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Protein measurement with the FOLIN phenol reagent
... The acid ninhydrin complex in toluene was used to measure the amount of free proline in fresh leaf tissue (Bates, 1973). The homogenate's protein content was measured using bovine serum albumin (BSA) as a reference in Tri Chloro Acetic Acid (TCA) precipitate (Lowery, 1951).Calorimetric measurements of sugars were made (Nelson, 1941). ...
... The hemolymph samples from the crabs showing different molt cycle stages were collected and the total protein from measured hemolymph was precipitated by adding 10% cold trichloroacetic acid (TCA) followed by centrifugation at 3000 rpm for 15 min. The precipitate was dissolved in 0.1 N NaOH and the resulted sample was analyzed for the total protein (after Lowry et al., 1951;Anilkumar and Adiyodi, 1980;Sudha and Anilkumar, 2007;Nagathinkal et al., 2017). Briefly, reagent A was prepared by mixing 2% sodium carbonate in 0.1 N sodium hydroxide solution. ...
... The protein content within exosomes was evaluated by the Lowry Protein Assay Kit (Bio Basic Inc, Canada [17] . 150 μg of exosomal protein (extracted from about 5 ml of cultured media) suspended in PBS (100 μl) was administered intravenously to each rat [18] . ...
... These samples were then homogenized with 5 mL of phosphate buffer (100 mM, pH 7.5) containing 1 mM EDTA and 0.01% Triton X-100. The soluble protein content (mg/g) was estimated by the technique of Lowry et al. [39] using bovine serum albumin as a protein standard. ...
Drought stress represents a major constraint with significant impacts on wheat crop globally. The use of plant growth-promoting bacteria (PGPB) has emerged as a promising strategy to alleviate the detrimental impacts of water stress and enhance plant development. We investigated 24 strains from diverse ecosystems, assessed for PGP traits and tolerance ability to abiotic stresses (drought, salinity, temperature, pH, heavy metals, pollutants, herbicides, and fungicides). The most effective bacterial strains Providencia vermicola ME1, Pantoea agglomerans Pa, Pseudomonas knackmussi MR6, and Bacillus sp D13 were chosen. Furthermore, these strains exhibited PGP activities under osmotic stress (0, 10, 20, and 30% PEG-6000). The impact of these osmotolerant PGPBs on wheat (Triticum durum L.) growth under drought stress was assessed at two plant growth stages. In an in vitro wheat seed germination experiment, bacterial inoculation significantly enhanced germination parameters. In pot experiments, the potential of these bacteria was evaluated in wheat plants under three treatments: Well-watered (100% field capacity), moderate stress (50% FC), and severe stress (25% FC). Results showed a significant decline in wheat growth parameters under increasing water stress for uninoculated seedlings. In contrast, bacterial inoculation mitigated these adverse effects, significantly improving morphological parameters and chlorophyll pigment contents under the stress conditions. While malondialdehyde (lipid peroxidation) and proline contents increased significantly with drought intensity, they decreased after bacterial inoculation. The antioxidant enzyme activities (GPX, CAT, and SOD) in plants decreased after bacterial inoculation. The increased root colonization capacity observed under water stress was attributed to their ability to favorable adaptations in a stressful environment. This study highlighted the potential of selected PGPB to alleviate water stress effects on wheat, promoting practical applications aimed at enhancing crop resilience under conditions of water shortage.
Beer lees (BL), a by-product of beer production, consist mainly of dead yeast cells with potential nutritional value. On the other hand, yeast extract (YE), obtained through the lysis of yeast cells, is commonly used as a nutrient-rich supplement for the growth of fastidious microorganisms such as lactic acid bacteria (LAB). However, YE is a high-cost ingredient. Therefore, the aim of this study was to optimize the use of BL as a low-cost alternative source of YE through different lysis treatments, evaluating its suitability to support the growth of UNQLpc 10 and UNQLp 11 strains in a whey permeate (WP)-based medium. Growth kinetics and cell viability were compared with those obtained in MRS broth. The best results were observed with sonicated BL, up to 10 logarithmic units, which supported LAB growth comparable to MRS. Although autolyzed BL promoted lower bacterial growth than sonicated BL, it showed greater cell disruption and higher levels of nitrogen, proteins, and amino acids (5.32%, 26.0%, and 277 nM, respectively). Additionally, autolyzed BL exhibited lower concentrations of reducing sugars and a higher presence of Maillard reaction products, as indicated by colorimetric analysis. These changes, which may be related to the formation of Maillard reaction products during the autolysis process, could have negatively affected the nutritional quality of the extract and, thus, reduced its effectiveness as a bacterial growth promoter.
