A cement material has been developed that has a controllable set time, compressive strength of about 50 MPa and shrinkage of under 2% when heated to 960 degrees centigrade. The main compound of this material has been shown to be the potassium analogue of struvite, in which potassium ion is substituted for the ammonium ion. The cement is formed by reacting a slurry made with potassium carbonate and monoammonium phosphate with magnesium oxide. Ihe amount and rate of addition of water to the mixture is controlled to determine the set time. It has been demonstrated that this cement may be used as a matrix in conjunction with cenospheres or cellulose fibres to form a low density composite material, and with silica powder to form a high strength composite. As well as optimising the cement mix for maximum strength and minimum shrinkage, a variety of properties of the K-cement were investigated. It was found that long term exposure of the cement to the atmosphere does not result in the formation of magnesium carbonate, but long term exposure to saline solutions can result in the formation of dypingite on the surface. No effect on strength was observed as a consequence of the age of the slurry or the cement, within 1-2 weeks / Doctor of Philosophy (PhD)

Secondary salinisation has rendered over 100 million hectares of land throughout the world, and over 5 million hectares in Australia, unsuitable for conventional agriculture. The utilization of salinised land and its associated water resources for mariculture is an adaptive approach to this environmental problem with many potential economic, social and environmental benefits. Despite this, inland mariculture is yet to develop into an industrial-scale, rural enterprise. The main aim of this study was therefore to identify and address some of the technical and biological limitations to the development of an inland finfish mariculture industry. Three technical aspects essential to the development of an Australian inland mariculture industry were reviewed; potential sources of water, the species suitable for culture in these water sources and the production systems available to produce them. Based on factors such as their quantity, quality and proximity to infrastructure, the most appropriate water sources were deemed to be groundwater obtained from interception schemes and waters from operational or disused mines. In terms of species, mulloway (Argyrosomus japonicus) were identified as having many positive attributes for inland mariculture, including being temperate and therefore having the ability to be cultured year-round in the regions where the majority of secondary salinity occurs. Seasonal production of barramundi (Lates calcarifer) in ponds in the temperate climatic zones has potential, but may be more appropriate for those salinised water sources located in the warmer parts of the country. Rainbow trout (Oncorhynchus mykiss) were also identified as having excellent potential provided water temperature can be maintained below the upper lethal limit and also have potential for seasonal production, perhaps in rotation with barramundi. In terms of production systems, pond-based culture methods were found to have many advantages specific to inland mariculture. Static ponds enable culture in areas with low groundwater yield and more cost-effective potassium supplementation compared with flow through ponds. Static ponds also largely overcome the issues associated with the disposal of salt-laden and eutrophied waste water; however yields from static ponds are typically low and limited by the nutrient input into the pond. In response to the yield constraints of static pond culture, a new culture technology known as the Semi-Intensive Floating Tank System (SIFTS) was designed, patented and constructed in collaboration with the aquaculture industry and tested in a static inland saline pond in the wheatbelt of Western Australia. This technology was designed to reduce nutrient input into ponds by the collection of settleable wastes and to provide large volumes of well-oxygenated water to the target species, to ameliorate the loss of fish from low dissolved oxygen during strong microalgal blooms. The three species identified above has having excellent potential for inland mariculture (mulloway, rainbow trout, and barramundi) were grown in SIFTS held within a 0.13 ha static, inland saline water body (salinity 14 ppt) over a period of 292 days, yielding the equivalent of 26 tonnes/ha/year (total for all three species). Rainbow trout were grown with an FCR of 0.97 from 83 to 697 grams over 111 days (SGR, 1.91%/day) between June and September, when average daily water temperatures ranged from 12.3„aC to 18.2„aC. Over the same time period, mulloway grew only from 100 to 116 grams, however, once temperatures increased to approximately 21„aC in October, feed intake increased and mulloway grew to an average size of 384 grams over 174 days with an SGR and FCR of 0.68 %/day and 1.39, respectively. Barramundi stocked in November with an average weight of 40 grams increased to 435 grams in 138 days (SGR 1.73%/day) with an