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Protein estimation with Folin phenol reagent
... The phosphate buffer soluble protein was estimated in the dried powdered leaves by the procedure of Lowry et al. (1951) [37] . For this, 200 mg of sample was crushed in 5 ml cold potassium phosphate buffer solution (pH 7.5; 0.1 M) on ice-bath using pestle and mortar. ...
... The phosphate buffer soluble protein was estimated in the dried powdered leaves by the procedure of Lowry et al. (1951) [37] . For this, 200 mg of sample was crushed in 5 ml cold potassium phosphate buffer solution (pH 7.5; 0.1 M) on ice-bath using pestle and mortar. ...
... To determine insoluble protein or complex protein modified procedure of Lowry et al., (1951) was used [37] . For this, the pellet obtained earlier after centrifugation of the crude extract of leaves in the phosphate buffer during extraction of soluble protein was digested (keeping in boiling water bath for 1 hr with 2 ml of 1N NaOH). ...
Herbal phytoresource have been recognized as a potential source of traditional therapeutics and nutrients. The genus Ipomea with 500-600 species comprises the largest number of species within the convolvulaceae. Since ancient time, this genus has been in continuous use for different purposes, such as-nutritional, medicinal, ritual and agricultural purposes. These rich sources of ethno medicinal and nutritional information demand successful chemical studies for effective selection of plants. With the aim in view, a field survey was conducted on 192 respondents of the Mog, Reang and Uchai of South Tripura. Using semi structured interviews and discussions on medicinal uses of plants, the investigator gathered information from the respondents in addition to some traditional health prescriptioners; viz; Ujha, Baidya, Sadhan, Kabiraj. A rigorous qualitative and quantitative analysis of collected data on 73 plants with their uses in curing 69 ailments, three species of Ipomea such as-I.aquatica Forsk., I. batatus Linn. and I. carnea Linn. were found as potential source of trado-medicines. In the present study, the leaves of these plants were targeted to analyze by standard chemical techniques to evaluate their primary and secondary metabolites. The analysis of the results provided evidences that leaf extracts of these tested plants contain moderate amount of moisture (12.27-22.73%), solid content (77.27-87.72%), total ash (4.85-12.25g%),carbohydrate (7.57-17.71g%), crude proteins (25.35-27.25 g%), alkaloids (17.66-19.26 mg g-1), phenolics (15.33-20.25 mg g-1), flavonoids (11.41-12.71 mg g-1), carotenoids (0.55-0.64 mg g-1), ascorbic acid (32-146 mg/100g) that justifies their use in the traditional medicines for the treatment of various common diseases. The outcome of the study also suggest that these three tested species of genus Ipomea have very good medicinal potentials, meet the standard requirements for drug formulation and serve as good sources of nutrients predominant in the leaf.
... The su per natant was sep a rated and used as leaf ex tract for the es ti ma tion of pro tein ( Mathur et al., 2015 ). Pro tein con tent pre sent in the sam ple has been es ti mated by the stan dard pro to col of Lowry et al. (1951) . Bovine Serum Al bu min (BSA) was used as a stan dard. ...
In the current study, Seaweed Liquid Fertilizer (SLF) prepared from the combination of two marine algae namely Turbinaria ornata and Ulva reticulata, was investigated on Raphanus sativus (Radish), Phaseolus vulgaris (Green Pea) and Vigna radiata (Mung). The potentiality was evaluated on the basis of seed germination, plant growth and various biochemical criterions in comparison with chemical fertilizer and plants without any supplement. The seeds of V. radiata, P. vulgaris, and R. sativus were supplemented with 5 different SLF concentrations (20, 40, 60, 80 and 100%) and allowed for germination. Seeds of all the plants showed 100% germination with both 80% and 100% SLF concentrations. It has been observed that 100% SLF treated plants was very effective in enhancing growth rate followed by 80% SLF. The root and shoot length, protein, amino acid, amino acid, carbohydrate and phenol concentration of the plants were found to be maximal at 100% SLF. Among the three plants, P. vulgaris has shown remarkable growth and yield followed by R. sativus and V. radiata. Additionally, the SLF also displayed antagonistic activity against plant phytopathogens such as Rhizoctonia solani, Macrophomina phaseolina, Sclerotium rolfsii and Alternaria solani. The GC-MS analysis of SLF indicated the presence of several bioactive compounds such as cyclopentasiloxane tetradeca-methyl, 3-acetonylcyclopentanone, etc. which are known for its role in plant growth and antifungal activity. The results suggest that the T. ornata and U. reticulata derived SLF could be an effective alternative to chemical fertilizers and warrant further research to investigate its possible use on field.
... Reaction was stopped by addition of 1.5 ml of 1 M trichloroacetic acid. After 15 min, the mixture was centrifuged at 10,000 rpm for 10 min and the protein concentration in supernatant was determined according to the method of Lowry et al. (1951). One unit (U) of protease activity is equivalent to μg of tyrosine liberated per ml of enzyme under prescribed conditions. ...
The diversity of actinobacteria associated with marine ascidian Phallusia nigra from Andaman Islands was investigated. A total of 10 actinobacteria were isolated and based on the biochemical and molecular characterization, the isolates were assigned to 7 different actinobacterial genera. Eight putatively novel species belonging to genera Rhodococcus, Kineococcus, Kocuria, Janibacter, Salinispora and Arthrobacter were identified based on 16S rDNA sequence similarity with the NCBI database. The organic extracts of ten isolates displayed considerable bioactivity against test pathogens, which were Gram-positive and Gram-negative in nature. PCR-based screening for type I and type II polyketide synthases (PKS-I, PKS-II) and nonribosomal peptide synthetases (NRPS) revealed that, 10 actinobacterial isolates encoded at least one type of polyketide synthases biosynthesis gene. Majority of the isolates found to produce industrially important enzymes; amylase, protease, gelatinase, lipase, DNase, cellulase, urease, phosphatase and l-asparaginase. The present study emphasized that, ascidians are a prolific resource for novel bioactive actinobacteria with potential for novel drug discovery. This result expands the scope to functionally characterize the novel ascidian associated marine actinobacteria and their metabolites could be a source for the novel molecules of commercial interest.
... The values are expressed as μmol of CDNB-GSH conjugate formed per min per mg of protein. Protein contents were measured according to the method ofLowry et al. (1951) using BSA. All spectrophotometric measurements were recorded using a microplate reader (BMG LABTECH GmbH SPECTROStarNano, Ortenberg, Germany). ...
The present study investigated the effect of culture extracts (CB08035-SCA and CB08035-SYP) from Marinobacter hydrocarbonoclasticus (strain CB08035) on cell viability and the potential protective effects attributed to molecular mechanisms underlying antioxidant response to survive oxidative stress injuries. Caco-2 cells were submitted to oxidative stress by treatment with tert-butylhydroperoxide (t-BOOH). Both extracts prevented cell damage and enhanced activity of antioxidant defenses (NQO1 and GST activities and GSH levels) reduced by treatment with t-BOOH. Increased ROS and caspase 3/7 activity induced by t-BOOH were dose-dependently prevented when cells were treated with the extracts. CB08035-SCA caused up-regulation of Nrf2, AKT1 and Bcl-2 gene expressions. Moreover, CB08035-SCA and CB08035-SYP treatments reduced significantly Bax, BNIP3, APAF1, ERK1, JNK1, MAPK1, NFκB1, TNFα, IL-6, IL-1β and HO-1 gene expressions of apoptosis, proinflammation and oxidative stress induced by t-BOOH. CB08035-SCA and CB08035-SYP CPE extracts confer a significant protection against oxidative insults to cells. Our results show that culture extracts CB08035-SCA and CB08035-SYP from M. hydrocarbonoclasticus (strain CB08035) appeared to have antioxidant potential, based on their ability to protect antioxidant enzymes and mRNA gene expressions linked to apoptosis/oxidative pathways. These results suggest that culture extracts CB08035-SCA and CB08035-SYP can be a potential ingredient in the pharmaceutical and cosmeceutical industries.