In this study, we aim to analyze whether supplementation with cactus has the potential to minimize the testicular damage caused by heat stress, assess which of the three cactuses would have the best potential, and suggest a possible pathway (oxidative or hormonal) for the action of cactus on the testicular parameter alterations caused by heat stress. Thirty-two male lambs, of the Santa Inês type, not castrated, approximately six months old, and averaging 21.0 ± 2.0 kg body weight, were divided into four groups, as follows: G1, which was fed an elephant grass hay diet (control); G2, with a diet with partial replacement using small cactus forage (Opuntia cochenillifera) (SMALL); G3, which was fed Mexican elephant ear (Opuntia stricta Haw) (MEE); and G4, which was fed IPA Sertania (Nopalea cochenillifera Salm Dyck) (IPA) for 63 days. After slaughter, blood and testicles were removed. The right testicles were fixed for histological analyses, and the left testicles were stored in the freezer for oxidative stress analyses. Serum testosterone, T3, and T4 levels were analyzed. The body weight of animals treated with cactus forage was higher than in the control group. However, the gonadosomatic index did not differ among experimental groups. Heat stress triggered the degradation of testis tissue in all experimental groups. The testicular degeneration process was characterized by tubular atrophy, reduction in germ epithelium height, germ cell vacuolization and necrosis, Sertoli cell vacuolization, germ cell scaling of the tubular fire, and increased intertubular space. The three different cactus forages used in this study had different weaknesses regarding their antioxidant defenses, hormonal levels, and histopathology. However, it is important to highlight that the IPA group had lower qualitative changes in the intertubular areas than the other experimental groups. The testosterone level increased in MEE (Opuntia stricta) and IPA groups, while T3 and T4 increased in SMALL (Opuntia cochenillifera) and IPA groups. The malondialdehyde, an important marker of lipid peroxidation, was reduced only in the IPA group. The testosterone level increased in MEE and IPA groups, while T3 and T4 increased in SMALL and IPA groups. In conclusion, heat stress triggers several histopathologies in testis tissue, and IPA cactus (Nopalea cochenillifera) was the most appropriate supplementation for reducing the damages, compared with an elephant grass hay diet or small cactus forage and Mexican elephant ear supplementation.
The work reported here investigates the effects of temperature on starch hydrolysis carried out by limit dextrinase (LD) and α-glucosidase (AGL) in diverse barley germplasm. The original activity of both the carbohydrases in barley genotypes declined after 10 min of heat treatment at 55 °C. The decline in thermostability was less than 50% for LD at 55 °C, while it was more than 50% of the original activity for AGL. This indicates the higher temperature optima (T50) for LD than AGL. We identified five genotypes for LD which had % remaining activity of more than 50% after heat treatment at 55 °C and six genotypes for AGL activity. One mutant, BL2096 was found to have high activity for both the studied enzymes. The identified genotypes with high LD and AGL activities and thermostability can be used profitably to find alleles or genetic factors encoding more thermostable enzymes in barley germplasm.
Diabetes is commonly associated with oxidative stress, a factor believed to contribute in its development and progression. Chlorogenic acid (CGA), a phenolic compound found in coffee and other food sources has been shown to have antioxidant and anti-inflammatory properties by scavenging free radicals. In the present study, CGA was assessed for its ability to reduce oxidative stress in the liver and kidney, as well as serum inflammatory response in streptozotocin (STZ)-induced diabetic rats.STZ (50 mg/kg)was administered intraperitoneally to male albino Wistar rats to induce experimental diabetes and were divided into five groups (n=5); control, diabetic control, diabetic/metformin-treated, diabetic/CGA (100 and 150 mg/kg) treated.Diabetic rats showed significant elevation in pro-inflammatory cytokines (TNF-α, IL-6, and IFN-γ) and decreased concentration of the anti-inflammatory cytokine IL-10. They also had reduced activity of antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST), and reduced glutathione (GSH), as well as increased levels of peroxidation marker in their liver and kidney tissues when compared to control rats. Oral administration of CGA (100 and 150 mg) for 28 days markedly (p<0.05) improved all of these inflammatory and oxidative stress parameters in diabetic rats. These findings were further corroborated through histopathological examination of hepatic and renal tissues.The observed results indicate that CGA possess antioxidant and anti-inflammatory property, as evidenced by reduction in redox imbalance, inflammation and lipid peroxidation to near normal, thus offering protection to the liver and kidneys tissues from damage inflicted due to oxidative stress and inflammation in diabetic rats.
Fascioliasis, which is caused by Fasciola hepatica and Fasciola gigantica ( F. gigantica ), is a public health problem worldwide. Stages infective to humans occur on aquatic plants and in water. The gold standard for diagnosing human infection depends on finding fluke eggs in stool samples. However, this method has low diagnostic sensitivity; eggs from other trematode species might be misidentified as those of Fasciola , and eggs are not voided in ectopic infections. Therefore, serological analysis can support the diagnosis of human fascioliasis. We evaluated whether a new fascioliasis immunochromatographic test kit, which detects specific IgG antibodies using F. gigantica excretory–secretory antigen, can be used for rapid diagnosis from whole blood. The kit (“the fascioliasis whole-blood test kit”) was evaluated under laboratory conditions using 250 whole-blood samples (WBSs), of which 41 were from fascioliasis cases. Results based on simulated WBSs and the corresponding serum samples were compared and showed almost perfect agreement (percent agreement = 97.2%; kappa value = 0.9145). The diagnostic sensitivity and specificity of the test using simulated WBSs were 97.6% and 96.2%, respectively. Comparable values when serum samples were tested were 100.0% and 93.3%, respectively. This kit represents a significant advance because it does not require extensive training of personnel, is easy to use, and can support diagnosis at the bedside or in local and remote hospitals with limited facilities. The kit may also contribute significantly to epidemiological surveys.