FCR of 0.90. The SIFTS significantly reduced nutrient input into the pond by removing settleable wastes as a thick sludge with a dry matter content of 5 to 10%. The total quantity of dry waste removed over the 292 day culture period was 527 kg (5 tonnes/ha/yr), which was calculated to contain 15 kg of nitrogen (144 kg/ha/yr) and 16 kg of phosphorus (153 kg/ha/yr). The release of soluble nutrients into the pond resulted in blooms of macro- and micro- algae which caused large and potentially lethal diurnal fluctuations in dissolved oxygen within the pond, however, comparatively stable levels of dissolved oxygen were maintained within each SIFT through the use of air lift pumps. It is well documented that saline groundwater is deficient in potassium which, depending on the extent of the deficiency, can negatively impact on the performance of marine species, including fish. The physiological effects of this deficiency on fish, however, have not been previously described. As such, I conducted a bioassay investigating the physiological effects of a hypersaline (45 ppt) groundwater source containing 25% of the potassium found in equivalent salinity seawater (i.e. 25% K-equivalence) on juvenile barramundi. Histopathological examination of moribund fish revealed severe degeneration and necrosis of skeletal muscles, marked hyperplasia of branchial chloride cells and renal tubular necrosis. Clinical chemistry findings included hypernatraemia and hyperchloridaemia of the blood plasma and lowered muscle potassium levels. It was concluded from this study that the principal cause of death of these barramundi was skeletal myopathy induced by unsustainable buffering of blood plasma potassium levels from the muscle. Although such hypokalaemic muscle myopathies have been previously described in mammals and birds, this was the first description of such myopathies in fish. It was hypothesized from the results described above that the physiological effects of potassium deficiency are dependent on salinity and that they would be ameliorated by potassium supplementation. These predictions were tested in a subsequent study which measured the effects of potassium supplementation between 25% and 100% K-equivalence on the growth, survival and physiological response of juvenile barramundi at hyperosmotic (45 ppt), near-isosmotic (15 ppt) and hyposmotic (5 ppt) salinities. Unlike those juvenile barramundi reared at 45 ppt and 25% K-equivalence in the previous study, those reared in 50% K-equivalence water at 45 ppt in this study survived for four weeks but lost weight; whereas at 75% and 100% K-equivalences fish both survived and gained weight. Homeostasis of blood plasma potassium was maintained by buffering from skeletal muscle. Fish reared in 50% K-equivalence at this salinity exhibited muscle dehydration, increased branchial, renal and intestinal (Na+-K+)ATPase activity and elevated blood sodium and chloride, suggesting they were experiencing osmotic stress. At 15 ppt, equal rates of growth were obtained between all K-equivalence treatments. Buffering of plasma potassium by muscle also occurred but appeared to be in a state of equilibrium. Barramundi at 5 ppt displayed equal growth among treatments. At this salinity, buffering of plasma potassium from muscle did not occur and at 25% K-equivalence blood potassium was significantly lower than at all other K-equivalence treatments but with no apparent effect on growth, survival or (Na+-K+)ATPase activities. These data confirmed the hypothesis that proportionally more potassium is required at hyperosmotic salinities compared to iso- and hypo- osmotic salinities and also demonstrated that barramundi have a lower requirement for potassium than other marine and estuarine species being investigated for culture in inland saline groundwater. In addition to ongrowing fish, saline groundwater has potential for hatchery production. Specific advantages include the vertical integration of inland saline farms and the production of disease-free certified stock through isolation from the pathogens and parasites found naturally in coastal water. To determine the potential of utilizing inland saline groundwater for hatchery production, barramundi larvae were reared from 2 to 25 days post hatch in 14 ppt saline groundwater with either no potassium supplementation (38% K-equivalence) or full potassium supplementation (100% K-equivalence). Growth, survival and swimbladder inflation of these larvae were compared against those grown in control treatments of seawater (32 ppt) and seawater diluted to 14 ppt. Those reared in saline groundwater with 38% K-equivalence exhibited complete mortality within 2 days, whilst those held in groundwater with full supplementation survived at a rate equal to both control treatments (pooled average 51.1 ¡Ó 0.5%). At 25 days post hatch, there was no significant difference in larval length or dry weight between those grown in the 14 ppt control treatment and those in the saline groundwater with full potassium supplementation. There were no significant differences in swim bladder inflation between any of the surviving