The translocation of organisms is becoming a frequently used tool in conservation biology. There are, however, a disproportionate number of unsuccessful attempts translocating populations of herpetofauna. Logistical and temporal limits of monitoring, combined with ambiguous metrics concerning "success," have led to few advances regarding reptile translocations. Successfully established and persistent populations are those in which both the founding population and subsequent generations show consistent or positive levels of survival and reproduction. A small population of the threatened Florida Sand Skink (Plestiodon reynoldsi) was translocated in 2007. Data collected from 2007 to 2009 confirmed survival and reproduction among the founding individuals, but the sampling did not include a long enough period to allow for the evaluation of the survival and reproduction of individuals born on the site. In this study, individuals were collected during two separate sampling events, one during the third spring and one during the sixth spring after the translocation occurred. Survival estimates, reproduction, population size and generation structure were calculated by combining and analyzing data from all years post-translocation. The numbers of both total and new individuals captured in the sixth year exceeded captures from every prior sampling event since monitoring began in 2008. Founding individuals represented only 14% of the total individuals captured, while the number of individuals born on site continued to increase. The proportion of recruits and increased number of hatchlings despite the loss of founders shows that the filial generations are producing offspring. The methods utilized in assessing this translocation effort will further the understanding of the population dynamics of the Florida Sand Skink and allow for more informed decisions in future management studies of this threatened species.

This thesis presents the results of an investigation into the uptake and distribution of cadmium (Cd) in two cultivars of potato (Solanum tuberosum L.) shown to contain different concentrations of Cd in the tuber at maturity. An initial glasshouse trial sought to determine whether differences in tuber Cd between these two cultivars resulted from differences in uptake from the soil, or were due to differences in the allocation of Cd to the various tissues within the plant. Total uptake of Cd from the soil did not differ between cultivars, nor did the yield of tubers. However, there were marked differences in Cd distribution within the plant. Most of the differences in tuber Cd concentration could be accounted for by a large (3-fold) retention of Cd in the roots of cultivar Wilwash. The concentration of Cd in the shoots of Wilwash was also higher than of Kennebec, although to a lesser extent than the roots. Further studies were conducted to trace the pathways of Cd uptake and movement within the plant. A split-pot trial, involving long-term growth of potatoes in 109Cd-labelled soil, was undertaken to determine the overall pattern of Cd distribution and the importance of the root system in supplying Cd to the tubers. The root system of the potato plant is different to many plants, in that the main root system (basal roots) is augmented after tuber initiation by roots extending from the stolon and from the tuber itself. The basal roots were found to be the dominant source of Cd to all tissues and accounted for approximately 85 % of tuber Cd. The remaining tuber Cd was sourced directly from the stolon and tuber roots. However, there was no evidence of a direct link between the main (basal) root system and the stolons. Although Cd was found to accumulate in the periderm of the tubers, there was no uptake into the tuber tissue itself. Isotopic studies were undertaken to investigate the short-term movement of newly absorbed Cd in the xylem and the phloem. Cadmium was found to be highly mobile in both the xylem and phloem, with added Cd being rapidly assimilated into all tissues following both root and foliar application. Newly absorbed Cd was rapidly sequestered by the stems when applied to either the soil or to a source leaf, suggesting that the stems may act as a transitional storage pool when rapid turnover of nutrients and other mineral elements is required during tuber bulking. Inhibition of Cd uptake by zinc (Zn), has been proposed as a method for reducing the concentration of Cd in various agricultural crops, including potatoes. The ability of Zn to reduce Cd uptake was found to be highly dependent upon cultivar and on the concentration of Cd in the external medium. Although competition between Zn and Cd was found for cultivar Wilwash when the external concentration of Cd was low, when the concentration of Cd in the external media was high, increasing Zn served to increase Cd uptake. Both synergistic and competitive responses were also noted for cultivar Kennebec. However, the patterns of response were opposite to those evident in Wilwash. The complexity of these interactions highlighted the possible shortcomings in using soil applied Zn to limit Cd uptake by potatoes. / Thesis (Ph.D.) -- University of Adelaide, School of Earth and Environmental Sciences, 2004.

Published works [representing] original research conducted during the various phases of [his] academic development--Pref / Includes references / 1 v. (various pagings) : / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (D. Sc.)--Faculty of Science, University of Adelaide, 1990

