Molecular and morphological description of a Hepatozoon species in reptiles and their ticks in the Northern Territory, Australia

Ticks, representing 3 species of Amblyomma, were collected from the water python (Liasis fuscus) and 3 additional reptile species in the Northern Territory, Australia, and tested for the presence of Hepatozoon sp., the most common blood parasites of snakes. In addition, blood smears were collected from 5 reptiles, including the water python, and examined for the presence of the parasite. Hepatozoon sp. DNA was detected in all tick and reptile species, with 57.7% of tick samples (n = 187) and 35.6% of blood smears (n=35) showing evidence of infection. Phylogenetic analysis of the 18S rRNA gene demonstrated that half of the sequences obtained from positive tick samples matched closest with a Hepatozoon species previously identified in the water python population. The remaining sequences were found to be more closely related to mammalian and amphibian Hepatozoon species. This study confirms that species of Amblyomma harbor DNA of the same Hepatozoon species detected in the water pythons. The detection of an additional genotype suggests the ticks may be exposed to 2 Hepatozoon species, providing further opportunity to study multiple host-vector-parasite relationships

Binding affinities of amino acid analogues at the charged aqueous titania interface : implications for titania-binding peptides

Despite the extensive utilization of biomolecule-titania interfaces, biomolecular recognition and interactions at the aqueous titania interface remain far from being fully understood. Here, atomistic molecular dynamics simulations, in partnership with metadynamics, are used to calculate the free energy of adsorption of different amino acid side chain analogues at the negatively-charged aqueous rutile TiO2 (110) interface, under conditions corresponding with neutral pH. Our calculations predict that charged amino acid analogues have a relatively high affinity to the titania surface, with the arginine analogue predicted to be the strongest binder. Interactions between uncharged amino acid analogues and titania are found to be repulsive or weak at best. All of the residues that bound to the negatively-charged interface show a relatively stronger adsorption compared with the charge-neutral interface, including the negatively-charged analogue. Of the analogues that are found to bind to the titania surface, the rank ordering of the binding affinities is predicted to be "arginine" > "lysine" ≈ aspartic acid > "serine". This is the same ordering as was found previously for the charge-neutral aqueous titania interface. Our results show very good agreement with available experimental data and can provide a baseline for the interpretation of peptide-TiO2 adsorption data.

In vitro and in situ screening systems for morphological and phytochemical analysis of Withania somnifera germplasms

We report, for the first time for Withania somnifera, the use of a modified in vitro system for morphological and phytochemical screening of true to type plants as compared with those grown in a conventional in situ system. Eleven germplasms of cultivated W. somnifera from different regions of India were collected to examine chemotypic variation in withaferin A (WA). Methods were developed to optimize WA extraction. The maximum concentration of WA was extracted from manually ground leaf and root material to which 60 % methanol was added followed by sonication in a water bath sonicator. Variation in WA concentration in whole plants was observed amongst the different germplasms. In the in vitro system, the concentration of WA ranged between 0.27 and 7.64 mg/g dry weight (DW) and in the in situ system, the range in concentration was between 8.06 and 36.31 mg/g DW. The highest amount of WA found in leaves was 7.37 and 41.42 mg/g DW in the in vitro and the in situ systems respectively. In roots, the highest WA concentration was 0.27 mg/g DW in the in vitro and 0.60 mg/g DW in the in situ system. There are distinct advantages in using the in vitro grown plants rather than those grown in the in situ system including the simplicity of design, efficient use of space and nutrition and a system which is soil and contaminant free. The proposed in vitro system is therefore ideal for utilization in molecular, enzymatic and biochemical studies.

Physiological and morphological responses of the temperate seagrass Zostera muelleri to multiple stressors: investigating the interactive effects of light and temperature

Understanding how multiple environmental stressors interact to affect seagrass health (measured as morphological and physiological responses) is important for responding to global declines in seagrass populations. We investigated the interactive effects of temperature stress (24, 27, 30 and 32°C) and shading stress (75, 50, 25 and 0% shade treatments) on the seagrass Zostera muelleri over a 3-month period in laboratory mesocosms. Z. muelleri is widely distributed throughout the temperate and tropical waters of south and east coasts of Australia, and is regarded as a regionally significant species. Optimal growth was observed at 27°C, whereas rapid loss of living shoots and leaf mass occurred at 32°C. We found no difference in the concentration of photosynthetic pigments among temperature treatments by the end of the experiment; however, up-regulation of photoprotective pigments was observed at 30°C. Greater levels of shade resulting in high photochemical efficiencies, while elevated irradiance suppressed effective quantum yield (ΔF/FM'). Chlorophyll fluorescence fast induction curves (FIC) revealed that the J step amplitude was significantly higher in the 0% shade treatment after 8 weeks, indicating a closure of PSII reaction centres, which likely contributed to the decline in ΔF/FM' and photoinhibition under higher irradiance. Effective quantum yield of PSII (ΔF/FM') declined steadily in 32°C treatments, indicating thermal damage. Higher temperatures (30°C) resulted in reduced above-ground biomass ratio and smaller leaves, while reduced light led to a reduction in leaf and shoot density, above-ground biomass ratio, shoot biomass and an increase in leaf senescence. Surprisingly, light and temperature had few interactive effects on seagrass health, even though these two stressors had strong effects on seagrass health when tested in isolation. In summary, these results demonstrate that populations of Z. muelleri in south-eastern Australia are sensitive to small chronic temperature increases and light decreases that are predicted under future climate change scenarios.