treatments (average 93.3 ¡Ó 2.5%). This is the first description of rearing barramundi larvae both in low salinity seawater and in saline groundwater, and demonstrates that the requirement for potassium by larval barramundi is higher than for juveniles of the same species. In addition to a deficiency in potassium, saline groundwater in Western Australia often contains an elevated concentration of manganese relative to seawater as a result of anaerobic reduction of manganese oxides or the pedogenic weathering of manganese-bearing rock. The effects of elevated manganese on marine or estuarine fish have not been described and a study was therefore conducted to determine if manganese, at a concentration typical of that found in saline groundwater, has any impact on fish. The effects of 5 mg/L of dissolved manganese on juvenile mulloway at salinities of 5, 15 and 45 ppt were determined by comparing the survival, growth and blood and organ chemistry with those grown at the same salinities without manganese addition. Survival of mulloway at 45 ppt in the presence of 5 mg/L of manganese (73 ¡Ó 13%) was significantly lower than all other treatments, which achieved 100% survival. Those fish grown in seawater without manganese exhibited rapid growth, which was not affected by salinity (SGR = 4.05 ¡Ó 0.29%/day). Those fish grown at 5 ppt and 45 ppt in the presence of manganese lost weight over the two week trial (SGR 0.17 ¡Ó 0.42 and -0.44 ¡Ó 0.83%/day, respectively), whilst those at 15 ppt gained only a small amount of weight (SGR 1.70 ¡Ó 0.20%/day). Growth was therefore affected by manganese and by the interaction of manganese and salinity, but not salinity alone. Manganese was found to accumulate in the gills, liver and muscle of the fish. No gill epithelial damage or other significant histological findings were found, however, significant differences in blood chemistry were observed. Blood sodium and chloride of manganese exposed fish were significantly elevated in hyperosmotic salinity (45 ppt) and depressed at hyposmotic salinity (5 ppt) compared with unexposed fish; consistent with manganese causing apoptosis or necrosis to chloride cells. Blood potassium was significantly elevated and liver potassium significantly reduced at all salinities in the presence of manganese. These findings are consistent with manganese interfering with carbohydrate metabolism. There were no differences in blood sodium, chloride or potassium across salinities in fish not exposed to manganese, demonstrating mulloway are capable of efficient osmoregulation across this salinity range.

Schizophrenia is a devastating psychiatric disorder that affects approximately 1% of the population. The main treatments for schizophrenia are antipsychotic drugs that target dopamine receptors, yet the underlying biological mechanisms through which they alleviate the symptoms of schizophrenia remain ill defined. In this study, we used microarray analysis to profile the expression changes of thousands of genes simultaneously, following antipsychotic drug treatment of mice. Mice were treated chronically (28 days), or for a novel intermediate time-point (7 days), with one of three antipsychotic drugs: clozapine, haloperidol or olanzapine. The use of three drugs enabled us to discern antipsychotic-specific effects co-regulated by multiple drugs, rather than the side effects of individual compounds. Transcript profiling and validation by quantitative PCR of whole brain tissue revealed antipsychotic drug regulation of genes in diverse biological pathways, including: dopamine metabolism, neuropeptide and second-messenger signalling, neurogenesis, synaptic plasticity, cell adhesion, myelination, and voltage-gated ion channels. The regulation of voltage-gated channels by antipsychotic drugs has been suggested previously by electrophysiological studies, although thorough analysis has not been undertaken in vivo. Therefore, the second aim of this study was to characterise the regional mRNA and protein expression of two genes altered by multiple APDs, the voltage-gated potassium channel ??-subunit (Kcna1) and voltage-gated potassium channel interacting protein (Kchip3). Regional characterisation and expression analyses were carried out by immunohistochemistry, in situ hybridisation, and Western blot analysis of mouse brain regions of interest to schizophrenia and its treatment. Following 7-day haloperidol treatment we observed up-regulation of Kcna1 in the striatum and dentate gyrus, with increased protein in the striatum, hippocampus and midbrain; and down-regulation of Kchip3 in the striatum, with decreased protein in the cortex, hippocampus and midbrain. These studies implicate voltage-gated potassium channels in the antipsychotic drug regulation of midbrain dopaminergic neuronal activity, adult neurogenesis and/or striatothalamic GABAergic neuronal inhibition. These findings indicate that regulation of potassium channels may underlie some of the mechanisms of action of antipsychotic drugs, and that voltage-gated ion channels may provide alternative drug targets for the treatment of schizophrenia.

