Collaborative Research: Linkages among Mitochondrial Form, Function and Thermal Tolerance of Antarctic Notothenioid Fishes

Antarctic notothenioid fishes have evolved in the Southern Ocean for 10-14 MY under an unusual set of circumstances. Their characteristics include the complete absence of the circulating oxygen-binding protein, hemoglobin (Hb) within the Channichthyid (Icefish) family of notothenioids. Moreover, some species within the 16 members of this family have also lost the ability to express the oxygen-binding and storage protein, myoglobin (Mb) in cardiac muscle. Our previous work has determined that the loss of Hb and/or Mb is correlated with significant increases in densities of mitochondria within oxidative tissues, and extensive remodeling of these vital organelles. To date, nothing is known about how modifications in mitochondrial architecture of icefishes affect organelle function, or more importantly, how they affect organismal-level physiology. Most critical for Antarctic fishes is that mitochondrial characteristics have been linked to how well ectotherms can withstand increases in temperature. This collaborative research project will address the hypothesis that the unusual mitochondrial architecture of Antarctic Channichthyids has led to changes in function that impact their ability to withstand elevations in temperature. Specifically, the research will (1) determine if the unusual mitochondrial architecture of icefishes affects function and contributes to organismal thermal sensitivity, (2) identify differences in organismal thermal tolerance between red- and white- blooded notothenioids, (3) identify molecular mechanisms regulating changes in mitochondrial structure in icefishes. The results may establish channichthyid icefishes as a sentinel taxon for signaling the impact of global warming on the Southern Ocean. Broad impacts of this project will be realized by participation of high school biology teachers in field work through cooperation with the ARMADA project at the University of Rhode Island, as well as graduate education

Evolution of an Oxygen-Binding Hemoprotein in a Unique Environment: Myoglobin in the Hemoglobinless Antarctic Icefishes

For approximately the last 40 million years, a unique fish fauna has been evolving in the frigid seas surrounding Antarctica. Physiological function of these water-breathing animals has demanded many adaptations to ensure proper metabolism and regulation of biochemical processes at cell temperatures of about OoC. Among the polar fishes, one family is particularly unusual, the Channichthyid icefishes. Species in this family lack hemoglobin in their circulating blood and, at least the majority, do not possess the intracellular respiratory protein, myoglobin, that is normally responsible for enhancing movement of oxygen through aerobic muscle tissues. The PI\u27s have recently found two species of icefish, Pseudochaenichthys georgianus and Chionodraco rastrospinosus, however, that express myoglobin, but only in one tissue - heart ventricle. The metabolism of all icefish species is highly aerobic and is based largely on the combustion of fatty fuels as energy sources. The field work will consist of a combination of trawling from R/V Polar Duke to capture icefish species and laboratory work at Palmer Station using tissues from these animals. At Palmer Station they will conduct experiments and prepare purified, fixed and or frozen material for shipment back to our CONUS laboratory for further analyses. Our overall goals are: 1) to test whether to determine myoglobin protein remains physiologically significant at the cold body temperatures of Antarctic fishes, 2) to reconstruct the evolution of the myoglobin gene within the icefish family and, 3) to identify the mechanisms controlling expression of myoglobin within icefish and related species

