Binding mechanism of affinity ligands for purification of plasmid DNA

Plasmid DNA for therapeutic and vaccination purposes must be highly purified. The high selectivity of affinity chromatography makes it ideal for the isolation of pDNA from complex biological feed stocks. Affinity chromatography makes use of the biological function and/or individual chemical structure of the interacting molecules. However, the success of any affinity purification protocol is dependent on the availability of suitable ligands. In this study, surface plasmon resonance (SPR) based Biacore system has been employed for the detection and quantification of the binding between lac operon (lacO) sequence contained in a pDNA and synthetic peptides based on the DNA-binding domain of the lac repressor protein, lad. The equilibrium dissociation constant (K D) and association and dissociation rate constants (ka, kd) for the interaction between plasmid DNA and designed peptides were determined.

A novel population balance model for the dilute acid hydrolysis of hemicellulose

Acid hydrolysis is a popular pretreatment for removing hemicellulose from lignocelluloses in order to produce a digestible substrate for enzymatic saccharification. In this work, a novel model for the dilute acid hydrolysis of hemicellulose within sugarcane bagasse is presented and calibrated against experimental oligomer profiles. The efficacy of mathematical models as hydrolysis yield predictors and as vehicles for investigating the mechanisms of acid hydrolysis is also examined. Experimental xylose, oligomer (degree of polymerisation 2 to 6) and furfural yield profiles were obtained for bagasse under dilute acid hydrolysis conditions at temperatures ranging from 110C to 170C. Population balance kinetics, diffusion and porosity evolution were incorporated into a mathematical model of the acid hydrolysis of sugarcane bagasse. This model was able to produce a good fit to experimental xylose yield data with only three unknown kinetic parameters ka, kb and kd. However, fitting this same model to an expanded data set of oligomeric and furfural yield profiles did not successfully reproduce the experimental results. It was found that a ``hard-to-hydrolyse'' parameter, $\alpha$, was required in the model to ensure reproducibility of the experimental oligomer profiles at 110C, 125C and 140C. The parameters obtained through the fitting exercises at lower temperatures were able to be used to predict the oligomer profiles at 155C and 170C with promising results. The interpretation of kinetic parameters obtained by fitting a model to only a single set of data may be ambiguous. Although these parameters may correctly reproduce the data, they may not be indicative of the actual rate parameters, unless some care has been taken to ensure that the model describes the true mechanisms of acid hydrolysis. It is possible to challenge the robustness of the model by expanding the experimental data set and hence limiting the parameter space for the fitting parameters. The novel combination of ``hard-to-hydrolyse'' and population balance dynamics in the model presented here appears to stand up to such rigorous fitting constraints.

Mechanisms underlying the effect of acupuncture on cognitive improvement: A systematic review of animal studies

Acupuncture has been reported to be beneficial in treating cognitive impairment in various pathological conditions. This review describes the effort to understand the signaling pathways that underlie the acupunctural therapeutic effect on cognitive function. We searched the literature in 12 electronic databases from their inception to November 2013, with full text available and language limited to English. Twenty-three studies were identified under the selection criteria. All recruited animal studies demonstrate a significant positive effect of acupuncture on cognitive impairment. Findings suggest acupuncture may improve cognitive function through modulation of signaling pathways involved in neuronal survival and function, specifically, through promoting cholinergic neural transmission, facilitating dopaminergic synaptic transmission, enhancing neurotrophin signaling, suppressing oxidative stress, attenuating apoptosis, regulating glycometabolic enzymes and reducing microglial activation. However, the quality of reviewed studies has room for improvement. Further high-quality animal studies with randomization, blinding and estimation of sample size are needed to strengthen the recognition of group differences.