Transactions of magnesium (Mg) along the gastrointestinal tract and the effect of change in potassium (K) intake were recorded in two in vivo experiments in sheep and cattle. Additional information on the sensitivity to K intake was obtained by comparing Mg transport and electrochemical properties of isolated rumen epithelia of sheep and cattle in 4 additional in vitro experiments. The experiment described in Chapter 2, and performed in sheep housed indoors in metabolic crates, investigated the compensatory capacity of the intestine to respond to the reduction in Mg absorption from the stomach as a consequence of increase in K intake. The animals were equipped with a ruminal cannula and two intestinal cannulae (duodenum and ileum) and flow of digesta was measured by the addition of two indigestible markers, chromium ethylenediaminetetra-acetic acid (Cr-EDTA) and ytterbium acetate (Yb). The animals were infused in a Latin square design for periods of 10 days with a solution of K bicarbonate that provided between 15 and 47 g of K/day. The diet consisted of a 50:50 combination of concentrates plus lucerne hay that provided around 3.7 g of Mg per day and 15 g of K per day. After 5 days of infusion samples of feed, faeces, urine and plasma were collected and analysed for Mg and K content. After 6 days of infusion, samples of duodenal and ileal flow were obtained. The treatments reproduced the detrimental effect of K on Mg absorption, especially in the rumen; a rise in K intake from 15 to 23 g/day reduced total Mg absorption from the gastrointestinal tract from 1.36 to 1.23 g/day, further increase in K intake to 38 and 47 g/day reduced absorption to 1.12 and 1.05 g/day, an overall reduction of around 50% in Mg apparent availability. Magnesium was mainly absorbed in the stomachs and large intestine with the small intestine a site of net secretion. Most of the reduction in Mg absorption with increase in K intake occurred in the stomachs, reducing from 1.86 to 1.11 g/day. A compensatory reduction in the net secretion of Mg from the intestines (small and large) was observed. This compensation was largely due to reduction in net secretion from the small intestine, from 0.85 to 0.22 g/day, rather than an increase in net absorption from the large intestine, although both segments acted synergistically. Results also suggested significant individual variation in plasma Mg concentration, urinary Mg excretion and in the flow and absorption of Mg along the gastrointestinal tract. It was suggested that most of that variability was due to genetic factors. Differences between species (cattle and sheep) were pursued during the course of the experiment described in Chapter three. Four triple cannulated rams and 3 triple cannulated dry cows were placed in metabolic crates, fed daily fresh-cut pasture and infused, in a total randomised design that provided the equivalent of an intake of 30,40 and 50 g of K per kg dry matter intake (DMI) per day. Solutions of K (as K bicarbonate) and markers (CrEDT A and Yb acetate) were infused continuously for a period of 10 days; after 5 days of infusion samples of pasture, faeces, urine and plasma were collected and analysed for Mg and K content. After 6 days of infusion, samples of duodenal and ileal flow were obtained. Total feed offered, refusals and water consumption were recorded daily. Results showed a greater sensitivity of cattle to the increase in K supply. A rise in K supply from 30 to 40 g per kg DMI/day reduced Mg absorption by almost 50% from 0.32 to 0.16 g per kg DMI/day, whereas only the highest treatment dose (50 g of K per kg DMI/day) produced the same deleterious effect in sheep. The absorption of Mg occurred mainly in the stomachs and large intestine; in contrast the small intestine was a site of net secretion in both species. The addition of K slightly reduced the rate of Mg absorption from the rumen, especially in cattle. Similarly, net Mg secretion within the intestines increased with increasing K intake in both species, only to be counterbalanced by a greater Mg absorption from the large intestine. The large intestine in both species (sheep and cattle) reduced faecal losses of Mg but was unable to fully compensate for the reduction in Mg absorption from the stomach or the greater net Mg secretion observed at the small intestine. Differences between species in water content of the faeces were observed to be mainly related to the moisture content of the digesta that reached the ileum rather than a result of differences in absorption in the large intestine. More evidence of species differences in Mg transport and of sensitivity to K intake were obtained by using isolated rumen epithelia and the Ussing chamber technique. Transport and electrophysiological properties of the tissues were observed in standard conditions and by adding different K concentrations to the mucosal side. Under standard conditions and open-circuit