The impact of quality of light on plant growth and development has been extensively studied. However, the interplay between photosynthesis and metabolic regulation in Artemisia annua remains largely unexplored. To address this gap, we investigated how various light qualities; monochromatic red (R), blue (B), a 1:1 red-blue combination (RB), and broad-spectrum white light (W); affect physiological parameters, photosynthetic activity, and metabolic processes. Plants were exposed to these light conditions at a photosynthetic photon flux density (PPFD) of 200 µmol·m⁻²·s⁻¹ for 10 days. Exposure to different light treatments resulted in significant changes in morphological attributes, chlorophyll and carotenoid content, stomatal conductance, intercellular CO₂ concentration, and transpiration rates. DPPH-scavenging activity and ascorbic acid levels increased under RB and B light, with increments of 20.76 %–25.6 % and 23 %–43.29 %, respectively, compared to W light. Gas chromatography-mass spectrometry (GC-MS) analysis showed the highest monoterpene concentration (36 %) under B light, followed by RB light (31 %), R light (28 %), and W light (27 %). Further, High-resolution mass spectrometry (HRMS) indicated elevated levels of flavonoids, terpenes, phenolics, and other organic compounds in RB light-exposed plants, with B light showing the following highest levels. Additionally, vital photosynthesis-regulating genes such as LHCII, CAO, TK, PsbA, PsaB, PsbD, RbcL, ndhB, RbcS, PetB, PetD, AtpA, and FBP demonstrated significant upregulation under various light conditions. Genes involved in artemisinin biosynthesis, including HMGS, MK, MCT, MPDC, CYP71AV1, and 1,8-cineol synthase, showed increases of 38.4 %–13.3 %, 20.4 %–22.4 %, 29.5 %–37.5 %, 15 %–16 %, 43 %–68 %, and 93.4 %–106.5 %, respectively, in response to B and RB light. These findings underscore the complex influence of various light qualities on the metabolic pathways of A. annua, providing a basis for future research.
The aim of this study was to evaluate mixtures of bioagents and resistance inducers for protection of cotton roots against root infecting fungal pathogens. Three biological control agents (BCAs); Trichoderma hamatum (TM), Trichoderma harzianum (TZ) and Paecilomyces lilacinus (PL) and two resistance inducers (RIs); Bion (benzo(1,2,3)thiadiazole-7-carbothioic acid S-methyl ester) (BTH), salicylic acid (SA) were applied individually or in combination to test their efficacy in controlling cotton root rot disease caused by Fusarium oxysporum (FO) and Pythium debaryanum (PD) under greenhouse and field conditions. In greenhouse experiments, all applied treatments protected cotton seedlings against FO root rot. Disease index percentage (DI%) was significantly reduced up to 78.8%, while germination percentage increased significantly up to 199.60% compared with the infected control. All treatments significantly reduced PD DI% compared to infected controls. In field experiments, the maximum protection of cotton roots against FO and PD resulted from application of TM þ PL þ SA þ BTH, where DI% was reduced to 72.3% and 69.3% relative to infected controls, respectively. Increase in cell wall fractions (cellulose, hemicelluloses and lignin) resulted from application of both BCAs and RIs in case of PD. Lignin content significantly increased 1.68-1.93 (FO) and 1.07-1.39 (PD) fold over the infected controls. A significant increase in free phenolics content was positively proportional to the degree of plant resistance against the two pathogens. The main conclusion of this study is that by combining BCAs with RIs there was increased consistency of suppression of root rot of cotton seedlings caused by either F. oxysporum or P. debaryanum.
Key message
ARG6 and ARG10 pea accessions exhibited better tolerance to drought by keeping drought-associated attributes stable and higher, that is, stable chlorophyll content, high antioxidant activity, and the presence of polymorphic bands with stress-responsive EST-SSR markers.
Abstract
Each year, a significant portion of crops is lost due to various abiotic stresses, and even pea (Pisum sativum) crop growth and yield are severely affected by the challenges posed by drought stress. Drought is a critical factor that limits crop growth and development, and its impact is exacerbated by changes in the magnitude of climatic conditions. Drought induces oxidative stress in plants, leading to the accumulation of high concentrations of reactive oxygen species that damage cell structures and vital functioning of cells. The primary objective was to identify stress-tolerant plants by evaluating different morphological and biochemical attributes, such as biomass, chlorophyll content, relative water content, ascorbate peroxidase (APX), superoxide dismutase (SOD), and DPPH scavenging activity, as well as protein, proline, and phenolic content. Our study revealed that pea accessions (ARG6 and ARG10) were more resilient to drought stress as their chlorophyll, relative water, protein, and proline contents increased under drought conditions. Antioxidant enzymes, such as SOD, APX, and DPPH activities, also increased under drought stress in ARG10 and ARG6, suggesting that these accessions could bolster the antioxidant defense system in response to drought stress. Based on putative (cellular, biological, and metabolic) functions, ten EST-SSR primers were selected for the amplification study. Three EST-SSR primers, AUMP06_110, AUMP18_300, and AUMP31_250, were used for ARG6 and ARG10. Based on the correlation between the presence or absence of specific EST-SSR alleles, various physiological and morphological traits, and DPPH scavenging activity, both ARG10 and ARG6 demonstrated resistance to drought stress.