... Protein estimation was carried out by method as described by Lowry et al. [30]. ...
Three-step purification technique (isopropanol precipitation, ion-exchange and size-exclusion chromatography) was used for the purification of an endoinulinase from the culture broth of Aspergillus tritici BGPUP6. The molecular mass of purified endoinulinase was found to be 53.45 kDa and 53.70 kDa by denatured protein gel (SDS-PAGE) and size-exclusion (Sephadex G-100) chromatographic analysis, respectively. Higher Km (1.02 mM), Vmax (19.60 mM/min·mg), Kcat (1.3 × 10-3/min) and Vmax/Km ratio (19.21/min·mg) of purified endoinulinase for inulin than stachyose depicts its higher affinity towards inulin. Purified enzyme was found stable in the pH range 4.0-7.0 with an optimal pH 5.5. The optimal temperature of purified biocatalyst was 55 °C with thermostability in the range of 50-70 °C. D-value and Z-value for endoinulinase at 55 °C was found to be 100.08 h and 11.62 °C, respectively. Thermodynamics inactivation parameters (ΔG, ΔH and ΔS) of endoinulinase shows its wide range thermal stability. Endoinulinase activity was enhanced by CaCl2 and MnSO4, while CuSO4, CoCl2, AgNO3, CdCl2, NiCl2, ZnSO4, BaCl2, HgCl2 and EDTA inhibited the activity of enzyme. Purified endoinulinase was successfully used for the production of fructooligosaccharides from inulin.
... The total phenolic content was found out by Lowry et al. [15] method where the phenolic compounds present in the extract reduce Folin-Ciocalteu reagent, forms a blue colored complex and can be quantified using visible light spectrophotometer at 760 nm. The concentration of phenolic compound in the extract was determined from the standard curve prepared from varying concentrations (20-100 μg/ml, R 2 =0.989aqueous extract and R 2 =0.989-80% methanol extract) of Gallic acid analyzed in the same manner as the extract samples. ...
... The PMS was used for the estimation of lipid peroxidation, glutathione (GSH), glutathione S-transferase (GST), glutathione reductase (GR), glutathione peroxidases (GPx), superoxide dismutase (SOD) and catalase (CAT). Total protein estimation in the samples was carried out by the method of Lowry et al. (1951). ...
The objective of the present study was to investigate the hepatoprotective effects of methanol extract of Meconopsis aculeata, an important medicinal plant of Kashmir Himalayas, against CCl 4 induced oxidative damage in rats. Acute hepatotoxicity was induced in Wistar albino rats by single intraperitoneal injection of CCl 4 at 1 ml/kg body weight dose. Methanol extract of M. aculeata was orally administered at the doses of 100, 200 and 300 mg/kg body weight/day for 14 days. The results revealed that administration of CCl 4 caused a significant increase in serum AST, ALT and LDH levels as compared to the control group (p < 0.001). Additionally, there was a significant decrease in the level of hepatic GSH, GPx, GST, SOD and CAT activities associated with a significant increase of MDA content in CCl 4 treated group compared to those of the control group. However, the treatment with methanol extract of M. aculeata prevented these alterations and maintained the antioxi-dant status. The hepatoprotective property of the extract was further confirmed by histopathological analysis, wherein the extract was shown to prevent neutrophil infiltration and tissue necrosis in the liver samples of treated animals. The results of the present investigation indicate that methanol extract of M. aculeata possesses potent hepatoprotective activity, possibly due to its antioxidant properties.
... Protein estimation has been carried out according to Lowry et al. [16]. The standard curve was established by using different concentrations of bovine serium albumin (BSA). ...
... Protein quantification of Penicillium sp. lectins was performed by the Lowry method [42]. Purified lectins were then evaluated for their antioxidant potential and antimicrobial activity. ...
Lectins are a diverse group of proteins of non-immune origin that interact specifically with glycans. Owing to their specificity, they can mediate various cellular and molecular recognition processes. To explore information on biological activities of lectins from Penicillium duclauxii, P. proteolyticum and P. griseoroseum, they were investigated for their antioxidant and antimicrobial activities. Penicillium sp. lectins exhibited moderate antioxidant activity. P. duclauxii, P. proteolyticum and P. griseoroseum lectins inhibited DPPH with an IC50 value of 71.42, 75.04 and 82.11 ?g/mL, respectively. P. duclauxii, P. proteolyticum and P. griseoroseum lectins inhibited the hydrogen peroxide radical with IC50 values of 198.57, 209.76 and 215.31 ?g/mL, respectively. P. duclauxii and P. proteolyticum lectins exhibited potent antibacterial activity against Gram-negative bacteria. P. griseoroseum lectin inhibited only Gram-positive bacteria. Penicillium sp. lectins did not exhibit antifungal activity. The biological potential of Penicillium sp. lectins will help to understand their biomedical applications. This is the first report on the antioxidant and antimicrobial activities of purified lectins from Penicillium sp.
... Estimation of total protein and total starch content -Total protein content was measured as described by Lowry et al. (1951). For analysis, 0.5 g fresh plant material was homogenized in 3 ml of Potassium Phosphate Buffer (pH 7.0) and centrifuged at 15,000×g for 15 min. ...
Jasmonates (JAs) are a rising class of lipid-derived signalling molecules with varied functions ranging from induction of abiotic and biotic stress-responsive genes to regulation of plant growth and development under natural condition. The rationale of present study was to investigate the nutritional importance of JAs in inducing and triggering accumulation of carbohydrates, sugars, nutritional pigments and vitamins in three varieties of Brassica oleracea L. (variety botrytis, capitata, italica) edible heads. Exogenous application of Jasmonic acid (JA) and methyl ester of Jasmonic acid (Me–JA), made edible heads of var. botrytis, capitata and italica nutritionally more accrete as compared to control untreated edible heads. Results showed that the treatment of JA and Me-JA enhanced growth of edible heads in terms of head diameter, head length, fresh weight, dry weight, moisture content, total protein, total starch, total carbohydrates, total soluble sugar, non-reducing sugar, vitamins C, A, E and B2 in var. botrytis but at the same time dramatic decline in reducing sugar, beta-carotene and lycopene was seen. In var. capitata certain reduction in vitamin E accumulation was recorded by the exogenous application of JA and Me–JA while overall growth of edible heads was accelerated which could also be supported by enhanced nutritional and productivity level. In variety italica, JAs reduced total starch and vitamin C, but overall growth was surged along with productivity. Both JA and Me-JA could be attributed to induce growth and enhance the nutritional potential of edible heads in three varieties of Brassica oleracea as compared to their untreated control edible heads. From the present results, it has been suggested that these two eco-friendly oxylipins can be further explored for enhancing the growth and nutritional value of these vegetables by exogenous application of JAs in μM to pM concentration.
