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Immune function plays an important role in an animal's defense against infectious disease. In reptiles, immune responses may be complex and counterintuitive, and diagnostic tools used to identify infection, such as induced antibody responses are limited. Recent studies using gene transcription profiling in tortoises have proven useful in identifying immune responses to various intrinsic and extrinsic stressors. As part of a larger experiment with Mojave desert tortoises (Gopherus agassizii), we facilitated the transmission of the pathogenic bacteria, Mycoplasma agassizii (Myag), to naïve adults and measured innate and induced immune reactions over time. Specifically, we evaluated clinical condition, presence of Myag in the nasal/oral cavity, induced antibody responses specific to Myag, and measured molecular reactions (gene transcript profiles) in 15 captive tortoises classified as naïve, exposed, or infected and 14 wild tortoises for comparison. Myag was confirmed inside the nasal/oral cavity in exposed tortoises within 30–60 days of introduction to infected animals, yet we did not detect Myag specific induced antibody responses in these individuals until 420–595 days post exposure. Surprisingly, we found no overall differences in the gene transcript profiles between our experimental treatment groups throughout this study. This work highlights the complexities in assessing immune function and diagnosing pathogen related infections in tortoises and other reptiles. We facilitated the transmission of pathogenic bacteria to Mojave desert tortoises in order to evaluate innate and induced immune responses over time. We evaluated the clinical condition, presence of bacteria, inducted antibody responses, and molecular reactions (gene transcription profiles). Pathogenic bacteria were detected within 30–60 days of introduction to infected animals, yet inducted antibody responses were not detected. Surprisingly, we found no differences in the gene transcript profiles among our experimental treatment groups throughout this experiment. This work highlights the complexities in assessing immune function and diagnosing pathogen related infection in tortoises and other reptiles. [ABSTRACT FROM AUTHOR]

Habitat disturbance from development, resource extraction, off-road vehicle use, and energy development ranks highly among threats to desert systems worldwide. In the Mojave Desert, United States, these disturbances have promoted the establishment of nonnative plants, so that native grasses and forbs are now intermixed with, or have been replaced by invasive, nonnative Mediterranean grasses. This shift in plant composition has altered food availability for Mojave Desert tortoises (Gopherus agassizii), a federally listed species. We hypothesized that this change in forage would negatively influence the physiological ecology, immune competence, and health of neonatal and yearling tortoises. To test this, we monitored the effects of diet on growth, body condition, immunological responses (measured by gene transcription), and survival for 100 captive Mojave tortoises. Tortoises were assigned to one of five diets: native forbs, native grass, invasive grass, and native forbs combined with either the native or invasive grass. Tortoises eating native forbs had better body condition and immune functions, grew more, and had higher survival rates (>95%) than tortoises consuming any other diet. At the end of the experiment, 32% of individuals fed only native grass and 37% fed only invasive grass were found dead or removed from the experiment due to poor body conditions. In contrast, all tortoises fed either the native forb or combined native forb and native grass diets survived and were in good condition. Health and body condition quickly declined for tortoises fed only the native grass (Festuca octoflora) or invasive grass (Bromus rubens) with notable loss of fat and muscle mass and increased muscular atrophy. Bromus rubens seeds were found embedded in the oral mucosa and tongue in most individuals eating that diet, which led to mucosal inflammation. Genes indicative of physiological, immune, and metabolic functions were transcribed at lower levels for individuals fed B. rubens, indicating potential greater susceptibility to disease or other health-related problems. This study highlights the negative indirect effects of invasive grasses, such as red brome, in desert ecosystems, and provides definitive evidence of a larger negative consequence to health, survival, and ultimately population recruitment for Mojave Desert tortoises than previously understood. [ABSTRACT FROM AUTHOR]