Reduction of cardiac myocyte loss and repair of the vasculature post myocardial infarction are important therapeutic goals because the potential for intrinsic repair is limited. Preclinical and limited clinical data support the possibility that bone marrow-derived mesenchymal stem cells may be a suitable cell type for cellular therapy. The goal of this research was to determine the effectiveness of using MSCs from aged mice in cellular therapy for the treatment of AMI. The central hypothesis for this research was that therapeutic potential of mesenchymal stem cells decreases with age. This research utilized global gene expression analysis to investigate molecular differences in MSCs harvested from three different age groups of mice. Microarray analysis was performed to investigate changes in gene expression with respect to aging. Furthermore, both in vitro and in vivo experiments were completed to analyze the functional and molecular characteristics of the MSCs. The data identified age-related defects in mouse MSCs as well as determined the molecular basis for these deficiencies. This study indicates that MSCs from 26m mice are severely deficient in the induction of angiogenesis and cardiac repair due to defective paracrine factor secretion caused by decreased expression of growth factor/cytokine genes. Hypoxia attenuates the deficiency in the aged mice, whereas in young mice low oxygen promotes secretion of paracrine growth factors. It was determined a dysfunction in HIF-1 alpha signaling was present in MSCs from 26m mice and is regulated by the PI3K/Akt signaling in MSCs. Furthermore, two novel and important and novel aspects of this study were the discovery that cell cycle regulation gene expression decreases with age and MSCs have increased insulin resistance with age. Increased insulin resistance in this cell type with aging is likely to have profound effects on the clinical outcomes of using these cells therapeutically. Likewise, loss of cell cycle regulation during proliferation could also lead to undesirable clinical effects. Gaining insight to the repair potential of these cells with respect to age will help to better define future trials of autologous stem cells not only for heart disease but for all of the many applications proposed for these cells.

Approximately 10% of the open reading frames in the genome of the Gram-negative bacterium E. coli encodes secretory proteins, and 20% encodes integral inner membrane proteins (IMPs). These proteins are sorted to their correct cellular compartments (the periplasm and the outer and inner membranes) by specialized targeting and translocation/insertion systems. So far, a very limited set of model proteins have been used to study proteins sorting requirements in E. coli. The main objective of all the papers presented in this thesis was to determine the targeting and translocation/insertion requirements of more E. coli proteins. In papers I and II, this was done using focused approaches. Selected model proteins (lipoproteins and putative outer membrane proteins) were expressed from plasmids and their targeting and translocation were analysed in vitro by crosslinking experiments and/or in vivo by pulse-chase analysis in different E. coli mutant strains. In papers III a comparative sub-proteome analysis was carried out to define the role of the cytoplasmic chaperone SecB in protein targeting. In paper IV, a similar approach was used to study how protein translocation and insertion is affected upon depletion of the essential Sec-translocon component SecE. The ‘global’ approach used in paper III and IV allowed us to study protein targeting and translocation/insertion requirements on a proteome level. This led to the identification of several novel SecB substrates and a large number of potential Sec-translocon independent IMPs.

The mammalian intestine contains a dense and diverse community of microorganisms. The resident microbiota makes contributions to host to promote proper immune system development and limit pathogen colonization. In this study, the effects of microbiota disruption with or without TLRs stimulation on intestinal permeability and immunity were examined in C57BL/6 mice receiving antibiotic treatment for 6 days and in antibiotics-treated mice received dead E. coli or S. aureus at day 4. The results showed that antibiotic treatment significantly decreased the total number of bacteria including specific aerobic group Enterobacteriaceae and Enterococcus, and specific anaerobic group Lactococcus/Bifidobacterium in intestinal mucosa and lumen. Although only a slight increase in the intestinal permeability and no change in caspase-3 activity of intestinal mucosa were observed after antibiotic treatment, the bacterial translocation (BT) to mesenteric lymph nodes (MLN) increased significantly. Subsequent experiments showed that antibiotic treatment decreased the mucosal killing activity and the expression of non-defensin family including RegIII£], RegIII£^, CRP-ductin and RELM£] but not the defensin family, and increased the translocation of pathogen K. pneumoniae significantly, suggesting that the increase of BT to MLN after antibiotic treatment is likely due to a reduction in gut immunity rather than an increase of intestinal permeability. Moreover, stimulation of TLR4 reversed the effect of antibiotic treatment, suggesting that the functioning of TLR4 in intestinal epithelium is required to prevent pathogenic invasion and maintain intestinal homeostasis.

Catalase-peroxidase (KatG) of Mycobacterium tuberculosis is a bifunctional heme enzyme that has been shown to play an important role in the activation of a first line drug, isoniazid (INH), used in the treatment of tuberculosis infection. Mutations in the katG gene have been found to be associated with INH resistance. The most commonly encountered mutation is the Ser315Thr point mutation. In this dissertation, the x-ray crystallographic structures of MtbKatG and the mutant enzyme KatG[S315T] are presented to explore the molecular basis of the INH activation and resistance. The structure is dimeric and contains a heme cofactor in each subunit of the dimer. The most important change in KatG[S315T] is due to the presence of the methyl group of the threonine 315 side chain, which is located at the narrowest part of the substrate channel. The protruding methyl group effectively constricts the accessibility to the heme by closing down the dimensions of the channel, constraining the substrate entrance. VirB11 of Brucella abortus is a hexameric ATPase that belongs to the type IV secretion system. The crystal structure of BaVirB11 was found to contain six molecules per asymmetric unit. The Walker A (P loop), His box, and Glu box from the C-terminal domain are located at the interface of the N- and C-terminal domain. A large conformational change was found in the linker region when compared with that of HP0525 structure, the VirB11 analogous from H. pylori. To elucidate the functional role of each domain, seven functional mutations were generated and used for biochemical studies. The GER motif and the linker region were found to be crucial for ATP hydrolysis activity of BaVirB11. Mutations in the GER motif (R101Q) and the linker region (R120E) of BaVirB11 completely abolish the ATP hydrolysis activity of the enzyme. The binding affinities of the two mutants to the ATP; however, are similar to that of the wild-type enzyme, indicating that mutation in the GER motif or the linker region has no effect on ATP binding.