Carcinogenic Effects in a Phenylketonuria Mouse Model

Phenylketonuria (PKU) is a metabolic disorder caused by impaired phenylalanine hydroxylase (PAH). This condition results in hyperphenylalaninemia and elevated levels of abnormal phenylalanine metabolites, among which is phenylacetic acid/phenylacetate (PA). In recent years, PA and its analogs were found to have anticancer activity against a variety of malignancies suggesting the possibility that PKU may offer protection against cancer through chronically elevated levels of PA. We tested this hypothesis in a genetic mouse model of PKU (PAHenu2) which has a biochemical profile that closely resembles that of human PKU. Plasma levels of phenylalanine in homozygous (HMZ) PAHenu2 mice were >12-fold those of heterozygous (HTZ) littermates while tyrosine levels were reduced. Phenylketones, including PA, were also markedly elevated to the range seen in the human disease. Mice were subjected to 7,12 dimethylbenz[a]anthracene (DMBA) carcinogenesis, a model which is sensitive to the anticancer effects of the PA derivative 4-chlorophenylacetate (4-CPA). Tumor induction by DMBA was not significantly different between the HTZ and HMZ mice, either in total tumor development or in the type of cancers that arose. HMZ mice were then treated with 4-CPA as positive controls for the anticancer effects of PA and to evaluate its possible effects on phenylalanine metabolism in PKU mice. 4-CPA had no effect on the plasma concentrations of phenylalanine, phenylketones, or tyrosine. Surprisingly, the HMZ mice treated with 4-CPA developed an unexplained neuromuscular syndrome which precluded its use in these animals as an anticancer agent. Together, these studies support the use of PAHenu2 mice as a model for studying human PKU. Chronically elevated levels of PA in the PAHenu2 mice were not protective against cancer

Turkdean Roman Villa, Gloucestershire: archaeological investigations 1997-1998

Before the transmission of the first ‘live’ Time Team television programme on 23 August 1997, the existence of a Roman villa near Chalkhill Barn in the parish of Turkdean, 12 miles north-east of Cirencester and 2 miles from the Fosse Way, was hardly known to the archaeological community (FIG. 1). That a Roman building did exist in this location had, however, been suspected for a number of years by the landowner, the late Mr Wilf Mustoe. Distinctive linear parchmarks suggestive of buildings had been clearly visible at ground level in the grass pasture each dry summer, and in 1976 Mr Mustoe made a measured sketch plan of them on the back of an envelope. Subsequently the sketch was drawn up into a scale plan entitled ‘Roman villa’ by Simon Goddard, a relation. There was little knowledge of the site outside of Mr Mustoe's family until it was independently ‘discovered’ by local archaeologist Roger Box in August 1996 whilst fortuitously flying over the site in a helicopter. In the evening light Mr Box instantly recognised the parchmarks of an unmistakable Roman villa and took a series of photographs (FIG. 2). Mr Box showed his photographs to Mr Mustoe, and with his agreement wrote to Time Team suggesting that this would be an excellent site for a television programme. Arrangements were duly set in place and the evidence of the cropmarks was confirmed by a trial geophysical survey in March 199

Kožna dekontaminacija živčanoga bojnog otrova sarina s apsorpcijskim pripravkom u uvjetima in vivo

Our Institute’s nuclear, biological, and chemical defense research team continuously investigates and develops preparations for skin decontamination against nerve agents. In this in vivo study, we evaluated skin decontamination efficacy against sarin by a synthetic preparation called Mineral Cationic Carrier (MCC®) with known ion exchange, absorption efficacy and bioactive potential. Mice were treated with increasing doses of sarin applied on their skin, and MCC® was administered immediately after contamination. The results showed that decontamination with MCC® could achieve therapeutic efficacy corresponding to 3 x LD50 of percutaneous sarin and call for further research.Istraživački tim NBKO (nuklearno-biološko-kemijske obrane) radi na pronalasku i razvoju pripravka za dekontaminaciju kože od živčanih bojnih otrova. Cilj ovog istraživanja bio je ispitati dekontaminacijska svojstva (adsorpcijska i/ili kemisorpcijska) pripravka MCC® rabeći živčani bojni otrov sarin kao kožni kontaminant u uvjetima in vivo. MCC® je sintetski pripravak koji je biokemijski aktivan i ima ionskoizmjenjivačka i adsorpcijska svojstva. Istraživanje u uvjetima in vivo napravljeno je na miševima aplikacijom rastućih doza sarina na kožu životinje. Pripravak MCC® uporabljen je kao kožni dekontaminant neposredno nakon kožne kontaminacije sarinom. Istraživanja su pokazala da pripravak MCC® posjeduje adsorpcijska svojstva, ujedno važna za dekontaminaciju živčanih bojnih otrova. Eksperimenti u uvjetima in vivo na miševima (NOD-soj) pokazali su da se dekontaminacijom pripravkom MCC® može postići terapijski učinak od 3 LD50 (perkutano, sarin)

