Tumor necrosis factor family member LIGHT acts with IL-1ß and TGF-ß to promote airway remodeling during rhinovirus infection.

BACKGROUND: Rhinovirus (RV) can exacerbate allergen-driven asthma. However, it has been suggested that serial infections with RV may also lead to asthma-like features in childhood without prior allergen exposure. AIM: We sought to test the effects of RV infection in the absence of allergen challenge on lung tissue remodeling and to understand whether RV induced factors in common with allergen that promote remodeling. METHODS: We infected C57BL/6 mice multiple times with RV in the absence or presence of allergen to assess airway remodeling. We used knockout mice and blocking reagents to determine the participation of LIGHT (TNFSF14), as well as IL-1ß and TGF-ß, each previously shown to contribute to lung remodeling driven by allergen. RESULTS: Recurrent RV infection without allergen challenge induced an increase in peribronchial smooth muscle mass and subepithelial fibrosis. Rhinovirus (RV) induced LIGHT expression in mouse lungs after infection, and alveolar epithelial cells and neutrophils were found to be potential sources of LIGHT. Accordingly, LIGHT-deficient mice, or mice where LIGHT was neutralized, displayed reduced smooth muscle mass and lung fibrosis. Recurrent RV infection also exacerbated the airway remodeling response to house dust mite allergen, and this was significantly reduced in LIGHT-deficient mice. Furthermore, neutralizing IL-1ß or TGF-ß also limited subepithelial fibrosis and/or smooth muscle thickness induced by RV. CONCLUSION: Rhinovirus can promote airway remodeling in the absence of allergen through upregulating common factors that also contribute to allergen-associated airway remodeling.

Polar and magnetic layered A-site and rock salt B-site-ordered NaLnFeWO6 (Ln = La, Nd) perovskites.

We have expanded the double perovskite family of materials with the unusual combination of layered order in the A sublattice and rock salt order over the B sublattice to compounds NaLaFeWO6 and NaNdFeWO6. The materials have been synthesized and studied by powder X-ray diffraction, neutron diffraction, electron diffraction, magnetic measurements, X-ray absorption spectroscopy, dielectric measurements, and second harmonic generation. At room temperature, the crystal structures of both compounds can be defined in the noncentrosymmetric monoclinic P2(1) space group resulting from the combination of ordering both in the A and B sublattices, the distortion of the cell due to tilting of the octahedra, and the displacement of certain cations. The magnetic studies show that both compounds are ordered antiferromagnetically below T(N) ≈ 25 K for NaLaFeWO6 and at ∼21 K for NaNdFeWO6. The magnetic structure of NaNdFeWO6 has been solved with a propagation vector k = ((1/2) 0 (1/2)) as an antiferromagnetic arrangement of Fe and Nd moments. Although the samples are potential multiferroics, the dielectric measurements do not show a ferroelectric response.

Crystal structure, phase transitions, and magnetic properties of iridium perovskites Sr2MIrO6 (M = Ni, Zn).

Double perovskites containing Ir(6+)/Ir(5+) with formula Sr2MIrO6 (M = Ni, Zn) have been synthesized under high oxygen pressure conditions. Their crystal structures have been studied by X-ray and neutron powder diffraction at room temperature (RT) and 2 K. At RT, these oxides crystallize in the monoclinic space group P2(1)/n with unit-cell parameters a ≈ √2a0, b ≈ √2a0, and c ≈ 2a0, and ß ≈ 90°. The thermal evolution of the structure of the Ni-containing compound shows the presence of two phase transition in the 373-673 K interval following the sequence P2(1)/n → I4/m → Fm3m. These materials have also been characterized by magnetic measurements, suggesting the onset of antiferromagnetic interactions at T(N) = 58 and 46 K, for M = Ni, Zn, respectively. X-ray absorption spectroscopy sheds light on the oxidation states of M and Ir ions within these double perovskites.

Biotic interactions in temporal trends (1992-2010) of organochlorine contaminants in the aquatic food web of Lake Laberge, Yukon Territory.

