-PBA treatment before seizures significantly inhibited PTZ-induced elevations of apoptosis-related indicators caspase-3 and caspase-12, and significantly reduced the number of histopathological lesions of the hippocampus region at 24th post seizures. These findings suggest that administration of VA plus 4-PBA prior to PTZ-induced seizures may be involved in the neuroprotective potential of these agents for seizures.
... At the late exponential phase, the total algal biomass was harvested by centrifugation at 6000 rpm for 5 min and dried at 60 °C overnight, then the dried biomass was used for protein and carbohydrates measurements. Protein content was determined following the Lowry method using bovine serum albumin as a standard [69]. Total carbohydrate content was determined following the phenol-sulfuric method and was quantified by a standard glucose curve at 490 nm using Spectramax M3 multimode spectrophotometer (Molecular Devices, San Jose, CA, USA) [70]. ...
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Synthesized astaxanthin (ASX), stereoisomers of 3S,3′R, 3R,3′R, and 3S,3′S, have over 95% market share and have relatively poor antioxidant and bioactivity properties, with persistent issues in terms of biological functions, health benefits, and biosafety if compared to natural ASX. Bioprospecting of new microalgal strains could be vital for a new source of powerful antioxidant (ASX). In this study, a new algal strain was isolated from the Indian foothills of the Himalayas. Its identity was discerned by morphological and DNA barcode studies. It is a unicellular spheroidal cell-shaped alga with 100–200 μm diameter. The isolate has 93.4% similarity to Dysmorphococcus globosus species based on 18S-rDNA phylogenetic analysis and named as D. globosus-HI (HI stands for Himalayan India). Its growth and major cellular components (carotenoids, carbohydrates, protein, lipids, fatty acid profile, and ASX) were optimized using the seven different culture media. The highest biomass (1.14 g L−1) was observed in the MBBM medium, with a specific growth rate (0.087 day−1), division/day (0.125), and cellular yield (6.16 x 106 cells/mL). The highest carotenoids (1.56 mg g−1), lipids (32.5 mg L−1), and carbohydrates (135.62 mg L−1) were recorded in the 3N-BBM medium. The maximum ω3-FAs (17.78%), ω6-FAs (23.11%), and ω9-FAs (7.06%) were observed in MBBM, JW, and BG-11 medium respectively. The highest amount of antioxidant ASX was accumulated in the 3N-BBM medium (391 mg L−1). It is more than any other known algal species used in the production of natural ASX. The optimized biochemical studies on the D. globosus-HI strain should fulfill the increasing demand for natural ASX for commercial application.
... The dry matter of microalgae was determined according to the AOAC standard methods [39]. The protein assay method of Lowry [40] was used by the combination of Folin with Biuret's reagents. The Lipids content was determined gravimetrically after the Soxhlet extraction of dried samples with hexane for 2 hours using Nahita Model 655 (Navarra, Spain). ...
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The saltern of Sfax is a thalasso haline paralic ecosystem were the salinity ranged from 45 to 450 PSU. The microalgae distribution of saltern showed a spatial ecological succession. The specific richness of microalgae decreased with the salinity, accounting 37, 17 and 5 species at three level of salinity from 40 to 80, 80 to 200 and 200 to 450 PSU, respectively. To better understand the behavior of the hyper-halo tolerant microalgae, three autotrophic species Halamphora sp. SB1 MK575516 (Diatom), Phormidium versicolor NCC-466 (Cyanophyceae) and Dunaliella salina (Chlorophyceae) were isolated from each level of salinity and they are grown in batch in artificial seawater at laboratory scale. Growth and metabolites synthesized by these microalgae were assessed. Salinity reacts on the physiology of these three species which possess mechanisms of resistance to more or less effective stresses and generally by the synthesis of different biomolecules such as pigments, sugars, proteins and fatty acids.
... Protein content of garlic peel was determined as described by Lowry, et al. [15]. ...
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The present study was designed to develop and evaluate the nutritional, functional, sensory properties and texture profile data of garlic peel powder fortified viz. 2%, 5% and 10% value added pukhelein and compared with control sample. Garlic peel showed crude fiber, carbohydrate and ash content viz. 60.57%, 21.71% and 16.34%, total phenolic content (TPC) of 108.79 µg/ml GAE, total flavonoid content (TFC) of 33.27 mg Quercetin/ml and antioxidant activity (33.38 mg/ml) respectively. 5% formulation exhibited the highest crude fiber, protein content, TPC (326.6 µg/ml GAE), TFC (91.38 mg Quercetin/ml) and total antioxidant content (54.68 µg/ml).Sensory evaluation data showed good result in 5% formulation in terms of appearance, texture and taste. Texture profile analysis showed that a considerable decrease in hardness, springiness, cohesiveness, gumminess and chewiness and resilience. The outcome of the study revealed that 5% garlic peel powder fortified pukhelein exhibited the best result among all formulations.