voltage, sheep isolated rumen epithelia had greater transmural potential difference (PDt), and lower conductance (Gt) but similar short-circuit current (Isc) than those from cattle. These results suggested that the rumen epithelium of cattle is leakier than that of sheep. Measurement of the transport of Mg showed that isolated rumen epithelia of cattle transported more Mg and was saturated at higher Mg concentrations (12 vs 4 mM) than sheep epithelia. These differences in Mg influx (transport from mucosa to serosa) were also observed in studies of Mg transport using stable isotopes. Magnesium influx (transport from mucosa to serosa) from the isolated rumen of cattle was greater than in sheep (57.5 ± 12.72 vs. 17.3 ± 12.72 nmol.cm⁻².h⁻¹); however this was counterbalanced by a greater Mg efflux (transport from serosa to mucosa) of 48.1 ± 12.72 vs. 9.9 ± 12.72 nmol.cm⁻².h⁻¹, for cattle and sheep respectively, when mucosal K concentrations were around 25 mM. A increase in K concentration on the mucosal side enhanced transmural potential difference (PDt) and short-circuit current (Isc) to a greater extent in sheep than in cattle, suggesting a greater effect of K on sheep than on cattle epithelia. On the other hand, the transport of Mg measured by stable isotopes suggested that net absorption of Mg (7.4 ± 12.72 vs. 11.1 ± 12.72 nmol.cm⁻².h⁻¹) in sheep epithelia was similar at 25 and 50 mM of K on the mucosal side, whereas net Mg influx in cattle was largely depressed as a consequence of a reduction in Mg influx (mucosa to serosa) from 57.7 ± 12.72 to 2.9 ± 12.72 nmol.cm⁻² h⁻¹ together with a constant Mg efflux (serosa to mucosa) 48.1 ± 12.72 and 41.2 ± 12.72 nmol.cm⁻².h⁻¹, presumably leaving through a paracellular shunt. However, this finding was based on date from a small size and caution should be applied to this conclusion. In conclusion, data collected from several comparative studies suggest differences in Mg apparent availability between sheep and cattle and also a greater sensitivity of cattle to an increase in K intake. This high sensitivity to K represents a great risk of hypomagnesaemia in dairy cattle in New Zealand where high K concentration is endemic in pastures. Most importantly, these results suggest that models for Mg metabolism in cattle should be based on measurements from cattle nutritional and physiological studies rather than on extrapolation from sheep studies.

Le plomb se distingue de ses homologues de la colonne IVB par une moindre stabilité des composés de degré d'oxydation 4+, la stabilité des composés de degré 2+ s'accroissant simultanément. C'est ainsi que les oxydes tétravalents du germanium et de l'étain possèdent une très grande stabilité thermique....

<p>Experimentally derived biomolecular structures were determined by Nuclear Magnetic Resonance (NMR). The properties of selected peptides and proteins in solution and in membrane mimicking micelles were observed by circular Dichroism (CD), mass spectrometry (MS), and other spectroscopic techniques.</p><p>The mDpl(1-30) peptide (30 residues) of the mouse Doppel protein was found to be positioned as an α-helix in a DHPC micelle. The same peptide can disrupt and cause leakage in small unilamellar vesicles.</p><p>Single D-amino acid isomers of Trp-cage (20 residues), the smallest peptide with a protein-like fold, were analyzed by CD spectroscopy and were found to have different secondary structures and melting temperatures. They were compared against MS measurements specially designed to reveal the secondary structure of proteins.</p><p>We studied a novel protein in E. coli of unknown structure that is encoded by the putative transcription factor ORF: ygiT (131 residues). This protein comprises a helix-turn-helix (HTH) domain in the C-terminus and contains two CxxC motives in the N-terminal domain, which binds Zn. This protein was named 2CxxC. We succeeded in overexpressing and purifying 2CxxC in E. coli with enough yield for a 13C, 15N uniformly labeled NMR sample. The chemical shift assignment was completed and the NMR structure was calculated in reducing, slightly acidic conditions (1mM DTT, pH 5.5). The determined HTH domain shows good similarity with structures predicted by a homology search, while the N-terminal domain has no other homologous structure in the Protein Data Bank (PDB).</p><p>The structure of the paddle region (27 residues) of the HsapBK(233-260) voltage and Ca+2 activated potassium channel, in DPC micelles, was determined by NMR. It shows a helix-turn-helix loop, which agrees well with the expected structure and could help to verify the proposed models of the voltage gating mechanism.</p><p>The C-repressor (dimer of 99 residues) of bacteriophage P2 was analyzed by NMR. We assigned the chemical shifts and NMR structure determination is under way.</p>

Graduation date: 1995

Based on a Ph.D. thesis in the Dept. of Electrical Engineering, 1963. / Bibliography: p. 81-83.

