H2O2 Assay Research Articles

In vitro antioxidant and antinociceptive properties of Porphyra vietnamensis.

Background: Current investigation explores the anti-oxidant and antinociceptive potential of edible seaweed namely Porphyra vietnamensis.Methods: Radical scavenging and antinociceptive potential of ethanolic (EE), aqueous (AE), acetone (ACE) and chloroform (CE) fractions were determined using various models and assays. Writhing, formalin, hot plate, acetic acid induce response models were performed to determine antinociceptive activity whereas different assays have been used to determine antioxidant potential.Results: Among the various fractions, ACE showed maximum biological activity. In DPPH assay half maximal inhibitory concentration (IC50) was found to be 0.470 ug/ml (DPPH assay), 0.381 jug/m/ (H2O2 assay), 0.470 ug/ml (super oxide assay), 0.591ug/ml (lipid peroxidation) and 0.430 ug/ml (nitric oxide assay). However, comparatively the TPC was more in EE (977.0 mg GAE/gm DW).Conclusion: It was concluded that acetone fraction of Porphyra showed marked antinociceptive and antioxidant activities, however pharmacological and chemical investigations are required to identify principle compounds responsible for activities and characterize their respective mechanism(s) for respective actions.

BENEFICIAL OUTCOMES AFTER COMBINED THERAPY WITH AZATHIOPRINE AND PIOGLITAZONE IN PRECLINICAL ARTHRITIC MODELS

Objectives: The present study was designed to investigate the antioxidant and anti-arthritic activities of azathioprine-pioglitazone combination therapy to check whether it has an add on benefit over monotherapy with azathioprine or pioglitazone on disease activity in rodent models.
 Methods: The antioxidant activity of test drugs and their combination was screened by H2O2 and nitric oxide scavenging assays. They were further evaluated for anti-arthritic activity using in vitro models such as protein denaturation and membrane stabilization and in vivo methods such as formaldehyde and complete Freund’s adjuvant (CFA)-induced arthritis.
 Results: The combination of test drugs showed better inhibition of free radicals in both H2O2 and nitric oxide scavenging assay than individual counterparts revealing its potential antioxidant activity. They also showed significant inhibition of protein denaturation and proliferation of the red blood cell in in vitro models. The test drugs showed significant inhibition of the paw volume in both the formaldehyde and CFA-induced arthritis along with reverting the altered biochemical parameters. These findings were corroborated by radiological and histopathological studies.
 Conclusions: Combination of azathioprine and pioglitazone exhibited better antioxidant and anti-arthritic effect than the individual drugs showing synergistic interaction between them.

Etched PtCu nanowires as a peroxidase mimic for colorimetric determination of hydrogen peroxide

Etched PtCu nanowires (NWs) were synthesized by a hydrothermal reaction and chemical etching process. The NWs are shown to be viable peroxidase (POx) mimics capable of catalyzing the oxidation of the substrate 3,3',5,5'-tetramethylbenzidine (TMB) in the presence of H2O2 to form a blue-green coloration. The mechanism of catalysis was investigated and the results demonstrated that H2O2 is decomposed to form hydroxyl radicals which oxidize TMB in the presence of the NWs. Under optimized conditions, a steady-state kinetic analysis revealed that the NWs possess a stronger affinity for H2O2 and TMB compared to the enzyme horseradish POx. Based on the high POx-like activity, a colorimetric assay for H2O2 was established. Absorbance at 652 nm increases linearly in the 0.1-300 μM H2O2 concentration range, and the detection limit is 0.06 μM (at S/N = 3). The assay was successfully applied to the determination of H2O2 in (spiked) milk and contact lens solution. Furthermore, a highly sensitive test strip was designed which represents a low cost and fast alternative for the visual determination of H2O2. Graphical Abstract Schematic presentation of the colorimetric detection of H2O2. PtCu nanowires (PtCu NWs) can catalyze 3,3',5,5'-tetramethylbenzidine (TMB) oxidation by H2O2 to produce blue-green oxidized 3,3',5,5'-tetramethylbenzidine (oxTMB). Based on the color change, test strips were designed for H2O2 detection.

