The feasibility and performance of Jicama peroxidase (JP) immobilized Buckypaper/polyvinyl alcohol (BP/PVA) membrane for methylene blue (MB) dye removal was investigated in a customized multi-stage filtration column under batch recycle mode. The effect of independent variables, such as influent flow rate, ratio of H2O2/MB dye concentration, and contact time on the dye removal efficiency, were investigated using response surface methodology (RSM). To capture the inherent characteristics and better predict the removal efficiency, a data-driven adaptive neuro-fuzzy inference system (ANFIS) is implemented. Results indicated that the optimum dye removal efficiency of 99.7% was achieved at a flow rate of 2 mL/min, 75:1 ratio of H2O2/dye concentration with contact time of 183 min. The model predictions of ANFIS are significantly good compared with RSM, thus resulting in R2 values of 0.9912 and 0.9775, respectively. The enzymatic kinetic parameters, Km and Vmax, were evaluated, which are 1.98 mg/L and 0.0219 mg/L/min, respectively. Results showed that JP-immobilized BP/PVA nanocomposite membrane can be promising and cost-effective biotechnology for the practical application in the treatment of industrial dye effluents.
A comparative study was conducted to evaluate three different permeabilization methods: FACS Permeabilizing Solution (FPerm), CytoFix/CytoPerm Kit (CFP) and Paraformaldehyde-Tween 20 (PFT) reagents, in cytoplasmic labeling of myeloperoxidase (MPO). Peripheral blood cells from 23 healthy subjects were fixed and permeabilized according to the proposed procedures, prior to direct immunofluorescence staining with CD14, CD45, IgG1, IgG2 and MPO monoclonal antibodies (McAb). Subsequent flow cytometric analysis was performed on FACSCalibur flow cytometer (Becton Dickinson, BD). As far as the antigenic expression of MPO in normal samples is concerned, FPerm and CFP demonstrated better cytoplasmic staining by inducing minor effects on light-scattering properties of the cell populations, whereas PFT-treated samples showed a diminished ability to distinguish the cell types. However, the simple and rapid FPerm method required an earlier processing of samples since the stored whole blood samples (for more than 8 hours) tended to show a significant decrease of fluorescence intensity. We also have demonstrated that P/N ratio possesses added value in evaluation of cell reactivity in immunophenotyping, based upon the apparent nonspecific cytoplasmic staining of MPO in the lymphocyte population.
Candidacidal assay was used to assess the phagocytic and killing activities of polymorphonuclear leukocytes from paediatric patients diagnosed to manifest with characteristics of atopic dermatitis. From this group of patients evaluated, all the 11 cases exhibited lower candidacidal activity in comparison to the normal children. However, there were no abnormalities observed in the engulfing abilities and myeloperoxidase activities of these leukocytes. The result indicated that lower killing capacity of polymorphonuclear leukocytes in these patients may contribute to the frequent bacterial infections in atopic dermatitis children.
Recent bioinspired efforts of designing novel nanoenzyme-based electrocatalysts are driven by the urgency of making bioelectrofuels more affordable and efficient. Unlike natural enzymes, nanoenzyme-modified electrodes with large surface areas enclose numerous biomimicking active sites to facilitate enhanced microbial growth followed by increased reactant-to-bioelectrofuel conversion.