The objective of this study was to apply response surface methodology to estimate the emulsifying capacity and stability of mixtures containing isolated and textured soybean proteins combined with pectin and to evaluate if the extrusion process affects these interfacial properties. A simplex-centroid design was applied to the model emulsifying activity index (EAI), average droplet size (D[4,3]) and creaming inhibition (CI%) of the mixtures. All models were significant and able to explain more than 86% of the variation. The high predictive capacity of the models was also confirmed. The mean values for EAI, D[4,3] and CI% observed in all assays were 0.173 ± 0.015 nm, 19.2 ± 1.0 μm and 53.3 ± 2.6%, respectively. No synergism was observed between the three compounds. This result can be attributed to the low soybean protein solubility at pH 6.2 (<35%). Pectin was the most important variable for improving all responses. The emulsifying capacity of the mixture increased 41% after extrusion. Our results showed that pectin could substitute or improve the emulsifying properties of the soybean proteins and that the extrusion brings additional advantage to interfacial properties of this combination.
The study aimed to determine the CCl4-induced liver fibrosis model in pinealectomized rats and biochemically, immunohistochemically, and histopathologically investigate the therapeutic effect of melatonin on liver fibrosis. The surgical procedure for pinealectomy was performed at the beginning of the study, and the sham and pinealectomized rats were administered CCl4 dissolved in corn oil (1:1) alone every other day to induce liver fibrosis or together with melatonin (10 mg/kg) therapy for 15 days. Melatonin is an essential therapeutic agent and offers an alternative therapeutic strategy in CCl4-induced liver fibrosis by suppressing inflammation, oxidative stress, and the TGF-β1 signaling pathway. Treatment with melatonin ameliorated CCl4-induced liver fibrosis by restoring hepatocellular damage and reducing plasma AST, ALT, and ALP values. Melatonin increases the activity of SOD and CAT, which are important enzymes for antioxidant defence, and raises GSH levels, which further enhances antioxidant function. Also, melatonin reduced hepatic inflammation (IL-6 and IL-1β) and oxidative stress indices. Moreover, histopathological changes and immunohistochemical expression of TGF-β1 were restored following melatonin supplementation in the CCl4-induced liver fibrosis model in pinealectomized rats. Our study shows that melatonin supplementation has a beneficial effect in protecting the liver fibrosis induced by CCl4 in pinealectomized rats.
Keratin waste has become an increasingly serious environmental and health hazard. Keratin waste is mainly composed of keratin protein, which is one of the most difficult polymers to break down in nature and is resistant to many physical, chemical, and biological agents. With physical and chemical methods being environment damaging and costly, microbial degradation of keratin using keratinase enzyme is of great significance as it is both environment friendly and cost-effective. The aim of this study was to extract and purify keratinase from bacterial species isolated from the soil. Among the organisms, an isolate of Bacillus velezensis, coded as MAMA could break down chicken feathers within 72 hours (h). The isolated strain produced significant levels of keratinase in mineral salt medium by supplying chicken feathers as the sole source of nitrogen and carbon. Feather deterioration was observed with the naked eye, and enzyme activity was evaluated using a spectrophotometric assay. Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and zymography results revealed that the keratinase protein produced by Bacillus velezensis had a molecular weight between 40 and 55 kilodalton (kDa).
Background
Cotton ( Gossypium hirsutum L.) is one of the most significant fibre and cash crops and plays an important role in Indian industrial and agricultural economies. However, over the years quantity and quality have been hampered by the pest leafhopper. Leafhopper alone has been shown to cause yield losses of up to 40%. In this study, screening and evaluation were performed to identify and categorize 100 cotton genotypes along with 5 checks as resistant, moderately resistant, sensitive and highly sensitive to leafhoppers.
Results
A total of hundred genotypes were evaluated along with five checks for leafhopper resistance. Based on the screening results, a total of 19 genotypes were resistant to leafhoppers, which was on par with the findings of the check KC 3. The contents of total soluble sugar, total soluble protein, and total free amino acids were significantly positively correlated with the mean grade, whereas total phenols content and trichome density were significantly negatively correlated with the susceptibility grade. However, based on screening and biochemical analysis, the genotypes KC 2, JR-23, Samaru-26-T, D 4, TCH 1728, RS 253, and B-61-1862 exhibited high resistance to leafhopper.