... Quantification of protein in the sample tissues was carried out using Lowry's method (Lowry et al., 1951). Tissue homogenate (0.1 ml) was taken in a test tube and precipitated using 1 ml of 10% TCA followed by centrifugation at 5,000 rpm for 20 min to collect the protein, which was further dissolved in 0.5 ml of 0.1 N Na0H. ...
A feeding trial was conducted with inclusion levels of Cajanus cajan leafmeal (CCLM)
in the diet of Labeo rohita to study the growth, haematological indices, digestive
enzyme, physio-metabolic changes and molecular expression of insulin-like growth
factor-1 (IGF-1), insulin-like growth factor-binding protein-1 (IGFBP-1), insulin-like
growth factor-1 receptor (IGF-1R) genes. Four practical diets with CCLM, control,
T20 (20%), T30 (30%) and T40 (40%) were prepared. Weight gain%, SGR, FER and
PER were significantly (p < 0.05) higher when 30% de-oiled rice bran (DORB) was replaced
by CCLM whereas the treatment group 40% registered with lower FCR value.
The protease and amylase activities were higher in the T30 group. T20 showed significantly
higher (p < 0.05) hepatic and muscular lactate dehydrogenase and serum glucose
was also higher, whereas, WBC content was found significantly (p < 0.05) higher
in control. The hepatic IGF-1 gene expression was significantly higher (p < 0.05) in
T40 followed by the T30 group but the weight gain was not significantly (p > 0.05)
higher in T30 from the T40 group. Results reveal that the response of IGF-1, IGF-1R
and IGFBP-1 genes was complementary to the biological growth parameters and
hence were used as an indicator to evaluating growth. The study also demonstrates
that CCLM can replace 100% DORB in the diet of L. rohita without any antinutritional
or adverse growth effects.
... Initial (0 h) and final (96 h) algae samples (15 mL) were collected for the estimation of protein content and pigment content. Protein content of experimental S. platensis samples was estimated as per Lowry et al. (1951). Sample preparation and estimation of total chlorophyll and carotenoids content of S. platensis were determined as per Gita et al. (2019), Moran (1982) and Chamovitz et al. (1993) respectively. ...
A concentration-dependent decrease in growth rate and pigment concentration of the blue-green alga Spirulina platensis was recorded after the exposure to graded (5–40 ppm) concentration of six textile dyes. The profile of vital elements (C, H, N, S) also showed a significant variation due to dye toxicity. The algal population showed up to 50% decrease in protein content after exposure to the dyes. Among the pigments, the dye exposure resulted in > 90% decreases in phycocyanin however, total chlorophyll and carotenoids exhibited up to a 50% decrease compared to control. The findings indicate that the unregulated discharge of textile dyes will directly impact the photoautotrophic organisms leading to ecological imbalance in aquatic ecosystems. Overall observations of the report provide baseline information about the toxicity of textile dyes and giving a better insight into the little-understood mechanisms of dye toxicity.
... The specific activity of APX was expressed as μmol (ascorbate oxidized) min -1 mg -1 (protein). Protein in each enzyme preparation was measured by Lowry's method using bovine serum albumin (BSA) as the standard (Lowry et al. 1951). ...
High-temperature stress severely impacts both yield and quality of tomato fruits, and therefore, it is required to develop stress-tolerant cultivars. In the present study, two tomato genotypes, H88-78-1 and CLN-1621, identified through preliminary phenotypic screening were characterized by analysis of molecular, physiological, and biochemical traits in comparison with a susceptible genotype Punjab Chhuhara. Phenotypic stress tolerance of both the genotypes was validated at biochemical level as they showed higher amount of relative water content, photosynthetic pigments, free cellular proline, and antioxidant molecules while less amount of H2O2 and electrolyte leakage. Expression analysis of 67 genes including heat shock factors, heat shock proteins, and other stress-responsive genes showed significant up-regulation of many of the genes such as 17.4 kDa class III heat shock protein, HSF A-4a, HSF30, HSF B-2a, HSF24, HSF B-3 like, 18.1 kDa class I HSP like, and HSP17.4 in H88-78-1 and CLN-1621 after exposure to high-temperature stress. These candidate genes can be transferred to cultivated varieties by developing gene-based markers and marker-assisted breeding. This confirms the rapid response of these genotypes to high-temperature stress. All these traits are characteristics of a stress-tolerance and establish them as candidate high-temperature stress-tolerant genotypes that can be effectively utilized in stress tolerance improvement programs.
Supplementary information:
The online version contains supplementary material available at 10.1007/s13205-020-02587-6.
... The amount of protein was quantified by the method of Lowry et al. (1951). ...
Present investigation describes immobilization efficiency of endoinulinase onto hetero-functionalized halloysite nanoclay using 3-aminopropyltriethoxysilane and glutaraldehyde as crosslinkers. Under optimal conditions (APTES 0.75%, sonication time 2.25 h, glutaraldehyde 0.75%, activation-time 65 min, immobilized endoinulinase load 60 IU and coupling-time 1 h), maximum yield in enzyme activity (70.65%) and immobilization (89.61%) was obtained. Developed immobilized biocatalyst shown maximum activity at 65 ◦C and pH 5.0 with wide range thermal (50–80 ◦C) and pH (4.0–9.0) stability. Increase in half-life (28.70-fold) of immobilized endoinulinase was observed as compared to free enzyme. An enhanced Km and reduced Vmax of endoinulinase for inulin was recorded after immobilization. Maximum FOSs production 98.42% was obtained, under optimized conditions (inulin 10%; immobilized endoinulinase load 85 IU; hydrolysis-time 10 h and agitation rate 130 rpm) containing kestose (36.26%), nystose (27.02%), fructofuranosylnystose (9.98%) and FOSs DP 5–9 (25.15%). Developed immobilized biocatalyst exhibited a splendid operational stability for 18 batch cycles.
... Folin and Ciocalteu's phenol reagent was used to determine concentration of protein in samples with bovine serum albumin as the standard (Lowry 1951). ...
Copper (Cu) is an extensively used heavy metal and an indispensible micronutrient for living beings. However, Cu is also toxic and exerts multiple adverse health effects when humans are exposed to high levels of this metal. We have examined the effect of single acute oral dose of copper chloride (CuCl2) on parameters of oxidative stress, cellular metabolism, membrane and DNA damage in rat intestine. Adult male Wistar rats were divided into four groups and separately administered a single oral dose of 5, 15, 30 and 40 mg CuCl2/kg body weight. Rats not administered CuCl2 served as the control. Oral administration of CuCl2 led to significant alterations in the activities of metabolic and membrane-bound enzymes; brush border enzymes were inhibited by 45–75% relative to the control set. Inhibition of antioxidant enzymes diminished the metal-reducing and free radical quenching ability of the cells. Oxidative damage caused cellular oxidation of thiols, proteins and lipids. Diphenylamine and comet assays showed that CuCl2 treatment enhanced DNA damage while DNA-protein crosslinking was also increased in the intestinal cells. Examination of stained sections showed that CuCl2 treatment led to marked histological changes in the intestine. All the changes seen were in a CuCl2 dose-dependent manner with more prominent alterations at higher doses of CuCl2. These results clearly show that oral administration of CuCl2 results in oxidative damage to the intestine which can impair its digestive and absorptive functions.