Declines in 6 organochlorine (OC) contaminant groups; chlordane (CHL), DDT, HCH, toxaphene (CHB), PCB and chlorinated benzenes (CBz) were measured in biota of a sub-Arctic lake (Lake Laberge, YT) following the closure of a commercial fishery in 1991. This study examined morphological (length, weight, age), biochemical (lipid content, δ(13)C, δ(15)N), population and OC data for 9 fishes and zooplankton between 1993 and 2003 (2010 for lake trout) to investigate causes for the OC declines. Growth dilution was a major factor influencing the decrease of OCs in lake trout, round whitefish and possibly zooplankton most notably in the early 2000s. A decline in lipids of most fish species also contributed to OC declines, although no such change was evident for zooplankton. It is suspected that increases in fish populations or climate variations over the 1990s, may have contributed towards a shift in plankton community composition. From 1991 to 1999, CPUE increased for 7 of the fish species and declined for 2 others. Concurrently, the zooplankton community shifted from an abundance of C. scutifer in 1993 to dominance by D. pribilofensis in 2001. Nitrogen and carbon stable isotope data suggested that food web interactions for most fish species have not changed over time. Although concentrations of OCs have declined in many fishes, the "rate" of OC transfer (using slopes of log OC vs. nitrogen isotope ratios) through the food web was greater in 2001 than in 1993. Overall, the declines in OC concentrations in the fish from Lake Laberge occurred concurrently with changes in their growth, lipid, and abundance, suggesting that ecosystem responses to the closure of the fishery were in part responsible for the lower contaminants in these fishes. As a result of this study, the Yukon government rescinded the health advisory for limiting the consumption of fish from Lake Laberge.

Magnetic and structural studies of the multifunctional material SrFe(0.75)Mo(0.25)O(3-δ).

SrFe0.75Mo0.25O3-δ has been recently discovered as an extremely efficient electrode for intermediate temperature solid oxide fuel cells (IT-SOFCs). We have performed structural and magnetic studies to fully characterize this multifunctional material. We have observed by powder neutron diffraction (PND) and transmission electron microscopy (TEM) that its crystal symmetry is better explained with a tetragonal symmetry (I4/mcm space group) than with the previously reported orthorhombic symmetry (Pnma space group). The temperature dependent magnetic properties indicate an exceptionally high magnetic ordering temperature (TN ∼ 750 K), well above room temperature. The ordered magnetic structure at low temperature was determined by PND to be an antiferromagnetic coupling of the Fe cations. Mössbauer spectroscopy corroborated the PND results. A detailed study, with X-ray absorption spectroscopy (XAS), in agreement with the Mössbauer results, confirmed the formal oxidation states of the cations to be mixed valence Fe(3+/4+) and Mo(6+).

Role of the New South Wales Department of Primary Industries' Laboratory Information Management System (LIMS) in the 2007 equine influenza emergency animal disease response.

A Laboratory Information Management System (LIMS) was used to manage the laboratory data and support planning and field activities as part of the response to the equine influenza outbreak in Australia in 2007. The database structure of the LIMS and the system configurations that were made to best handle the laboratory implications of the disease response are discussed. The operational aspects of the LIMS and the related procedures used at the laboratory to process the increased sample throughput are reviewed, as is the interaction of the LIMS with other corporate systems used in the management of the response. Outcomes from this tailored configuration and operation of the LIMS resulted in effective provision and control of the laboratory and laboratory information aspects of the response. The extent and immediate availability of the information provided from the LIMS was critical to some of the activities of key operatives involved in controlling the response.

Phenotype of autosomal dominant cone-rod dystrophy due to the R838C mutation of the GUCY2D gene encoding retinal guanylate cyclase-1.

AIMS: To describe the phenotype of members of a large Caucasian British family affected by autosomal dominant cone-rod dystrophy due to an R838C mutation in the guanylate cyclase 2D (GUCY2D) gene encoding retinal guanylate cyclase-1 (RETGC-1). METHODS: Retrospective review of 29 patients from four generations of the same family. RESULTS: Visual symptoms usually commenced in childhood. Only two patients, aged 14 and 25 years, had visual acuity compatible with driving. Of the 12 patients aged over 40 years, eight (66%) had vision of counting fingers or worse and were eligible for blind registration in the UK. Of the 29 patients, 18 (62%) had myopia greater than 5 D in at least one eye. Most had discernible macular changes on biomicroscopy, which varied from subtle RPE change to gross macular atrophy. All patients who underwent computerised perimetry exhibited a central or paracentral scotoma with normal peripheral field of vision. Of the 21 patients who underwent electrodiagnostic testing, all exhibited decreased cone function, but rod function was normal in 12 (57%) patients. CONCLUSION: We believe this report highlights the importance of phenotype-genotype correlation in cone and cone-rod dystrophies. Increased understanding of the varying phenotypes associated with different genetic mutations allows appropriate counselling of patients. In addition, the phenotypic characterisation of the natural history of these conditions may prove valuable in the future should therapeutic interventions become available.