The onset of 21st century witnessed the awareness among the masses regarding the diet-health linkages. The researchers attempted to explore traditional products/plants were in the domain of pharmacy and nutrition focussing on their health benefits. In the present research intervention, we investigate the role of Nigella sativa fixed oil (NSFO) and essential oil (NSEO) in improving antioxidant status and modulation of enzymes. The National Institute of Health (NIH) provided us 30 Sprague Dawley rats that were equally placed in three groups. The groups were fed on their respective diets (56 days) two experimental diets i.e. D2 (NSFO @ 4.0%) and D3 (NSEO @ 0.30%) and control. The indices pertaining to antioxidant status, antioxidant enzymes, and parameters pertaining to immunity were evaluated at 4 weeks interval. The experimental diets (NSFO@ 4.0% & NSEO@ 0.30%) modulated the activities of antioxidant enzymes i.e., catalase (CAT), superoxide dismutase (SOD), glutathione transferase (GST), glutathione reductase (GR) and glutathione peroxidase (GPx), positively. Indices of antioxidant status like tocopherols and glutathione were in linear relationship with that of GPx, GR and GST (P<0.01). Myeloperoxidase activities were in negative correlation with GST (P<0.01) but positive correlation with some other parameters. In the nutshell, the fixed and essential oil of Nigella sativa are effective in improving the indices pertaining to antioxidant status, however, the immune boosting potential needs further clarification. However, authors are of the view that there is need to explore the molecular targets of Nigella sativa fixed and essential oils. Findings from such studies would be useful to validate this instant study for health promoting potential against diabetes mellitus and cardiovascular disorders.
Chemical synthesis of Ag-NPs was carried out using reduction method. The reduction mechanistic approach of silver ions was found to be a basic clue for the formation of the Ag-NPs. The nanoparticles were characterized by UV-vis, FT-IR and TEM analysis. We had designed some experiments in support of our hypothesis, "low concentrations of novel nanoparticles (silver and gold) increases the activity of plant peroxidases and alter their structure also", we had used Ag-NPs and HRP as models. The immobilization/interaction experiment had demonstrated the specific concentration range of the Ag-NPs and within this range, an increase in HRP activity was reported. At 0.08 mM concentration of Ag-NPs, 50% increase in the activity yield was found. The U.V-vis spectra had demonstrated the increase in the absorbance of HRP within the reported concentration range (0.06-0.12 mM). Above and below this concentration range there was a decrease in the activity of HRP. The results that we had found from the fluorescence spectra were also in favor of our hypothesis. There was a maximum increase in ellipticity and α-helix contents in the presence of 0.08 mM concentration of Ag-NPs, demonstrated by circular dichroism (CD) spectra. Finally, incubation of a plant peroxidase, HRP with Ag-NPs, within the reported concentration range not only enhances the activity but also alter the structure.
Biomarkers play a pivotal role in the diagnosis and management of patients with acute coronary syndrome. This study aimed to investigate the differences in level of several biomarkers, i.e. C-reactive protein, myeloperoxidase, soluble CD40 ligand and placental growth factor, between acute coronary syndrome and chronic stable angina patients. The relationship between these biomarkers in the coronary circulation and systemic circulation was also investigated.
A study was undertaken to evaluate the ability of flow cytometric analysis of intracellular myeloperoxidase (MPO) in differentiating populations of lymphocytes (L), monocytes (M) and granulocytes (G), by means of lysed whole blood method. Anticoagulated blood from 23 normal individuals was lysed with FACS lysing solution and permeabilized with FACS permeabilizing solution before subjected to direct immunofluorescence staining. The geometric means of the fluorescence intensity were measured using FACSCalibur flow cytometer (Becton Dickinson). Populations of L, M and G were gated based on their light scatter characteristics and expression of CD14 and CD45. Then, the fluorescence intensity of MPO expression was studied in these individual cell populations. The results showed that fluorescence intensity of MPO was the strongest in G and weakest in L, whereas M showed intermediate fluorescence intensity. Our findings reveal that discrimination of these three cell types is achievable based upon the sole expression of intracellular MPO.
The specificity and sensitivity of an indirect and two (an 'ordinary' and a 'rapid') double sandwich enzyme-linked immunosorbent assay (ELISA) procedures for the quantitation of Calloselasma rhodostoma (Malayan pit viper) venom were examined. The three assays were equally sensitive and the accuracy of the assays was not substantially affected by individual variation in the venom composition. The specificity of the assays was examined against 26 venoms from snakes of the families Viperidae and Elapidae. While the double sandwich ELISA procedures were sufficiently specific to be used in the clinical immunodiagnosis of C. rhodostoma bite in Malaysia, the indirect ELISA procedure exhibited extensive cross-reactivity with other Malaysian pit viper venoms. Attempts were made to improve the specificity of the indirect ELISA procedure for the quantitation of C. rhodostoma venom. A 'low ELISA cross-reactivity' venom fraction (termed VF52) was isolated from C. rhodostoma venom by repeated Sephadex G-100 gel filtration chromatography. The indirect ELISA procedure using antibodies to VF52 as immunoreagent showed an improvement in specificity. The use of the indirect ELISA procedure for the detection of C. rhodostoma antibodies was also examined and the results show that the assay was sufficiently specific to be used for retrospective diagnosis of C. rhodostoma bite in Malaysia, in particular when VF52 was used as the coating antigen.