... Protein estimation was doneusing bovine serum albumin (BSA) as standard [15] . Optical density was measured at 670 nm. ...
... The fishes of both groups were sacrificed and the desired tissues were dissected out, homogenized and mixed with 5ml of deionized water then centrifuged at 3000 rpm for 10 minutes to obtain supernatant. The supernatants were filtered and the filtrates were used for analysis of the glycogen, protein and free amino acids content by standard methods of Carroll et al. (1956), Lowry et al. (1951) and Rosen (1957) respectively. ...
... The OMVs were purified using the method described by MacDonald and Kuehn (2013). The protein concentration of OMVs was estimated using modified Lowry's method (Lowry et al., 1951). The yield of OMV was expressed in terms of protein concentration. ...
Background: The non-typhoidal Salmonella causes gastroenteritis in humans that makes its way to the food chain mainly through the animal products. The multiple drug resistance imposes one of the major hurdle in the treatment of the disease. The vaccination appears to be the most important method for prevention of the disease. Unfortunately, there is no liscenced vaccine available against non-typhoidal Salmonellae. The use of outer membrane vesicles (OMVs) of Salmonella as a vaccine candidate has attained significant centre-stage in the recent years given to its protective immunogenicity. However, the large scale production of OMVs is difficult owing to low yield per liter of culture. Methods: In the present study, we have optimized the culture conditions viz. pH, phase of growth and presence of oxidative stress for maximum production of OMVs from Salmonella Typhimurium. The OMVs were characterized based on yield based on protein concentration, lipopolysaccharide concentration and zeta size. Result: In the present study, it was found that incubation of Salmonella Typhimurium up to peak of the growth phase at pH 7 in presence of oxidative stress was found to be the most suitable condition for maximum production of OMVs.
... The whole OMPs were extracted from all the three strains as per the method described by Choi-Kim et al. (1991). The protein was quantified by the method described by Lowry et al., (1951). The presence of proteins were documented by one dimensional SDS-PAGE, using 5% stacking gel and 12% separating gel (Sambrook and Russel, 2001). ...
Background: Swine pasteurellosis, caused by Pasteurella multocida capsular types A and D, causes heavy economic loss to the pig farmers. The vaccine presently used is a bacterin of Pasteurella multocida capsular type B that is proven to be effective against bovine pasteurellosis. However, its efficacy against swine pasteurellosis is questionable. Methods: The present study was carried out to evaluate the efficacy of calcium phosphate nanoparticle adjuvanted bivalent subunit vaccine prepared from Pasteurella multocida capsular types A and D along with a monovalent subunit vaccine prepared from Pasteurella multocida capsular type B in mice. The Alum precipitated bacterin vaccine was used as the control. Result: The bivalent subunit vaccine showed significantly higher serum IgG response than either of the other two vaccines. The calcium phosphate nanoparticle adjuvanted vaccines could elicit 100% protection in mice against homologous challenges but the aluminum hydroxide adjuvanted bacterin vaccine could not elicit significant protection. Based on this preliminary work, it was concluded that the bivalent subunit vaccine would be a better option for immunization of swine against swine pasteurellosis.
... Protein concentration was measured according to the study of Lowry et al. (1951) with bovine serum albumin as standard. ...
In the present study, the amylase enzyme producing potential of four different Aspergillus species was analyzed. The extracted amylase enzyme was purified by diethyl amino ethyl (DEAE) cellulose and Sephadex G-50 column chromatography and the enzyme activity was measured by using synthetic substrate starch. The partially purified enzyme exhibits maximum activity at the optimum pH (7.0), temperature (60 to 70°C) and substrate concentration (1.5 to 2.0%) under standard assay conditions. Among the four different Aspergillus species examined, Aspergillus flavipes showed maximum production of amylase. The characteristics of the partially purified enzyme such as optimum pH and temperature were also favourable for industrial applications.
... Protein quantification was carried out by Lowry et al. (Lowry et al., 1951) protocol. ...
Current work describes the enhancement of immobilization efficacy of Aspergillus tritici endoinulinase onto halloysite nanoclay using crosslinker glutaraldehyde. Under statistical optimized immobilization conditions, viz. glutaraldehyde 1.50% (v/v), enzyme coupling-time 2.20 h, glutaraldehyde activation-time 1.00 h and endoinulinase load 50 IU, maximum activity yield (65.77%) and immobilization yield (82.45%) was obtained. An enhancement of 1.15- and 1.23-fold in both enzyme activity yield and immobilization yield of endoinulinase was observed, when compared with APTES-functionalized halloysite nanoclay immobilized endoinulinase. Immobilized biocatalyst showed maximum activity at pH 5.0 and temperature 60 °C with broad pH (4.0–8.5) and temperature (50–75 °C) stability. Further, optimal hydrolytic conditions (inulin concentration 8.0%; endoinulinase load 80 IU; agitation 125 rpm and hydrolysis-time 13 h) supported fructooligosaccharides yield (95.44%) in a batch system. HPTLC studies blueprint confirmed 95.44% fructooligosaccharides containing 35.41% kestose, 26.19% nystose and 9.69% fructofuranosylnystose. The developed immobilized biocatalyst shown good stability of 8 cycles for inulin hydrolysis.
... The total body protein of D. busckii flies was estimated through Lowry Method (Lowry 1951). Three replicates of 10 flies each (control vs acclimated) were homogenized in a 1 ml lysis buffer, and the homogenate was subjected to sonication for 5 s. ...
The wild populations of Drosophila show highly adaptive body metabolism in response to changing environments. The
traits associated with starvation and desiccation stress vary with the geographical locations as well as climatic conditions.
The energy storage and utilization during these stresses varies among diferent species. We examined the efects of physiological stress i.e., desiccation and starvation stress in wild populations of Drosophila busckii collected from the Lalpur
region including Indira Gandhi National Tribal University (IGNTU) campus (22.67°N 81.75°E), Amarkantak. We found
a diference in the level of metabolites between control vs desiccation hardened and starvation acclimated fies along with
sexual dimorphism. This study focused on the abilities of wild D. busckii to tolerate physiological stress (desiccation and
starvation) by altering the level of energy metabolites. In the present work, the levels of lipid and protein metabolites showed
an interesting diference between control vs. hardened fies. Hardening or pre-treatment enhanced the survival against non-lethal stress, while climbing activity, cuticular lipids and total body lipids decreased with the increase in desiccation hardening and starvation acclimation period. We found a signifcant increase in the level of total body proteins in 3 h desiccation
acclimated, and 16 h starvation acclimated fies as compared to control. The female Drosophila busckii were more resistant
to sub-lethal stress. The compensatory changes in the levels of total body lipids, cuticular lipids and proteins suggest possible
energetic homeostasis in D. busckii.
... malondialdehyde (MDA) measurement was carried in compliance with Ohkawa et al. [26] the organic layer was separated and its absorbance was measured at 532 nm by micro plate spectrophotometry. Protein estimation was conducted according to Lowry et al. [27]. The data were expressed as nmol/mg protein. ...