... Protein estimation and ph determination: Cell protein was estimated according to standard procedure (Lowry et al., 1951). bovine Serum Albumin (bSA) was used as standard. ...
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Microorganisms and their enzymes are important part of industrial biotechnology because of their high adaptability, versatile metabolic machinery and simple genetic constitution which can be easily manipulated to meet different needs. A mesophilic novel strain of Serratia marcescens-VT1 was found to produce extracellular protease (13.173 U/ml) and cold active and stable psychrophilic lipase (23.17 U/ml). Characterization of protease showed the enzyme to be active over a wide range of pH 4-11, optimum pH 10, and optimum temperature 50°C. SMVT1 lipase was found to be active from pH 7-9 with optimum pH 7 and good stability for 90 minutes, optimum temperature 30°C and to retain almost 80 % activity at lower temperatures (20 and 10°C). SMVT 1 protease was stable in hydrophilic solvents like methanol and ethanol. Lipase showed stability in both hydrophobic and hydrophilic solvents, hexane-92.67%, methanol-87.4% and acetone-85.23%. The wide range pH active protease and psychrophilic and organic solvent stable lipase can be made use in various sectors like bioremediation, waste management, and chemical synthesis, food processing and detergent additives.
... The change of pH in the solution was determined by the bromine thymol blue indicator, which is blue at pH 7.6 and yellow at pH 6. Protein amounts in the purified enzyme solution and hemolysate were determined by modified Lowry method using the PierceTM BCA protein determination kit [52,53]. ...
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Magnetic nanoparticles (MNPs) have been used for purification of specific biomolecules form mixtures. The aim of this study is to develop a new, cheap, reusable, and magnetic-based material to purify the carbonic anhydrase (CA) enzyme in a short time with high efficiency. In the first part of this study, silica-coated iron oxide magnetic nanoparticles (Fe3O4@SiO2 MNPs) were obtained. Surface modification of Fe3O4@SiO2 MNPs was accomplished with 3-(4-Hydroxyphenyl) propionic acid (PA) and sulfanilamide (SA), respectively. SA is a selective inhibitor of CA, and it selectively binds to CA. The final particle was named Fe3O4@SiO2-PA-SA MNPs and characterized by SEM, TEM, XRD, and FT-IR. It was determined that the produced MNPs contained multicore, were smaller than 100 nm in size, and had a spherical morphology. The CA was purified from bovine blood hemolysate in a short time such as 2.5 h and in a simple manner. The maximum enzyme purifying capacity of MNPs was calculated as 13.87 ± 3.27 mg CA/g MNP. SDS-PAGE analysis was confirmed that high CA purification success was achieved.
... 11 Total protein content of the algal culture was estimated by Lowry's method. 12 Total chlorophyll content was estimated by warm extraction using methanol. 13 Four replications were maintained for all the experiments. ...
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Eco-friendly anti-algal agents are in demand for preventing the growth of unwanted algae. Green synthesized nanoparticles exhibit antimicrobial properties and have been used as a better alternative against chemical and physical processes. In the present study, treatment of silver nitrate with leaf extracts (5% w/v) of Tinospora cordifolia, a plant with proven antimicrobial effects, exhibited UV-visible absorption maxima between 440-460 nm after 1h indicating bioreduction of silver to nanoparticles. The green synthesised silver nanoparticles (5 mgl-1) exhibited inhibition zones against Chlamydomonas reinhardtii in in vitro agar assays. Treatment with green synthesised silver nanoparticles during exponential phase of algal growth resulted in significant reduction in algal population, carbohydrate, protein and chlorophyll contents confirming the anti-algal potential. This is the first report on the growth inhibitory potential of green synthesised silver nanoparticles against green algae.
... GSH and GPx were measured in the supernatant fraction of the samples after centrifugation (10,000 g for 10 min 4 • C). GSH concentrations and GPx activity were calculated to the total protein content of the supernatant measured by the Folin-Ciocalteu reagent with the protocol of Lowry et al. (1951). ...