Investigating the Effects of Stove Emissions on Ocular and Cancer Cells

More than a third of the world’s population relies on solid fuels for cooking and heating, with major health consequences. Although solid fuel combustion emissions are known to increase the prevalence of illnesses such as chronic obstructive pulmonary disease and lung cancer, however, their effect on the eyes is underexplored. This study assesses the acute toxicity of solid fuel combustion emissions on healthy ocular cells and a cancer cell line. Three healthy ocular cell lines (corneal, lens, and retinal epithelial cells) and a cancer cell line (Chinese hamster ovary cells) were exposed to liquid and gas phase emissions from applewood and coal combustion. Following the exposure, real-time cell attachment behavior was monitored for at least 120 hours with electrical cell impedance spectroscopy. The viability of the cells, amount of apoptotic cells, and generation of reactive oxygen species (ROS) were quantified with MTT, ApoTox-Glo, and ROS-Glo H2O2 assays, respectively. The results showed that coal emissions compromised the viability of ocular cells more than applewood emissions. Interestingly, the cancer cells, although their viability was not compromised, generated 1.7 to 2.7 times more ROS than healthy cells. This acute exposure study provides compelling proof that biomass combustion emissions compromise the viability of ocular cells and increase ROS generation. The increased ROS generation was fatal for ocular cells, but it promoted the growth of cancer cells.

Apoptotic effects of cigarette smoke extracts on mouse embryonic stem cells via oxidative stress.

Previous studies have reported that cigarette smoke and cigarette smoke extract (CSE) have negative effects on embryonic development. However, no studies have investigated the mechanism through which CSE affects the cellular signaling pathway leading to apoptosis and oxidative stress in embryonic cells, or how the two pathways are cross-linked. Thus, we studied the effects of CSE on apoptosis and oxidative stress in mouse embryonic stem cells (mESCs). Specifically, we measured changes in cell viability in response to CSEs (3R4F and two domestic cigarettes CSE 1 and 2) using a water soluble tetrazolium (WST) assay and a neutral red uptake (NRU) assay, which revealed that cell viability decreased in a concentration-dependent manner. Western blot analysis revealed that the expression of cyclin D1 and cyclin E1 was decreased and that of p21 and p27 was increased by CSE. Additionally, the number of terminal deoxynucleotidyl transferase (TUNEL)-stained cells was increased by CSE, while the levels of Bax and Caspase-3 increased and Bcl-2 decreased. Moreover, a 2',7'-dichlorofluorescin diacetate (DCF-DA) assay and reactive oxygen species (ROS)-Glo H2 O2 assay confirmed that ROS were generated in response to CSE and that they were associated with up-regulated Keaf-1 and CHOP. Overall, the results revealed that cigarette smoke extract (CSE) inhibited cell proliferation by regulating cell cycle-related protein expression and increased oxidative stress by regulating the expression of Kelch-like ECH-associated protein 1 (Keap-1) and CCAAT/enhancer-binding protein homologous protein (CHOP), resulting in apoptosis in mESCs.

CuO nanoparticles as haloperoxidase-mimics: Chloride-accelerated heterogeneous Cu-Fenton chemistry for H2O2 and glucose sensing

CuO nanoparticles (CuO NPs) were demonstrated as a novel nanoenzyme, i.e., haloperoxidase mimic, which catalyzes the oxidation of halide ions to form reactive halogen species (RHS). Two functions of CuO NPs have been demonstrated in the work. First, RHS halogenate bacterial signaling molecules, thus preventing biofilm formation. Second, CuO is used in sensitive colorimetric assays for H2O2 and glucose.

Bifunctional gold nanoclusters enable ratiometric fluorescence nanosensing of hydrogen peroxide and glucose