A preliminary OSL chronology for coastal dunes on Moreton island, Queensland, Australia – Marginal deposits of A large-scale quaternary shelf sediment system

Moreton Island and several other large siliceous sand dune islands and mainland barrier deposits in SE Queensland represent the distal, onshore component of an extensive Quaternary continental shelf sediment system. This sediment has been transported up to 1000 km along the coast and shelf of SE Australia over multiple glacioeustatic sea-level cycles. Stratigraphic relationships and a preliminary Optically Stimulated Luminance (OSL) chronology for Moreton Island indicate a middle Pleistocene age for the large majority of the deposit. Dune units exposed in the centre of the island and on the east coast have OSL ages that indicate deposition occurred between approximately 540 ka and 350 ka BP, and at around 96±10 ka BP. Much of the southern half of the island has a veneer of much younger sediment, with OSL ages of 0.90±0.11 ka, 1.28±0.16 ka, 5.75±0.53 ka and <0.45 ka BP. The younger deposits were partially derived from the reworking of the upper leached zone of the much older dunes. A large parabolic dune at the northern end of the island, OSL age of 9.90±1.0 ka BP, and palaeosol exposures that extend below present sea level suggest the Pleistocene dunes were sourced from shorelines positioned several to tens of metres lower than, and up to few kilometres seaward of the present shoreline. Given the lower gradient of the inner shelf a few km seaward of the island, it seems likely that periods of intermediate sea level (e.g. ~20 m below present) produced strongly positive onshore sediment budgets and the mobilisation of dunes inland to form much of what now comprises Moreton Island. The new OSL ages and comprehensive OSL chronology for the Cooloola deposit, 100 km north of Moreton Island, indicate that the bulk of the coastal dune deposits in SE Queensland were emplaced between approximately 540 ka BP and prior to the Last Interglacial. This chronostratigraphic information improves our fundamental understanding of long-term sediment transport and accumulation on large-scale continental shelf sediment systems.

Understanding past climate variation and environmental change for the future of an iconic landscape – K'gari Fraser Island, Queensland, Australia

The unique combination of landscapes and processes that are present and operate on Fraser Island (K'gari) create a dynamic setting that is capable of recording past environmental events, climate variations and former landscapes. Likewise, its geographic position makes Fraser Island sensitive to those events and processes. Based on optically stimulated luminescence dating, the records archived within the world's largest sand island span a period that has the potential to exceed 750 ka and contain specific records that are of extremely high resolution over the past 40,000 years. This is due to the geographic position of Fraser Island, which lies in the coastal subtropical region of Queensland Australia. Fraser Island is exposed to the open ocean currents of the Coral Sea on the east coast and the waters of Hervey Bay on its western margin and is positioned to receive moisture from the Indo-Australian monsoon, southeast trade winds and experiences occasional tropical and ex-tropical cyclones. We review literature that presents the current level of understanding of sea level change, ecological variation and environmental change on Fraser Island. The previous works illustrate the importance of Fraser Island and may link processes, environments and climates on Fraser Island with global records.

Large variations in the Holocene marine radiocarbon reservoir effect reflect ocean circulation and climatic changes

Accurate radiocarbon dating of marine samples requires knowledge of the marine radiocarbon reservoir effect. This effect for a particular site/region is generally assumed constant through time when calibrating marine 14C ages. However, recent studies have shown large temporal variations of several hundred to a couple of thousand years in this effect for a number of regions during the late Quaternary and Holocene. Here we report marine radiocarbon reservoir correction (ΔRΔR) for Heron Reef and Moreton Bay in southwestern (SW) Pacific for the last 8 ka derived from 14C analysis of 230Th-dated corals. Most of our ΔRΔR for the last ∼5.4 ka agree well with their modern value, but large ΔRΔR variability of ∼410 yr (from trough to peak) with possible decadal/centennial fluctuations is evident for the period ∼5.4–8 ka. The latter time interval also has significant variations with similar features in previously published ΔRΔR values for other sites in the Pacific, including southern Peru–northern Chile in southeastern (SE) Pacific, the South China Sea, Vanuatu and Papua New Guinea, with the largest magnitude of ∼920 yr from SE Pacific. The mechanisms for these large ΔRΔR variations across the Pacific during the mid-Holocene are complex processes involving (1) changes in the quantity and 14C content of upwelled waters in tropical east Pacific (TEP) (frequency and intensity of ocean upwelling in the TEP, and contribution of Subantarctic Mode Water to the upwelled waters, which is influenced by the intensity and position of southern westerly winds), and (2) variations in ocean circulation associated with climate change (La Niña/El Niño conditions, intensity of easterly trade winds, positions of the Intertropical Convergence Zone and the South Pacific Convergence Zone), which control the spreading of the older upwelled surface waters in the TEP to the western sites. Our results imply the need for employing temporal changes in ΔRΔR values, instead of constant (modern) values, for age calibration of Holocene marine samples not only for the SW Pacific sites but also for other tropical and subtropical sites in the Pacific.