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Selenium is an essential trace element; therefore, soils are often amended with selenium fertiliser in selenium poor regions to increase the selenium consumption of the population. The amendment form can be sodium selenate and sodium selenite, depending on crop type. Climate change increases the mean temperature and also the frequency of extreme events such as drought and extreme rain. Therefore, testing the effects of different selenium forms outside the range of standard conditions can help predicting the effects of such amendments or selenium pollution. Effects of sublethal concentrations of selenite and selenate were tested on parameters of mortality, reproduction, growth, and oxidative stress parameters of Folsomia candida (Collembola) in case of different climate scenarios. The standard 20 °C and the increased 25 °C temperatures were combined with three different soil moisture conditions: drought, standard water content and increased water content. Selenite was found to be more toxic than selenate. However, the collembolan treated with selenite tolerated the increase of temperature better than those treated with selenate. The temperature increase was additive with selenium treatment in most cases. Drought was found to increase selenium toxicity in general. Also, different water contents had interactions with selenium treatments causing differences in concentration-response curves. Juvenile and egg numbers were decreased in the case of suboptimal water contents; especially drought decreased juvenile and egg numbers. No significant mortality was found under standard conditions, but mortality increased with the different climate scenarios. Results were not conclusive in the case of measurement of lipid peroxidation and glutathione status. In standard conditions, selenium did not have an effect on the juvenile number, but in combination with climate scenarios, from 1 mg Se kg⁻¹ a decrease was found in the juvenile number. Therefore, soil selenium amendments should be carefully planned regarding the effects of changing climate, especially drought.
... The carbohydrate and protein contents of the freeze-dried biosurfactant samples obtained from bioreactor studies were estimated via the colorimetric methods of Dubois [31] and Lowry [32] respectively. The functional groups of biosurfactants were also identified via thin layer chromatography (TLC) technique using pre-developed plates (TLC Silica gel 60 F 254 , Sigma-Aldrich) in chloroform: methanol (2:1 v/v). ...
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Microbial surfactants have induced research and industrial interest recently, owing to their significant advantages over the chemically produced counterparts. The pursuit of novel biosurfactant producers includes first the isolation and screening, followed by studies on fermentation strategies. The utilization of non-pathogenic strains, like lactobacilli, will expand the potential for targeted and innovative food-oriented applications. Hence, this study undertakes the evaluation of different bioprocessing parameters and schemes for biosurfactant production using Lactobacillus strains selected from a previous screening study. Cheese whey permeate (CWP) was used as a low-cost nutrient supplement, in the context of valorizing food industry by-products towards the development of biorefinery concepts. Microplates and shake flasks experiments were performed to assess the effect of pH, temperature and CWP supplementation with micronutrients and exogenous nitrogen sources. Two different media were applied in bioreactor studies, along with several fermentation conditions and significant biosurfactant production was evidenced via surface tension reduction (< 30 mN/m). Among the Lactobacillus strains studied, Limosilactobacillus fermentum ACA-DC 0183 demonstrated the maximum surface tension reduction (34.9 mN/m) after 32 h, observed even at the early hours of fermentation (4 h). Notably, the majority of the experiments indicated an inverse correlation between biosurfactants and lactic acid production. The produced biosurfactants were also extracted and partially characterized. The results presented hereof communicate promising findings in the context of mitigating the costs associated with biosurfactant production and foster the development of food industry by-products refining to enhance the circularity and sustainability of food systems.
... The level of malondialdehyde (MDA) in tissue was determined colorimetrically according to the methods of Placer, Cushman and Johnson [24]. The protein content of the liver and kidney tissue homogenates were determined using the [25]. ...
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Excessive fluoride intake has been reported to cause toxicities to brain, thyroid, kidney, liver and testis tissues. Hesperidin (HSP) is an antioxidant that possesses anti-allergenic, anti-carcinogenic, anti-oxidant and anti-inflammatory activities. Presently, the studies focusing on the toxic effects of sodium fluoride (NaF) on heart tissue at biochemical and molecular level are limited. This study was designed to evaluate the ameliorative effects of HSP on toxicity of NaF on the heart of rats in vivo by observing the alterations in oxidative injury markers (MDA, SOD, CAT, GPX and GSH), pro-inflammatory markers (NF-κB, IL-1β, TNF-α), expressions of apoptotic genes (caspase-3, -6, -9, Bax, Bcl-2, p53, cytochrome c), levels of autophagic markers (Beclin 1, LC3A, LC3B), expression levels of PI3K/Akt/mTOR and cardiac markers. HSP treatment attenuated the NaF-induced heart tissue injury by increasing activities of SOD, CAT and GPx and levels of GSH, and sup- pressing lipid peroxidation. In addition, HSP reversed the changes in expression of apoptotic (caspase-3, -6, -9, Bax, Bcl-2, p53, cytochrome c), levels of autophagic and inflammatory parameters (Beclin 1, LC3A, LC3B, NF-κB, IL-1β, TNF-α), in the NaF-induced cardiotoxicity. HSP also modulated the gene expression levels of PI3K/Akt/mTOR signaling pathway and levels of cardiac markers (LDH, CK-MB). Overall, these findings reveal that HSP treatment can be used for the treatment of NaF-induced cardiotoxicity.