Temporal trends of organochlorine contaminants in burbot and lake trout from three selected Yukon lakes.

Historical studies have demonstrated that organochlorine (OC) concentrations in top predators can vary considerably from lake to lake within a small geographic region but temporal trends of these contaminants have rarely been monitored in a sub-Arctic area for a long period of time. This study examined OC concentrations, including chlordane (CHL), DDT, hexachlorocyclohexane (HCH), toxaphene (CHB), PCB and chlorinated benzenes (CBz) in lake trout and burbot, from three Yukon lakes (Laberge, Kusawa, Quiet), over a span of 11 years (1992-2003). Temporal and spatial differences continue to exist in the OC concentrations of burbot and lake trout between these lakes. There is strong evidence that these contaminants are declining at various rates in lake trout (Salveninus namaycush) in Laberge, Kusawa and Quiet Lakes. For example, SigmaDDT concentrations have decreased 39%, 85% and 84% in Kusawa, Quiet and Laberge Lakes, respectively. Conversely, no consistent trends were observed in OC concentrations for burbot (Lota lota). For example, there is no evidence of a decline in toxaphene concentrations of Kusawa burbot yet a 58% decrease was observed in Laberge samples. Increases were also observed in the SigmaHCH levels of Kusawa Lake burbot, as well as increases in all OC groups (except SigmaHCH) for the Quiet Lake burbot samples. Decreases in burbot were evident in SigmaHCH and SigmaCHB for Lake Laberge fish and in SigmaCHL for Kusawa Lake samples. Spatial variations in OC levels are quite evident as Lake Laberge trout and burbot continued to maintain the highest levels over the eleven-year period from 1992 to 2003 followed by Kusawa Lake and then Quiet Lake. These differences were related to a variety of factors especially the species morphological characteristics such as log age, log weights and fish lipid content. A decreasing trend in Quiet and Laberge Lake trout lipid content, coupled with fluctuating condition factors and increases in body masses, suggest biotic changes may be occurring within the food webs due to fish population variations related to the cessation of commercial fishing or potentially an increase in lake plankton productivity related to annual climate variation. It is suspected that biotic factors rather than atmospheric inputs are the primary factors affecting the contaminant concentrations in lake trout and burbot in the study lakes.

Accommodative function in rhesus monkeys: effects of aging and calorie restriction.

Numerous degenerative changes in the visual system occur with age, including a loss of accommodative function possibly related to hardening of the lens or loss of ciliary muscle mobility. The rhesus monkey is a reliable animal model for studying age-related changes in ocular function, including loss of accommodation. Calorie restriction (CR) is the only consistent intervention to slow aging and extend lifespan in rodents, and more recently the beneficial effects of CR have been reported in nonhuman primates. The goal of the present study was to evaluate age-related changes in ocular accommodation and the potential effect of long-term (>8 years) CR on accommodation in male and female rhesus monkeys. Refraction, accommodation (Hartinger coincidence refractometer), and lens thickness (A-scan ultrasound) were measured in 97 male and female rhesus monkeys age 8-36 years under Telazol/acepromazine anesthesia. Refraction and accommodation measurements were taken before and after 40% carbachol corneal iontophoresis to induce maximum accommodation. Half the animals were in the control (CON) group and were fed ad libitum. The CR group received 30% fewer calories than age- and weight-matched controls. Males were on CR for 12 years and females for eight years. With increasing age, accommodative ability declined in both CON and CR monkeys by 1.03 ± 0.12 (P = 0.001) and 1.18 ± 0.12 (P = 0.001) diopters/year, respectively. The age-related decline did not differ significantly between the groups (P = 0.374). Baseline lens thickness increased with age in both groups by 0.03 ± 0.005 mm/year (P = 0.001) and 0.02 ± 0.005 mm/year (P = 0.001) for the CON and CR groups, respectively. The tendency for the for the lens to thicken with age occurred at a slower rate in the CR group vs. the CON group but the difference was not statistically significant (P = 0.086). Baseline refraction was -2.8 ± 0.55 and -3.0 ± 0.62 diopters for CON and CR, respectively. Baseline refraction tended to become slightly more negative with age (P = 0.070), but this trend did not differ significantly between the groups (P = 0.587). In summary, there was no difference in the slope of the age-related changes in accommodation, lens thickness, or refraction in the carbachol-treated eyes due to diet. These data are consistent with previous findings of decreased accommodative ability in aging rhesus monkeys, comparable to the age-dependent decrease in accommodative ability in humans. This study is the first to indicate that the accommodative system may not benefit from calorie restriction.