Analysis of phasic and tonic electromyographic signal characteristics: electromyographic synthesis and comparison of novel morphological and linear-envelope approaches

The pattern of tonic and phasic components in an EMG signal reflects the underlying behaviour of the central nervous system (CNS) in controlling the musculature. One avenue for gaining a better understanding of this behaviour is to seek a quantitative characterisation of these phasic and tonic components. We propose that these signal characteristics can range between unvarying, tonic and intermittent, phasic activation through a continuum of EMG amplitude modulation. In this paper, we present two new algorithms for quantifying amplitude modulation: a linear-envelope approach, and a mathematical morphology approach. In addition we present an algorithm for synthesising EMG signals with known amplitude modulation. The efficacy of the synthesis algorithm is demonstrated using real EMG data. We present an evaluation and comparison of the two algorithms for quantifying amplitude modulation based on synthetic data generated by the proposed synthesis algorithm. The results demonstrate that the EMG synthesis parameters represent 91.9% and 96.2% of the variance of linear-envelopes extracted from lumbo-pelvic muscle EMG signals collected from subjects performing a repetitive-movement task. This depended, however, on the muscle and movement-speed considered (F=4.02, p<0.001). Coefficients of determination between input and output amplitude modulation variables were used to quantify the accuracy of the linear-envelope and morphological signal processing algorithms. The linear-envelope algorithm exhibited higher coefficients of determination than the most accurate morphological approach (and hence greater accuracy, T=8.16, p<0.001). Similarly, the standard deviation of the coefficients of determination was 1.691 times smaller (p<0.001). This signal processing algorithm represents a novel tool for the quantification of amplitude modulation in continuous EMG signals and can be used in the study of CNS motor control of the musculature in repetitive-movement tasks.

Tuning the mechanical and morphological properties of self-assembled peptide hydrogels via control over the gelation mechanism through regulation of ionic strength and the rate of pH change

Hydrogels formed by the self-assembly of peptides are promising biomaterials. The bioactive and biocompatible molecule Fmoc-FRGDF has been shown to be an efficient hydrogelator via a π-β self-assembly mechanism. Herein, we show that the mechanical properties and morphology of Fmoc-FRGDF hydrogels can be effectively and easily manipulated by tuning both the final ionic strength and the rate of pH change. The increase of ionic strength, and consequent increase in rate of gelation and stiffness, does not interfere with the underlying π-β assembly of this Fmoc-protected peptide. However, by tuning the changing rate of the system's pH through the use of glucono-δ-lactone to form a hydrogel, as opposed to the previously reported HCl methodology, the morphology (nano- and microscale) of the scaffold can be manipulated.

Morphological and molecular investigations of the holocephalan elephant fish nephron: the existence of a countercurrent-like configuration and two separate diluting segments in the distal tubule

In marine cartilaginous fish, reabsorption of filtered urea by the kidney is essential for retaining a large amount of urea in their body. However, the mechanism for urea reabsorption is poorly understood due to the complexity of the kidney. To address this problem, we focused on elephant fish (Callorhinchus milii) for which a genome database is available, and conducted molecular mapping of membrane transporters along the different segments of the nephron. Basically, the nephron architecture of elephant fish was similar to that described for elasmobranch nephrons, but some unique features were observed. The late distal tubule (LDT), which corresponded to the fourth loop of the nephron, ran straight near the renal corpuscle, while it was convoluted around the tip of the loop. The ascending and descending limbs of the straight portion were closely apposed to each other and were arranged in a countercurrent fashion. The convoluted portion of LDT was tightly packed and enveloped by the larger convolution of the second loop that originated from the same renal corpuscle. In situ hybridization analysis demonstrated that co-localization of Na(+),K(+),2Cl(-) cotransporter 2 and Na(+)/K(+)-ATPase α1 subunit was observed in the early distal tubule and the posterior part of LDT, indicating the existence of two separate diluting segments. The diluting segments most likely facilitate NaCl absorption and thereby water reabsorption to elevate urea concentration in the filtrate, and subsequently contribute to efficient urea reabsorption in the final segment of the nephron, the collecting tubule, where urea transporter-1 was intensely localized.