VIP Barcoding: Composition vector-based software for rapid species identification based on DNA barcoding

Species identification based on short sequences of DNA markers, that is, DNA barcoding, has emerged as an integral part of modern taxonomy. However, software for the analysis of large and multilocus barcoding data sets is scarce. The Basic Local Alignment Search Tool (BLAST) is currently the fastest tool capable of handling large databases (e.g. >5000 sequences), but its accuracy is a concern and has been criticized for its local optimization. However, current more accurate software requires sequence alignment or complex calculations, which are time-consuming when dealing with large data sets during data preprocessing or during the search stage. Therefore, it is imperative to develop a practical program for both accurate and scalable species identification for DNA barcoding. In this context, we present VIP Barcoding: a user-friendly software in graphical user interface for rapid DNA barcoding. It adopts a hybrid, two-stage algorithm. First, an alignment-free composition vector (CV) method is utilized to reduce searching space by screening a reference database. The alignment-based K2P distance nearest-neighbour method is then employed to analyse the smaller data set generated in the first stage. In comparison with other software, we demonstrate that VIP Barcoding has (i) higher accuracy than Blastn and several alignment-free methods and (ii) higher scalability than alignment-based distance methods and character-based methods. These results suggest that this platform is able to deal with both large-scale and multilocus barcoding data with accuracy and can contribute to DNA barcoding for modern taxonomy. VIP Barcoding is free and available at http://msl.sls.cuhk.edu.hk/vipbarcoding/.

Biosorption of metals using nonliving biomass - A review

A review of published literature on the biosorption of metals using nonliving biomass is presented. Factors such as pH, temperature, initial metal concentration, biomass loaning, the presence of co-ions and the pretreatment of biomass influence the metal uptake by biomass. Although few generalizations are possible, unified theor ies regarding the mechanism of uptake are not available. Therefore, the above aspects of metal biosorption have to be defined individually for each biomass and metal-ion pair.

ELISA for the detection of venoms from four medically important snakes of India

A double antibody sandwich enzyme linked immunosorbent assay (ELISA) was developed to detect Echis carinatus venom in various organs (brain, heart, lungs, liver, spleen and kidneys) as well as tissue at the site of injection of mice, at various time intervals (1, 6, 12, 18, 24 h and 12 h intervals up to 72 h) after death. The assay could detect E. carinatus venom levels up to 2.5 ng/ml of tissue homogenate and the venom was detected up to 72 h after death. A highly sensitive and species-specific avidin-biotin microtitre ELISA was also developed to detect venoms of four medically important Indian snakes (Bungarus caeruleus, Naja naja, E. carinatus and Daboia russelli russelli) in autopsy specimens of human victims of snake bite. The assay could detect venom levels as low as 100 pg/ml of tissue homogenate. Venoms were detected in brain, heart, lungs, liver, spleen, kidneys, tissue at the bite area and postmortem blood. In all 12 human victim cadavers tested the culprit species were identified. As observed in mice, tissue at the site of bite area showed the highest concentration of venom and the brain showed the least. Moderate amounts of venoms were found in liver, spleen, kidneys, heart and lungs. Development of a simple, rapid and species-specific diagnostic kit based on this ELISA technique useful to clinicians is discussed.

