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Protein Measurement With Folin Fenol Reagent
Abstract
Since 1922 when Wu proposed the use of the Folin phenol reagent for
the measurement of proteins (l), a number of modified analytical pro-
cedures ut.ilizing this reagent have been reported for the determination
of proteins in serum (2-G), in antigen-antibody precipitates (7-9), and
in insulin (10).
Although the reagent would seem to be recommended by its great sen-
sitivity and the simplicity of procedure possible with its use, it has not
found great favor for general biochemical purposes.
In the belief that this reagent, nevertheless, has considerable merit for
certain application, but that its peculiarities and limitations need to be
understood for its fullest exploitation, it has been studied with regard t.o
effects of variations in pH, time of reaction, and concentration of react-
ants, permissible levels of reagents commonly used in handling proteins,
and interfering subst.ances. Procedures are described for measuring pro-
tein in solution or after precipitation wit,h acids or other agents, and for
the determination of as little as 0.2 y of protein.
... The protein concentrations of coproantigen, oocyst antigen, affinity purified antigen and the hyperimmune serum were estimated as described by Lowry et al. [32]. ...
... Protein was measured as described byLowry et al. (1951). b Activity unit (Au): defined as the amount of required protein to give one well of agglutination. ...
CRYPTOSPORIDIOSIS is a significant disease that causes diarrhea in humans and animals with relatively high morbidity and mortality. The present study aimed to adopt a purified and potent antigen for the accurate diagnosis of cryptosporidiosis. A total of 278 animal hosts (60 newborn calves and 218 buffaloes) were used in the current study. Sixty fecal samples were collected from newborn calves aged less than one month raised in the Beni-Suef and Qalyubia governorates. The samples were examined under a microscope after modified Ziehl-Neelsen staining, and Cryptosporidium oocysts were isolated from naturally infected calves. These oocysts were used in mice experimental infection. The oocyst antigen and coproantigen were prepared from the mice feces. The diagnostic efficacy of the two prepared antigens was evaluated using an Enzyme Linked Immunosorbent Assay (ELISA) with experimentally infected mice sera. The crude oocyst antigen proved to have higher diagnostic potential than coproantigen, so, it was chosen for purification using Cyanogen Bromide-activated Sepharose-4B affinity chromatography coupled with rabbit hyperimmune serum raised against oocyst antigen. The affinity purified fraction and its crude Cryptosporidium antigen were evaluated using the ELISA. The resulting purified fraction was 6733 fold increase in binding activity compared with its crude antigen. Characterization of the isolated fraction was performed using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), western blot, and amino acid analysis. SDS-PAGE clarified that the fraction contained three polypeptides of 94.7, 65, and 50 kDa, which were identified as immune-reactive components using a western blot analysis. The isolated fraction exhibited 17 amino acids and was rich in tyrosine, alanine, and phenylalanine. The affinity purified Cryptosporidium oocyst antigen effectively detected Cryptosporidium antibodies in experimentally infected mice sera and naturally infected buffalo sera with a sensitivity of 94.4% and 95.24 %, and a specificity of 100% and 93.33%, respectively. The purified fraction succeeded in diagnosis of cryptosporidiosis in 182 random serum samples collected from buffaloes with an incidence of 57.14 %. In conclusion, the affinity purified fraction of the Cryptosporidium oocyst antigen might be a good diagnostic candidate for cryptosporidiosis diagnosis and seroepidemiological surveillance.
... A comparative account of traditional feed (commonly used in the study area) and seaweed based formulated feed is given (Table 7.3.1). The proximate composition of the seaweed meal was determined using the methods of Lowry for protein (Lowry et al., 1951), Soxhlet for lipid (Tecator, 1983) and Anthrone method for carbohydrate (Trevelyan and Harrison, 1952). Astaxanthin was estimated as per the standard spectrophotometric method (Schuep and Schierle, 1995). ...
... Astaxanthin in the shrimp tissue was estimated as per the standard spectrophotometric method (Schuep and Schierle 1995) and body pigmentation of the cultured shrimp was assessed (for each treatment) after boiling the shrimp for 5 min in water and comparing the orangered coloration with Roche SalmoFan TM color score. The protein content of shrimp was estimated by Lowry's method (Lowry et al., 1951). ...
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... The amount of protein was measured according to Lowry's (1951) method with different concentrations of Bovine serum albumin (BSA) as a standard. 17 .The concentration was calculated in response to the absorbance at 650 nm in a spectrophotometer. ...
... The amount of protein was measured according to Lowry's (1951) method with different concentrations of Bovine serum albumin (BSA) as a standard. 17 .The concentration was calculated in response to the absorbance at 650 nm in a spectrophotometer. ...
Crustaceans have been recognized as rich sources of bioactive compounds with valuable nutraceutical and pharmaceutical potentials. Fresh water crab Oziotelphusa senex senex is the abundantly existing crab with unknown health benefits Cellular damage caused by reactive oxygen species has been implicated in several diseases; hence antioxidants have significant importance in human health Antioxidants play an important role as health protecting factor.Scientific evidences suggests that antioxidants reduce the risk for chronic diseases including cancer and heart diseases .So the.the antioxidant activity of the hemolymph from the fresh water Crab Oziotelphusa senex senex was. evaluated .Hemolymph of Oziotelphusa senex senex a fresh water crab, were subjected for its antioxidant activity using DPPH,ABTS and hydrogen peroxide scavenging assays. In antioxidant assay, the percentage of 2, 2-diphenyl-1-picrylhydrazyl scavenging activity was recorded as 57.5%. ABTS scavenging activity (68%) hydrogen peroxide scavenging (64%).The antioxidant activity of the hemolymph increased in a concentration dependent manner. Hence, the present study revealed that the hemolymph from the fresh water crab Oziotelphusa senex senex can be used as an accessible source of natural antioxidants with consequent health benefits.
... A comparative account of traditional feed (commonly used in the study area) and seaweed based formulated feed is given (Table 7.3.1). The proximate composition of the seaweed meal was determined using the methods of Lowry for protein (Lowry et al., 1951), Soxhlet for lipid (Tecator, 1983) and Anthrone method for carbohydrate (Trevelyan and Harrison, 1952). Astaxanthin was estimated as per the standard spectrophotometric method (Schuep and Schierle, 1995). ...
... Astaxanthin in the shrimp tissue was estimated as per the standard spectrophotometric method (Schuep and Schierle 1995) and body pigmentation of the cultured shrimp was assessed (for each treatment) after boiling the shrimp for 5 min in water and comparing the orangered coloration with Roche SalmoFan TM color score. The protein content of shrimp was estimated by Lowry's method (Lowry et al., 1951). ...
Oceans, seas, bays and estuaries are potential reservoirs of minerals which can be categorized into aggregates, placer deposits, phosphorite, evaporite deposits, polymetallic sulphides, polymetallic nodules, hydrocarbons and gas hydrates. These mineral resources can upgrade the economy of the nation if they are rationally explored, extracted and used for generating various usable products through circular economy. There is a limitation/boundary of every nation to explore and extract these resources, which is referred to as Exclusive Economic Zone (EEZ). The present paper highlights these non-living resources of the sea along with their distribution pattern.
... Among enzymes Lactate Dehydrogenase (LDH), Alanine amino tranferase (ALT) and Aspartate amino tranferase (AAT), were studied. Total proteins were estimated according to the method of Lowry et al. (1951) [9] . Cholesterol was estimated according to the method of Hanel and Dam (1955) [6] . ...
... Among enzymes Lactate Dehydrogenase (LDH), Alanine amino tranferase (ALT) and Aspartate amino tranferase (AAT), were studied. Total proteins were estimated according to the method of Lowry et al. (1951) [9] . Cholesterol was estimated according to the method of Hanel and Dam (1955) [6] . ...
... The homogenate was kept on ice for 30 min, and centrifuged at 5,000 × g for 20 min. The 136 supernatant of soluble peptides was evaluated according to the procedure of Lowry et al. (1951). 137 ...
... The moisture and ash contents were determined by the method of AOAC, 1990. The total protein and water-soluble protein were determined by the micro-Kjeldhal method by Jayaraman (1981) and spectrophotometrically (Lowry et al., 1951) respectively. Lipid contents were determined colorimetrically as per Bligh and Dyer, 1959. ...