Australian survey of acrylamide in carbohydrate-based foods.

A method was developed and validated for the determination of acrylamide in carbohydrate-based foods. Solid-phase extraction employing a mixed-bed anion and cation exchange cartridge in series with a C18 extraction disk was used to clean-up water extracts of food samples before analysis by liquid chromatography coupled with tandem mass spectrometry detection. The limit of detection was calculated as approximately 25 microg kg(-1) and the limit of reporting was 50 microg kg(-1). The average method recovery for 84 samples from a range of matrices reporting was 99% with a relative standard deviation of 11.2%. A survey was conducted of 112 samples of carbohydrate-based foods composited from 547 products available in the Australian market. The analytical results were used in conjunction with Australian food consumption data derived from the 1995 National Nutrition Survey (NNS) to prepare preliminary dietary exposure estimates of Australians to acrylamide through only the food groups examined. Mean dietary exposure to acrylamide resulting from consumption of the foods tested, for Australians aged 2 years and above, was estimated as 22-29 microg day(-1) (equivalent to 0.4-0.5 microg kg(-1) bodyweight day(-1)) and between 73 and 80 microg day(-1) (1.4 and 1.5 microg kg(-1) bodyweight day(-1)) for 95th percentile consumers. Young children (2-6 years) consuming acrylamide-containing foods had a higher acrylamide exposure on a per kilogram bodyweight basis (mean 1.0-1.3 microg kg(-1) bodyweight day(-1)). The estimated exposure of Australians to acrylamide is similar to that estimated for other countries.

Circumvention of MHC class II restriction by genetic immunization.

The fate of T cell responses to peptide-based vaccination is subject to constraints by the major histocompatibility complex (MHC), MHC restriction. Using as a model system of T and B cell epitopes from the circumsporozoite protein of Plasmodium falciparum malaria parasite, we show that vaccination by somatic transgene immunization readily primes Balb/c mice (H-2(d)) a strain previously reported to be non-responder to immunization with a synthetic peptide vaccine encompassing these epitopes. Following genetic vaccination Balb/c mice developed a primary T cell response comparable to that of the responder strain C57Bl/6 (H-2(b)). Following booster immunization on day 45 Balb/c mice responded with a typical T cell memory response. Priming induced the formation of specific antibodies, which rose sharply after booster immunization. These findings suggests that genetic immunization can circumvent MHC class II restriction.

OX40 promotes Bcl-xL and Bcl-2 expression and is essential for long-term survival of CD4 T cells.

It is important to understand which molecules are essential for long-lived immunity. We show that OX40 (CD134) is required with CD28 for the survival of CD4 T cells following antigen-driven expansion. In contrast to CD28-/- T cells, which show defects early, OX40-/- T cells are relatively unimpaired in IL-2 production, cell division, and expansion. However, OX40-/- T cells fail to maintain high levels of Bcl-xL and Bcl-2 4-8 days after activation, and undergo apoptosis. Conversely, OX40 stimulation promotes Bcl-xL and Bcl-2 and suppresses apoptosis. Moreover, retroviral transduction of OX40-/- T cells with Bcl-xL or Bcl-2 reverses their survival defect. Thus, a temporal relationship exists between CD28 and OX40, with OX40 being a critical regulator of antigen-driven T cell survival.

Signaling through OX40 (CD134) breaks peripheral T-cell tolerance.

Peripheral T-cell tolerance is a mechanism to limit autoimmunity, but represents a major obstacle in diseases such as cancer. Tolerance is due to limited accumulation of antigen-specific T cells accompanied by functional hypo-responsiveness, and is induced by antigen encounter in a non-inflammatory environment. In contrast to advances in preventing induction of T-cell tolerance, there has been little progress in defining targets to reverse established tolerance. Here we show that signals from a single dose of an agonistic antibody against OX40 (CD134, a member of the tumor necrosis-factor family of receptors) can break an existing state of tolerance in the CD4+ T-cell compartment. OX40 signals promote T-cell expansion after the hypo-responsive phenotype is induced and restore normal functionality. These data highlight the potent costimulatory capacity of OX40, and indicate OX40 as a target for therapeutic intervention in a variety of related diseases.

Ultrasound biomicroscopy of the aging rhesus monkey ciliary region.