Background: Tuberoinfundibular Peptide of 39 (TIP39) is a neuroendocrine hormone, potentially acting through parathyroid hormone receptor 2 receptor (PTH2R) abundantly expressed in brain. Objective: This study aimed to evaluate the neuroendocrine role of TIP39 in chronic unpredictable mild stress (CUMS) induced depression and to elucidate its underlying mechanism. Method: The depression was induced in rats by CUMS for a period of four weeks. TIP39 was administered through intracerebroventricular (ICV) route at doses (1 & 10 nmol/rat) for four weeks on alternate days, parallel with the daily exposure of stress. At the end of the treatment period, animals were evaluated for sucrose preference, behavioral, biochemical and oxidative changes. Further the molecular mechanism of anti-stress activity of TIP39 confirmed through gene expression study. Results: TIP39 administration significantly reversed the CUMS induced increased immobility time in de-pressive rats and increased plasma corticosterone as well as decreased open-field activity and sucrose consumption. CUMS lowers the activity of superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) and elevated the production of malondialdehyde (MDA) in hippocampus and prefrontal cortex, which was reversed by the administration of TIP39. Moreover, TIP39 could effectively reverse alteration in interleukin 6 (IL-6), interleukin-1 (IL-1) and tumor necrosis factor-(TNF-) in brain tissue. Conclusion: Chronic ICV administration of TIP39 alleviated the behavioral deficits of chronic unpredictable mild stress, consanguinity to its concurrent modulatory repercussion on hypothalamic pituitary adrenal axis, inflammation, and oxidative courses.
... The values were expressed as μmol of CDNB-GSH conjugate formed per min per mg of protein. Protein contents were measured according to the method of Lowry et al. (1951) using BSA. All spectrophotometric measurements were recorded using a microplate reader (BMG LABTECH GmbH SPECTROStarNano, Ortenberg, Germany). ...
Gongolaria baccata (S.G. Gmelin) is marine brown seaweed mainly found on the coasts of the Baltic Sea south to the Mediterranean Sea, Canary Islands, Mauritania and Western Sahara. Herein, we report the cell viability and protective effects attributed to molecular mechanisms underlying antioxidant response to survive oxidative stress injuries. Caco-2 cells were submitted to oxidative stress by treatment with tert-butylhydroperoxide (tert-BOOH). The extract prevented cell damage and enhanced activity of antioxidant defenses (NQO1 and GST activities and GSH levels) reduced by treatment with tert-BOOH. The increases of MDA levels, the amount of intracellular ROS and caspase 3/7 activity induced by tert-BOOH were prevented when cells were treated with the G. baccata extract. Moreover, G. baccata extract caused up-regulation of GSTM2, Nrf2, and AKT1 gene expressions, as well as G. baccata extract reduced significantly Bax, BNIP3, APAF1, ERK1, JNK1, MAPK1, P38, P53, NFκB1, TNFα, IL-6, IL-1β and HO-1 gene expressions related to apoptosis, proinflammation and oxidative stress induced by tert-BOOH. These results suggest that G.baccata extract protected the cells against oxidative damage and inflammation; protective effects that could be linked to their bioactive constituents. Hence, this brown seaweed G.baccata extract could be used for the development of functional foods and/or nutraceuticals.
... Cell extracts were prepared by sonication and centrifugation as described previously [105]. Bovine serum albumin (Sigma Aldrich) was used as a reference for analyses of protein concentrations in cell extracts [106]. ...
Background
In most fungi, quinone-dependent Class-II dihydroorotate dehydrogenases (DHODs) are essential for pyrimidine biosynthesis. Coupling of these Class-II DHODHs to mitochondrial respiration makes their in vivo activity dependent on oxygen availability. Saccharomyces cerevisiae and closely related yeast species harbor a cytosolic Class-I DHOD (Ura1) that uses fumarate as electron acceptor and thereby enables anaerobic pyrimidine synthesis. Here, we investigate DHODs from three fungi (the Neocallimastigomycete Anaeromyces robustus and the yeasts Schizosaccharomyces japonicus and Dekkera bruxellensis ) that can grow anaerobically but, based on genome analysis, only harbor a Class-II DHOD.
Results
Heterologous expression of putative Class-II DHOD-encoding genes from fungi capable of anaerobic, pyrimidine-prototrophic growth ( Arura9, SjURA9, DbURA9 ) in an S. cerevisiae ura1Δ strain supported aerobic as well as anaerobic pyrimidine prototrophy. A strain expressing DbURA9 showed delayed anaerobic growth without pyrimidine supplementation. Adapted faster growing DbURA9 -expressing strains showed mutations in FUM1 , which encodes fumarase. GFP-tagged SjUra9 and DbUra9 were localized to S. cerevisiae mitochondria, while ArUra9, whose sequence lacked a mitochondrial targeting sequence, was localized to the yeast cytosol. Experiments with cell extracts showed that ArUra9 used free FAD and FMN as electron acceptors. Expression of SjURA9 in S. cerevisiae reproducibly led to loss of respiratory competence and mitochondrial DNA. A cysteine residue (C265 in SjUra9) in the active sites of all three anaerobically active Ura9 orthologs was shown to be essential for anaerobic activity of SjUra9 but not of ArUra9.
Conclusions
Activity of fungal Class-II DHODs was long thought to be dependent on an active respiratory chain, which in most fungi requires the presence of oxygen. By heterologous expression experiments in S. cerevisiae , this study shows that phylogenetically distant fungi independently evolved Class-II dihydroorotate dehydrogenases that enable anaerobic pyrimidine biosynthesis. Further structure–function studies are required to understand the mechanistic basis for the anaerobic activity of Class-II DHODs and an observed loss of respiratory competence in S. cerevisiae strains expressing an anaerobically active DHOD from Sch. japonicus .
... Roots of Z. mays were homogenized in phosphate buffer (PO 4 3-, 0.1 M, pH = 7.0), and the homogenate was sieved and centrifuged (15,000×g; 4°C; 30 min). The protein content in the supernatant was determined as per Lowry et al. (1951) and supernatant was stored at 4°C prior to enzymatic assays. Superoxide dismutase (SOD) was quantified at λ=560 nm based on photochemical inhibition of nitroblue tetrazolium chloride (Beauchamp and Fridovich 1971). ...
Heavy metals’ amassment in the soil environment is a threat to crop and agricultural sustainability and consequentially the global food security. For achieving enhancement of crop productivity in parallel to reducing chromium (Cr) load onto food chain demands continuous investigation and efforts to develop cost-effective strategies for maximizing crop yield and quality. In this context, we investigated the amelioration of Cr(VI) toxicity through β-pinene in experimental dome simulating natural field conditions. The protective role of β-pinene was determined on physiology, morphology and ultrastructure in Zea mays under Cr(VI) stress (250 and 500 μM). Results exhibited a marked reduction in the overall growth (shoot and root length and dry matter) of Z. mays plants subjected to Cr(VI) stress. Photosynthetic pigments (chlorophyll and carotenoids) were evidently reduced, and there was a loss of membrane integrity. Supplementation of β-pinene (100 μM), however, declined the toxicity induced by Cr(VI). Interestingly, Cr-tolerant abilities were improved in relation to plant growth, photosynthetic pigments and membrane integrity with the combined treatment of Cr(VI) and β-pinene. β-Pinene also reduced the root-mediated uptake of Cr(VI) and translocation to shoots. Moreover, significant ultrastructural damages recorded in roots and shoots under Cr(VI) stress were partially reverted upon addition of β-pinene. Our analyses revealed that β-pinene mitigates Cr(VI) toxicity in Z. mays, either by membrane stabilization or serving as a barrier to the uptake of Cr from soil. Thus, exogenous supply of β-pinene can be an effective alternative to mitigate Cr toxicity in soil. However, it is deemed essential to investigate further the responses throughout the life cycle of the plant on β-pinene supplementation under natural conditions.