Historically, the collared peccary (Tayassu tajacu) occurred throughout much of Texas including the northern portion of the Texas Hill Country. Remaining peccary populations were extirpated in much of their former range due to over harvest and habitat loss. In 2004, efforts to restore peccary populations to the Texas Hill Country began when Texas Parks and Wildlife Department biologists translocated 29 collared peccaries into the 2,157 ha, Mason Mountain Wildlife Management Area (MMWMA). I evaluated the success of peccary translocations for mixed and intact family groups by comparing survival, ranges, and dispersal of translocated, radio-tagged peccaries. In addition, I evaluated two release methods (soft versus hard) to determine differences in population demographics. I found that peccary ranges and dispersal patterns did not differ (P > 0.05) between intact and mixed groups or release method (soft versus hard). However, I did find that peccary fidelity to release sites was greater for soft releases of family groups. Individuals from the soft release group dispersed the shortest distance and stayed on MMWMA. Only 2 individuals from the hard releases stayed on MMWMA while the rest (19 individuals) dispersed 4-8 km. Future peccary translocations should emphasize the release method employed and family structure of individuals released to improve translocation effectiveness in establishing populations in target areas.

Coral reefs thrive in nutrient-deficient environments yet function among the most productive ecosystems on Earth as a consequence of the symbiosis between coral hosts and their symbiotic zooxanthellae. The symbiotic unit (holobiont) can utilize both inorganic and organic sources of nutrients for the accumulation of carbon and nitrogen required for metabolism, growth, and reproduction. An iterative model was created to describe the flux of carbon and nitrogen between a host and its algae. The model design is based on a previously published conceptual model of algal symbioses; functions and values of input parameters are based on published studies of the coral species Stylophora pistillata. The model is designed to simulate responses of the coral, zooxanthellae and the holobiont to different environmental variables, either one at a time or changing simultaneously. Simulations presented are for default values based on previously published data for S. pistillata adapted to high-light (shallow-euphotic) and low-light (deep-euphotic) environments, and for single-variable manipulations of rates of a) host feeding, b) photosynthesis, and c) dissolved inorganic nitrogen (DIN) uptake. Simulations examining feeding rates between 0% and 6.5% of host biomass indicate that biomass of both high-light and low-light adapted holobionts increase exponentially with increased feeding, with benefit to the high-light holobiont ~8 times greater than to the low-light holobiont. Increasing rates of photosynthesis illustrated that a low-light holobiont is carbon limited, is primarily dependent upon host feeding, and can benefit from a small increase in photosynthesis rate. Simulations examining rates of DIN input indicate that the high-light holobiont functions optimally when inorganic nitrogen input is very low. Increase in DIN up to 0.5% resulted in benefit to the holobiont, but more resulted in unrealistically excessive growth by the zooxanthellae until a function to maintain a fixed range for the host-zooxanthellae biomass ration function was included in the model. Simulations for the low-light holobiont did not indicate any benefit from DIN input. The model was originally designed using a spreadsheet-based program which frequently became overloaded when testing multiple variables. Modification of the model in software better designed for modeling is recommended for future work.

The GTPase elongation factor G (EF-G) is an important player in the complex process of protein synthesis by bacterial ribosomes. Although extensively studied much remains to be learned about this fascinating protein. In the elongation phase, after incorporation of each amino acid into the growing peptide chain, EF-G translocates the ribosome along the mRNA template. In the recycling phase, when the synthesis of a protein has been completed, EF-G, together with ribosome recycling factor (RRF), splits the ribosome into its subunits. We developed the first in vitro assay for measuring the average time of a complete translocation step at any position along the mRNA. Inside the open reading frame, at saturating EF-G concentration and low magnesium ion concentration, translocation rates were fast and compatible with elongation rates observed in vivo. We also determined the complete kinetic mechanism for EF-G- and RRF-dependent splitting of the post-termination ribosome. We showed that splitting occurs only when RRF binds before EF-G and that the rate and GTP consumption of the reaction varies greatly with the factor concentrations. The antibiotic fusidic acid (FA) inhibits bacterial protein synthesis by binding to EF-G when the factor is ribosome bound, during translocation and ribosome recycling. We developed experimental methods and a theoretical framework for analyzing the effect of tight-binding inhibitors like FA on protein synthesis. We found that FA targets three different states during each elongation cycle and that it binds to EF-G on the post-termination ribosome both in the presence and absence of RRF. The stalling time of an FA-inhibited ribosome is about hundred-fold longer than the time of an uninhibited elongation cycle and therefore each binding event has a large impact on the protein synthesis rate and may induce queuing of ribosomes on the mRNA. Although ribosomes in the elongation and the recycling phases are targeted with similar efficiency, we showed that the main effect of FA in vivo is on elongation. Our results may serve as a basis for modelling of EF-G function and FA inhibition inside the living cell and for structure determination of mechanistically important intermediate states in translocation and ribosome recycling.