Composition, physicochemical properties and enzyme inactivation kinetics of coconut water were compared between immature (IMC), mature (MC) and overly-mature coconuts (OMC). Among the samples studied, pH, turbidity and mineral contents for OMC water was the highest, whereas water volume, titratable acidity, total soluble solids and total phenolics content for OMC water were the lowest. Maturity was found to affect sugar contents. Sucrose content was found to increase with maturity, and the reverse trend was observed for fructose and glucose. Enzyme activity assessment showed that polyphenol oxidase (PPO) in all samples was more heat resistant than peroxidase (POD). Compared to IMC and MC, PPO and POD from OMC water showed the lowest thermal resistance, with D83.3°C=243.9s (z=27.9°C), and D83.3°C=129.9s (z=19.5°C), respectively.
Exercise is beneficial to health, but during exercise the body generates reactive oxygen species (ROS) which are known to result in oxidative stress. The present study analysed the effects of vitamin E (Tri E®) on antioxidant enzymes; superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (Cat) activity and DNA damage in rats undergoing eight weeks exercise.
The efficiency of advanced oxidation processes (AOPs), enzymatic treatment and combined enzymatic/AOP sequences for the colour remediation of soda and chemimechanical pulp and paper mill effluent was investigated. The results indicated that under all circumstances, the AOP using ultraviolet irradiation (photo-Fenton) was more efficient in the degradation of effluent components in comparison with the dark reaction. It was found that both versatile peroxidase (VP) from Bjerkandera adusta and laccase from Trametes versicolor, as pure enzymes, decolorize the deep brown effluent to a clear light-yellow solution. In addition, it was found that in the laccase treatment, the decolorization rates of both effluents were enhanced in the presence of 2, 2'-azinobis (3-ethylbenzthiazoline-6-sulfonate), while in the case of VP, Mn(+2) decreased the efficiency of the decolorization treatment. The concomitant use of enzymes and AOPs imposes a considerable effect on the colour remediation of effluent samples.
Myeloperoxidase (MPO) is present in azurophilic granules which appear in the promyelocyte stage of differentiation and is expressed in granulomonocytic cells. MPO is usually detected by cytochemistry. The demonstration of peroxidase in at least 3% of bone marrow blasts defines an acute leukaemia as acute myeloblastic leukaemia (AML). MPO is important in distinguishing acute myeloblastic leukaemia (AML) from acute lymphoblastic leukaemia (ALL). It is difficult to diagnose AML with minimal evidence of myeloid differentiation (AML- M0) by conventional light microscopy. However, these AML-M0 blasts can be detected by monoclonal antibodies. Anti-MPO recognizes the enzymatically inactive precursor forms of MPO. There are a few commercially available monoclonal antibodies against MPO. In this study, we evaluated two monoclonal antibodies against MPO from different commercial sources.