Aegle marmelos L. leaves were analyzed at different maturity levels for their physico-chemical compositions and change in contents of some hydrolytic and oxidative enzymes. The pH of the Aegle marmelos L. leaves was in the acidic ranges at all the maturity stages and the acidity of the leaves increased gradually with the advancement of maturity. The moisture content decreased gradually while ash content increased remarkably with age. Protein, total sugar, reducing sugar, sucrose and vitamin contents increased rapidly with the advancement of maturity while lipid and starch content decreased with maturation. The activity of amylase and invertase increased up to mature stage and thereafter decreased rapidly at the ripen stage. Mineral contents increased up to the mature stage and then decreased in ripen stage. Polyphenol oxidase and peroxidase activity high in immature stage but decreased dramatically in mature stage and thereafter increased in ripen stage while the activity of protease and lipase increased all the maturity stage. In this study, ripen Aegle marmelos L. leaves contained the highest amount of protein, carbohydrate, b-Carotene, vitamin B1, vitamin B2 and vitamin C whereas mature and immature Aegle marmelos L. leaves are rich sources of minerals and starch respectively.
... GI tract homogenates (10%) prepared with phosphate buffered saline (PBS; 0.1 M, pH 7.4, 0.89% NaCl) were centrifuged (10,000×g, 4°C, 30 min), and the resultant supernatants were used as the source of enzyme extracts. The protein content of the enzyme extract was determined using the standard solution of bovine serum albumin (BSA) (Lowry et al., 1951). Amylase activity was analyzed by the dinitro-salicylic acid method using soluble starch as the substrate (Bernfeld, 1955). ...
Introduction
Linseed or flaxseed (Linum usitassimum L.) contains a prospective source of protein and energy to be utilized in animal feed. This study aimed at re-cycling and value-addition of Linseed Oil Cake (LOC) for formulation of non-conventional carp diets.
Methods
The LOC was bio-processed through solid state fermentation (SSF) with a fish gut bacterium, Bacillus pumilus (KF640221). Nine experimental sets of diets were formulated using raw (R1-R4) and SSF-processed (F1-F4) LOC at 10%, 20%, 30% and 40% levels substituting fishmeal as well as other ingredients in a reference diet, and rohu, Labeo rohita fingerlings (2.08±0.03 g) were fed for 70 days feeding trial. Growth, carcass composition, activities of digestive enzymes, digestibility and haemato-biochemical parameters were studied following standard methodologies.
Results
SSF significantly (P< 0.05) improved crude protein along with amino acids, whereas crude fibre and antinutritional factors were reduced considerably. Experimental diets were isocaloric (4.8 kcal) and isonitrogenous (36%). Diets with bio-processed LOC had significantly better performance than the raw LOC. Fish fed diet F3 with 30% fermented LOC resulted in the highest weight gain (6.25 ± 0.09 g), specific growth rate (% day ⁻¹) and carcass protein deposition (16.77±0.34%). Activities of the digestive enzymes (amylase, lipase and protease) were also significantly (P<0.05) higher in fish receiving diets containing fermented LOC. Analyses of blood parameters revealed that haemoglobin, erythrocytes, leukocytes, plasma lipid, total plasma protein, albumin and globulin contents were increased, while plasma glutamic-oxaloacetic transaminase (GOT) and glutamic-pyruvic transaminase (GPT) levels were decreased in fish fed bio-processed LOC supplemented diets.
Conclusion
The present study might propose substitution of fish meal along with other conventional ingredients by incorporation of 30% SSF-processed LOC in the diets of rohu with no negative effect to the growth performance, carcass composition and feed utilization.
... The protein content of the cellulase produced was determined by Lowry's method [18] using bovine serum albumin (BSA) as standard. The crude cellulase (1 mL) was taken in a test tube along with 1-mL BSA solution (1 mg/mL) in another test tube. ...
In the current investigation, numerous fungi were isolated from diverse sources of soil, and the isolate with the strongest hydrolytic capacity was selected by the Congo red test. The selected fungal isolate was evaluated using the Basic Local Alignment Search Tool for nucleotide (BLASTN) and identified by comparing it with the National Center for Biotechnology Information (NCBI) database. It was further used for screening several cellulosic materials for cellulase production with the best activity by solid-state fermentation. The selected fungi and substrate were used to optimize the culture conditions for cellulase production using solid-state fermentation by response surface methodology (RSM) selecting incubation temperature (20–40 °C) incubation time (96–192 h), nitrogen concentration as ammonium sulfate (0.8–2.0 g/L), and phosphate concentration as di-potassium hydrogen phosphate (1.0–3.0 g/L) as independent variables and their effect on cellulase activity and protein content as the dependent variables. Using analysis of variance (ANOVA), on the responses, the optimum culture conditions were found to be an incubation temperature of 35 °C, incubation time of 120 h, nitrogen concentration of 1.1 g/L, and phosphate concentration of 1.5 g/L. On conducting the fermentation at optimum levels of each parameter, the cellulase activity obtained was 1.43 IU/mL against a predicted value of 1.33 IU/mL, and the protein content of cellulase obtained was 6.57 mg/mL against a predicted value of 6.50 mg/mL. Therefore, it can be concluded from the present study that the fungus Aspergillus foetidus isolated from soil can be used for cellulase production with high activity and protein content, and cauliflower stalk residue can be used as a suitable substrate for cellulase production for various industrial purposes.
... Ash was determined by muffling the sample at 6000-7000°C to dry ash. [21]. First, cleaned porcelain crucibles and heated them in a muffle furnace at 6000°C. ...
This paper reports the isolation, characterization, and GC-MS analysis of Mugil parsia fish oil collected from the Rupsha River in Khulna. The oil was extracted from the dried fish by using the Soxhlet apparatus with n-hexane, with an oil yield of 17± 4.0 %. The moisture, ash, and protein content of the raw fish were found to be 75.45 %, 1.258 %, and 20.00 % respectively. The physicochemical compositions of the extracted oil like color, specific gravity, peroxide value, saponification value, free fatty acid (FFA), acid value, and iodine value were determined and were found to be yellowish brown in color, 0.913±0.31, 9.9±0.1 meq/kg, 295.4±3.0 mg KOH/g, 11.9±2.0%, 5.7±0.11mg KOH/g, and 150.3±0.12 mg/100g respectively. Fatty acid composition was identified by GC-MS analysis of the Mugil parsia fish oil, showed the Phthalic acid, mono-(2-ethylhexyl) ester, palmitic acid, oleic acid, stearic acid, icosanoic anhydride, glycidyl stearate were identified as the major constituents.
... The moisture and ash contents were determined by the method of AOAC (1990). The total protein and water-soluble protein were determined by the micro-Kjeldhal method by Jayaraman (1981) and spectrophotometrically (Lowry et al., 1951) respectively. Lipid contents were determined colorimetrically as per Bligh and Dyer (1959). ...
Context: Coccinia Cordifolia Lin. is an important tropical vegetable and it belongs to the Cucurbitaceae family. The fruits of C. cordifolia as a rich source of different nutrients were analyzed. Objective: Studies were conducted to investigate the changes of nutritional compositions and hydrolytic and oxidative enzymes of C. cordifolia fruits at different maturity levels. Materials and Methods: The pH was measured by pH meter. The moisture and ash contents were determined by the method of AOAC. Total and water-soluble proteins were determined by the micro-Kjeldhal method and spectrophotometrically respectively. Lipid contents were determined by Bligh and Dyer. Total sugar and starch content were estimated by Anthrone method. Thiamin and riboflavin were estimated by Anonymous and -carotene was estimated by Jensen. Vitamin-C content was determined by the titrimetric method. Calcium, iron, sodium, potassium, cupper and magnesium content were determined by Atomic Absorption Spectroscopic method. Phosphorus was determined by colorimetric means. The protease and amylase activity were measured by Kunitz and Jayaraman respectively. Invertase activity was assayed by Mahadevan and Sridhar. Results: The pH was acidic. The moisture content decreased and ash content increased with age. Protein, total sugar, reducing sugar, lipid and vitamin contents increased rapidly while starch content decreased with maturation. Mineral contents increased up to the mature stage and decreased in ripen stage. The activity of amylase and invertase increased up to mature and thereafter decreased. Polyphenol oxidase and peroxidase activity were high in immature stage but decreased in matured stage and thereafter increased in ripen stage while the activity of protease and lipase increased all the maturity stage. Conclusion: In this study, ripen C. cordifolia fruits contained the highest amount of protein, total sugar, reducing sugar, -Carotene, vitamin B1, vitamin B2 and vitamin C whereas matured and immatured C. cordifolia fruits are rich sources of minerals and starch respectively.