The accurate quantification of hydrogen peroxide (H2O2) and glucose is essential significance in clinical diagnosis. Herein a selective and sensitive ratiometric fluorescent nanosensor was developed for the determination of H2O2 and glucose by integrating peroxidase–like catalytic and fluorescent bifunctional properties of glutathione protected gold nanoclusters (GSH–AuNCs). The GSH–AuNCs exhibit inherent peroxidase–like activity and accelerate the decomposition of H2O2 into hydroxyl radicals. The produced hydroxyl radicals oxidize terephthalic acid (TA), a typical non–fluorescent substrate of peroxidase, to a highly fluorescent product hydroxyterephthalate (TAOH). Upon excitation with single–wavelength at 315 nm, dual–emission fluorescence peaks were recorded at 430 and 600 nm simultaneously. The fluorescence signal of TAOH at 430 nm continuously increased with increasing the concentration of H2O2 while the fluorescence signal of GSH–AuNCs at 600 nm remained unchangeable. Based upon on these facts, a ratiometric fluorescent nanosensor was fabricated for H2O2 assay with TAOH as response unit and GSH–AuNCs as reference, respectively. By converting glucose into H2O2 with catalytic oxidation of glucose oxidase (GOx), this nanosensor was further exploited for glucose assay. Under the optimum conditions, the detection limits of 10 nmol/L H2O2 and 20 nmol/L glucose were acquired. The relative standard deviations were less than 5% for both H2O2 and glucose (5.0 μmol/L solution, n = 11). The practicability of the nanosensor was verified by the determination of glucose in human serum samples. This nanosensor can be easily expanded as a general platform for the detection of other substances involving H2O2 produced or consumed.

Synthesis of zinc oxide nanoparticles (ZnO NPs) using pure bioflavonoid rutin and their biomedical applications: antibacterial, antioxidant and cytotoxic activities

The present study reports an eco-friendly green synthesis of zinc oxide nanoparticles (ZnO NPs) using the bioflavonoid rutin. The synthesized ZnO NPs were characterized by UV–visible spectroscopy, XRD, FE-SEM, EDX, FTIR and zeta potential analyses. FE-SEM image showed that the synthesized ZnO NPs were rod shaped. FTIR spectral studies confirmed the role of bioflavonoid rutin for the reduction, capping and synthesis of ZnO NPs. The green synthesized ZnO NPs showed significant antibacterial activity against both Gram-positive (B. subtilis and S. aureus) and Gram-negative (K. pneumoniae and E. coli) bacterial pathogens. However, the synthesized ZnO NPs were more effective against Gram-negative bacteria compared to Gram-positive bacteria. The in vitro antioxidant ability of ZnO NPs was investigated spectrophotometrically using DPPH and H2O2 assay. The percentage of antioxidant activity increased with increasing concentration of ZnO NPs. Furthermore, synthesized ZnO NPs showed effectiveness in inhibiting the human MCF-7 breast cancer cells. In conclusion, the synthesized ZnO NPs using bioflavonoid rutin might be used as a strong biocidal and antioxidant agent in biomedical and pharmaceutical industries.

Ethnobotanical, Phytochemical, and Biological Study of Tamarix aphylla and Aerva javanica Medicinal Plants Growing in the Asir Region, Saudi Arabia

In this study, Tamarix aphylla and Aerva javanica, plants used in folkloric medicine in the Asir region of Saudi Arabia were studied ethnobotanically, chemically, and biologically to assess their medicinal uses, phytochemical constituents, and biological activities. Ethnobotanical data were collected using semistructured interviews and the use values were calculated. A total of 61 informants were interviewed and results indicated high-value uses (0.9 and 0.7) for the plants. Phytochemical investigation indicated that the plants contained most of the phytochemicals that were tested for. The antioxidant activity was examined by assaying total phenolic content, 1,1-Diphenyl-2-picrylhydrazyl (DPPH), and hydrogen peroxide (H2O2) radical scavenging activity. .The bark extract of T. aphylla showed the best antioxidant effect in total phenolic content, DPPH, and H2O2 assays (278.02 ± 0.16 mg GAE/100 g; The half-maximal inhibitory concentration (IC50): DPPH, 18.39 ± 0.62 μg/mL; H2O2, 252.94 ± 1.86 μg/mL), followed by extract of the A. javanica aerial parts (228.60 ± 2.09 mg GAE/100 g; IC50: DPPH, 28.54 ± 0.53 μg/mL; H2O2, 154.17 ± 0.78 μg/mL) compared with ascorbic acid (DPPH: 27.27 ± 0.11 μg/mL; H2O2: 164.9 ± 0.37 μg/mL). The disc diffusion method performed for antimicrobial activity revealed weak activities in T. aphylla and A. javanica extracts (100 μg/mL) against six human pathogens. It was concluded that these plants possess therapeutic potential in the treatment of 18 types of ailments, which are important phytochemical constituents and antioxidant activities that justify their therapeutic uses in traditional medicine. Thus, this study laid sufficient background for further pharmacological research on extracts of these plants.