Conclusion
According to the findings of this study, choosing genotypes with high total phenolics content together with high trichome density and low contents of total soluble sugar, total soluble protein, and free amino acids may aid in the development of resistant genotypes.
This study evaluated Streptomyces rochei strain NAM-19 solid-state fermentation of agricultural wastes to produce alkaline protease. Alkaline protease production increased with flaxseed, rice bran, and cheese whey fermentation reaching 147 U/mL at 48 h. Statistical optimization of alkaline protease production was performed using the central composite design (CDD). Results of CDD and the optimization plot showed that 4.59 g/L flaxseed, 4.31 g/L rice bran, 4.17 mL cheese whey, and a vegetative inoculum size of 7.0% increased alkaline protease production by 27.2% reaching 186 U/mL. Using the 20–70% ammonium sulfate fractionation method, the optimally produced enzyme was partially purified to fivefold. The partially purified alkaline protease was then covalently immobilized on a biopolymer carrier, glutaraldehyde-polyethylene-imine-κ-carrageenan (GA-PEI-Carr), with 90% immobilization efficiency. Characterizations revealed that immobilization improved thermostability, reusability, optimum temperature, and sensitivity towards metal ions of the free enzyme. The optimal temperature for free and immobilized enzymes was 40 and 50 °C, respectively. Both enzymes had the same optimum pH of 10. Immobilization increased Km from 19.73 to 26.52 mM and Vmax from 56.7 to 62.5 mmol min−1L−1. The immobilized enzyme retained 35% of its initial activity at 70 °C, while the free enzyme retained only 5%. The immobilized enzyme kept 80% of its initial activity at the 20th cycle. After 7 weeks of storage, the free enzyme lost all its initial activity, whereas the immobilized enzyme retained 50%. The free and immobilized enzymes were able to hydrolyze gelatin, and azo-casein demonstrating different relative activity, 85, 80, 90 and 95%, respectively, compared to casein (100%).
Phycocyanin is a highly valued pigment present in Spirulina platensis biomass with applications in the food industry in terms of biorefinery concepts; specifically, its antioxidant and antimicrobial capacity are an advantage that could be incorporated into a food matrix. This study aims to use rice husk as an alternative culture medium for S. platensis biomass growth and phycocyanin extraction by ohmic heating processing using a 3D-printed reactor. S. platensis was cultivated in rice husk extract (RHE) from 0–100% (v/v). The highest content of microalgal biomass was 1.75 ± 0.01 g/L, with a specific growth rate of 0.125 ± 0.01 h⁻¹. For the phycocyanin extraction under an ohmic heating process, a 3D-printed reactor was designed and built. To optimize phycocyanin extraction, a central composite rotatable design (CCDR) was evaluated, with three factors: time (min), temperature (°C), and pH. The highest phycocyanin content was 75.80 ± 0.98 mg/g in S. platensis biomass grown with rice husk extract. Ohmic heating is a promising method for rapid phycocyanin extraction, and rice husk as a culture medium is an alternative for the growth of S. platensis biomass in the integration of second- and third-generation biorefineries.
This study explored the impacts of supplementation of different levels of coated methionine (Met) in a high-plant protein diet on growth, blood biochemistry, antioxidant capacity, digestive enzymes activity and expression of genes related to TOR signaling pathway in gibel carp (Carassius auratus gibeilo). A high-plant protein diet was formulated and used as a basal diet and supplemented with five different levels of coated Met at 0.15, 0.30, 0.45, 0.60 and 0.75%, corresponding to final analyzed Met levels of 0.34, 0.49, 0.64, 0.76, 0.92 and 1.06%. Three replicate groups of fish (initial mean weight, 11.37 ± 0.02 g) (20 fish per replicate) were fed the test diets over a 10-week feeding period. The results indicated that with the increase of coated Met level, the final weight, weight gain (WG) and specific growth rate initially boosted and then suppressed, peaking at 0.76% Met level (P< 0.05). Increasing dietary Met level led to significantly increased muscle crude protein content (P< 0.05) and reduced serum alanine aminotransferase activity (P< 0.05). Using appropriate dietary Met level led to reduced malondialdehyde concentration in hepatopancreas (P< 0.05), improved superoxide dismutase activity (P< 0.05), and enhanced intestinal amylase and protease activities (P< 0.05). The expression levels of genes associated with muscle protein synthesis such as insulin-like growth factor-1, protein kinase B, target of rapamycin and eukaryotic initiation factor 4E binding protein-1 mRNA were significantly regulated, peaking at Met level of 0.76% (P< 0.05). In conclusion, supplementing optimal level of coated Met improved on fish growth, antioxidant capacity, and the expression of TOR pathway related genes in muscle. The optimal dietary Met level was determined to be 0.71% of the diet based on quadratic regression analysis of WG.