... GABA concentration (pmol/ml) in eluted solution was appraised at 509 nm and by the standard curve of GABA (Swamy et al. 2013). Protein was estimated by the scheme elaborated by Lowry et al. (1951). ...
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Cerebral hypoperfusion (CH) is a common underlying mechanism of dementia disorders linked to aberrations in the neurovascular unit. Hemodynamic disturbances adversely affect cellular energy homeostasis that triggers a sequence of events leading to irrevocable damage to the brain and neurobehavioral discrepancies. Theobromine is a common ingredient of many natural foods consumed by a large population worldwide. Theobromine has shown health benefits in several studies, attributed to regulation of calcium homeostasis, phosphodiesterase, neurotransmission, and neurotrophins. The current study evaluated the neuroprotective potential of theobromine against CH in the permanent bilateral common carotid artery occlusion (BCCAO) prototype. Wistar rats were distributed in Sham-operated (S), S + T100, CH, CH + T50, and CH + T100 groups. Animals received permanent BCCAO or Sham treatment on day 1. Theobromine (50, 100 mg/kg) was given orally in animals subjected to BCCAO for 14 days daily. CH caused neurological deficits (12-point scale), motor dysfunction, and memory impairment in rats. Treatment with theobromine significantly attenuated neurological deficits and improved sensorimotor functions and memory in rats with CH. In biochemistry investigation of the entire brain, findings disclosed reduction in brain oxidative stress, inflammatory intermediaries (tumor necrosis factor-α, interleukin-1β and − 6, nuclear factor-κB), markers of cell demise (lactate dehydrogenase, caspase-3), acetylcholinesterase activity, and improvement in γ-aminobutyric acid quantity in rats that were given theobromine for 14 days daily after CH. Histopathological analysis substantiated attenuation of neurodegenerative changes by theobromine. The findings of this study indicated that theobromine could improve neurological scores, sensorimotor abilities, and memory in CH prototype.
... The supernatant was re-prepared after the supernatant was repeatedly frozen and thawed up for 2 times during the enzymatic activity assay. The activity of intestinal protease was determined using the Folin phenol reagent (Lowry et al. 1951). Lipase activity was determined using 4-nitrophenyl-myristate (Gjellesvik et al. 1992). ...
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Nowadays, both pelleted feed (PF) and extruded feed (EF) have been widely adopted in the aquaculture industry. However, limited information is available comparing their utilization efficiencies and meanwhile interpreting the underlying mechanisms. This study aimed to compare the utilization efficiencies of both PF and EF by blunt snout bream (Megalobrama amblycephala) based on growth performance, digestive capacities, and endocrine functions. Two feeds with identical formulas were prepared and named PF and EF. Fish were randomly distributed into two groups, including one that fed the PF continuously, and one that offered the EF continuously. The whole feeding trail lasted 8 weeks. The results showed that the protein efficiency (PER), retention of nitrogen and energy (NRE and ERE), viscera index (VSI), apparent digestibility of dry matter, protein, carbohydrate, and gross energy, whole-body crude protein and energy contents, intestinal enzymatic activities of protease, amylase, and Na⁺,K⁺-ATPase, intestinal villi length, crypt depth, muscular layer thickness, and the transcriptions of leptin (LEP) and cholecystokinin (CCK) of the EF group were all significantly higher than those of the PF group, while the opposite was true for feed intake and feed conversion ratio. These findings suggested that compared with PF, EF could improve the feed utilization and nutrient retention of blunt snout bream by enhancing the intestinal digestive and absorptive functions but reduce the feed intake through the stimulation of both LEP and CCK.
... The specific activity of sMMO was expressed as nanomoles of naphthol formed per milligram of cell protein per minute. Protein estimation was carried out according to Lowry [21] using bovine serum albumin as standard and Methylobacteria grown broth used for sMMO activity was used for the enumeration of colony forming units. ...
... The total sugar content in the raw seaweed waste and treated residue after microwave treatment at lower temperatures (i.e., bioactive peptide and protein extraction) was determined by acid hydrolysis using the method provided by the National Renewable Energy Laboratory method [32]. Subsequently, 15 mL of 72% (w/w) H 2 SO 4 was added to 1 g of dry sample and the solution was left at room temperature for 4 h. ...