From Waste to Profit : Assessment of α-Amylase Inhibitory and Antioxidant Activities Along with the development of an Active Packaging Film for Valorization of Pea peels

The present study aimed to valorize pea peels (Pisum sativum) using new unusual aspects. In other words, to identify and quantify the phenolic profile of pea peels 80 % ethanol extract (PEE), to estimate the antioxidant and anti-α-amylase activities of successive extraction solvent fractions of the previously mentioned extract, to evaluate chitosan(Cs) / polyvinyl alcohol (PVA) / PEE edible film as antioxidant agent. The results exposed that ethyl acetate fraction had the highest amounts of polyphenols and flavonoids where their values were 165.43 mg GAE/g and 26.63 mg QE/g ,respectively. Hesperidin was documented as the most abundant polyphenol in pea peels ethanolic extract with remarkably high amount (2017.2µg/g). Butanol fraction recorded considerable anti-α-amylase activity (IC50 = 1.61 mg/ml). The present study extremely evidenced the antioxidant activity of pea peel extracts. Besides , ethyl acetate fraction had the lowest IC50 values for DPPH, NO and H2O2 assays with the values of 167.23, 302.36 and 317.66 μg/ml, respectively. Interestingly, a significant and negative correlation was remarked between total flavonoids content and IC50 for the scavenging DPPH radical. Finally , it could be suggested that Cs/PVA film containing PEE can be used for development of active food packaging materials.

Microfabricated needle for hydrogen peroxide detection.

A microfabricated needle-like probe has been designed and applied for hydrogen peroxide (H2O2) sampling and detection using a commercial, single-step fluorescent H2O2 assay. In this work, droplets of the assay reagent are generated and sent to the needle tip using a mineral-oil carrier fluid. At the needle tip, the sample is drawn into the device through 100 μm long hydrophilic capillaries by negative pressure. The sampled fluid is immediately merged with the assay droplet and carried away to mix and react, producing a sequence of droplets representing the H2O2 concentration as a function of time. We have characterized the assay fluorescence for small variations in the sample volume. With the calibration, we can calculate the concentration of H2O2 in the sampled liquid from the size and intensity of each merged droplet. This is a microfluidic data-logger system for on-site continuous sampling, controlled reaction, signal storage and on-line quantitative detection. It is a useful tool for monitoring dynamic chemical reactions in analytical chemistry and biological applications.

Magnetic-core@dual-functional-shell nanocomposites with peroxidase mimicking properties for use in colorimetric and electrochemical sensing of hydrogen peroxide.

A self-sacrificing catalytic method is described for the preparation of magnetic core/dual-functional-shell nanocomposites composed of magnetite, gold and Prussian Blue (type Fe3O4@Au-PB). Two reaction pathways are integrated. The first involves chemical dissolution of Fe3O4 (the self-sacrificing step) by acid to release ferrous ions which then reacts with hexacyanoferrate(IV) to generate PB in the proximity of the magntic nanoparticles (MNPs). The second involves the reduction of tetrachloroaurate by hydroxylamine to generate gold under the catalytic effect of the MNPs. At the end, the MNPs@Au-PB nanocomposite is formed. This method exploits both the chemical reactivity and catalytic effect of the MNPs in a single step. The multi-function material was applied (a) in an optical assay for H2O2; (b) in an amperometric assay for H2O2; (c) in an enzymatic choline assay using immobilized choline oxidase. The limit of electrochemical detection of H2O2 (at a potential as low as 50mV) is 1.1μM which is comparable or better than most analogous methods. The sensors display superior performance compared to the use of conventional core@single-shell (MNPs@PB) nanomaterials. Graphical abstract A self-sacrificing catalytic method is described to prepare magnetic core/dual-functional-shell nanocomposites composed of magnetic nanoparticle, gold and Prussian Blue (type MNP@Au-PB). The nanocomposites work well as candidates to develop colorimetric and electrochemical sensors of H2O2 with superior performance to analogues.