The present study was carried out to determine the effect of dietary cadmium exposure on growth performance, changes in manganese, zinc, copper, calcium, magnesium, iron, selenium, and cadmium metals in liver and muscle tissue, liver antioxidant enzymes, and the histology of the fish. Rainbow trout (Oncorhynchus mykiss) with weights of 39.45 ±1.13 g was used in the experiment conducted in 2 groups and three replicates. The Control group was fed a cadmium-free diet, and the Cadmium group was fed a diet containing 5.03 µg/kg of cadmium twice a day until satiation. It was determined that cadmium intake through the diet affected growth rate and the feed evaluation performance negatively. In fish exposed to cadmium, manganese, zinc, copper, calcium, magnesium, iron, selenium, superoxide dismutase, catalase and glutathione peroxidase values in both muscle and liver tissues were significantly (p<0.05) decreased, whereas cadmium levels in muscle and liver and malondialdehyde levels in liver were significantly (p<0.05) increased. The histopathological examination of the liver revealed that cadmium caused liver damage. These results showed that rainbow trout exposed to dietary cadmium were highly sensitive to the metal, and the decreased levels of metals such as copper, zinc, manganese, and selenium in the liver tissue, which are involved in the antioxidant defence system, can be considered an indicator of the weakening of the antioxidant defence system.
The nitrilase enzymes are able to convert nitriles to their corresponding carboxylic acid. The present study reports the purification and characterisation of nitrilase (EC 3.5.5.1) which is able to convert acrylonitrile to acrylic acid and ammonia from Bacillus subtilis AGAB 2 (GenBank Ascension number- MW857547) in an optimized mineral medium. The purified nitrilase showed maximum enzyme activity of 5874.82 U ml⁻¹ and, specific enzyme activity of 2097.92 U mg⁻¹ with a 9.95-fold purification. The purified nitrilase showed high specificity for acrylonitrile (Vmax-344.82 µM s⁻¹; Kcat-1603.84 s⁻¹; Km-2.8 mM). The characterization showed that purified nitrilase is most stable and works best under the pH range of 7–8, and temperature range of 28 ºC—37 ºC temperature. The presence of heavy metal like—Co⁺², Cu⁺², Fe⁺², Hg⁺², Mg⁺², Mn⁺², Ni⁺², Ca⁺², Pb⁺², Ba⁺² and Zn⁺² negatively affected the enzyme activity. Among the tested cosolvents (Methanol, Ethanol, Hexane, DMSO, IPA, Acetone, Ethyl Acetate, Benzene, Chloroform, and Butanol), only Methanol negatively affected nitrilase activity. Hydroxylamine competitively inhibited nitrilase against acrylonitrile, with an IC50 value of 5.5 mM. The biotransformation of acrylonitrile to acrylic acid achieved 95% yield with high purity and selectivity in 48 h, confirmed by HPLC and GCMS analysis.
Graphical Abstract
An experiment was conducted with drought-tolerant and susceptible genotypes to find out the key traits that
determine drought tolerance in cassava. The drought-tolerant clone 8S501 and the two susceptible clones H-97
and CR43-11 were planted in the field in a Randomized Block Design and the drought stress was imposed when
the plants were three months old. The observations were measured at the beginning of the stress period (0-
stress), 30 and 75 days of stress. The repeated measures ANOVA stated that there were significant differences
among the genotypes, stress duration and their interaction for the majority of the traits. Growth, development
and tuber yield of cassava was not significantly affected up to 30 days of drought stress. There was a greater
decline in LA, TBM, CGR, RGR and NAR between 30 and 75 days of drought stress suggesting that measures are
to be taken if drought stress prolongs for more than 30 days to realize optimum growth and yield of cassava.
There was no increment in TY and PSR after 30 days of stress. A greater reduction was observed from 30 to 75
days of stress in leaf TC and CP content. There was a considerable increase in proline and total phenols content in
the leaf from 30 to 75 days of stress. The inherent free radical scavenging activity measured as DPPH, FRAP and
ABTS assays also demonstrated a notable increase in antioxidant activity at 75 days of drought stress. The early
vigour in terms of initial high leafiness, leaf area, tuber yield, proportion of storage roots and total biomass in the
first three months after planting facilitated the clone 8S501 to effectively tolerate the drought stress conditions.
Saharan population in Algeria still depending on bovine milk, which suffers from serious constraints undermining its sustainability. Camelus dromedarius milk has experienced growing demand following the emerging market requirements for livestock production and dairy farming over the past decade. The present work aimed at analysing the effect of nutritional regime on milk quality. The differences in pH, Acidity D°, Ash and Fats were significant. The pH was negatively influenced by the intensification conditions such as the much higher use of concentrates. The major constituents of milk were strongly and positively correlated with barley, wheat bran, TN/Kg.DM (Total Nitrogen/ Kg. Dry Matter), Kg.DM, Concentrates and daily watering. The results showed that a good energy-protein balance around 73 g PDI/UFL (Protein Digestible in the Intestine/Energetic Forage Unit for milk production) was beneficial for a better milk protein ratio. The use of corn, soybeans, palm dates and VM-premix (Vitamin Mineral) supplementation were also favourable to the synthesis of fats. Crude fiber and cell walls were better valued in the synthesis of fats with the availability of concentrates and the increasing of TN /Kg.DM and VM-premix rate in dietary regime. The vitamin C content elevate following high ratio of UFL /Kg.DM and PDI/UFL. For thus, the influence of nutritional status can lead to major improvements that need also more advanced and detailed studies.