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Continuous extraction production process by microwave-assisted extraction (MAE) using brown macroalgae Undaria pinnatifida waste was investigated to obtain proteins and a sulfated polysaccharide, fucoidan, at low (55–160 °C) and high (150–170 °C) temperatures. At the lower temperatures, protein extraction increased with increasing extraction condition severity. All extracted proteins exhibited angiotensin-converting enzyme inhibitory activity. The proteins extracted at 100 and 120 °C for 20 and 5 min, respectively, displayed the highest activities (evaluated as IC50 of protein concentration), which were 0.03 and 0.05 g/L, respectively. Subsequently, the highest fucoidan yields were attained after extraction at 150 °C for 30 min (12.3% based on the raw material) during the higher temperature extraction process. The yield was equivalent to that extracted directly by the MAE treatment (without treatment at a lower temperature). The results indicated that the continuous extraction of protein and fucoidan using MAE is possible. Moreover, U. pinnatifida waste could potentially be used as feedstock to provide valuable chemicals.
... Both alkali treatment and boiling of biomass aim to dissolve protein into the liquor well. The protein concentration of the liquor was measured according to Lowry protein assay (Lowry et al., 1951) at 550 nm. The standard protein solution was prepared by bovine serum albumin, using 1 M NaOH instead of DI water for gradient dilution. ...
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... Total protein was quantified after 7 days of culture, according to the modified method of Lowry et al. 30 After removing the culture medium, the wells were washed three times with PBS at 37 C and filled with 2 ml of 0.1% sodium lauryl sulfate (Sigma-Aldrich) for protein extraction. ...
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The objective was to synthesize and characterize fine polycaprolactone (PCL) fibers associated with a new 58S bioglass obtained by the precipitated sol–gel route, produced by the electrospinning process in order to incorporate therapeutic ions (Mg and Li). In PCL/acetone solutions were added 7% pure bioglass, bioglass doped with Mg(NO3)2 and Li2CO3 and were subjected to electrospinning process. The fibers obtained were characterized morphologically, chemically and biologically. The results showed the presence of fine fibers at the nanometric scale and with diameters ranging from 0.67 to 1.92 μm among groups. Groups containing bioglass showed particles both inside and on the surface of the fibers. The components of the polymer, bioglass and therapeutic ions were present in the fibers produced. The produced fibers showed cell viability and induced the formation of mineralization nodules. It was observed the applicability of that methodology in making an improved biomaterial, which adds the osteoinductive properties of the bioglass to PCL and to those of therapeutic ions, applicable to guided bone regeneration.
... The oxidation of the glutamate was measured in both microbial fractions (protozoal and bacterial) by the decrease in optical density by measuring the rate of change in absorbance at 340 nm caused by reduction of di-phospho pyridine nucleotide (DPNH). Protein in the enzyme extracts was estimated by the method of Lowry [49] to work out the specific activities of the assayed enzymes. Proteolytic activity of the bacteria was studied by estimating undigested protein from casein as described by Blackburn and Hobson [50] with a modification used by Kumar and Singh [51]. ...
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... The resulting homogenate was centrifuged in a cooling centrifuge at 14,000 × rpm for 20 min. Protein concentration was estimated in the lysates obtained through Lowry's protein assay [31] and the samples were processed for western blotting. Lysates were mixed with 2× SDS reducing SDS buffer, and the mixture was boiled at 95 °C for 5 min. ...