Experimentally derived biomolecular structures were determined by Nuclear Magnetic Resonance (NMR). The properties of selected peptides and proteins in solution and in membrane mimicking micelles were observed by circular Dichroism (CD), mass spectrometry (MS), and other spectroscopic techniques. The mDpl(1-30) peptide (30 residues) of the mouse Doppel protein was found to be positioned as an α-helix in a DHPC micelle. The same peptide can disrupt and cause leakage in small unilamellar vesicles. Single D-amino acid isomers of Trp-cage (20 residues), the smallest peptide with a protein-like fold, were analyzed by CD spectroscopy and were found to have different secondary structures and melting temperatures. They were compared against MS measurements specially designed to reveal the secondary structure of proteins. We studied a novel protein in E. coli of unknown structure that is encoded by the putative transcription factor ORF: ygiT (131 residues). This protein comprises a helix-turn-helix (HTH) domain in the C-terminus and contains two CxxC motives in the N-terminal domain, which binds Zn. This protein was named 2CxxC. We succeeded in overexpressing and purifying 2CxxC in E. coli with enough yield for a 13C, 15N uniformly labeled NMR sample. The chemical shift assignment was completed and the NMR structure was calculated in reducing, slightly acidic conditions (1mM DTT, pH 5.5). The determined HTH domain shows good similarity with structures predicted by a homology search, while the N-terminal domain has no other homologous structure in the Protein Data Bank (PDB). The structure of the paddle region (27 residues) of the HsapBK(233-260) voltage and Ca+2 activated potassium channel, in DPC micelles, was determined by NMR. It shows a helix-turn-helix loop, which agrees well with the expected structure and could help to verify the proposed models of the voltage gating mechanism. The C-repressor (dimer of 99 residues) of bacteriophage P2 was analyzed by NMR. We assigned the chemical shifts and NMR structure determination is under way.

Voltage-gated ion channels are fundamental for electrical signaling in living cells. They are composed of four subunits, each holding six transmembrane helices, S1-S6. Each subunit contains a voltage-sensor domain, S1-S4, and a pore domain, S5-S6. S4 contains several positively charged amino-acid residues and moves in response to changes in membrane voltage. This movement controls the opening and closing of the channel. The structure of the pore domain is solved and demonstrates principles of channel selectivity. The molecular mechanism of how the voltage sensor regulates the opening of the channel is still under discussion. Several models have been discussed. One of the models is the paddle model where S3b and S4 move together. The second one is the helical-twist where S4 makes a small rotation in order for the channel to open. The third one is the helical-screw model where S4 twists around its axis and moves diagonally towards the extracellular side of the channel. The aim of this PhD project was to study the molecular movement of the voltage sensor in the depolarization-activated Shaker K channel. Cloned channels were expressed in Xenopus laevis oocytes, and investigated with several electrophysiological techniques. 1. We show that S4 moves in relation to both S3b and S5. The formation of some disulfide bonds between S4 and neighboring positions, in only the open state, shows that the paddle model cannot be correct. Furthermore, electrostatic and steric effects of residues in S3b suggest that S3b is tilted, with the intracellular part close to S4. 2. We show that the relatively Mg-sensitive Shaker K channel is changed into the less Mg-sensitive Kv2.1 K channel with respect to its sensitivity to extracellularly applied Mg2+ by changing the charge of three extracellularly positioned amino acid residues. One of the residues, F425C, mediates its effect through the neighboring residue K427. 3. We show that oxaliplatin, an anti-cancer drug, has no effect on the Shaker K channel. It has been suggested that a negatively charged monochloro complex of oxaliplatin is the active substance, and also causes the neurotoxic side effects. Neither this complex shows any effect on the channel. Our experiments point towards the helical-screw model. The other models for voltage-sensor movements are incompatible with the results in this study.