A New Synthesis Of Dibenzo[Aj]Xanthene Derivatives

Oxidation of mu-H/alkylbisnaphthols (4a-g) gives the 14-substituted dibenzo[aj]xanthenes (5a-g) as the sole product while that of mu-arylbisnaphthols (4h-j) gives the xanthenes (5h-j) along with the corresponding spironaphthalenones (1h-j). A probable mechanism for the formation of the products has been suggested.

Growth and attachment of Thiobacillus ferrooxidans during sulfide mineral leaching

The growth of Thiobacillus ferrooxidans, their attachment to sulfide minerals and detachment during bacterial leaching are discussed in this paper. Growth of the bacteria has been measured by cell count of the supernatants of the mineral suspensions while attachment to minerals and detachment were measured by periodic protein estimations for both the solid and liquid phases, Even in the absence of the nutrients, bacterial growth occurs and increases the available cell population during leaching; such growth was greater in sphalerite suspensions than in galena suspensions, The bacterial attachment studies suggest that more cells are attached onto galena mineral surface than to sphalerite surface. The mechanisms of bacterial attachment and detachment are discussed.

Studies on interaction of Paenibacillus polymyxa with iron ore minerals in relation to beneficiation

Interaction between Paenibacillus polymyxa with minerals such as hematite, corundum, quartz and kaolinite brought about significant surface chemical changes on all the minerals. Quartz and kaolinite were rendered more hydrophobic, while hematite and corundum, became more hydrophilic after biotreatment. The predominance of bacterial polysaccharides on interacted hematite and corundum and of proteins on quartz and kaolinite was responsible for the above surface-chemical changes. Bio-pretreatment of the above iron ore mineral mixtures resulted in the selective separation of silica and alumina from iron oxide, through bioflotation and bioflocculation. The utility of bioprocessing in the beneficiation of iron ores is demonstrated.

Surface chemical studies of Thiobacillus ferrooxidans with reference to copper tolerance

A strain of Thiobacillus ferrooxidans was adapted to grow at higher concentrations of copper by single step culturing in the presence of 20 g/L (0.314 mol/L) cupric ions added to 9K medium. Exposure to copper results in change in the surface chemistry of the microorganism. The isoelectric point of the adapted strain (pI=4.7) was observed to be at a higher pH than that of the wild unadapted strain(pI=2.0). Compared to the wild strain, the copper adapted strain was found to be more hydrophobic and showed enhanced attachment efficiency to the pyrite mineral. The copper adsorption ability of the adapted strain was also found to be higher than that of the wild strain. Fourier transform infrared spectroscopy of adapted cells suggested that a proteinaceous new cell surface component is synthesized by the adapted strain. Treatment of adapted cells with proteinase-K, resulted in complete loss of tolerance to copper, reduction in copper adsorption and hydrophobicity of the adapted cells. These observations strongly suggest a role played by cell surface modifications of Thiobacillus ferrooxidans in imparting the copper tolerance to the cells and bioleaching of sulphide minerals.

B- and T-cell epitopes of tropomyosin, the major shrimp allergen

Seafood allergy is often encountered on ingestion of crustaceans such as shrimp, lobster, crab, and crayfish (1). On eating cooked shrimp, sensitive individuals experience a wide spectrum of reactions ranging from abdominal discomfort to anaphylaxis. The presence of cross-reacting heat-stable allergens in crustacean food was first recognized by Hoffman et al. (2) and Lehrer et al. (3). Subsequently, the major allergen was isolated and characterized from the shrimp species Paneaus indicus (Pen i 1) (4) and I? aztecm (Pen a 1) (5). Pen i 1 (originally designated Sa-TI) and Pen a 1, with mol. mass of 34 and 36 kDa, respectively, contain 301 and 312 amino-acid residues with a predominance of gluta- mate/glutamine and asparatate/asparagine.