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Parkinson’s disease (PD) is a progressive motor neurodegenerative disorder significantly associated with protein aggregation related neurodegenerative mechanisms. In view of no disease modifying drugs, the present study was targeted to investigate the therapeutic effects of pharmacological agent 4-phenylbutyric acid (4PBA) in PD pathology. 4PBA is an FDA approved monocarboxylic acid with inhibitory activity towards histone deacetylase and clinically treats urea cycle disorder. First, we observed the significant protective effects of 4PBA on PD specific neuromuscular coordination, level of tyrosine hydroxylase, α-synuclein level and neurotransmitter dopamine in both substantia nigra and striatal regions of the experimental rat model of PD. Further results revealed that treatment with 4PBA drug exhibited significant protection against disease related oxidative stress and augmented nitrite levels. The disease pathology-related depletion in mitochondrial membrane potential and augmented level of calcium as well as mitochondrion membrane located VDAC1 protein level and cytochrome-c translocation were also significantly attenuated with 4PBA administration. Inhibited neuronal apoptosis and restored neuronal morphology were also observed with 4PBA treatment as measured by level of pro-apoptotic proteins t-Bid, Bax and cleaved caspase-3 along with cresyl violet staining in both substantia nigra and striatal regions. Lastly, PD-linked astrocyte activation was significantly inhibited with 4PBA treatment. Altogether, our findings suggest that 4PBA exerts broad-spectrum neuroprotective effects in PD animal model. Graphical Abstract
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Myocardial ischemia/reperfusion (I/R) injury leads to significant impairment of cardiac function and remains the leading cause of morbidity and mortality worldwide. Activation of peroxisome proliferator-activated receptor β/δ (PPARβ/δ) confers cardioprotection via pleiotropic effects including antioxidant and anti-inflammatory actions; however, the underlying mechanisms are not yet fully elucidated. The aim of this study was to investigate the effect of PPARβ/δ activation on myocardial mitochondrial respiratory function and link this effect with cardioprotection after ischemia/reperfusion (I/R). For this purpose, rats were treated with the PPARβ/δ agonist GW0742 and/or antagonist GSK0660 in vivo. Mitochondrial respiration and ROS production rates were determined using high-resolution fluororespirometry. Activation of PPARβ/δ did not alter mitochondrial respiratory function in the healthy heart, however, inhibition of PPARβ/δ reduced fatty acid oxidation (FAO) and complex II-linked mitochondrial respiration and shifted the substrate dependence away from succinate-related energy production and towards NADH. Activation of PPARβ/δ reduced mitochondrial stress during in vitro anoxia/reoxygenation. Furthermore, it preserved FAO-dependent mitochondrial respiration and lowered ROS production at oxidative phosphorylation (OXPHOS)-dependent state during ex vivo I/R. PPARβ/δ activation was also followed by increased mRNA expression of components of FAO -linked respiration and of transcription factors governing mitochondrial homeostasis (carnitine palmitoyl transferase 1b and 2-CPT-1b and CPT-2, electron transfer flavoprotein dehydrogenase -ETFDH, peroxisome proliferator-activated receptor gamma co-activator 1 alpha- PGC-1α and nuclear respiratory factor 1-NRF-1). In conclusion, activation of PPARβ/δ stimulated both FAO-linked respiration and PGC-1α/NRF -1 signaling and preserved mitochondrial respiratory function during I/R. These effects are associated with reduced infarct size.
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Background Salinized soils negatively affect plant growth, so it has become necessary to use safe and eco-friendly methods to mitigate this stress. In a completely randomized design, a pot experiment was carried out to estimate the influence of the inoculation with endophytic bacterial isolates Azospirillum brasilense , Pseudomonas geniculata and their co-inoculation on growth and metabolic aspects of flax ( Linum usitatissimum ) plants that already grown in salinized soil. Results The results observed that inoculation of salinity-stressed flax plants with the endophytes A. brasilense and P. geniculata (individually or in co-inoculation) increases almost growth characteristics (shoot and root lengths, fresh and dry weights as well as number of leaves). Moreover, contents of chlorophylls and carotenoids pigments, soluble sugars, proteins, free proline, total phenols, ascorbic acid, and potassium (K ⁺ ) in flax plants grown in salinized soil were augmented because of the inoculation with A. brasilense and P. geniculata . Oppositely, there are significant decreases in free proline, malondialdehyde (MDA), hydrogen peroxide (H 2 O 2 ), and sodium (Na ⁺ ) contents. Regarding antioxidant enzymes, including superoxide dismutase (SOD), peroxidase (POD), and ascorbate peroxidase (APX), the inoculation with the tested endophytes led to significant enhancements in the activities of antioxidant enzymes in stressed flax plants. Conclusions The results of this work showed that the use of the endophytic bacterial isolates Azospirillum brasilense , Pseudomonas geniculata (individually or in co-inoculation) could be regarded as an uncommon new model to alleviate salinity stress, especially in salinized soils.