ABSTRACT Wildfires burned 24,254 ha of critical habitat designated for the recovery of the threatened Mojave desert tortoise ( Gopherus agassizii) in southern Nevada during 2005. The proliferation of non-native annual grasses has increased wildfire frequency and extent in recent decades and continues to accelerate the conversion of tortoise habitat across the Mojave Desert. Immediate changes to vegetation are expected to reduce quality of critical habitat, yet whether tortoises will use burned and recovering habitat differently from intact unburned habitat is unknown. We compared movement patterns, home-range size, behavior, microhabitat use, reproduction, and survival for adult desert tortoises located in, and adjacent to, burned habitat to understand how tortoises respond to recovering burned habitat. Approximately 45% of home ranges in the post-fire environment contained burned habitat, and numerous observations ( n = 12,223) corroborated tortoise use of both habitat types (52% unburned, 48% burned). Tortoises moved progressively deeper into burned habitat during the first 5 years following the fire, frequently foraging in burned habitats that had abundant annual plants, and returning to adjacent unburned habitat for cover provided by intact perennial vegetation. However, by years 6 and 7, the live cover of the short-lived herbaceous perennial desert globemallow ( Sphaeralcea ambigua) that typically re-colonizes burned areas declined, resulting in a contraction of tortoise movements from the burned areas. Health and egg production were similar between burned and unburned areas indicating that tortoises were able to acquire necessary resources using both areas. This study documents that adult Mojave desert tortoises continue to use habitat burned once by wildfire. Thus, continued management of this burned habitat may contribute toward the recovery of the species in the face of many sources of habitat loss. Published 2015. This article is a U.S. Government work and is in the public domain in the USA. [ABSTRACT FROM AUTHOR]

Wildlife translocation is increasingly used to mitigate disturbances to animals or habitat due to human activities, yet little is known about the extent to which translocating animals causes stress. To understand the relationship between physiological stress and translocation, we conducted a multiyear study (2007-2009) using a population of desert tortoises ( Gopherus agassizii) near Fort Irwin, California. Blood samples were collected from adult tortoises in three treatment groups (resident, translocated and control) for 1 year prior to and 2 years after translocation. Samples were analyzed by radioimmunoassay for plasma total corticosterone ( CORT), a glucocorticoid hormone commonly associated with stress responses in reptiles. CORT values were analyzed in relation to potential covariates (animal sex, date, behavior, treatment, handling time, air temperature, home-range size, precipitation and annual plant production) among seasons and years. CORT values in males were higher than in females, and values for both varied monthly throughout the activity season and among years. Year and sex were strong predictors of CORT, and translocation explained little in terms of CORT. Based on these results, we conclude that translocation does not elicit a physiological stress response in desert tortoises. [ABSTRACT FROM AUTHOR]

The method of in situ time-resolved high-energy X-ray diffraction, using the intrinsically highly collimated X-ray beam generated by the European Synchrotron Radiation Facility, is demonstrated. A specially designed cell, which allows the addition of liquid components, has been used to study the reaction mechanisms of a foamed bioactive calcia–silica sol–gel glass immersed in simulated body fluid. Analysis of the X-ray diffraction data from this experiment provides atomic distances, via the pair correlation functions, at different stages of the dissolution of the glass and of the associated calcium phosphate, and ultimately hydroxyapatite, i.e. bone mineral, formation. Hence, changes in the atomic scale structure can be analysed as a function of reaction time, giving an insight into the evolution of the structure of both the glass matrix and the hydroxyapatite surface growth. [ABSTRACT FROM AUTHOR]

In areas of the country where the U.S. Department of Energy has classified the available wind resources as Class 3 or greater, the use of wind turbines to provide power to relatively small remediation systems such as groundwater circulation wells may be technically and economically feasible. Groundwater circulation wells are a good candidate technology to couple with renewable energy, because the remediation system removes contamination from the subject aquifer with no net loss of the groundwater resource, while the wind turbine does not create potentially harmful air emissions. Wind data collected in the vicinity of the former Nebraska Ordnance Plant Superfund site were used to select a wind turbine system to provide a portion of the energy necessary to power a groundwater circulation well located in an area of high trichloroethylene groundwater contamination. Because utility power was already installed at the remediation system, a 10 kW grid inter-tie wind turbine system supplements the utility system without requiring batteries for energy storage. The historical data from the site indicate that the quantity of energy purchased correlates poorly with the quantity of groundwater treated. Preliminary data from the wind turbine system indicate that the wind turbine provides more energy than the remediation system treatment components and the well submersible pump require on a monthly average. The preliminary results indicate that the coupling of wind turbines and groundwater circulation wells may be an attractive alternative in terms of the system operation time, cost savings, and contaminant mass removal. © 2004 Wiley Periodicals, Inc. [ABSTRACT FROM AUTHOR]