... The detoxified waste bread was subjected to sequential fractionation into four protein fractions based on the solubility (Monteiro et al., 1982) i.e.salt soluble fraction or globulin, water soluble fraction or albumin, alcohol soluble fraction or prolamin and buffer soluble fraction or glutelin. The protein content of fractions was estimated (Lowry et al., 1951). Data were analysed by one way ANOVA (Snedecor and Cochran, 1994) by using SPSS (2009) Version 16 and the differences in means by using Tukey's b test. ...
Wadhwa, M. and Bakshi, M.P.S. 2023. Effect of feeding detoxified waste bread in total mixed ration on nutrient utilization and rumen fermentation pattern in adult buffaloes. Animal Nutrition and Feed Technolog, 23: 221-231. The present study was undertaken to detoxify the stale waste bread (WB) and evaluated its nutritional worth in the total mixed ration of adult male buffaloes. The aflatoxin B1 content in the WB before and after detoxification was 55 and 8 ppb, respectively. The in vitro gas production studies revealed that the net gas production (NGP), digestibility of nutrients and availability of metabolizable energy (ME) from WB was higher (P<0.05) than that from maize and wheat. The digestion kinetic parameters for DM and CP assessed by in-sacco technique revealed that the effective and true digestibility was higher (P<0.05) in WB followed by that in wheat and maize grains. For in vivo studies 12 adult male buffaloes (BW 443.0±3.60 kg) divided in to three groups were offered total mixed ration (TMR) containing concentrate mixture in which maize grains were replaced with WB at 0, 50 or 100% on N basis. The complete replacement of maize grains by WB did not show any adverse effect on the daily DM intake, digestibility of nutrients, N-retention, efficiency of N utilization and urinary excretion of purine derivatives in adult male buffaloes. For assessing the impact of these TMRs on the biochemical changes in the rumen, one diet was fed at a time for 30 days to three rumen fistulated male buffaloes. Different N-fractions in the rumen and fluid outflow rate from the rumen were also not affected by the level of WB in the ration of rumen fistulated male buffaloes. It was concluded that detoxified WB can be fed safely and that complete replacement of maize grains in the ration of adult buffaloes did not show any adverse effect on nutrient utilization and health of adult buffaloes.
... Protein Assay: The protein was assayed following the method of Lowry et al., [30] using bovine serum albumin as standard. ...
Abstract: In a hypogonadal rat model, calcium ATPase (Ca2+-ATPase) activity and
membrane fluidity were examined in plasma membrane of duodenal enterocytes under two
different stress conditions developed by chemical change (lipid saturation) and physical
change (temperature). Data generated show that, under both these stress conditions, calcium
ATPase activity was decreased. Membrane fluidity (phase transition temperature) study by
fluorescence polarization measurement indicated that, compared to corresponding control
enterocyte membranes, differences were seen in the phase transition temperature (Tc) of the
oophorectomized rats’ enterocyte membrane under both the conditions of stress, which
suggested a decrease in membrane fluidity under both the experimental situations. It is
proposed that the degree of alteration in calcium ATPase activity in oophorectomized rats’
duodenal enterocyte plasma membrane under two different stress conditions was nearly alike,
which was influenced by changes in membrane fluidity induced by chemical change (lipid
saturation) or physical change (temperature).
Key words: Plasma membrane; Ca2+-ATPase; membrane fluidity, high-lipid diet; cold stress
... The corneal tissue samples were studied immediately. The protein levels in the supernatants were determined by the Lowry method (21). The VEGF and PDGF levels in the supernatants were measured using the Enzyme-Linked Immuno Sorbent Assay (ELISA) method. ...
Aim: This study aims to investigate the neovascularization-inhibiting effect of topical nilotinib and to determine the effective dose of nilotinib. Material and Method: In this study, 42 healthy Wistar albino rats were randomly divided into six groups. The left corneas of all rats except group 1 were cauterized with silver nitrate. Group 1 was the healthy control, with no corneal vascularization, which did not receive any treatment; Group 2 (sham) did not receive treatment, only topical DMSO; Groups 3, 4, and 5 received topical nilotinib at doses of 10, 20, and 40 μM three times a day, respectively; Group 6 received 5 mg/dL topical bevacizumab three times for a day for seven days. On the 8th day, photographs of the corneas were taken, and the percentage of corneal neovascularization area was calculated. Following all rats being killed via anesthesia, the corneas were removed to determine the levels of vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF) ELISA and corneal immune staining. Results: Other than Group 3, the percentage of neovascular corneal area was lower in the treatment groups compared to Group 2 (p
... Chemical oxygen demand (COD) and soluble COD (sCOD, after filtering sample through 0.45 µm membrane filter) was determined according to USEPA approved digestion method (Jirka and Carter, 1975) in triplicate. Soluble protein and carbohydrate in 0.45 µm filtered samples were analyzed in triplicate using Folin-Ciocalteu's Protein Measurement Method (Lowry et al., 1951) and Dubois Method (Dubois et al., 1956), respectively. ...
Plastics are resilient, hard to degrade materials that can persist in nature for centuries. Microplastics (MPs) exhibit similar tough character and hold the potential to harm marine and terrestrial ecosystems upon their release into the environment. Most modern wastewater treatment plants remove MPs from wastewater with over 90% efficiency but unfortunately concentrate them in sludge. Recent studies have reported MPs’ impact on the performance of sludge treatment systems, including anaerobic digesters. Despite its resilience, polyethylene terephthalate (PET) has inherent weaknesses against alkaline and thermal conditions and becomes more prone to further degradation if exposed to such stress conditions. Sludge pretreatment practices aiming to increase biogas production by disrupting floc structure show great similarity with the stress factors mentioned. Thus, this study aims to integrate pretreatment with anaerobic digestion and investigate the fate and effects of PET MPs during these processes. For this purpose, waste activated sludge samples spiked with different doses of PET (0, 1, 3, 6 mg/g TS) in sizes of 250–500 µm were pretreated by 0.5 M alkali for two days and then thermally hydrolyzed at 127 °C for 120 min. Pretreated and unpretreated sludges were digested in a 60-day biochemical methane potential test. The results showed that the spiking of PET MPs into sludge posed a positive impact on the methane yield of unpretreated reactors at statistically significant levels. Integrating pretreatment increased the methane yield by 22.0% and made the impact of MPs on digester efficiency no longer observable. Also, PET exposed to pretreatment and 60-day digestion experienced remarkable changes in surface morphology, crystallinity and carbonyl index, which can further impact their fate and effects on the environment.
... The amount of Malondialdehyde (MDA) was estimated according to the protocol of Devasagayam and Tarachand, 1987 18 . Protein content of ventricular tissue of heart was measured by the method of Lowry et al, 1951 19 , with Bovine Serum Albumin as the standard. ...
Bisphenol-A (BPA) is a chemical produced in large volumes worldwide and has large market diffusion in many consumer products. BPA is used mainly as a monomer in the production of polycarbonate plastics and as a precursor of epoxy resins. So the present study was undertaken to examine the toxic effect of BPA on heart function in pregnant rat models. BPA (50 mg/kg.b.wt/day and 500 mg/kg.b.wt/day) was administered for 8 th to 15 th day by oral-gavage. The rats were sacrificed by cervical dislocation after last treatment. Paraffin embedded heart tissue sections were stained with hematoxylin-eosin. Significant degenerative changes were observed in H&E stained heart tissue sections of test rats. The activities of antioxidant enzymes like-superoxide dismutase, catalase and glutathione peroxidase were seen to be decreased significantly in treated rats; while, the level of malondialdehyde, a biomarker of lipid peroxidation, was increased than treated rats. Up-regulation of proapoptotic protein has been reported in heart failure and myocardial infarction, to determine whether caspase-3 protein can affect cardiac function in treated pregnant rats. In conclusion, BPA inhibits ventricular function in rat presumably by producing oxidative stress induced apoptosis and causes potential cardio toxicity.