This study aimed to investigate the quality and quantity of organic matter (OM) using biochemical components (protein, carbohydrate and lipids) in the sediment cores collected from the Central Indian Ocean (CIOB) under the program Equatorial Indian Ocean Process Study Dynamics and Biogeochemistry (EIOPS). Total organic carbon (TOC) and biochemical parameters (carbohydrate, lipid, protein, Tannin and lignin) were determined in core I and core II, respectively. Total carbohydrates varied from 4.66 to 2557.32 mg/kg (average 459.31 mg/kg) and 142.23 to 821.56 mg/kg (average 380.01 mg/kg) in core I and II, respectively. In core I, PRT varied from 70.95 to 107.05 mg/kg, and the minimum and maximum content of CHO was 143.23 and 822.56 mg/kg. The maximum and minimum concentrations of BPC in core II were 786.32 and 381.07 at 0–10 cm depth, respectively, which corresponded to the concentrations of PRT, CHO, LPD, and Tannin. The results showed that PRT was statistically significant with the TOC and negatively correlated with the LPD and CHO, while LPD was highly significant with clay and silt grains. In most of the samples, the ratio of LPD to CHO ratio was > 1, which indicated higher productivity of benthic organisms inhabiting the CIOB.
Mycoplasma gallisepticum (MG) is responsible for chronic respiratory disease in avian species, characterized by symptoms like respiratory rales and coughing. Existing vaccines for MG have limited efficacy and require multiple doses. Certain MG cytoadherence proteins (GapA, CrmA, PlpA, and Hlp3) play a crucial role in the pathogen’s respiratory tract colonization and infection. Plant-based proteins and therapeutics have gained attention due to their safety and efficiency. In this study, we designed a 21.4-kDa multi-epitope peptide vaccine (MEPV) using immunogenic segments from cytoadherence proteins. The MEPV’s effectiveness was verified through computational simulations. We then cloned the MEPV, introduced it into the plant expression vector pSiM24-eGFP, and expressed it in Nicotiana benthamiana leaves. The plant-produced MEPV proved to be immunogenic when administered intramuscularly to chickens. It significantly boosted the production of immunoglobulin Y (IgY)-neutralizing antibodies against cytoadherence protein epitopes in immunized chickens compared to that in the control group. This preliminary investigation demonstrates that the plant-derived MEPV is effective in triggering an immune response in chickens. To establish an efficient poultry health management system and ensure the sustainability of the poultry industry, further research is needed to develop avian vaccines using plant biotechnology.
This Reflection article begins with my family background and traces my career through elementary and high school, followed by time at the University of Illinois, Vanderbilt University, the University of Michigan, and then for 98 semesters as a Vanderbilt University faculty member. My research career has dealt with aspects of cytochrome P450 enzymes, and the basic biochemistry has had applications in fields as diverse as drug metabolism, toxicology, medicinal chemistry, pharmacogenetics, biological engineering, and bioremediation. I am grateful for the opportunity to work with the Journal of Biological Chemistry not only as an author but also for 34 years as an Editorial Board Member, Associate Editor, Deputy Editor, and interim Editor-in-Chief. Thanks are extended to my family and my mentors, particularly Profs. Harry Broquist and Minor J. Coon, and the more than 170 people who have trained with me. I have never lost the enthusiasm for research that I learned in the summer of 1968 with Harry Broquist, and I have tried to instill this in the many trainees I have worked with. A sentence I use on closing slides is “It’s not just a laboratory—it’s a fraternity.”
Schistosomiasis is a tropical disease that widely neglected. Schistosoma mansoni reproduce asexually within the freshwater snail, Biomphlaria alexandrina. Sodium hypochlorite (NaOCl) is a widely used disinfectant, so its effect against gainst B. alexandrina snails was evaluated. The present results showed that NaOCl has a molluscicidal activity against adult B. alexandrina snails at LC50 1.25 ppm. Hemocytes displayed varied morphological forms after being exposed to the LC10 and LC25 concentrations of NaOCl in B. alexandrina snails, and the phagocytic index of B. alexandrina snail's hemocytes significantly increased. The phagocytic potency of exposed hemocytes to charcoal showed ruptured plasma membrane, engulfed particles, vacuolation in the cytoplasm and degeneration of nuclei. When B. alexandrina snails were treated with sublethal concentrations of NaOCl, transaminases (AST & ALT), alkaline and acid phosphatase activities were significantly increased. In contrast, the total protein, albumin concentrations, Testosterone (T) and 17β Estradiol (E) showed a significant decrease (p ≤ 0.05) as compared to the control groups. The molecular docking interaction showed high efficiency for the ligand, NaOCl against the receptor binding sites of the acid phosphatase, alanine aminotransferase, estrogen and testosterone. The present results showed that NaOCl could be used as an effective molluscicide against B. alexandrina snails but more attention should be paid to investigate the side effects on the non-target organisms living in the freshwater environment.
