High-performance liquid chromatography with post-column 2,2′-diphenyl-1-picrylhydrazyl radical scavenging assay : methodological considerations and application to complex samples

An improved post-column 2,2´-diphenyl-1-picrylhydrazyl (DPPH•) radical scavenging assay for the screening of antioxidants in complex matrices was developed. Experimental parameters believed to be influential to DPPH• response were studied in a univariate approach. Optimum conditions were found to be: 5×10⁻⁵M DPPH• reagent prepared in a 75% methanol: 25% 40mM citric acid–sodium citrate buffer (pH 6) solution, degassed with nitrogen; reaction coil of 2m×0.25mm i.d. PEEK tubing; detection at 521 nm; analysis at room temperature. The analytical utility of this protocol was evaluated by screening for antioxidants in thyme and green tea, in comparison with two commonly employed methodologies.

Toad's tongue for breakfast : exploitation of a novel prey type, the invasive cane toad, by scavenging raptors in tropical Australia

Although interest in the ecological impacts of invasive species has largely focused on negative effects, some native taxa may benefit from invader arrival. In tropical Australia, invasive cane toads (Bufo marinus) have fatally poisoned many native predators (e.g., marsupials, crocodiles, lizards) that attempt to ingest the toxic anurans, but birds appear to be more resistant to toad toxins. We quantified offtake of dead (road-killed) cane toads by raptors (black kites (Milvus migrans) and whistling kites (Haliastur sphenurus)) at a site near Darwin, in the Australian wet-dry tropics. Raptors readily took dead toads, especially small ones, although native frogs were preferred to toads if available. More carcasses were removed in the dry season than the wet season, perhaps reflecting seasonal availability of alternative prey. Raptors appeared to recognize and avoid bufotoxins, and typically removed and consumed only the toads’ tongues (thereby minimizing toxin uptake). The invasion of cane toads thus constitutes a novel prey type for scavenging raptors, rather than (as is the case for many other native predators) a threat to population viability.

Bio-conjugation of antioxidant peptide on surface-modified gold nanoparticles: a novel approach to enhance the radical scavenging property in cancer cell

BACKGROUND: Functionalized gold nanoparticles are emerging as a promising nanocarrier for target specific delivery of the therapeutic molecules in a cancer cell, as a result it targeted selectively to the cancer cell and minimized the off-target effect. The functionalized nanomaterial (bio conjugate) brings novel functional properties, for example, the high payload of anticancer, antioxidant molecules and selective targeting of the cancer molecular markers. The current study reported the synthesis of multifunctional bioconjugate (GNPs-Pep-A) to target the cancer cell. METHODS: The GNPs-Pep-A conjugate was prepared by functionalization of GNPs with peptide-A (Pro-His-Cys-Lys-Arg-Met; Pep-A) using thioctic acid as a linker molecule. The GNPs-Pep-A was characterized and functional efficacy was tested using Retinoblastoma (RB) cancer model in vitro. RESULTS: The GNPs-Pep-A target the reactive oxygen species (ROS) in RB, Y79, cancer cell more effectively, and bring down the ROS up to 70 % relative to control (untreated cells) in vitro. On the other hand, Pep-A and GNPs showed 40 and 9 % reductions in ROS, respectively, compared to control. The effectiveness of bioconjugate indicates the synergistic effect, due to the coexistence of both organic (Pep-A) and inorganic phase (GNPs) in novel GNPs-Pep-A functional material. In addition to this, it modulates the mRNA expression of antioxidant genes glutathione peroxidase (GPX), superoxide dismutase (SOD) and catalase (CAT) by two-threefolds as observed. CONCLUSIONS: The effects of GNPs-Pep-A on ROS reduction and regulation of antioxidant genes confirmed that Vitis vinifera L. polyphenol-coated GNPs synergistically improve the radical scavenging properties and enhanced the apoptosis of cancer cell.

Copper in disorders with neurological symptoms: Alzheimer`s, Menkes, and Wilson Diseases

Copper is an essential element for the activity of a number of physiologically important enzymes. Enzyme-related malfunctions may contribute to severe neurological symptoms and neurological diseases: copper is a component of cytochrome c oxidase, which catalyzes the reduction of oxygen to water, the essential step in cellular respiration. Copper is a cofactor of Cu/Zn-superoxide-dismutase which plays a key role in the cellular response to oxidative stress by scavenging reactive oxygen species. Furthermore, copper is a constituent of dopamine-β-hydroxylase, a critical enzyme in the catecholamine biosynthetic pathway. A detailed exploration of the biological importance and functional properties of proteins associated with neurological symptoms will have an important impact on understanding disease mechanisms and may accelerate development and testing of new therapeutic approaches. Copper binding proteins play important roles in the establishment and maintenance of metal-ion homeostasis, in deficiency disorders with neurological symptoms (Menkes disease, Wilson disease) and in neurodegenerative diseases (Alzheimer’s disease). The Menkes and Wilson proteins have been characterized as copper transporters and the amyloid precursor protein (APP) of Alzheimer’s disease has been proposed to work as a Cu(II) and/or Zn(II) transporter. Experimental, clinical and epidemiological observations in neurodegenerative disorders like Alzheimer’s disease and in the genetically inherited copper-dependent disorders Menkes and Wilson disease are summarized. This could provide a rationale for a link between severely dysregulated metal-ion homeostasis and the selective neuronal pathology.