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Backround This study was conducted to evaluate whether sildenafil effectively treats necrotizing enterocolitis (NEC). Material and Methods 38 rat pups were divided into 4 groups: control, sildenafil-control, NEC, and sildenafil-NEC (Sil-NEC). NEC was induced by hypoxia/reoxygenation and cold stress. The pups were treated by administering 1 mg/kg sildenafil by intraperitoneal injection once a day until the fourth postnatal day. The tissues were stained with hematoxylin/eosin staining and examined with the TUNEL test for apoptosis. The intestinal levels of malondialdehyde (MDA), interleukin 1β (IL-1β), inducible nitric oxide synthase (iNOS), caspase-3, and glutathione peroxidase (GSH-px) activity were quantified. Results TUNEL positivity (p=0.002) and intestinal damage grade (p<0.001) were found to be significantly lower in the Sil-NEC group. In addition, MDA, IL-1β, iNOS, caspase-3 levels, and GSH-px activity were also found to be significantly lower in the Sil-NEC group (p<0.001, p=0.004, p=0.011, p=0.026, p=0.002 respectively). Conclusion In this study, sildenafil has been shown to reduce intestinal damage and prevent the development of necrosis biochemically and histopathologically, with its antioxidant, anti-apoptotic, and anti-inflammatory effects, in the treatment of the experimental necrotizing enterocolitis model. This may suggest that sildenafil can be used to treat necrotizing enterocolitis, but further clinical studies are required.
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Introduction: CDKL5 deficiency disorder (CDD) is a rare neurodevelopmental condition, primarily affecting girls for which no cure currently exists. Neuronal morphogenesis and plasticity impairments as well as metabolic dysfunctions occur in CDD patients. The present study explored the potential therapeutic value for CDD of FRAX486, a brain-penetrant molecule that was reported to selectively inhibit group I p21-activated kinases (PAKs), serine/threonine kinases critically involved in the regulation of neuronal morphology and glucose homeostasis. Methods: The effects of treatment with FRAX486 on CDD-related alterations were assessed in vitro (100 nM for 48 h) on primary hippocampal cultures from Cdkl5-knockout male mice (Cdkl5-KO) and in vivo (20 mg/Kg, s.c. for 5 days) on Cdkl5-KO heterozygous females (Cdkl5-Het). Results: The in vitro treatment with FRAX486 completely rescued the abnormal neuronal maturation and the number of PSD95-positive puncta in Cdkl5-KO mouse neurons. In vivo, FRAX486 normalized the general health status, the hyperactive profile and the fear learning defects of fully symptomatic Cdkl5-Het mice. Systemically, FRAX486 treatment normalized the levels of reactive oxidizing species in the whole blood and the fasting-induced hypoglycemia displayed by Cdkl5-Het mice. In the hippocampus of Cdkl5-Het mice, treatment with FRAX486 rescued spine maturation and PSD95 expression and restored the abnormal PAKs phosphorylation at sites which are critical for their activation (P-PAK-Ser144/141/139) or for the control cytoskeleton remodeling (P-PAK1-Thr212). Conclusions: Present results provide evidence that PAKs may represent innovative therapeutic targets for CDD.
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Some analytical methods can be miniaturized to mitigate waste generation, cost, analysis time, and analysts’ exposure to toxic compounds. Spectrophotometric methods generally require analyte derivation to form chromophores, thus, increasing the quantity of reagents and residues. Miniaturization of the quantification step, which is carried out by reducing the reaction volume, maintaining the stoichiometric relation of the reaction, and absorbing characteristic of chromophores, is an interesting strategy to make routine analytical activities friendlier. Therefore, this study aimed at establishing a reliable protocol to miniaturize the quantification step in methods used for determining reducing sugars, total starch and soluble proteins in wheat-based samples. The greener of the miniaturized methods was evaluated by a metric tool. Miniaturization enabled reduction of up to 87% of waste and 7.5-fold the analysis cost without affecting reliability of methods, by comparison with original protocols (similarities > 98% and coefficients of correlation > 96%) and reaction stoichiometry (quantification of reference samples) with no significant difference (p > 0.05) from original methods and applicability to wheat-based samples. Resulting protocols of each method maintained their fundamentals and made them “greener” (scores > 0.60 by the Analytical GREEnness calculator), with low cost, lower analyst’s exposure and contributing reducing the environmental impact of these routine analyses.
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To improve the utilization value of chicken by-products, we utilized the method of step-by-step hydrolysis with bromelain and flavourzyme to prepare low molecular weight chicken skin collagen peptides (CCP) (<5 kDa) and characterized the amino acids composition of the CCP. Then, we prepared novel CCP-chelated zinc (CCP–Zn) by chelating the CCP with ZnSO 4 . We found that the bioavailability of CCP–Zn is higher than ZnSO 4 . Besides, CCP, ZnSO 4 , or CCP–Zn effectively repressed the tumor growth, invasion, and migration in a Drosophila malignant tumor model. Moreover, the anti-tumor activity of CCP–Zn is higher than CCP or ZnSO 4 . Furthermore, the functional mechanism studies indicated that CCP, ZnSO 4 , or CCP–Zn inhibits tumor progression by reducing the autonomous and non-autonomous autophagy in tumor cells and the microenvironment. Therefore, this research provides in vivo evidence for utilizing chicken skin in the development of zinc supplements and cancer treatment in the future.