... The concentration of protein was determined using the Lowry method and Bovine serum albumin (BSA) as standard. 23 Purification of Aspergillus fumigatus ASF4 GOx The crude GOx was brought to 60% saturation using (NH4)2SO4 salt as described by Chilaka et al. 24 The precipitated enzyme (20ml) was dialyzed for 12 hr against sodium phosphate buffer (0.05 M, pH 7.0). The dialysate was loaded into a (2.0 x 14 cm) DEAE chromatographic column equilibrated with 50 mM sodium acetate buffer (pH 5.5). ...
... Soluble protein content of the enzyme source was measured after Lowry et al. (1951) and quantitative enzyme activities were expressed as unit (U). Identification of isolates by 16S rRNA partial gene sequence analysis: Three promising extracellular enzyme producing strains were identified through 16S rRNA partial gene sequence analysis after isolation and PCR amplification following the methods described in Das et al. (2014). ...
The present study was carried out to screen autochthonous gut bacteria in freshwater air breathing walking catfish, Clarias batrachus Linnaeus. Altogether, 100 extracellular enzyme-producing bacteria were isolated from the foregut (FG) and hindgut (HG) regions. Data were presented as log viable counts g-1 gut (LVC). The occurrence of heterotrophic bacterial population was higher in the FG region (LVC = 8.25) than the HG (LVC= 7.3). Similarly, proteolytic, amylolytic and lipolytic bacteria in FG outnumbered (LVC=7.25, 6.77 and 5.23 respectively) the HG (LVC=6.38, 5.58 and 4.04 respectively). However, occurrence of cellulolytic bacteria in both, FG and HG was less (LVC=2.1 and 1.34 respectively) in comparison to the other extracellular enzyme-producing bacteria. Out of the 100 bacterial isolates, 22 isolates were primarily selected through qualitative assay of extracellular enzyme activities. Among them, 3 promising isolates were chosen as potent extracellular enzyme producers on the basis of cumulative scores (≥11) of the qualitative assay and quantitative enzyme assay. Maximum protease activity was revealed by the strain FG10 (201±2.40U), while FG43 exhibited maximum amylase (208.3±10.8U) and lipase (4.73±0.05U) activities. Among the strains isolated from the HG, the highest protease (188.3±1.2U), amylase (97.6±0.46U) and lipase (3.7±0.11U) activities were recorded with the strain HG01.
... Leaf total soluble protein (TSP) was measured by Lowry's method (Lowry et al., 1951) using Folin-Ciocalteu reagent. The optical density was measured using a spectrophotometer at 750nm and TSP was expressed as mg/g FW. ...
... The serum alkaline phosphatase activity was estimated by the method of [7] . Protein was estimated by the method of [8] . ...
Diabetes mellitus is a metabolic disorder of multiple aetiology characterized by chronic hyperglycaemia with disturbances of carbohydrate, fat and protein metabolism resulting from defects in insulin secretion, insulin action, or both. Liver is the central metabolic organ in body responsible for glucose homeostasis, diabetes leads to hepatic dysfunction. In the present study to investigate the protective effect of Mollugo cerviana extract on liver markers in alloxan induced diabetic rats. Hepatospecific enzymes (ALP, ALT and AST), protein and albumin content were activated when hepatocellular damage gave rise to abnormalities of liver function and these enzymes are remarkably increased in diabetic rats. Further confirmed in the histopathological studies of liver. Mollugo cerviana helps in parenchymal cell regeneration in liver, thus protecting membrane integrity and thereby minimizing enzyme leakage. Histopathological studies also supported the biochemical parameters. This result suggested that Mollugo cerviana possess potential hepato regenerating activity
... For the specific enzymatic activity, the protein content was determined by the method of Lowry et al. (1951). ...
... After blood collection, within one-hour samples were centrifuged at 3000 rpm for 10 min, and then serum was separated. Serum biochemical parameters including Alanine aminotransferase (ALT) and Aspartate aminotransferase (AST) by Reitman and Frankel (1957) [11]; by Kind and King's (1954) [12] method for Alkaline phosphatase (ALP) [13] method for total protein (TP), Urea [14], Creatine [15], Total bilirubin [16], Albumin [17,18] method for Albumin globulin ratio (A/G ratio) and Triglycerides (TG), High density lipoprotein (HDL) [19] and Total cholesterol; Low density lipoprotein (LDL) [20] were determined by Microlab-300 autoanalyzer (Merk Pvt. Ltd., Mumbai, India) were analysed. ...
Background: Hepatocellular carcinoma plays an inadequate mention in the second cause of death because of cancer worldwide. An alternative therapy with high rate of prognosis and also without side effects. Several data indicated the therapeutic efficacy of Enhalus acoroides. There were no scientific studies on chemopreventive and antioxidant potential of Enhalus acoroides against hepatocellular carcinoma.
Purpose: To investigate the hepatoprotective efficacy of ethanolic extract of Enhalus acoroides (EEEA) against DEN induced hepatocellular carcinoma using wistar albino rats.
Study design: Animals were divided into five groups each comprising six rats. Normal saline given to Group I- Control rats. By using DEN, liver cancer was induced to Group II, III, IV and V rats as single intraperitoneally injection (100 mg/kg body weight). At the beginning of 6th week, Groups III rats received EEEA (200mg/kg body weight/day) upto 16 weeks. Group IV rats received EEEA for one week before the administration of DEN and continued till the 16th week. After the administration of DEN, Group V positive control rats received Silymarin (100 mg/kg body weight) at the beginning of 6th week and continued upto 16 weeks. The efficacy of Enhalus acoroides for its Hepatoprotective and antioxidant properties during its simultaneous treatment against DEN induced liver damage was evaluated in rats.
Methods: The hepatoprotective efficacy of EEEA (200 mg/kg) was investigated against DEN (100 mg/kg/b.w) induced hepatotoxicity, was measured by evaluating serum liver markers levels (ALT, AST, GGT and ALP), Kidney markers (Urea and Creatinine), Lipid profile (TG, HDL, LDL & Total cholesterol) and Serum tumor markers (DNA, RNA, AFP and CEA). EEEA-aided antioxidant defence against hepatotoxic insult of DEN was measured by evaluating various Antioxidant biomarkers (GSH, SOD, CAT, GPx, Vit C and Vit E) Morphometric gross analysis and Histopathological studies were done to support the outcomes of the present study.
Results: A significant increased antioxidant defence and reduced MDA levels in the serum of EEEA treated animals compared to the DEN induced animals. The resulting data showed that the administration of EEEA decreased the serum liver markers levels, kidney markers, Lipid profile and serum tumor markers when compared to the untreated rats. The histopathological anomalies were altered on administration of EEEA indicating its protective effects on hepatocytes when compared with untreated rats.
Conclusions: Our consequences established that crude ethanolic extract of Enhalus acoroides shown an effective impact against DEN-induced hepatocellular carcinoma, and serves as a better option for chemopreventive treatments.
... A protein concentration was measured in the gills, skin, and muscles of both types of fish by following the method mentioned by Lowry (Randall, 1951), using Follin reagent and making a standard curve using (BSA). Protein concentrations were measured based on the absorbance using UV Spectroscopy and expressed Result, mg / mL, the percentage calculated from dry weight. ...
This study is conducted during the period between October 2019 till July 2020 to measure the ratios of some chemical parameters in two species of fish prevalent in Al-Haffar Drainage. The concentrations of fats and proteins measured in three parts (gills, skin, and muscles) of Tilapia Zilli and Liza Abu fish. The percentages of protein in Liza Abu fish were 14.41, 24.5, 14.47%, while in Tilapia Zilli were as: 14.6, 16.74, and 11.2% for gills, skin, and muscles separately. The levels of protein in tilapia were relatively higher, and the highest levels were recorded in the skin, while in Liza Abu, The highest rate of protein is noticing in the skin and muscles, Fat percentages in Liza. A fish were 1.33, 7.01, 2.09%, while in Tilapia.z f were 2.7, 5.19, and 2.3%, in gills, skin, and muscles, the highest fat ratio is registered in the skin of both species, The lowest levels of fat in Liza. A recorded in the gills, the lowest percentage of Fat in tilapia .z was recorded in muscles.