Effects of purified herbal extract of Saliva miltiorrhiza on ischemic rat myocardium after acute myocardial infarction

In the current study, we compared purified Salvia miltiorrhiza extract (PSME) with Angiotensin-converting enzyme inhibitor, Ramipril, in in vitro experiments and also in vivo using animal model of myocardial infarction. PSME was found to have a significantly higher trolox equivalent antioxidant capacity which indicated a great capacity for scavenging free radicals. PSME could also prevent pyrogallo red bleaching and DNA damage.

After 2 weeks treatment with PSME or Ramipril, survival rates of rats with experimental myocardial infarction were marginally increased (68.2% and 71.4%) compared with saline (61.5%). The ratios of infarct size to left ventricular size in both PSME-and Ramipril-treated rats were significantly less than that in the saline-treated group. Activity of cardiac antioxidant enzyme superoxide dismutase (SOD) was significant higher while level of Thiobarbituric acid-reactive substances (TBARs) was lower in the PSME treated group. Purified and standardized Chinese herb could provide an alternative regimen for the prevention of ischemic heart disease.

Antioxidant treatment with N-acetylcysteine regulates mammalian skeletal muscle Na+-K+-ATPase ∝ gene expression during repeated contractions

Exercise increases Na+–K+ pump isoform gene expression and elevates muscle reactive oxygen species (ROS). We investigated whether enhanced ROS scavenging induced with the antioxidant N-acetylcysteine (NAC) blunted the increase in Na+–K+ pump mRNA during repeated contractions in human and rat muscle. In experiment 1, well-trained subjects received saline or NAC intravenously prior to and during 45 min cycling. Vastus lateralis muscle biopsies were taken pre-infusion and following exercise. In experiment 2, isolated rat extensor digitorum longus muscles were pre-incubated without or with 10 mm NAC and then rested or stimulated electrically at 60 Hz for 90 s. After 3 h recovery, muscles were frozen. In both experiments, the muscles were analysed for Na+–K+ pump α1, α2, α3, β1, β2 and β3 mRNA. In experiment 1, exercise increased α2 mRNA by 1.0-fold (P = 0.03), but α2 mRNA was reduced by 0.40-fold with NAC (P = 0.03). Exercise increased α3, β1 and β2 mRNA by 2.0- to 3.4-fold (P < 0.05), but these were not affected by NAC (P > 0.32). Neither exercise nor NAC altered α1 or β3 mRNA (P > 0.31). In experiment 2, electrical stimulation increased α1, α2 and α3 mRNA by 2.3- to 17.4-fold (P < 0.05), but these changes were abolished by NAC (P > 0.07). Electrical stimulation almost completely reduced β1 mRNA but only in the presence of NAC (P < 0.01). Neither electrical stimulation nor NAC altered β2 or β3 mRNA (P > 0.09). In conclusion, NAC attenuated the increase in Na+–K+ pump α2 mRNA with exercise in human muscle and all α isoforms with electrical stimulation in rat muscle. This indicates a regulatory role for ROS in Na+–K+ pump α isoform mRNA in mammalian muscle during repeated contractions.

A simple 96-well microplate method for estimation of total polyphenolic content in seaweeds

Seaweed polyphenols are potent antioxidants and have also been shown to have α-glucosidase inhibiting activity. In our continuous efforts to develop new marine-based nutraceuticals and functional food ingredients, we have investigated many algal species collected on the Atlantic coast of Canada. A simple method for estimating the total polyphenol content in seaweeds and their extracts was developed based on the classic Folin-Ciocalteau colorimetric reaction. By using the 96-well microplate and a microplate reader, this new method saves experimental time, significantly reduces the amount of sample required, handles large number of samples in one experiment, and also improves the repeatability of the results.

A number of algal samples collected on the seashore of Nova Scotia, Canada, were analyzed for their levels of polyphenol content using this microplate-based method. The antioxidant activity of these samples was also assessed by using DPPH (2, 2-diphenyl-1-picrylhydrazyl) radical scavenging assay. The results showed that there is a strong correlation between the total polyphenol content and the potency of antioxidant effect.