... gill, liver, gonads, brain, kidney, intestine and muscles were processed for the biochemical estimations. Protein content was estimated by Follin phenol reagent method (Lowry et al., 1951). ...
Puntius ticto, a freshwater fish exposed to lethal (5.012 ppm) and two sublethal concentrations of dimethoate (2.506 ppm and 1.253 ppm) for 96h and 60 days and protein content was observed from different tissues after the exposure period. Acute exposure (5.012 ppm) results in significant decrease in the level of protein in testis, ovary and brain and slight decrease in intestine, muscles, liver and gills; whereas increased protein level was observed in kidney. Chronic toxicity results showed decrease in
the level of protein content in ovary, brain, intestine, muscles, gills and liver to 2.506 and 1.253 ppm exposure; whereas in testis protein level was increased to 1.253 ppm and decrease protein content was observed in 2.506 ppm exposure
... gill, liver, gonads, brain, kidney, intestine and muscles were processed for the biochemical estimations. Protein content was estimated by Follin phenol reagent method (Lowry et al., 1951). ...
Puntius ticto, a freshwater fish exposed to lethal (5.012 ppm) and two sublethal concentrations of dimethoate (2.506 ppm and 1.253 ppm) for 96h and 60 days and protein content was observed from different tissues after the exposure period. Acute exposure (5.012 ppm) results in significant decrease in the level of protein in testis, ovary and brain and slight decrease in intestine, muscles, liver and gills; whereas increased protein level was observed in kidney. Chronic toxicity results showed decrease in
the level of protein content in ovary, brain, intestine, muscles, gills and liver to 2.506 and 1.253 ppm exposure; whereas in testis protein level was increased to 1.253 ppm and decrease protein content was observed in 2.506 ppm exposure.
Desugarization is an important technique in industrial egg processing to maintain the color and shelf life of egg white powder. Its effect on proteolysis of egg white protein is essential in understanding the functionality changes in the obtained hydrolysate. Two desugarization methods, including yeast fermentation and enzyme desugarization using glucose oxidase are commonly employed in industrial scale egg white processing but the suitable method which can improve egg white protein hydrolysates' functionality has not yet been reported. Thus, the present work compares the physicochemical, functional, and surface properties of egg white protein hydrolysates obtained through both the desugarization methods. Alcalase enzyme (2% w/w) was used for enzyme hydrolysis. The degree of hydrolysis of desugared egg white hydrolysate was higher than the non-desugared samples by 34.5%. The desugared samples (especially yeast fermented) had higher foaming and emulsifying ability than non-desugared samples; however, it showed emulsion activity index and surface hydrophobicity. Compared to native protein, the in-vitro trypsin digestibility of desugared hydrolysate samples was significantly (p<0.05) higher. The antioxidant properties increased with hydrolysis, with yeast desugared hydrolysate samples having the highest antioxidant activity (63.8%).
We have examined the regulation of repressible acid phosphatase (APase; orthophosphoric-monoester phosphohydrolase [acid optimum], EC 3.1.3.2) in Saccharomyces cerevisiae at the physiological and molecular levels, through a series of repression and derepression experiments. We demonstrated that APase synthesis is tightly regulated throughout the growth phase and is influenced by exogenous and endogenous Pi pools. During growth in a nonlimiting Pi medium, APase is repressed. When external Pi becomes limiting, there is a biphasic appearance of APase mRNA and enzyme. Our data on APase mRNA half-lives and on the flux of intracellular Pi and polyphosphate during derepression are consistent with a mechanism of transcriptional autoregulation for the biphasic appearance of APase mRNA. Accordingly, preculture concentrations of Pi control the level of corepressor generated from intracellular polyphosphate degradation. When cells are fully derepressed, APase mRNA levels are constant, and the maximal linear accumulation rate of APase is observed. A scheme to integrate phosphorus metabolism and phosphatase regulation in S. cerevisiae is proposed.
Aquaculture is one of the fastest growing food producing sectors in the world accounting for approximately 50% of fisheries products. The protein component in aquaculture diet is the single most expensive portion and important dietary nutrient. Generally, feed ingredients of animal origin having higher protein content than the plant protein source. Protein conversion efficiency expresses the growth pattern of fish. Therefore, in the present study, experiment was conducted to evaluate Protein Conversion efficiency (PCE) of freshwater fish Labeo rohita fed on 100%, 75%, 50% 25% non conventional formulated feed i.e. blood of bovine animals obtained from slaughter house and 100% conventional feed i.e. Groundnut oil Cake. During experiment, the fishes were fed at the rate of 2% of the total body weight per day. After experiment, the fish shows maximum values of PCE in formulated feed as compared to conventional feed.
Bombyx mori is a beneficial and environment friendly insect reared commercially for silk. In today's scenario, the silk farming shows a great downfall due to the promotion of synthetic materials. Thereby it is necessary to upsurge the production of silk fibres. The efficacy of silk is achieved through the growth and development of silkworms at a higher rate through supplementing their feed with antioxidant from the plant sources. Later the same worms were exposed to low dose radiation from the gamma source. These are considered as experimental groups while the worms without the supplement of antioxidant and radiation exposure, control group. The adaptability of the insects in terms of growth and economic parameters viz., silk filament, cocoon weight, shell weight and pupal weight was recorded. The total proteins in hemolymph and silk gland were assessed. The present study results indicated the increase in the efficacy of silk as well as economic traits in all the experimental groups compared to the control group. The assessed total proteins showed an increment in the silk gland and hemolymph was statistically significant. The larvae exposed to gamma radiation at low doses reflects hormesis that has exerted stimulatory and beneficial effects on the efficacy of silk production.
In the present study, using neuroblast‐enriched cultures derived from three‐day‐old chick embryos (E3WE), we examined the morphological effects of ethanol and/or GABA, as well as the developmental profile of the cholinergic and GABAergic neuronal phenotypes, as assessed by the activities of choline acetyltransferase (ChAT) and glutamate decarboxylase (GAD). Cultures exposed to ethanol (50 mM) exhibited smaller and fewer aggregates than controls with a neuritic network that lacked fasciculation. In cultures treated with GABA (10⁻5 M) alone or ethanol + GABA the size and number of the neuronal aggregates was increased and also neuritic arborization and fasciculation was enhanced. Thus, addition of GABA restored the normal growth pattern in the ethanol‐treated cultures. As previously shown, E3WE culture treated with ethanol alone showed a decrease in both ChAT and GAD activities compared to controls. Both cholinergic and GABAergic neuronal phenotypes were enhanced in cultures treated with GABA as assessed by increases in ChAT and GAD activities, respectively, compared to controls. Moreover, in cultures treated concomitantly with ethanol and GABA both ChAT and GAD activities were higher than in ethanol‐alone‐treated cultures. Thus, the presence of GABA in the ethanol‐treated cultures counteracted the decline in ChAT and GAD activities observed in the ethanol‐alone‐treated cultures. We concluded that GABA through its neuronotrophic actions can rescue neuroblasts from ethanol insult and restore neuronal phenotypes.