Pralidoxime in Acute Organophosphorus Insecticide Poisoning-A Randomised Controlled Trial

Background: Poisoning with organophosphorus (OP) insecticides is a major global public health problem, causing an estimated 200,000 deaths each year. Although the World Health Organization recommends use of pralidoxime, this antidote's effectiveness remains unclear. We aimed to determine whether the addition of pralidoxime chloride to atropine and supportive care offers benefit. Methods and Findings: We performed a double-blind randomised placebo-controlled trial of pralidoxime chloride (2 g loading dose over 20 min, followed by a constant infusion of 0.5 g/h for up to 7 d) versus saline in patients with organophosphorus insecticide self-poisoning. Mortality was the primary outcome; secondary outcomes included intubation, duration of intubation, and time to death. We measured baseline markers of exposure and pharmacodynamic markers of response to aid interpretation of clinical outcomes. Two hundred thirty-five patients were randomised to receive pralidoxime (121) or saline placebo (114). Pralidoxime produced substantial and moderate red cell acetylcholinesterase reactivation in patients poisoned by diethyl and dimethyl compounds, respectively. Mortality was nonsignificantly higher in patients receiving pralidoxime: 30/121 (24.8%) receiving pralidoxime died, compared with 18/114 (15.8%) receiving placebo (adjusted hazard ratio HR] 1.69, 95% confidence interval CI] 0.88-3.26, p = 0.12). Incorporating the baseline amount of acetylcholinesterase already aged and plasma OP concentration into the analysis increased the HR for patients receiving pralidoxime compared to placebo, further decreasing the likelihood that pralidoxime is beneficial. The need for intubation was similar in both groups (pralidoxime 26/121 21.5%], placebo 24/114 21.1%], adjusted HR 1.27 95% CI 0.71-2.29]). To reduce confounding due to ingestion of different insecticides, we further analysed patients with confirmed chlorpyrifos or dimethoate poisoning alone, finding no evidence of benefit. Conclusions: Despite clear reactivation of red cell acetylcholinesterase in diethyl organophosphorus pesticide poisoned patients, we found no evidence that this regimen improves survival or reduces need for intubation in patients with organophosphorus insecticide poisoning. The reason for this failure to benefit patients was not apparent. Further studies of different dose regimens or different oximes are required

Developing Health-Based Pre-Planning Clearance Goals for Airport Remediation Following Chemical Terrorist Attack: Introduction and Key Assessment Considerations

In the event of a chemical terrorist attack on a transportation hub, post-event remediation and restoration activities necessary to attain unrestricted facility reuse and re-entry could require hours to multiple days. While restoration timeframes are dependent on numerous variables, a primary controlling factor is the level of pre-planning and decision-making completed prior to chemical terrorist release. What follows is the first of a two-part analysis identifying key considerations, critical information, and decision criteria to facilitate post-attack and post-decontamination consequence management activities. A conceptual site model and human health-based exposure guidelines are developed and reported as an aid to site-specific pre-planning in the current absence of U.S. state or Federal values designated as compound-specific remediation or re-entry concentrations, and to safely expedite facility recovery to full operational status. Chemicals of concern include chemical warfare nerve and vesicant agents and the toxic industrial compounds phosgene, hydrogen cyanide, and cyanogen chloride. This work has been performed as a national case study conducted in partnership with the Los Angeles International Airport and The Bradley International Terminal. All recommended guidelines have been selected for consistency with airport scenario release parameters of a one-time, short-duration, finite airborne release from a single source followed by compound-specific decontamination