Wild type rat lens main intrinsic protein (MIP) and MIP mutated (F73I, F75L) to resemble the glycerol facilitator of Escherichia coli in the region of the NPA1 box were used to investigate the topology of MIP in the membrane of Spodoptera frugiperda (Sf21) cells using the baculovirus expression system and expression in mouse erythroid leukaemia cells (MEL C88). Differential fixation for staining was used, with paraformaldehyde for externally exposed antigenic sites, and acetone for both externally and internally exposed protein antigenic sites. Immunofluorescence using antibodies to synthetic MIP peptides showed that wild type MIP had a six transmembrane topography. The N- and C-termini were intracellular in both expression systems, and both NPA boxes were found to be extracellular. These results show that residues around the NPA1 box can influence the folding of the MIP in the membrane, and provide structural evidence for the poor water transport properties of MIP, as the NPA boxes lie outside the plane of the membrane. [Copyright &y& Elsevier]

Wild-type rat lens main intrinsic protein (MIP) was heterologously expressed in the membrane of Spodoptera frugiperda (Sf21) cells using the baculovirus expression system and in mouse erythroid leukaemia cells (MEL C88). Both MEL and Sf21 cell lines expressing wild-type MIP were investigated for the conductance of ions using a whole cell patch clamp technique. An increase in conductance was seen in both expression systems, particularly on lowering the pH to 6.3. In Sf21 cells, addition of antibodies to the NPA1 box resulted in a reduction of current flow. These results suggest that MIP has pH-dependent ion channel activity, which involves the NPA1 box domain. [Copyright &y& Elsevier]

Preschool children were taught four two-choice match-to-sample conditional discriminations with 10 arbitrary visual stimuli. For 6 participants, 2 of the 10 stimuli served as the sample, or conditional, stimuli in all discriminations. For 5 additional participants, the same pair of stimuli served as the discriminative, or comparison, stimuli in all discriminations. Equivalence classes were established with more participants in the latter group, replicating prior research with participants with retardation. Four participants, in whom equivalence classes were established and who were available for further participation, were exposed to new conditional discriminations without trial-by-trial feedback and involving some novel and some familiar stimuli. Consistent conditional responding was observed, and tests for inclusion of the novel stimuli in the original classes showed class expansion. Training to reverse the unreinforced conditional performances produced a reversal of class membership in 3 of 4 participants, an outcome not consistent with other studies. The results are discussed with respect to the interaction of class structure and size.

Experimental methods and apparatus were developed to measure platelet concentration in an ultrafiltering hollow fiber with blood and ultrafiltrative flow rates that are characteristic of current hemofiltration practice. Values are presented for the adjusted local platelet concentration, which allows for the expected increase due to loss of ultrafiltrate volume. Platelet concentrations in fiber samples are of interest since ultra-filtration can create platelet polarization in a fashion similar to the concentration polarization that occurs for proteins. For many combinations of inlet flow rate, ultrafiltrative flow rate, and fiber dimension, there are significant elevations in adjusted fiber platelet concentration. The adjusted platelet concentrations of the outflow samples are nominally equal to those of the feed (reservoir) samples; thus, reaction and adhesion of platelets is not a significant source of fiber concentration. Experiments with fibers of different diameters indicate that, as expected in concentration polarization, the excess platelets are located near the wall. Data for the fiber platelet concentration depend in a complex fashion upon the inlet wall shear rate and ultrafiltrative velocity. For certain fixed ultrafiltrative velocities, a maximum fiber platelet concentration occurs at an intermediate inlet wall shear rate. The data are qualitatively explained by an extension of a published model involving diffusive, ultrafiltrative, and wall-induced fluxes of red cells and platelets.

Extracorporeal membrane oxygenation (ECMO) can support neonates with severe respiratory failure. Currently, the most common application of ECMO requires venoarterial access. Venovenous (VV) ECMO is desirable to avoid common carotid artery ligation. However, the best technique of venous access for VV ECMO is not established. Using a single cannula with a double-lumen (DLC) in the right atrium for simultaneous drainage and infusion of blood, VV ECMO provided total respiratory support for six apneic puppies for 3 h each. Mean systemic arterial oxygenation was lower with DLC VV (50 torr) compared to VA ECMO (247 torr), but a physiologic pH (mean 7.34) was maintained on DLC VV bypass. Higher mean bypass flow was required on DLC VV (124 ml/kg/min) compared to VA flow (101 ml/kg/min) because of recirculation of oxygenated blood. The position of the DLC in the right atrium needed to be closely monitored. Hemorrhage was noted in the myocardium after use of DLC VV ECMO.