Fe3 O4 nanoparticles mediated synthesis of novel spirooxindole-dihydropyrimidinone molecules as Hsp90 inhibitors.

Heat shock protein 90 (Hsp90) is a validated molecular chaperone considered as the new key recipient for cancer intervention. The current study illustrates the synthesis of novel spirooxindole-dihydropyrimidinones (4a-j) by Fe3 O4 nanoparticles intervened synthesis and their Hsp90 ATPase inhibitory activity was investigated by the malachite green assay. All the compounds in the study demonstrated a moderate to potent ATPase inhibitory profile, with IC50 values ranging from 0.18 to 6.80 μM. Compounds 4j, 4h, 4f, and 4i exhibited maximum inhibitory potential with IC50 values of 0.18, 0.20, 0.35, and 0.55 μM, respectively. They were found to be better than the standard drug, geldanamycin (Hsp9 ATPase inhibition IC50 = 0.90 μM). Compounds 4h and 4j with IC50 values of 22.82 ± 0.532, 20.78 ± 0.234 and 21.32 ± 0.765, 28.43 ± 0.653 µM showed significantly greater potencies against the MCF-7 and HepG2 cell lines, respectively. Compound 4j showed good antioxidant activities in the DPPH test and H2 O2 assay (IC50 = 20.13.23 ± 0.32 and 23.27 ± 0.32 μg/mL) when compared with the standard ascorbic acid (IC50 = 19.16 ± 0.20 and 20.66 ± 1.09 μg/mL). A molecular docking study was performed to observe the binding efficiency and steric interactions of the lead moiety.

Evaluation of the Cytotoxic and Antioxidant Activity of Phyto-synthesized Silver Nanoparticles Using Cassia angustifolia Flowers

The present study reports an eco-friendly phyto-synthesis of silver nanoparticles (AgNPs) using aqueous flower extract of Cassia angustifolia. Preliminarily, the synthesis of AgNPs from flower extract was visually confirmed by color change. Further formation, shape, size, and stability of the synthesized AgNPs were characterized by UV-visible spectroscopy, SEM, EDX, XRD, FT-IR, DLS, and zeta potential analyses. SEM images showed that the synthesized AgNPs were spherical in shape with an average size of 10–80 nm. Phyto-chemical analysis and FT-IR studies confirmed the role of phyto-compounds in the flower extract for the capping, formation, reduction, and stabilization of AgNPs. The antioxidant ability of AgNPs and plant extract was evaluated by DPPH, H2O2, and FRAP assays. The percentage of antioxidant activity was increased with increasing concentration of AgNPs. In addition, cytotoxic activity of the AgNPs was evaluated in human breast cancer cells (MCF 7). Phyto-synthesized AgNPs showed dose-depended manner (IC50 − 73.82 ± 0.50 μg/mL) of cytotoxic activity against MCF 7 cancer cells.

Enhanced Peroxidase-Like Activity of MoS₂ Quantum Dots Functionalized g-C₃N₄ Nanosheets towards Colorimetric Detection of H₂O₂.

MoS2 quantum dots (QDs) functionalized g-C3N4 nanosheets (MoS2@CNNS) were prepared through a protonation-assisted ion exchange method, which were developed as a highly efficient biomimetic catalyst. Structural analysis revealed that uniformly-dispersed MoS2 QDs with controllable size and different loading amount grew in-situ on the surface of CNNS, forming close-contact MoS2@CNNS nanostructures and exhibiting distinct surface properties. Compared to MoS2 QDs and CNNS, the MoS2@CNNS nanocomposites exhibited a more than four times stronger peroxidase-like catalytic activity, which could catalyze the oxidation of 3,3’,5,5’-tetramethylbenzidine (TMB) in the presence of H2O2 to generate a blue oxide. Among the MoS2@CNNS nanocomposites, MoS2@CNNS(30) was verified to present the best intrinsic peroxidase-like performance, which could be attributed to the more negative potential and larger specific surface area. A simple, rapid and ultrasensitive system for colorimetric detection of H2O2 was thus successfully established based on MoS2@CNNS, displaying nice selectivity, reusability, and stability. The detection limit of H2O2 could reach as low as 0.02 μM. Furthermore, the kinetic and active species trapping experiments indicated the peroxidase-like catalytic mechanism of MoS2@CNNS. This work develops a novel, rapid, and ultrasensitive approach for visual assay of H2O2, which has a potential application prospect on clinical diagnosis and biomedical analysis.