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Abstract Nude mice carry an autosomal recessive mutation in the Foxn1 gene and therefore are homozygous recessive animals (Foxn1 -/-). The fertility rate of homozygous male (Foxn1-/-) is low, which seems to be related to the delay in the production of gametes at the beginning of sexual maturity. The present study evaluated the structural and organizational aspects of the testicles of homozygous and heterozygous offspring related to the Foxn1 gene in mice, describing its implications on spermatogenesis. Adult males Balb/c, Foxn1+/- and Foxn1-/- mice were used. Testes and epididymis were harvested for histological, biochemical, and sperm transit analyses. Gonadal weight was significantly lower in Foxn1+/-and Foxn1-/-animals, the same behavior was noticed for the activity of antioxidant enzymes. In addition, tubular parameters such as epithelial proportion, length, and area, as well as germ and Leydig cell’s populations were significantly reduced in the aforementioned groups, leading to lower sperm production. In conclusion, our results indicate the importance of the Foxn1 in Leydig cell’s function, reflecting in the preservation of spermatogenesis, thus in germ cell’s population and sperm cell production.
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Schizophrenia is a common mental disorder affecting patients’ thoughts, behavior, and cognition. Recently, the NRG1/ErbB4 signaling pathway emerged as a candidate therapeutic target for schizophrenia. This study investigates the effects of aripiprazole and sertindole on the NRG1/ErbB4 and PI3K/AKT/mTOR signaling pathways in ketamine-induced schizophrenia in rats. Young male Wistar rats received ketamine (30 mg/kg, intraperitoneally) for 5 consecutive days and aripiprazole (3 mg/kg, orally) or sertindole (2.5 mg/kg, orally) for 14 days. The proposed pathway was investigated by injecting LY294002 (a selective PI3K inhibitor) (25 μg/kg, intrahippocampal injection) 30 min before the drugs. Twenty-four hours after the last injection, animals were subjected to behavioral tests: the open field test, sucrose preference test, novel object recognition task, and social interaction test. Both aripiprazole and sertindole significantly ameliorated ketamine-induced schizophrenic-like behavior, as expected, because of their previously demonstrated antipsychotic activity. Besides, both drugs alleviated ketamine-induced oxidative stress and neurotransmitter level changes in the hippocampus. They also increased the gamma-aminobutyric acid and glutamate levels and glutamate decarboxylase 67 and parvalbumin mRNA expression in the hippocampus. Moreover, aripiprazole and sertindole increased the NRG1 and ErbB4 mRNA expression levels and PI3K, p-Akt, and mTOR protein expression levels. Interestingly, pre-injecting LY294002 abolished all the effects of the drugs. This study reveals that the antipsychotic effects of aripiprazole and sertindole are partly due to oxidative stress reduction as well as NRG1/ErbB4 and PI3K/AKT/mTOR signaling pathways activation. The NRG1/ErbB4 and PI3K signaling pathways may offer a new therapeutic approach for treating schizophrenia in humans.
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The current study explorated the hepatoprotective and immunomodulatory effects of Linalool (Lin) against carbon tetrachloride (CCl4)‐induced toxicity in mice. Four study groups (n = 8 each) were used: (1) a negative control group; (2) a toxicity control group (single dose of CCl4 administered at day 14 as 1ml/kg of CCL4 in 1% olive oil. Intraperitoneally (i.p)); and two experimental groups where mice were treated with either (3) Lin (25 mg/kg b.w, orally, daily for 15 days) or (4) pretreated with Lin (25 mg/kg b.w., orally, daily for 14 days) and intoxicated with CCl4 (1ml/kg of CCL4 in 1% olive oil. i.p) on day 14. The levels of the anti‐inflammatory cytokines interleukin 10 (IL‐10), pro‐inflammatory cytokines TNF‐ α, IL‐6 and TGF‐1β, and the histopathology of the liver were accessed. According to our findings, IL‐10 concentrations were significantly increased in Lin‐treated groups, while other cytokine levels were marked by a considerable decrease in the toxicity model group (CCl4‐treated group). Histopathological examinations for liver tissues showed that Lin treated groups had an almost normal structure. The current findings showed that Lin could inhibit the CCl4‐induced liver injury in mice, which warrants further investigation of Lin as a potential protective and therapeutic agent against hepatotoxicity. This article is protected by copyright. All rights reserved
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