1. The hypotensive effect of cross-fostering in spontaneously hypertensive rats (SHR) is thought to involve adjustments in renal function. However, its association with renal anti-oxidant/oxidant balance during cross-fostering is not known. 2. The present study examined the effect of cross-fostering and in-fostering of 1-day-old offspring between SHR and Wistar-Kyoto (WKY) dams on renal anti-oxidant/oxidant status and systolic blood pressure (SBP). Renal anti-oxidant/oxidant status and SBP were determined in the offspring from 4-16 weeks of age. 3. Cross-fostered SHR had significantly lower SBP than in-fostered SHR at 6, 8 and 12 weeks, but not at 16 weeks (127 ± 1 vs 144 ± 2, 138 ± 1 vs 160 ± 1, 174 ± 2 vs 184 ± 2 and 199 ± 2 vs 194 ± 3 mmHg at 6, 8, 12 and 16 weeks, respectively). No differences in SBP were evident between cross-fostered and in-fostered WKY rats. There were no significant differences in levels of thiobarbituric acid-reactive substances (TBARS), protein carbonyl and total anti-oxidant status (TAS) or superoxide dismutase, catalase, glutathione peroxidase (GPx), glutathione S-transferase and glutathione reductase activity between cross-fostered and in-fostered SHR or WKY offspring. However, compared with WKY rats, catalase activity was higher at 6 and 16 weeks, TAS was higher at 16 weeks and GPx activity and TBARS were lower at 16 weeks in SHR. 4. It appears that cross-fostering of SHR offspring to WKY dams during the early postnatal period causes a transient delay in the rise in blood pressure in SHR and that this does not involve the renal anti-oxidant/oxidant system.
This study was conducted to investigate the influence of formulation development methods on the stability (secondary structure, aggregation, and biological activity) of protein drugs embedded in lipid matrices. Catalase, horseradish peroxidase, and α-chymotrypsin were employed as model proteins, while Precirol® AT05 (glyceryl palmitostearate) was used as lipid matrix. Protein-loaded lipid matrices were prepared using melting and mixing and wet granulation methods. Attenuated total reflectance Fourier transform infrared (ATR FT-IR) spectroscopy, size exclusion chromatography (SEC) and biological activity analyses were performed. ATR FT-IR analysis indicated significant interference of the lipid with the protein amide-I band, which was eliminated using spectral subtraction. Wet granulation method induced more changes in protein secondary structure compared to melting and mixing method. SEC analysis gave evidence of protein aggregation for catalase upon adopting the wet granulation method. The biological activity of catalase was found to reduce significantly than other two proteins upon using wet granulation method, which might be ascribed to both secondary structure alterations and the formation of aggregates. Horseradish peroxidase and α-chymotrypsin did not form any soluble aggregates. In conclusion, melting and mixing method emerged as a better incorporation method compared to wet granulation because of better stability shown by the formulated proteins.
Collar rot of chili (Capsicum annuum L.) is a very destructive disease caused by a soil-borne fungal pathogen Sclerotium rolfsii Sacc. Generally, chemical fungicides are used to combat the menace but this practice is being discouraged because of health and environmental concerns. In the present study, an alternative environment friendly strategy was used to manage this disease by using farmyard manure (FYM) and two commercial biofertilizers namely Biopower and Feng Shou. S. rolfsii inoculated pot soil was amended with 1% and 2% FYM and the two commercial biofertilizers. Inoculation of soil with S. rolfsii only (positive control) resulted in the highest disease incidence (73%) and plant mortality (60%). Biopower and Feng Shou application reduced disease incidence to 20% and 7%, respectively and plant mortality to 0%. Likewise, 1% and 2% FYM amendment reduced disease incidence to 33% and plant mortality to 26% and 7%, respectively. Under biotic stress of S. rolfsii, FYM and biofertilizers applications, either alone or in combination, significantly enhanced root and shoot growth over positive control. S. rolfsii inoculation significantly increased peroxidase and polyphenol oxidase activities in chili plants which were further increased by application of either of the two biofertilizers. The present study concludes that biofertilizers Biopower and Feng Shou alone or in combination with 2% FYM can be effectively utilized to manage southern blight of chili.