Exploring Planococcus sp. TRC1, a bacterial isolate, for carotenoid pigment production and detoxification of paper mill effluent in immobilized fluidized bed reactor

Pulp and paper industry is one of the high priority sectors that generates large amount of solid (sludge) and liquid (paper mill effluent) wastes. Making use of this huge quantity of paper mill sludge (PMS) in an environment friendly manner is challenging and cost involving. Similarly the present wastewater treatment strategies based on physical and chemical methods encounter certain serious setbacks including secondary pollution generation with expensive mode of abatement processes. The present study aims at a “waste treats waste” strategy for the bioremediation of paper mill effluent in fluidized bed reactor (FBR) using another waste of the same industry, PMS as immobilizing matrix for Planococcus sp. TRC1, a wastewater bacterial isolate. This study simultaneously explores this isolate for the yellowish orange pigment it produces (2.3 ± 0.2 mg/gm of dry bacterial biomass) and characterizes it as a member of the pharmacologically important carotenoid pigment family via UV-Vis spectrophotometry, TLC, FT-IR and 13C NMR. The antioxidant potential of this pigment was studied by DPPH assay (IC50 = 33 ± 0.4 μg/ml) and H2O2 assay (IC50 = 147.4 ± 2.2 μg/ml). In FBR, the PMS immobilized bacteria showed removal of phenol, lignin, colour and COD from the effluent by 96%, 74%, 81% and 85% respectively after 60hr of treatment. The experimental data on immobilization fitted well with pseudo second-order (R2 = 0.955) and Freundlich adsorption isotherm (R2 = 0.996) models. The alterations in PMS before and after bacterial immobilization, as revealed by SEM and FT-IR, depicted the success of PMS as immobilization matrix. Phytotoxicity (90% seed germination) and mutagenicity studies confirmed that the treated effluent was substantially less toxic than its raw state. This study highlights a novel utilization possibility of PMS in an eco-friendly and economic way as immobilization matrix for Planococcus sp.TRC1 for paper pulp mill effluent treatment along with production of carotenoid pigment from this potential bacterial isolate as value added product.

Hollow and porous nickel sulfide nanocubes prepared from a metal-organic framework as an efficient enzyme mimic for colorimetric detection of hydrogen peroxide.

Hollow, porous NiS nanocubes were prepared by a hydrothermal method starting from Ni-Co Prussian blue analogue nanocubes as the template. The morphology and structure of the NiS nanocubes were tuned by adjustment of the ion-exchange rate and the degree of chemical etching, and they were characterized by scanning electron microscopy, energy-dispersive X-ray spectroscopy, transmission electron microscopy, X-ray diffraction, and nitrogen sorption measurements. The NiS nanocubes are shown to act as a peroxidase mimic that can catalyze the oxidization of 3,3',5,5'-tetramethylbenzidine by hydrogen peroxide (H2O2), producing a visible color change, for which the absorbance is best measured at 652 nm. The outstanding activity may result from the unique structure of the NiS nanocubes. The catalytic oxidation follows Michaelis-Menten kinetics and shows a ping-pong mechanism of enzyme action. The findings were used to develop a rapid, sensitive, and selective colorimetric H2O2 assay with a response that is linear in the 4-40 μM range with a detection limit of 1.72 μM (signal-to-noise ratio of 3). Graphical abstarct ᅟ.

Phytochemical, Free Radical Scavenging and Antifungal Profile of Cuscuta campestris Yunck. Seeds.