... A protein concentration was measured in the gills, skin, and muscles of both types of fish by following the method mentioned by Lowry (Randall, 1951), using Follin reagent and making a standard curve using (BSA). Protein concentrations were measured based on the absorbance using UV Spectroscopy and expressed Result, mg / mL, the percentage calculated from dry weight. ...
This study is conducted during the period between October 2019 till July 2020 to measure the ratios of some chemical parameters in two species of fish prevalent in Al-Haffar Drainage. The concentrations of fats and proteins measured in three parts (gills, skin, and muscles) of Tilapia Zilli and Liza Abu fish. The percentages of protein in Liza Abu fish were 14.41, 24.5, 14.47%, while in Tilapia Zilli were as: 14.6, 16.74, and 11.2% for gills, skin, and muscles separately. The levels of protein in tilapia were relatively higher, and the highest levels were recorded in the skin, while in Liza Abu, The highest rate of protein is noticing in the skin and muscles, Fat percentages in Liza. A fish were 1.33, 7.01, 2.09%, while in Tilapia.z f were 2.7, 5.19, and 2.3%, in gills, skin, and muscles, the highest fat ratio is registered in the skin of both species, The lowest levels of fat in Liza. A recorded in the gills, the lowest percentage of Fat in tilapia .z was recorded in muscles.
... Lowry assay was used for the determination of released protein content in rehydration water at defined time points during rehydration (Lowry et al., 1951). Aliquots of 200 µL of appropriately diluted rehydration water were mixed with 1 mL of Lowry's solution. ...
This study investigated the effect of pulsed electric fields (PEF) pretreatment on rehydration kinetics, firmness, and release of intracellular components of dried chickpeas during rehydration at 35 to 65°C. After soaking preconditioning, chickpeas were subjected to PEF treatments (2.5 and 3.3 kV/cm, 0.2 to 12.0 kJ/kg, 15 µs pulse width, 20 Hz frequency). PEF treated and untreated chickpeas were dried in crossflow air dryer and their rehydration at constant seed/water ratio of 1:5 was studied for 24 hr. During rehydration, moisture, firmness, and concentration of released proteins, carbohydrates and raffinose family oligosaccharides (RFO) were determined and described using appropriate mathematical models. PEF treatment led to up to 70% higher rehydration rates of dried chickpeas. This increase corresponds to rehydration time of approximately 1.5 hr, as opposed to 5 hr for untreated samples. Firmness of PEF treated chickpeas (for energy inputs higher than 3 kJ/kg) during rehydration decreased up to 30% compared to untreated samples. The firmness of untreated samples after 300 min of rehydration was achieved at much shorter times (up to 30 min) for PEF treated samples. At the end of 300 min of rehydration, more than 47.7%, 76.1%, and 86.6% of total raffinose, stachyose, and verbascose, respectively has been extracted, but only 0.03% of nutritionally valuable proteins from PEF treated chickpeas. Consequently, this study demonstrates that PEF processing could be implemented in dried chickpeas processing as pretreatment, for the reduction of rehydration time prior to cooking and of intestinal discomfort caused by RFO.
... Afterward, the supernatant was recovered from each tube and the protein content of the supernatant was measured by following the protocol of Lowry et al. (1951). ...
Biofilm, an aggregated form of microbial existence has been a major area of concern in the healthcare units. These sessile microbes not only protect themselves from the host immune system but also exhibit high resistance against several antimicrobials. One such widely reported Gram-positive pathogen is Staphylococcus aureus . This human commensal is known to cause severe harmful diseases like bacteremia, sepsis, pneumonia, etc. Thus, strategies need to be undertaken to deal with such biofilm challenges. In this respect, we aimed to inhibit microbial biofilm formation of Staphylococcus aureus under the influence of a natural compound, piperine. Our study revealed that the higher concentrations of piperine exhibited considerable antimicrobial activity against Staphylococcus aureus . Hence, lower concentrations of piperine were tested to examine its antibiofilm activity. Several experiments like crystal violet (CV) assay, total biofilm protein assay, and fluorescence microscopy observation established that lower concentrations (8 µg/mL and 16 µg/mL) of piperine showed efficient antibiofilm activity against Staphylococcus aureus . It was also noticed that the lower concentrations of piperine did not compromise the microbial growth of Staphylococcus aureus while exhibiting antibiofilm activity. In this connection, we also noticed that the lower concentrations of piperine showed a considerable reduction in microbial metabolic activity. Furthermore, we observed that the compound was found to accumulate reactive oxygen species in the bacterial cells that could play an important role in the inhibition of biofilm formation. Thus, piperine could be considered as a potential antibiofilm agent against the biofilm formation caused by Staphylococcus aureus .
... Contents of the entire vial were reconstituted using 10 ml sterile water provided by the manufacturer and stored at 2-8ºC. According to the manufacturer, 1 ml of reconstituted ASV could neutralize 0.6 mg of N.N venom and contained 2.2 mg protein by Lowry's method (Randall and Lewis, 1951). For the estimation of PT, aPTT and TT, aliquots of reconstituted ASV were used in the range of 5 µl-17 µl containing 11 µg-37.4 ...
Increasing evidence suggests a sizable involvement of hemotoxins in the morbidity associated with envenomation by the Indian spectacled cobra, Naja naja (N.N). This study investigates the ability of Indian polyvalent anti-snake venom (ASV), methanolic extract of Andrographis paniculata (MAP) and their combination in reversing the hemostatic abnormalities, viz. activated partial thromboplastin time(aPTT), prothrombin time(PT) and thrombin time(TT) in citrated plasma. These parameters were assessed in 2 groups of experiments. Group 1: Without the prior incubation of plasma with venom and Group 2: With prior incubation of plasma with venom for 90 min at 37°C. Venom caused significant ( p < 0.001) prolongation in aPTT (175%), PT (49%) and TT (34%) in Group 1 and ASV could completely bring them back to normal. MAP showed a concentration-dependent reversal in aPTT, normalization of PT and prolongation of TT. When low concentration of ASV was supplemented with MAP, their combined effect in normalizing aPTT and PT improved by 37% and 26% respectively when compared to ASV alone. In Group 2, venom caused significant ( p < 0.001) prolongation in aPTT (231%), PT (312%) and TT (245%). ASV had limited effect in reversing aPTT (52%), TT (31%) but completely normalized PT. MAP was marginally effective in reversing the prolonged aPTT and PT but caused further prolongation of TT. Combination of ASV and MAP was more effective than ASV alone in reversing venom-induced increase in aPTT (52%) and PT (29%). The study proved that, a drastic reduction of ASV by 70%, could be effectively supplemented by MAP in combating hemostatic abnormalities induced by NN venom.
... Eye tissue samples were homogenized, and the supernatants were separated and studied immediately. The protein levels in the supernatants were determined by the Lowry method [18]. The TNF-a and IL-6 levels in the supernatants were measured using the ELISA method. ...
PurposeTo investigate the protective effect of filgotinib in endotoxin-induced uveitis model in rats.Materials and methodThis study used 24 Wistar Albino rats. Group I (control group) included the healthy controls; in Group II (sham group), only 300 µg/kg intraperitoneal (ip) lipopolysaccharide (LPS) was administered; and in Group III (treatment group), 3 mg/kg/day filgotinib was administered orally for 10 days followed by 300 µg/kg ip LPS. In all groups, clinical activity scores were evaluated after 24 h. Moreover, histopathological and immunological examinations were performed.ResultsIn Groups I, II, and III, the mean clinical activity and histopathological examination scores were 0.00, 3.25 ± 0.70, and 1.89 ± 0.60 and 0.00, 2.88 ± 1.12, and 1.44 ± 0.52, respectively. The clinical activity and histopathological examination scores were significantly increased in the sham group compared to the control group (p < 0.05); these findings were significantly reduced in the treatment group (p < 0.05). The mean TNF-α and IL-6 ELISA levels in all groups were 50.20 ± 3.24, 59.87 ± 2.98, and 54.34 ± 4.62 and 30.88 ± 1.79, 36.77 ± 1.21, and 33.66 ± 1.86, respectively. The TNF-α and IL-6 ELISA levels were significantly decreased in the treatment group compared to the sham group (p < 0.05); there was no significant difference between the treatment group and the control group (p = 0.105, p = 0.067, respectively)Conclusion
Filgotinib may be an alternative treatment option in preventing the development of noninfectious uveitis.