Ultrasound biomicroscopy of the living rhesus monkey ocular ciliary region was undertaken to identify age-dependent changes that might relate to the progression of presbyopia. Monkeys were anesthetized and pharmacologically cyclopleged, the eyelids were held open with a lid speculum, and sutures were placed beneath the medial and lateral rectus muscles. Ultrasound biomicroscopy imaging of the nasal and temporal quadrants of the eye were performed, and the live images were recorded to videotape. Subsequent image analysis was performed to obtain objective morphometric measurements of the ciliary body region. The ciliary body inner radius of curvature, outer radius of curvature, inner arc length, area, thickness, perimeter, zonular fiber length, and circumlental space were measured. Zonular space was calculated. The circumlental space decreased with increasing age in the temporal quadrant. The other morphologic measurements were not significantly correlated with age or body weight. Most morphologic measurements were significantly different comparing temporal vs. nasal quadrants. Bifurcation of the posterior zonular fibers was frequently observed. Although temporal circumlental space was the only measurement found to change with age, ultrasound biomicroscopy of the living rhesus ciliary region did identify distinct nasal vs. temporal asymmetries, which may reflect anatomical requirements for convergence-associated accommodation.

Microchimerism does not induce tolerance and sustains immunity after in utero transplantation.

BACKGROUND: To date, over 40 in utero transplants have been performed in humans; the only successes were documented in the treatment of severe combined immunodeficiency syndromes. Hemoglobinopathies and metabolic disorders are candidate diseases for this approach; however, when applied clinically, the results have been discouraging. To address the role of the fetal immune system in the outcome of in utero transplantation, we have developed a murine model of in utero transplantation in immunologically intact murine recipients and have studied chimerism and tolerance/immunity to allogeneic donor cells through the lives of the animals. METHODS: We have performed experiments in which purified murine sca-1+/lin- cells and c-kit+/lin- cells of C57BL/6 (H2b) mice were injected into Balb/c (H2d) fetal recipients at early gestational ages. Chimerism was tested by highly sensitive semiquantitative polymerase chain reaction assay and tolerance/immunity to donor cells was studied by in vivo (skin grafts, responses to postnatal boosts) and in vitro (mixed lymphocyte culture, cytotoxicity, and cytokine release) assays. RESULTS: One hundred percent (10/10) of mice transplanted with c-kit+ cells and 44% (4/9) of mice transplanted with sca+ cells showed circulating donor cells within the first 6 months of life (P=0.031). Mice in the sca+ group rejected donor skin grafts at a mean time of 9.1+/-0.2 days, whereas mice in the c-kit+ group rejected donor skin grafts at a mean time of 15.1+/-0.7 days (P=0.001). The difference between the transplanted groups and non-transplanted controls was also significant (P<0.05). All mice transplanted with sca+/lin- cells showed greater response to donor cells than to third-party cells at all effector to target ratios (P=0.002). Differences in response to donor alloantigen between sca+ and c-kit+ groups were significant (P=0.003). Cytokine quantification demonstrated higher TH1 than TH2 cytokine release in all groups, and the response to donor cells was higher in the sca+ compared with c-kit+ mice (P=0.031). CONCLUSION: These results demonstrate a low level of chimerism and tolerance in mice transplanted in utero with sca+/lin- and c-kit+/lin- cells. The possibility of active in utero immunization to donor cells is supported by accelerated skin graft rejection in mice transplanted with sca+ cells and enhanced in vitro immune responses in mice with persistent microchimerism.

Development of allergic inflammation in a murine model of asthma is dependent on the costimulatory receptor OX40.

Asthma is thought to result from an abnormal expansion of CD4 T cells reactive with airborne allergens, and pathology is controlled by several cytokines of the T helper type 2 (Th2) family. The exact molecules which are involved in generating allergen-reactive T cells are not clear. Studies with blocking reagents or knockout animals have shown that the CD28/B7 interaction partially controls development of allergic asthma in mouse models, but may not be the sole molecule involved. In this report, we have investigated the role of the tumor necrosis factor receptor family member OX40 in allergic inflammation using OX40-deficient mice. OX40 has been shown to participate in regulating clonal expansion and memory development of CD4 T cells and may synergize with CD28. Our studies demonstrate that OX40(-/)- mice, primed with the model allergen ovalbumin and challenged through the airways with aerosolized antigen, are severely impaired in their ability to generate a Th2 response characterized by high levels of interleukin (IL)-5, IL-4, and immunoglobulin E. Moreover, OX40(-/)- mice exhibit diminished lung inflammation, including an 80-90% reduction in eosinophilia and mucus production, less goblet cell hyperplasia, and significantly attenuated airway hyperreactivity. These studies highlight the potential importance of OX40 in development of allergic asthma and suggest that targeting OX40 may prove useful therapeutically.