Day by day water scarcity is increasing and simultaneously waste water generation is also increasing. Therefore, proper management and recycling of waste water is required in order to minimise its ill-effects. One of the productive uses of the industrial waste water is its application in agricultural lands but not every type of industrial waste water is found fit for such application. Thus, in this study comparative analyses of soil, water and vegetable produced (growth and biochemical parameters) by irrigating using tap water and oil refinery waste water were done to bring out the differences between the cultivated vegetables. Numerous parameters from physicochemical properties of water, soil, germination, morphology, growth rate to biochemical aspects of the vegetables were analysed. It was found that epicotyl, hypocotyl and the growth of root were better when using the effluents in irrigation. Nutritional value of effluent-irrigated plants was found to be higher, as fruits contained appreciable amounts of protein, ascorbic acid, calcium, magnesium, sodium, potassium and phosphorus.
Studies from our laboratory have established that ethanol exerts morphological and biochemical neurotoxic effects during early neuroembryogenesis in the chick brain both in ovo and in culture. In the present study, we further localized the critical period for ethanol effects on cholinergic neuronal expression using neuroblast‐enriched cultures derived from 3‐day‐old chick embryos. Moreover, we report that NGF attenuated the cholinotoxic effects of ethanol. We used the following experimental paradigms: cultures treated with ethanol alone either C0‐C3 or C4‐C10; NGF alone CO‐C4 or C4‐C10; ethanol and NGF given concomitantly; ethanol given first then replaced with NGF in the medium; or NGF given first then replaced with ethanol in the medium. The results revealed: (1) the cholinotoxic effect of ethanol occurs between culture days CO and C4 with day 3 appearing to be most critical. (2) similarly, the critical period for the cholinotoxic effects of NGF is during early neuroblast differentiation, culture days CO‐C4. (3) NGF can prevent the cholinotoxic effects of ethanol only if both ethanol and NGF are given concomitantly or if ethanol is given first, then culture is replaced with NGF‐containing medium.
The mechanism of toxic action for organophosphates (OPs) is the persistent inhibition of acetylcholinesterase (AChE) resulting in accumulation of acetylcholine and subsequent hyperstimulation of the nervous system. Organophosphates display a wide range of acute toxicities. Differences in the OP's chemistries results in differences in the compound's metabolism and toxicity. Acute toxicities of OPs appear to be principally dependent on compound specific efficiencies of detoxication, and less dependent upon efficiencies of bioactivation and sensitivity of AChE. Serine esterases, such as carboxylesterase (CaE) and butyrylcholinesterase (BChE), play a prominent role in OP detoxication. Organophosphates can stoichiometrically inhibit these enzymes, removing OPs from circulation thus providing protection for the target enzyme, AChE. This in vitro study investigated age-related sensitivity of AChE, BChE and CaE to twelve structurally different OPs in rat tissues. Sensitivity of esterases to these OPs was assessed by inhibitory concentration 50s (IC50s). The OPs displayed a wide range of inhibitory potency toward AChE with IC50s in the low nM-μM range with no differences among ages; however, the CaE IC50s generally increased with age reflecting greater protection in adults. These results suggest age-related differences in acute toxicities of OPs in mammals are primarily a result of their detoxication capacities.
Inhibition kinetics assays were conducted with 16 commercial organophosphate (OP) pesticides or their metabolites on acetylcholinesterase (AChE) in erythrocyte “ghost” preparations from 18 individual humans (both sexes; adults, juveniles and cord blood samples; mixed races/ethnicities) and pooled samples from adult rats (both sexes). A well established spectrophotometric assay using acetylthiocholine as substrate and a chromogen was employed. The kinetic parameters bimolecular rate constant (ki), dissociation constant (KI) and phosphorylation constant (kp) were calculated for each compound. As expected, a wide range of potencies were displayed among the tested compounds. Statistical analysis of the resultant data indicated no differences in sex, age or race/ethnicity among the human samples that are unexpected based on chance (4.2% statistically significant out of 48 parameters calculated) and no differences between the sexes in rats. The bimolecular rate constants for 10 of the compounds were not statistically different between rats and humans. The data indicate that, consistent with the high level of conservation of AChE among species and the fact that AChE at different locations within a species arises from the same gene, the inhibition kinetic parameters calculated from rat erythrocyte ghost preparations should be useful in estimating potencies of OP compounds on target AChE in humans. Additionally the data indicate that differences in sensitivities among individual humans were not apparent. Impact Statement: These data are expected to be useful in consideration of the intraspecies and interspecies uncertainty factors in OP pesticide risk assessment.
The activities of the proline-specific permease (PUT4) and the general amino acid permease (GAP1) of Saccharomyces cerevisiae vary 70- to 140-fold in response to the nitrogen source of the growth medium. The PUT4 and GAP1 permease activities are regulated by control of synthesis and control of activity. These permeases are irreversibly inactivated by addition of ammonia or glutamine, lowering the activity to that found during steady-state growth on these nitrogen sources. Mutants altered in the regulation of the PUT4 permease (Per-) have been isolated. The mutations in these strains are pleiotropic and affect many other permeases, but have no direct effect on various cytoplasmic enzymes involved in nitrogen assimilation. In strains having one class of mutations (per1), ammonia inactivation of the PUT4 and GAP1 permeases did not occur, whereas glutamate and glutamine inactivation did. Thus, there appear to be two independent inactivation systems, one responding to ammonia and one responding to glutamate (or a metabolite of glutamate). The mutations were found to be nuclear and recessive. The inactivation systems are constitutive and do not require transport of the effector molecules per se, apparently operating on the inside of the cytoplasmic membrane. The ammonia inactivation was found not to require a functional glutamate dehydrogenase (NADP). These mutants were used to show that ammonia exerts control of arginase synthesis largely by inducer exclusion. This may be the primary mode of nitrogen regulation for most nitrogen-regulated enzymes of S. cerevisiae.
A nutritional characteristic of trypanosomatid protozoa is that they need a heme compound as a growth factor. Because of the
cytotoxic activity of heme and its structural similarity to cobalamins, we have investigated the in vitro and in vivo effect of vitamin B12 (or cyanocobalamin) on the different forms of Trypanosoma cruzi. Cyanocobalamin showed a marked antiparasitic activity against epimastigotes (50% inhibitory concentration [IC50], 2.42 μM), amastigotes (IC50, 10.69 μM), and trypomastigotes (IC50, 9.46 μM). Anti-epimastigote and -trypomastigote values were 1.7 to 4 times lower than those obtained with the reference
drug benznidazole (Bnz). We also found that B12 and hemin do not interact with each other in their modes of action. Our results show that B12 increases intracellular oxidative activity and stimulates both superoxide dismutase (50%) and ascorbate peroxidase (20%)
activities, while the activity of trypanothione reductase was not modified. In addition, we found that the antioxidants dithiothreitol
and ascorbic acid increase the susceptibility of the parasite to the cytotoxic action of B12. We propose that vitamin B12 exerts its growth-inhibitory effect through the generation of reactive oxygen species. In an in vivo assay, a significant reduction in the number of circulating parasites was found in T. cruzi-infected mice treated with cyanocobalamin and ascorbic acid. The reduction of parasitemia in benznidazole-treated mice was
improved by the addition of these vitamins. According to our results, a combination of B12 and Bnz should be further investigated due to its potential as a new therapeutic modality for the treatment of Chagas' disease.