... The total reducing sugars were determined by method described by Miller [9]. and protein was determined by Lowry's method [10]. ...
Biodegradation of lignocellulosic forest waste by Aspergillus niger F7 under solid state fermentation was explored. Different pretreatments were given to render forest wastes readily accessible to the enzymatic attack. SSF of pretreated forest biomass was found to be superior over untreated forest biomass. Pretreated forest biomass has emerged as a suitable substrate for cellulase production by A. niger F7 reaching upto 146.14 U/g in acid+ steam pretreated P. roxburghii needles. It has also been noticed that though forest lignocellulosics when used as carbon source yielded fairly good amount of cellulase. The ultimate goal of the study is the efficient utilization of wastes so as to recover cellulase in concentrations that make purification feasible. The data gathered in this study provides us a glimpse of some of the dynamics of the production of cellulase.
... Control blank solution on reagents and equipment revealed insignificant contamination of samples. Values were expressed as ng of single metal/mg of total proteins assayed by the Folin phenol reagent method (Lowry et al., 1951) using bovine serum albumin as standard. ...
h i g h l i g h t s Risks related to metal contamination in Antarctica. Effects of waterborne Cd and Cu on fish oogenesis. Oocyte degeneration and changes in glycan composition. Cd downregulation of alpha and beta estradiol receptors. Cd and Cu reduce fecundity in T. bernacchii by impairing stock recruitment. a b s t r a c t Antarctica has long been considered a continent free from anthropic interference. Unfortunately, recent evidence indicate that metal contamination has gone so far and that its effects are still unknown. For this reason, in the present work, the potential endocrine disrupting effect of two highly polluting metals, copper and cadmium, were examined in the Antarctic teleost Trematomus bernacchii. After a 10 days waterborne exposure, ovarian metal uptake was determined by atomic absorption; in parallel, classical histological approaches were adopted to determine the effects on oocyte morphology, carbohydrate composition and presence and localization of progesterone and estrogen receptors. Results show that both metals induce oocyte degeneration in about one third of the previtellogenic oocytes, no matter the stage of development. In apparently healthy oocytes, changes in cytoplasm, cortical alveoli and/or chorion carbohydrates composition are observed. Cadmium but not copper also induces significant changes in the localization of progesterone and beta-estrogen receptors, a result that well correlates with the observed increase in ovarian metals concentrations. In conclusion, the acute modifications detected are suggestive of a significantly impaired fecundity and of a marked endocrine disrupting effects of copper and cadmium in this teleost species.
Caffeine is known to induce chromosomal aberrations in proliferating cells when they are incubated during G 2 and mitotic prophase. In the present paper, this caffeine effect has been analyzed in Allium cepa root meristems growing at different culture temperatures under steady-state kinetics. Caffeine (1-10 mM) induces chromosomal aberrations in a dose-dependent manner, and the treatment efficiency correlates linearly with the square of caffeine concentration. The efficiency of caffeine incubations, within the range 5-25°C during equivalent cycle time periods has also been studied. It has been found that the lower the culture temperature, the higher the level of chromosomal aberrations. Moreover, at different temperatures, the level of chromosomal aberrations is a simple function of caffeine concentration and the ATP level. Therefore, the efficiency of caffeine treatment appears to be determined by some interaction between caffeine concentration and cellular ATP level. Our present results demonstrate that the influence of growth temperature on the chromosome-breaking effect of caffeine can be, at least partially, explained by the ATP levels during the incubation periods. In short, under different kinetics of plant cell proliferation, the ATP level, and/or something correlating with it, could explain the efficiency of caffeine in inducing chromosomal aberrations: the lower the ATP level, the higher the caffeine efficiency.
India is a habitat for nearly one thousand four hundred forty-seven species of spiders under three hundred and sixty-five genera
and sixty families. Our initial survey on toxic bite by spider revealed severe edema, itching, acute pain, and hemorrhage following
tissue necrosis, which are the general symptoms of envenomation, but there are no reports of mortality. Significantly, Hippasa
partita spider, commonly called “funnel web spider,” which is endemic in hilly regions of the Western Ghats is responsible for
envenomation. In this study, a nonenzymatic neurotoxin has been purified from H. partita venom gland extract. Gel filtration
and ion exchange chromatography were used to purify the toxin into homogeneity as shown by SDS-PAGE, RP-HPLC, and
MALDI-TOF. Neurotoxin is devoid of enzymatic activities but causes intense neurotoxic symptoms. Neurotoxin is found to inhibit
the twitch response of sciatic nerve gastrocnemius muscle preparation and is found to be postsynaptic in action. Neurotoxin
is devoid of coagulant activity, edema, and hemorrhage and is nonlethal to mice (up to 5mg/kg body weight). In conclusion,
a neurotoxin, which is a principle agent in whole venom responsible for induced neurotoxic symptoms, has been purified and
characterized.
Stress is one of the basic factors in the etiology of number of diseases. The present study was aimed to investigate the effect of Triphala (Terminalia chebula, Terminalia belerica and Emblica officinalis) on noise-stress induced alterations in the antioxidant status and on the cell-mediated immune response in Wistar strain male albino rats. Noise-stress employed in this study was 100 dB for 4 h/d/15 days and Triphala was used at a dose of 1 g/kg/b.w/48 days. Eight different groups of rats namely, non-immunized: control, Triphala, noise-stress, Triphala with noise-stress, and corresponding immunized groups were used. Sheep red blood cells (5 x 10(9) cells/ml) were used to immunize the animals. Biochemical indicators of oxidative stress namely lipid peroxidation, antioxidants superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), ascorbic acid in plasma and tissues (thymus and spleen) and SOD, GPx and corticosterone level in plasma were estimated. Cell-mediated immune response namely foot pad thickness (FPT) and leukocyte migration inhibition (LMI) test were performed only in immunized groups. Results showed that noise-stress significantly increased the lipid peroxidation and corticosterone level with concomitant depletion of antioxidants in plasma and tissues of both non-immunized and immunized rats. Noise-stress significantly suppressed the cell-mediated immune response by decreased FPT with an enhanced LMI test. The supplementation with Triphala prevents the noise-stress induced changes in the antioxidant as well as cell-mediated immune response in rats. This study concludes that Triphala restores the noise-stress induced changes may be due to its antioxidant properties.
In this study we have modified DNA by exposing it to ultraviolet light in the presence of hydrogen peroxide. The modified DNA was probed for binding to the antibodies present in the sera of patients suffering from various types of cancer. Higher recognition of modified DNA, as compared to native DNA, by antibodies from cancer patients has got far reaching significance.
Acid sphingomyelinase (Asm) and acid ceramidase (Ac) are parts of the sphingolipid metabolism. Asm hydrolyzes sphingomyelin to ceramide, which is further metabolized to sphingosine by Ac. Ceramide generates ceramide-enriched platforms that are involved in receptor clustering within cellular membranes. However, the impact of cell-intrinsic ceramide on T cell function is not well characterized. By using T cell-specific Asm- or Ac-deficient mice, with reduced or elevated ceramide levels in T cells, we identified ceramide to play a crucial role in T cell function in vitro and in vivo. T cell-specific ablation of Asm in Smpd1 fl/fl /Cd4 cre/+ (Asm/CD4cre) mice resulted in enhanced tumor progression associated with impaired T cell responses, whereas Asah1 fl/fl /Cd4 cre/+ (Ac/CD4cre) mice showed reduced tumor growth rates and elevated T cell activation compared to the respective controls upon tumor transplantation. Further in vitro analysis revealed that decreased ceramide content supports CD4 ⁺ regulatory T cell differentiation and interferes with cytotoxic activity of CD8 ⁺ T cells. In contrast, elevated ceramide concentration in CD8 ⁺ T cells from Ac/CD4cre mice was associated with enhanced cytotoxic activity. Strikingly, ceramide co-localized with the T cell receptor (TCR) and CD3 in the membrane of stimulated T cells and phosphorylation of TCR signaling molecules was elevated in Ac-deficient T cells. Hence, our results indicate that modulation of ceramide levels, by interfering with the Asm or Ac activity has an effect on T cell differentiation and function and might therefore represent a novel therapeutic strategy for the treatment of T cell-dependent diseases such as tumorigenesis.