The genus Streptomyces demonstrates enormous promise in promoting plant growth and protecting plants against various pathogens. Single and consortium treatments of two selected Streptomyces strains (Streptomyces shenzhenensis TKSC3 and Streptomyces sp. SS8) were evaluated for their growth-promoting potential on rice, and biocontrol efficiency through induced systemic resistance (ISR) mediation against Xanthomonas oryzae pv. oryzicola (Xoc), the causal agent of rice bacterial leaf streak (BLS) disease. Seed bacterization by Streptomyces strains improved seed germination and vigor, relative to the untreated seed. Under greenhouse conditions, seed bacterization with consortium treatment TKSC3 + SS8 increased seed germination, root length, and dry weight by 20%, 23%, and 33%, respectively. Single and consortium Streptomyces treatments also successfully suppressed Xoc infection. The result was consistent with defense-related enzyme quantification wherein single and consortium Streptomyces treatments increased peroxidase (POX), polyphenol oxidase, phenylalanine ammonia-lyase, and β,1-3 glucanase (GLU) accumulation compared to untreated plant. Within all Streptomyces treatments, consortium treatment TKSC3 + SS8 showed the highest disease suppression efficiency (81.02%) and the lowest area under the disease progress curve value (95.79), making it the best to control BLS disease. Consortium treatment TKSC3 + SS8 induced the highest POX and GLU enzyme activities at 114.32 μmol/min/mg protein and 260.32 abs/min/mg protein, respectively, with both enzymes responsible for plant cell wall reinforcement and resistant interaction. Our results revealed that in addition to promoting plant growth, these Streptomyces strains also mediated ISR in rice plants, thereby, ensuring protection from BLS disease.
Though fresh-cut products save our time, but they are very much prone to enzymatic browning that drastically affects product's quality and marketability. Drumstick pods are considered as super food due to high nutritional contents. However, the fresh-cut pods are prone to brown discoloration. The enzyme activities promote the softening and cut-surface browning of pods, thus deteriorates their texture, decreases consumer appeal and shortens the shelf life. So, we aimed to assess the effect of citric (1%) and ascorbic (1%) acid treatments on quality attributes of fresh-cut drumsticks at 3-d interval during storage (5 ± 1 °C). In general there was an increase in lignin and quinone contents, while phenolic content was decreased during storage. However, samples subjected to ascorbic acid dip had higher phenolic content, lower rate of lignin formation, and reduced membrane permeability. Enzyme activities (polyphenol oxidase and peroxidase) were found to increase during storage, however, samples treated with ascorbic acid showed lower activities than that of the control and citric acid treated samples. The reduced enzyme activities resulted in the reduced browning incidence and maintained the quality. Therefore, postharvest dip of fresh-cut drumstick in to ascorbic acid (1%) could be suggested to increase the shelf life with reduced browning during low temperature storage.
Treatment with hypovirulent binucleate Rhizoctonia (HBNR) isolates induced systemic resistance against anthracnose infected by Colletotrichum orbiculare in cucumber, as there were no direct interaction between HBNR and C. orbiculare. This is because of the different distances between HBNR and C. orbiculare, where the root was treated with HBNR isolate and C. orbiculare was challenged and inoculated in leaves or first true leaves were treated with HBNR isolate and C. orbiculare was challenged and inoculated in second true leaves. The use of barley grain inocula and culture filtrates of HBNR significantly reduced the lesion diameter compared to the control (p = 0.05). The total lesion diameter reduction by applying barley grain inoculum of HBNR L2, W1, W7, and Rhv7 was 28%, 44%, 39%, and 40%, respectively. Similar results was also observed in treatment using culture filtrate, and the reduction of total lesion diameter by culture filtrate of HBNR L2, W1, W7, and Rhv7 was 45%, 46%, 42%, and 48%, respectively. When cucumber root was treated with culture filtrates of HBNR, the lignin was enhanced at the pathogen penetration, which is spread along the epidermis tissue of cucumber hypocotyls. Peroxidase activity in hypocotyls in the treated cucumber plant with culture filtrates of HBNR significantly increased before and after inoculation of pathogens as compared to the control. Significant enhancement was also observed in the fast-moving anodic peroxidase isozymes in the treated plants with culture filtrates of HBNR. The results showed the elicitor(s) contained in culture filtrates in HBNR. The lignin deposition as well as the peroxidase activity is an important step to prevent systemically immunised plants from pathogen infection.