Numerous technologies for cleaning the environment without producing hazardous waste products have arisen in response to the rise in environmental pollution. Industrialization and the growing population have put society to a hard test. Continuous discharge of heavy metals (chromium, nickel, copper, mercury etc.) in water bodies is increasing by anthropogenic activities; these cause bio accumulation and shift towards a resistant microbial community. These hazardous metals enter the human body through water and food, creating a slew of health issues. Chromium (VI) is one such toxic pollutant and is more toxic than chromium (III). Several environment‐friendly technologies have been developed for decontaminating wastewater by using microorganisms. Microalgae, cyanobacteria, bacteria and fungi are used for heavy metals eradication from the water due to their metal tolerance, bioaccumulation and biotransformation capacity. In this study two cyanobacteria strains were compared for their physiological response and metal tolerance for Cr (VI) stress. Leptolyngbya lignicola and Phormidium tenue were grown in a medium containing various concentrations of chromium (1 ppm, 2 ppm, 3 ppm, 4 ppm, 5 ppm, and 10 ppm), and biochemical parameters (chlorophyll, carotenoids, protein, proline, amino acid, carbohydrate, reducing sugar and lipid content) were measured. The findings revealed that P. tenue was more sensitive to metal stress than L. lignicola . P. tenue showed a decreasing trend for all biochemical parameters except proline at higher ad lower doses of Cr (VI) stress. However L. lignicola showed an increase in biomass (6.09%) and other biochemical parameters over control at lower metal stress concentration (1 ppm), whereas, a gradual decrease in all parameters except proline was observed at higher concentrations. Further L. lignicola showing high tolerance index can be preferred for metal remediation from the water bodies of heavy metal contaminated area.
Land snails are the most harmful pests in agricultural fields. Eobania vermiculata is a widespread snail species that causes massive damage to all agricultural crops. Thus, the molluscicidal activity of calcium borate nanoparticles (CB-NPs) against Eobania vermiculata was evaluated and compared with metaldehyde (Gastrotox® E 5% G). The amorphous phase of CB-NPs was obtained after thermal treatment at a low temperature (500 °C) which conformed by X-ray diffraction (XRD) analysis. CB-NPs are composed of aggregated nano-sheets with an average thickness of 54 nm which enhanced their molluscicidal activity. These nano-sheets displayed meso-porous network architecture with pore diameters of 13.65 nm, and a 9.46 m2/g specific surface area. CB-NPs and metaldehyde (Gastrotox® E 5% G) exhibited molluscicidal effects on Eobania vermiculata snails with median lethal concentrations LC50 of 175.3 and 60.5 mg/l, respectively, after 72 h of exposure. The results also showed significant reductions of Eobania vermiculata snails hemocytes' mean total number, the levels of Testosterone (T) and Estrogen (E), alkaline phosphatase, acid phosphatase, albumin, and protein concentrations, succinate dehydrogenase, glucose, triglycerides and phospholipids levels, while significant increases in the phagocytic index and mortality index, both transaminases (ALT and AST) and glycogen phosphorylase concentration were observed after the exposure to LC50 of CB-NPs or metaldehyde (Gastrotox® E 5% G) compared to the control group. Therefore, CB-NPs could be used as an alternative molluscicide for controlling Eobania vermiculata, but further studies are needed to assess their effects on non-target organisms.
Since centuries, Cyperus rotundus L. has been used against gastric ailments in traditional Indian medicine, especially in Ayurveda and Siddha. Therefore, it is very obvious that this plant will have a greater potential to treat gastric ulcers. For this reason, in this study, we mainly focused on the ulcer-preventive role of C. rotundus in rats treated with non-steroidal anti-inflammatory drugs.
Seventy percent methanolic extract of the plant was prepared and fed to 36-h fasted rats. Ulcer was induced in these rats by single oral administration of aspirin (400 mg/kg) 1 h after the administration of the plant extract. After 4 h, the rats were sacrificed, ulcer index was calculated, and antioxidant activity of the extract in gastric mucosa was evaluated by determining the levels of superoxide dismutase, glutathione, glutathione peroxidase, and tissue lipid peroxidation.
Oral administration of different doses of C. rotundus rhizome methanolic extract (CME; 250 mg/kg and 500 mg/kg) significantly inhibited aspirin-induced gastric ulceration in animals in a dose-dependent manner (49.32% and 53.15%, respectively), which was also comparable with the standard gastric ulcer drug ranitidine. Administration of CME also significantly increased the activity of superoxide dismutase, cellular glutathione and glutathione peroxidase, and inhibited the lipid peroxidation in the gastric mucosa of ulcerated animals in a dose-dependent manner.
Our results showed that C. rotundus extract has the capacity to significantly inhibit aspirin-induced gastric ulcers through an antioxidant defense mechanism. This study warrants further examination of this plant for its gastroprotective activities.
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