Ancient literature reveals that snake shed skin had been frequently used to treat several diseases like glaucoma, hernia, psoriasis, hemorrhoids, etc. Previously, it was reported that Naja naja shed skin extract causes temporary cessation of the estrous cycle and altered female reproductive hormones in rodents, and the probable bioactive molecule behind such alterations was also identified. Based on this information, the present study was designed to establish the effect of Naja naja shed skin extract on the male reproductive system of Swiss albino mice. The extract was prepared with physiological saline and was injected intraperitoneally in the male mice for 7 consecutive days @ 10 mg kg-1 body weight. The short-term histopathological studies were performed by Haematoxylin and Eosin staining and the sperm morphology was analyzed by using bright-field microscopy. The snake skin extract markedly changed the gross histological architecture of the seminiferous tubules including disorganization of germinal epithelial cells, accumulation of exfoliated cells in the lumen, tailless sperm in the epididymis, disorganized basal cells in vas deferens as well as marked alteration of sperm morphology. The present work with snake shed skin strongly indicates that it is not simply a biological waste, but it might be a treasure house of many bioactive compounds.
The stability of heat-moisture treated (HMT) canna starches (starches with moisture contents of 15, 18, 20, 22, and 25%, treated at 1008C for 16 h) against acid, high shearing forces, and enzyme digestion was investigated. Micrographs of starch gels taken from a Rapid Visco Analyzer showed that granules of untreated native starch subjected to pH values of 7.2 and 4.6 were highly swelled, whereas granules exposed to higher acidic pH (3.0) values fragmented into small pieces. Yet, less swelling was found for the HTN15%, HMT18%, and HMT20% samples. Even at pH 3.0, HMT22% and HMT25% granules were still in an intact granular form. Similar findings were observed when native and HMT canna starches were agitated at various rates (160, 240, 320, and 480 rpm). Starch samples treated under higher moisture levels exhibited a higher tolerance to shearing forces acting on them. Comparative investigation of HMT and chemically crosslinked starches showed that pasting properties of HMT22% and HMT25% were equivalent to those of canna starches crosslinked by sodium trimetaphosphate at 0.2 and 0.5% (dwb), respectively. In regards to the enzyme digestibility of treated canna starches, there was no recognizable trend. The extent of starch hydrolysis (at 24 h) as a function of moisture level during HMT conformed to the following order: HMT25% > HTN15% > HMT18% % native > HMT20% > HMT22%.
The aim of the study was to evaluate the relationship between cigarette smoking and periodontal damage in terms of the levels of free radicals and antioxidants.
Thirty-five healthy subjects in the age group 25-56 yr and with chronic moderate inflammatory periodontal disease (attachment loss of 3-4 mm) were selected. All subjects were matched with respect to the clinical parameters plaque index, gingival index and attachment loss. Of the 35 subjects, 25 were smokers (smoking a minimum of 15 cigarettes/day) and 10 were nonsmokers. Smokers were subdivided into three subgroups: group I (10 subjects smoking 15-20 cigarettes/day); group II (10 subjects smoking 21-30 cigarettes/day) and group III (five subjects smoking > 50 cigarettes/day). Gingival tissue (obtained during Modified Widman surgery) and blood samples were collected from each of the subjects and analyzed for the following parameters: lipid peroxide, superoxide dismutase, catalase, glutathione and total thiol.
The level of lipid peroxide was lowest in nonsmokers (2.242 +/- 0.775 in tissue and 1.352 +/- 0.414 in blood) and highest in smokers smoking > 50 cigarettes/day (6.81 +/- 1.971 in tissue and 4.96 +/- 0.890 in blood), both in tissue and in blood. The increase was statistically significant in all groups, except in tissue of group I smokers. Catalase showed a similar trend, where the levels increased from 0.245 +/- 0.043 in controls to 0.610 +/- 0.076 in group III smokers for tissue, and from 0.231 +/- 0.040 in controls to 0.568 +/- 0.104 in group III smokers for blood. The increase was statistically significant for all groups. Total thiol levels were also higher in smokers than in controls (0.222 +/- 0.050 in controls vs. 0.480 +/- 0.072 in group III smokers in tissue; 0.297 +/- 0.078 in controls vs. 0.617 +/- 0.042 in group III smokers in blood). Except for group I in both tissue and blood, the increase was statistically significant. The superoxide dismutase (SOD) level was higher in nonsmokers (2.406 +/- 0.477 in tissue and 2.611 +/- 0.508 in blood) than in group III smokers (1.072 +/- 0.367 in tissue and 0.938 +/- 0.367 in blood), both in tissue and in blood, but this was significant only in the case of blood and for group III smokers in tissue. The glutathione level in tissue was consistently lower in smokers than in controls, showing a decrease from 121.208 +/- 37.367 in controls to 46.426 +/- 14.750 in group III smokers, but the decrease was not significant in group I smokers. In the case of blood, the glutathione level dropped from 262.074 +/- 68.751 in controls to 154.242 +/- 51.721 in group III smokers, but was statistically significant only for group III smokers.
The study results show that smoking increases the level of free radicals in periodontal tissues, which in turn may be responsible for the destruction seen in periodontal diseases.
In order to determine the toxic effect of chromium Cr(VI) on the seed germination, the root and shoot length, the root-cotyledonary leaves, the fresh and dry weight in eight-day-old seedlings Brassica oleracea L. var. acephala DC (kale) were treated with various concentrations of Cr in the growth medium. The accumulation of chromium in the tissues was determined in the cotyledons and the roots of the kale seedlings. High rate of Cr uptake was observed in the roots. But the organs could not accumulate large amount Cr. The effect of Cr on B. oleracea var. acephala was evaluated by changes in chlorophyll a, b, lipid peroxidation, proline, ascorbate, protein carbonyl groups, non-protein thiols and peroxidase activity. There were significant decreases in chlorophylls a, b content of the plants treated with Cr. Chromium treated kale seedlings had higher lipid peroxidation and the protein carbonyl groups in cotyledonary leaves than the roots. The changes refer to toxic effects of Cr. There were increases in the non-protein thiol, the total ascorbate, and proline content in the cotyledons and the roots of the seedlings grown on the media containing 0.1 and 0.15 mM Cr. The guaiacol peroxidase activity was higher in the roots of the seedlings than their cotyledons.
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.
Tissue engineering has opened up a new therapeutic avenue promising a revolution in regenerative medicine. Considerable attention has been given to chitosan composite materials and their applications in the field of the bone graft substitutes. We evaluated the antioxidative properties of chitosan-doped bioactive glass (BG-CH) with 17 wt% chitosan, and their applications in the guided bone regeneration. BG-CH was produced by a freeze-drying process and implanted in the femoral condyles of ovariectomized rats. Grafted bone tissues were carefully removed to evaluate the oxidative stress analysis, histomorphometric profile and mineral bone distribution by using inductively coupled plasma optical emission spectrometry (ICP-OES). A significant decrease of thiobarbituric acid-reactive substances (TBARs) was observed after BG-CH implantation. Superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) activities significantly increased in ovariectomized group implanted with chitosan-doped bioactive glass (OVX-BG-CH) as compared to ovariectomized group implanted with bioactive glass (OVX-BG). The histomorphometric analysis showed that bone/tissue volume (BV/TV), osteoblast number (N.Ob) and osteoblast surface/bone surface (Ob.S/ BS) were significantly higher in OVX-BG-CH group than in OVX-BG group. On the other hand, a rise in Ca and P ion concentrations in the implanted microenvironment was shown to lead to the formation/deposition of CaP phases. Trace elements such as Sr and Fe were detected in the newly formed bone and involved in bone healing. These results suggested that BG-CH composites could become clinically useful as a therapeutic and implantable material.