Ascophyllum compositions and methods

The invention provides extracts of brown seaweed of the genus Ascophyllum that have valuable pharmacological properties. In particular, compositions of the invention are useful for e.g.: inhibiting alpha-glucosidase activity; preventing or treating conditions mediated by alpha-glucosidase activity; reducing blood glucose levels; preventing or treating diabetes; modulating glucose uptake in adipocytes; preventing or treating obesity; scavenging free radicals; stimulating the immune system; activating macrophages; preventing or treating condition mediated by macrophage activation; and modulating nitric oxide production by macrophages. Methods for using the Ascophyllum extracts of the invention are provided, as are kits comprising Ascophyllum extracts of the invention and instructions for using the extracts.

Design and analysis of an antenna for wireless energy harvesting in head-mountable DBS device

This paper presents design and simulation of a circular meander dipole antenna at the industrial, scientific, and medical band of 915 MHz for energy scavenging in a passive head-mountable deep brain stimulation device. The interaction of the proposed antenna with a rat body is modeled and discussed. In the antenna, the radiating layer is meandered, and a FR-4 substrate is used to limit the radius and height of the antenna to 14 mm and 1.60 mm, respectively. The resonance frequency of the designed antenna is 915 MHz and the bandwidth of 15 MHz at a return loss of -10 dB in free space. To model the interaction of the antenna with a rat body, two aspects including functional and biological are considered. The functional aspect includes input impedance, resonance frequency, gain pattern, radiation efficiency of the antenna, and the biological aspect involves electric field distribution, and SAR value. A complete rat model is used in the finite difference time domain based EM simulation software XFdtd. The simulated results demonstrate that the specific absorption rate distributions occur within the skull in the rat model, and their values are higher than the standard regulated values for the antenna receiving power of 1W.

Human passive motions and a user-friendly energy harvesting system

Energy harvesting for wireless sensors and consumer electronic devices can significantly improve reliability and environmental sustainability of the devices. This is achieved by eliminating the dependency of these devices on rechargeable batteries, using clean and/or renewable energy sources. Energy harvesting from various energy sources is widely discussed among researchers and entrepreneurs, including harvesting energy from microscale phenomena. This topic is receiving increasing attention due to the rising numbers of low-power consumer electronic devices and wireless sensors, but also the increasing demand for more convenient and available devices. This article presents a feasibility study for an energy harvesting system based on a human's breathing motion. The system is based on a modified pants belt that is integrated with an array of piezoelectric films and a harvesting circuit. The proposed energy harvester generates electricity from reciprocal abdominal motions of the human subject. In comparison with existing breathing-based energy harvesters, the proposed system allows for safe and convenient energy harvesting with no influence on the natural movement of the lungs. Stomach pressure analysis and measurement, as well as the design and simulations of the proposed harvester, are presented. © 2013 The Author(s).

Do the numbers and locations of road-killed anuran carcasses accurately reflect impacts of vehicular traffic?

Road-killed animals are easy and inexpensive to survey, and may provide information about species distributions, abundances, and mortality rates. As with any sampling method, however, we need to explore methodological biases in such data. First, how does an animal's behavior (e.g., use of the center vs. periphery of the road) influence its vulnerability to vehicular traffic? Second, how rapidly do post-mortem processes (scavenging by other animals, destruction or displacement by subsequent vehicles) change the numbers and locations of roadkills? Our surveys of anurans on a highway in tropical Australia show that different anuran species are distributed in different ways across the width of the road, and that locations of live versus dead animals sometimes differ within a species. Experimental trials show that location on the road affects the probability of being hit by a vehicle, with anurans in the middle of the road begin hit 35% more often than anurans on the edges; thus, center-using species are more likely to be hit than edge-using taxa. The magnitude of post-mortem displacement and destruction by subsequent vehicles depended on anuran species and body size. The mean parallel displacement distance was 122.7 cm, and carcasses of thin-skinned species exhibited greater post-mortem destruction. Scavenging raptors removed 73% of carcasses, most within a few hours of sunrise. Removal rates were biased with respect to size and species. Overall, our studies suggest that investigators should carefully evaluate potential biases before using roadkill counts to estimate underlying animal abundances or mortality rates.