Background
Traditional soy protein isolate (SPI)‐based gel products, such as tofu, are generally produced by heating and by addition of metal salt ions to adjust the hydrophobicity and electrostatic force of soybean protein to facilitate the formation of a uniform network structure. However, the gelation rate of the soy protein gel network structure is difficult to control. Theoretically, epigallocatechin‐3‐gallate (EGCG) could be used to alter the surface hydrophobicity of thermally‐induced SPI to improve their gelation rate and form a more uniform network structure, thus improving SPI‐based gel properties (hardness, water‐holding capacity and rheological properties).
Results
An SPI‐EGCG complex (SPIE) was prepared, and properties of the resulting gel, following induction of transglutaminase (TG), were evaluated. Results showed that EGCG are bound to thermally‐induced SPI primarily via hydrophobic and hydrogen bonding, thus altering the secondary structure composition and reducing surface hydrophobicity of proteins in thermally‐induced SPI. Furthermore, the optimum amount of EGCG required to improve the gel strength, water‐holding capacity and rheological properties was ≤0.04:1 (SPI 1 g L ⁻¹, EGCG:SPI, w/w). Thermal stability analysis further indicated that EGCG in SPIE was more stable than free EGCG after heating.
Conclusion
This study demonstrated that EGCG can improve the gel properties of TG‐cross‐linked SPIE, while EGCG in SPIE exhibits enhanced thermal stability. Additionally, the results of this study provide a novel strategy for the development of SPI‐based gel foods with improved gel properties and that are enriched with bioactive compounds.
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Kinins are important inflammatory mediators that have potent effects on a variety of cell types via B1 and B2 receptors. This thesis aimed to examine kinin receptor expression and regulation in rat bladder smooth muscle cells. The pharmacological profile of the responses of these cells to B1 and B2 agonists in a functional assay, measuring 45Ca efflux, in conjunction with radioligand binding experiments, was consistent with the cells expressing both B1 and B2 receptors. In addition, the rat specific kinin, T-kinin, and its putative breakdown product, des-Arg11-T-kinin, were potent and selective B2 and B1 agonists respectively, with selectivity for rat over human kinin receptors. The pro-inflammatory cytokine IL-I upregulated the responsiveness of the rat bladder cells to B1 agonists, but had no effect on responses to B2 agonists. The effect of IL-1 was inhibited by agents that inhibit protein synthesis such as cycloheximide and dexamethasone. This reflected increased expression of the B1 receptor as IL-1 increased B1 receptor mRNA and specific binding of [3H]- des-Arg10-kallidin. A novel mechanism by which B1 responses can be modulated was elucidated. Treatment with dibutyryl cAMP also lead to an increase in B1 receptor-evoked 45Ca efflux with no effect on B2 receptor-mediated responses. Although the increased responsiveness induced by dibutyryl cAMP was inhibited by cycloheximide and dexamethasone, there was no increase in B1 receptor mRNA or in [3H]-des-Arg10-kallidin binding. This suggests that dibutyryl cAMP stimulates the expression of a factor that acts downstream of the B1 receptor to increase agonist-evoked 45Ca efflux. The results suggest that, during inflammation, the expression and function of B1 receptors in the bladder may be increased by pro-inflammatory cytokines, whilst agents that elevate cAMP may increase B1 receptor-mediated responses. In contrast B2 mediated responses do not appear to be upregulated by inflammatory mediators in bladder smooth muscle cells.
Among the biochemical processes associated with the atherogenic process are increased aortic cholesteryl ester (CE) accumulation and altered prostaglandin (PG) production. The precise physiological role of PG, particularly prostacyclin (PGI2), in the control of CE metabolism in intact aortic smooth muscle cells remains to be fully elucidated. We report here that cytosolic neutral cholesteryl ester hydrolytic activity (NCEH) in intact cultured aortic smooth muscle cells is significantly increased by 75-250 nM PGI2 at the end of a 2-hr incubation period. The effect was mediated by increased intracellular cAMP levels since the effect of PGI2 on NCEH activity was abolished in the presence of an inhibitor of adenylate cyclase activity, viz., dideoxyadenosine (DDA0. Although the addition of 20-100 microM dibutyryl cAMP (Bt2cAMP) and 50-100 microM sodium arachidonate also increased NCEH activity twofold, 6-keto PGF1 alpha, PGE1, and PGE2 did not increase the activity of this enzyme. In contrast to these findings, 75-250 nM PGE2 significantly inhibited CE synthetic activity (ACAT) approximately 60%. Arachidonate or Bt2cAMP did not affect ACAT activity. This decrease in ACAT activity induced by PGE2 does not appear to be mediated by cAMP. Taken together, these findings suggest that PGI2, a well known potent vasodilator and inhibitor of platelet aggregation, and PGE2 may have an important regulatory role in aortic CE metabolism.
Polyaromatic hydrocarbons are highly stable and can easily permeate through biological membranes due to their lipophilic nature. They are characterized by their ability to act at the site of application and even at sites distant to the zone of application. Any contamination associated with aquatic bodies due to the presence of PAHs can be assessed by investigating biochemical changes. The most common polycyclic aromatic hydrocarbon (PAH) pollutant naphthalene was selected for the bioassay experiment using Indian climbing perch Anabas testudineus. The biochemical response in the muscle tissue was investigated after exposure to varying concentrations of naphthalene by estimating protein, glycogen, Alanine Aminotransaminase, and Aspartate Aminotransaminase. The results showed inhibition of protein and glycogen levels due to naphthalene stress. Similarly, a decrease in the activity of transaminase in muscle tissues was noticed. The results have been computed using star plots for interpretation of integrated biomarker response (IBR). The results clearly support the role of biochemical parameters in assessing the impact of naphthalene stress on fish health. IBR index can be developed as a useful tool in monitoring quantitative as well as the qualitative effect of naphthalene toxicity in fishes and other aquatic animals.
Biological reduction of selenium oxyanions is widely used for selenium removal from wastewater. The process is, however, limited by the availability of a suitable, efficient and low cost electron donor. In this study, selenite and selenate reduction by waste activated sludge using hydrogen as the electron donor was investigated. Both selenite and selenate (80 mg/L) were completely removed using H2 within 8 days of incubation. In the presence of sulfate in the medium, the Se removal efficiency decreased to 77.8–95.4% (for selenite) and 88.2–99.4% (for selenate) at different temperatures and initial sulfate concentrations. Thermophilic conditions (50 °C) were better suited for both selenite and selenate reduction using H2 as electron donor with a 0.8–13.5% increase in overall Se removal. Similarly, sulfate reduction also increased from 69.1– 88% at 30 °C to 72–94.6% at 50 °C. Most of the H2 utilized was diverted towards Se and sulfate reduction with minimal production of byproducts such as methane (<0.32 mM) or volatile fatty acids (<0.92 mg/L). The elemental Se produced from selenite and selenate reduction ranged between 33.9 and 52.1 mg/L. The elemental selenium nanoparticles produced as a result of selenite and selenate reduction were characterized using transmission electron microscopy (TEM), energy-dispersive X-ray spectroscopy (EDX) and dynamic light scattering (DLS) spectroscopy. Furthermore, characterization of the biomass using Fourier-transform infrared spectroscopy (FTIR) and excitation emission matrix (EEM) spectra of the extracellular polymeric substances (EPS) produced by the waste activated sludge were performed to elucidate the mechanism of selenium oxyanion reduction to elemental selenium nanoparticles.
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