The two parameters of the active [methyl-3H]choline uptake into isolated rat forebrain microvessels, K, and V,,, , were determined for 1-, 3-, lo-, and 24-month-old Charles River male rats and compared with the activities of the enzymes choline acetyltransferase (ChAT), acetylcho-linesterase (AChE), and butyrylcholinesterase (BuChE) in these microvessels over the same time course. The value of K, remained constant over the entire period, but that of V,, increased from 8.5 f 1.0 to 80.6 f 16.4 nmol g-I (mean k SEM) over the first 3 months of life. Over the same period, the increase in ChAT activity, from an initial value of 7.1 f 1.6 to 10.2 f 0.3 nmol g-' min-l, was not proportional to that of choline uptake. Levels of BuChE activity (0.9-1.3 pmol g-' min-I) were almost unchanged throughout the entire 24-month period, but those of AChE showed a steady and significant increase from 1 to 24 months, remaining relatively high at senescence (4.7 pmol g-' min-I), when choline uptake had decreased to one-third of its optimal value. Selective inhibition of AChE with 1,5-bis(4-allyldimethylammonium-phenyl)pentan-3-one dibromide (0.5 p M) in unruptured capillaries from 3-month-old rats resulted in a decrease in V,, of choline uptake from-8 1 to 59 nmol g-I min-l or with 9-amino-l,2,3,4-tetrahydroacridine (10 p M) in capil-laries from 2-month-old rats from-30 to 15 nmol g-' min-I. Selective inhibition of BuChE with tetraisopropyl pyrophos-phoramide (100 p M) resulted in an increase in V,,, from-8 1 to 96 nmol g-l min-'. It is possible that the two vascular enzyme systems are coupled to a hypothetical endothelial choline transporter, but with an action opposite to each other.
The active uptake of [methyl-3H]choline into isolated rat brain microvessel suspension was studied as a likely guide to the transport of choline across the blood-brain barrier. The method consisted primarily of incubation of the suspension with a fixed concentration of labeled choline in the presence of increasing concentrations of unlabeled choline or any other inhibitor (I) of active uptake, defined as the difference in uptake at 37 degrees and 0 degrees C. From the linear regression of (1/V) against [I], the following values of Vmax (nmol g-1 min-1) and Km (microM) were obtained for choline: 2-month-old males, 10.6 +/- 3.8 and 6.1 +/- 0.9; 3-month old random females, 28.4 +/- 5.9 and 12.6 +/- 4.0; females at metaestrus, 17.8 +/- 10.3 and 8.3 +/- 5.0; at diestrus, 31.1 +/- 9.3 and 13.0 +/- 2.6; at proestrus, 54.9 +/- 2.2 and 14.0 +/- 1.5; at estrus, 19.2 +/- 2.2 and 2.6 +/- 1.7. The differences between males and random females (p less than 0.018) and between females at proestrus and estrus (p less than 0.005) are significant. It is suggested that these inter- and intrasex variations in choline uptake reflect a dynamic adjustment of supply in accordance with brain demand for choline at the time of assay. Hemicholinium-3 was an effective inhibitor of choline uptake, Ki = 14.0 +/- 8.5 microM; dimethylaminoethanol was much less effective; and imipramine had no measurable effect.
The dietary lecithin enhances thermal tolerance of Chanos chanos fingerlings reared under low dose endosulfan stress.•The critical thermal minima and lethal minima (CTmin and LTmin) were significantly enhanced in C. chanos fingerlings fed with lecithin supplementation and reared under low dose endosulfan stress.•The critical thermal maxima and lethal maxima (CTmax and LTmax) were also significantly enhanced in C. chanos fingerlings fed with lecithin supplementation and reared under low dose endosulfan stress.•Dietary lecithin also protects against antioxidative cellular stress (antioxidative enzymes) reared under low dose endosulfan stress for five weaks.•It also protectS neurotransmitter enzymes (AChE).
Growing experimental evidences have suggested the reciprocal correlation between sleep deprivation and pain. Inflammation and oxidative stress are among the key pathways underlying this correlation. Therefore, the present study was aimed to assess the effect of antioxidant and anti-inflammatory compound naringenin (NGN) against chronic sleep deprivation (CSD)-induced mechanical and thermal hyperalgesia in female Swiss albino mice. In this study, mice were chronically sleep-deprived for 8 h a day for five days a week with the weekend as a free sleep period and continued for nine weeks using a modified multiple platform method. The pain behavioral tests were conducted at the end of the fourth week to assess the development of hyperalgesia followed by the administration of NGN and a combination of NGN with Sirtinol (SIR, a sirtuin1 inhibitor) till the end of the study. After nine weeks, pain behavioral tests, along with oxidative stress and inflammatory parameters in cortex and striatum, were assessed. Results indicated that CSD-induced hyperalgesia in mice accompanied by increased oxidative stress and inflammatory markers in cortex and striatum of the brain. NGN combatted the hyperalgesic response and also decreased levels of oxidative stress and inflammatory markers. Furthermore, the pharmacological effect of NGN was mitigated with SIR. Thus, the findings of the present study reveal that NGN is acting via sirtuin1 to exert its antinociceptive activity against CSD-induced hyperalgesia.
The paper presents the results of our long-term (August 2014–October 2016) observations of changes in some properties of the poisonous secretion of eastern steppe vipers of the nominative subspecies Vipera renardi renardi (Christoph, 1861) during their postembryonic ontogenesis. The poisonous secretion of newborn vipers differed from the venom of adult snakes by an increased protease activity and the absence of any L-amino acid oxidase activity; all newborns had colorless venom. Adults produce venom either colorless, where no L-amino acid oxidase activity is detected, or yellow, where it is detected. It was found that the enzymatic activity of the venom of young vipers between their first and second winterings corresponded to the level of adults. After the second wintering, young vipers showed statistically insignificant seasonal changes in the activity of proteases and L-amino acid oxidase.
Biosynthetic pathways of phosphatidylcholine and triglyceride were studied in proliferating hepatic endoplasmic reticulum of rats pretreated with phenobarbital. Phosphatidylcholine accounted for the major increment in membrane phospholipid.
In vitro measurements of hepatic microsomal enzymes which catalyze phosphatidylcholine biosynthesis revealed a significant increase in specific activity of the enzyme governing phosphatidylcholine synthesis by sequential methylation of phosphatidylethanolamine. The specific activity of phosphorylcholine-glyceride transferase, which catalyzes phosphatidylcholine synthesis from d-1,2-diglyceride and CDP-choline, was not altered. Specific activity of diglyceride acyltransferase, which catalyzes triglyceride biosynthesis, was increased to a degree comparable to the increase in specific activity found in the phenobarbital-induced drug-metabolizing enzyme which oxidatively demethylates aminopyrine.
In vivo incorporation of methyl-³H from l-methionine-methyl-³H into microsomal phosphatidylcholine was significantly increased, resulting in an increased methyl-³H to choline-1,2-¹⁴C incorporation ratio of more than three times that found in control animals. A comparable increase in this incorporation ratio was noted in serum phospholipids.
The in vitro enzyme studies, in agreement with in vivo incorporation data, indicate that the increase in phosphatidylcholine content of phenobarbital-induced proliferating endoplasmic reticulum is related to increased activity of the pathway of phosphatidylcholine biosynthesis involving the sequential methylation of phosphatidylethanolamine.
The aim of the present study was to evaluate the effect of troxerutin (TXN) on Nickel (Ni) toxicity by using rats and in-vitro model. Ni toxicity induced in male albino wistar rats (20 mg/kg body weight (b.w) was administered orally for 20 days). TXN was administered orally (100 mg/kg (b.w) for 20 days with administration of Ni. The toxic effect of Ni and the action of TXN was measure by determining the lipid peroxidation markers and antioxidant levels in plasma and various in-vitro antioxidant systems. TXN exhibited a significant (p < 0.05) antioxidant activity in Ni induced toxicity by reversing the changes observed in TBARS, HP, Vitamin C, E and GSH. The free radical scavenging properties of TXN at different concentrations (10-50ug/mL) were investigated with various in-vitro methods such as 2, 2'-diphenyl-1- picrylhydrazyl radical (DPPH), 2, 2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) radical (ABTS•+), hydroxyl radical, superoxide anion scavenging activity and reducing power. Among the different concentrations, 50 μg/mL of TXN was more effective compared to other concentrations in all in-vitro assays. The above study conclude that TXN possesses potent in-vivo and in-vitro antioxidant activity with effective free radical scavenger for potential therapeutic value.
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