Invasive carnivores alter ecological function and enhance complementarity in scavenger assemblages on ocean beaches

Species composition is expected to alter ecological function in assemblages if species traits differ strongly. Such effects are often large and persistent for nonnative carnivores invading islands. Alternatively, high similarity in traits within assemblages creates a degree of functional redundancy in ecosystems. Here we tested whether species turnover results in functional ecological equivalence or complementarity, and whether invasive carnivores on islands significantly alter such ecological function. The model system consisted of vertebrate scavengers (dominated by raptors) foraging on animal carcasses on ocean beaches on two Australian islands, one with and one without invasive red foxes (Vulpes vulpes). Partitioning of scavenging events among species, carcass removal rates, and detection speeds were quantified using camera traps baited with fish carcasses at the dune–beach interface. Complete segregation of temporal foraging niches between mammals (nocturnal) and birds (diurnal) reflects complementarity in carrion utilization. Conversely, functional redundancy exists within the bird guild where several species of raptors dominate carrion removal in a broadly similar way. As predicted, effects of red foxes were large. They substantially changed the nature and rate of the scavenging process in the system: (1) foxes consumed over half (55%) of all carrion available at night, compared with negligible mammalian foraging at night on the fox-free island, and (2) significant shifts in the composition of the scavenger assemblages consuming beach-cast carrion are the consequence of fox invasion at one island. Arguably, in the absence of other mammalian apex predators, the addition of red foxes creates a new dimension of functional complementarity in beach food webs. However, this functional complementarity added by foxes is neither benign nor neutral, as marine carrion subsidies to coastal red fox populations are likely to facilitate their persistence as exotic carnivores.

Functional replacement across species pools of vertebrate scavengers separated at a continental scale maintains an ecosystem function

Summary: The composition of species pools can vary in space and time. While many studies are focused on understanding which factors influence the make-up of species pools, the question to which degree biogeographic variation in species composition propagates to biogeographic variation in ecological function is rarely examined. If different local species assemblages operate in ways that maintain specific ecological processes across continents, they can be regarded as functionally equivalent. Alternatively, variation in species assemblages might result in the loss of ecological function if different species fulfil different functions, and thereby fail to maintain the ecological process. Here, we test whether ecological function is affected by differences in the composition of species pools across a continental scale, comparing a tropical with a temperate pool. The model systems are assemblages of vertebrates foraging on ocean beaches, and the ecological function of interest is the consumption of wave-cast carrion, a pivotal process in sandy shore ecosystems. We placed fish carcasses (n = 179) at the beach-dune interface, monitored by motion-triggered cameras to record scavengers and quantify the detection and removal of carrion. Scavenging function was measured on sandy beaches in two distinct biogeographic regions of Australia: tropical north Queensland and temperate Victoria. The composition of scavenging assemblages on sandy beaches varied significantly across the study domain. Raptors dominated in the tropics, while invasive red foxes were prominent in temperate assemblages. Notwithstanding the significant biogeographic change in species composition, ecological function - as indexed by carcass detection and removal - was maintained, suggesting strong functional replacement at the continental scale. Species pools of vertebrate scavengers that are assembled from taxonomically distinct groups (birds vs. mammals) and located in distinct climatic regions (temperate vs. tropical) can maintain an ecological process via replacement of species with comparable functional traits.

Limited functional redundancy in vertebrate scavenger guilds fails to compensate for the loss of raptors from urbanized sandy beaches

Aim: Globally, urbanization is one of the most widespread, intense and ecologically destructive forms of landscape transformation, and it is often concentrated in coastal areas. Theoretically, species losses attributable to urbanization are predicted not to alter overall ecosystem function if functional redundancy (i.e. replacement of function by alternative species) compensates for such losses. Here, we test this expectation by measuring how coastal urbanization affects scavenger guilds on sandy beaches and whether changes in guild composition result either in an overall loss of scavenging efficiency, or in functional compensation under alternative guild structures, maintaining net ecosystem functioning. Location: Fourteen beaches along the east coast of Australia with variable levels of urbanization. Methods: Scavenging communities and rates of carrion removal were determined using motion-triggered cameras at the beach-dune interface. Results: A substantial shift in the community structure of vertebrate scavengers was associated with gradients in urbanization. Iconic and functionally important raptors declined precipitously in abundance on urban beaches. Importantly, other vertebrates usually associated with urban settings (e.g. dogs, foxes, corvids) did not functionally replace raptors. In areas where < 15% of the abutting land had been developed into urban areas, carcass removal by scavengers was often complete, but always > 70%. Conversely, on beaches bordering coastal cities with < 40% of natural vegetation remaining, two-thirds of fish carcasses remained uneaten by scavengers. Raptors removed 70-100% of all deployed fish carcasses from beaches with < 8% urban land cover, but this number dropped significantly with greater levels of urbanization and was not compensated by other scavenger species in urban settings. Main conclusions: There is limited functional redundancy in vertebrate scavenger communities of sandy beach ecosystems, which impacts the system's capacity to mitigate the ecological consequences of detrimental landscape transformations.