This work was conceptualized with the goal to investigate the phytochemical, free radical scavenging and antifungal profile of Cuscuta campestrisYunck. seeds. Total phenolics, amino acid and carbohydrate contents were evaluated in ethanolic, acetone and chloroform extract. Effective antioxidant activity was evaluated throughout seven antioxidant methods. The antifungal activity was assessed against eight fungal strains and Candida albicans. The results showed total phenol, flavonoid, flavonols and phenolic acids contents in amount of 1.51-6.35mg GAE/mL, 78-425μg RU/mL, 1.04-2.98mg QU/g and 12.01-30.58μg CAE/mL, respectively. The total amino acids and carbohydrates content ranged from 8.29 to 185.45μg Gly/mL and from 0.05 to 0.12μg Glu/mL. The ethanolic extract showed the best antioxidant activity in phosphomolybdenum, DPPH free radical scavenging, ferric reducing power and lipid peroxidation assays. The best activity in ferrous ion chelating and H2 O2 assays had the acetone extract, whereas the best hydroxyl radical scavenging activity was observed with the chloroform extract. The ethanolic extract at a concentration of 6mg/mL proved to be the most effective antimycotic, since it inhibited the growth of all tested fungi except Penicillium verrucosum. The obtained results indicate that C.campestris seeds could be attributed to a potential source of natural antioxidants in food and pharmaceutical products.

Phytochemical Screening for Various Secondary Metabolites, Antioxidant, and Anthelmintic Activity of Coscinium fenestratum Fruit Pulp: A New Biosource for Novel Drug Discovery.

Coscinium fenestratum (Gaertn.) Colebr. (CF, Family: Menispermaceae) is an important endangered woody climber in India. CF contains various major secondary metabolites for the treatment of various disease conditions. The present study aimed to establish the antioxidant and anthelmintic activity of Coscinium fenestratum fruit pulp. The dried fruit pulp was subjected to aqueous, methanol, and mixed aqueous and methanol (1:1) solvent extraction followed by phytochemical investigations, estimations of alkaloids, phenolics, flavonoids, antioxidant potentiality (DPPH and hydrogen peroxide scavenging methods), and anthelmintic activity tests were carried out. Preliminary phytochemical screening of CF fruit extracts revealed the presence of alkaloids phenols, flavonoids, tannins, steroids, and resins, which are responsible for biologic properties. The combined aqueous and methanol extract resulted in significant anthelmintic and antioxidant properties in a dose-dependent manner. The DPPH free radical scavenging assay and H2O2 assay exhibited IC50 values of 42.38±0.012 µg/mL and 46.80±0.011 µg/mL, respectively. Thereafter, the anthelmintic activity test was carried out against Pheretima posthuma and Taenia solium with the extract at varying concentrations of 25, 50, 100 and 150 mg/mL and compared with standard albendazole (25 and 50 mg/mL) and saline (0.9%) as a control. All the extracts exhibited concentration-dependent paralytic effect, followed by death on the test organism, but significant activity was observed with the combined methanol and aqueous extract. The study was conducted in order to find possible isolated compounds as a biosources for future novel antioxidants in food and pharmaceutical formulations. Our findings indicate for the first time that the CF fruit pulp has therapeutic values with prominent antioxidant and anthelmintic properties.

Effects of cigarette smoke extracts on apoptosis and oxidative stress in two models of ovarian cancer in vitro

Cigarette smoke contains thousands of harmful components, many of which exert toxic effects on reproductive organs, including the ovaries, and their development. In this study, we investigated the effects of cigarette smoke extract (CSE) on cell proliferation, apoptosis and oxidative stress in the SKOV3 and OVCAR3 ovarian cancer cell lines. A water soluble tetrazolium (WST) assay revealed that the cell proliferation of both ovarian cells was significantly inhibited by three types of CSE in a concentration-dependent manner. Western blot analysis showed that the inhibition of cell proliferation was related to regulation of cell cycle-related protein expression. To determine the effects of cigarette smoke on the death of ovarian cells, we conducted a TUNEL assay, which revealed a time-dependent increase in TUNEL-stained cells in response to CSE exposure. Moreover, the protein expression of the Bcl-2 family signaling pathway observed upon Western blot analysis were consistent with the results of the TUNEL assay. We also examined the reactive oxygen species (ROS) generation in two ovarian cell lines using DCF-DA (5(6)-Carboxy-2′,7′-dichlorofluorescein diacetate) assay and ROS-Glo™ H2O2 assay to investigate the oxidative stress caused by CSE. Both ROS detection assays demonstrated an increase in the amount of ROS by CSE in ovarian cells, which was consistent with the results observed for the Keap1-Nrf2 pathway upon Western blot analysis. Taken together, these results suggest that CSE may induce cell proliferation inhibition and oxidative stress in ovarian cells.