Distinct roles for IL-4 and IL-10 in regulating T2 immunity during allergic bronchopulmonary mycosis

Pulmonary Cryptococcus neoformans infection of C57BL/6 mice is an established model of an allergic bronchopulmonary mycosis that has also been used to test a number of immunomodulatory agents. Our objective was to determine the role of IL-4 and IL-10 in the development/manifestation of the T2 response to C. neoformans in the lungs and lung-associated lymph nodes. In contrast to wild-type (WT) mice, which develop a chronic infection, pulmonary clearance was significantly greater in IL-4 knockout (KO) and IL-10 KO mice but was not due to an up-regulation of a non-T cell effector mechanism. Pulmonary eosinophilia was absent in both IL-4 KO and IL-10 KO mice compared with WT mice. The production of IL-4, IL-5, and IL-13 by lung leukocytes from IL-4 KO and IL-10 KO mice was lower but IFN-gamma levels remained the same. TNF-alpha and IL-12 production by lung leukocytes was up-regulated in IL-10 KO but not IL-4 KO mice. Overall, IL-4 KO mice did not develop the systemic (lung-associated lymph nodes and serum) or local (lungs) T2 responses characteristic of the allergic bronchopulmonary C. neoformans infection. In contrast, the systemic T2 elements of the response remained unaltered in IL-10 KO mice whereas the T2 response in the lungs failed to develop indicating that the action of IL-10 in T cell regulation was distinct from that of IL-4. Thus, although IL-10 has been reported to down-regulate pulmonary T2 responses to isolated fungal Ags, IL-10 can augment pulmonary T2 responses if they occur in the context of fungal infection.     

KChIP3 rescues the functional expression of Shal channel tetramerization mutants

KChIP proteins regulate Shal, Kv4.x, channel expression by binding to a conserved sequence at the N terminus of the subunit. The binding of KChIP facilitates a redistribution of Kv4 protein to the cell surface, producing a large increase in current along with significant changes in channel gating kinetics. Recently we have shown that mutants of Kv4.2 lacking the ability to bind an intersubunit Zn(2+) between their T1 domains fail to form functional channels because they are unable to assemble to tetramers and remain trapped in the endoplasmic reticulum. Here we find that KChIPs are capable of rescuing the function of Zn(2+) site mutants by driving the mutant subunits to assemble to tetramers. Thus, in addition to known trafficking effects, KChIPs play a direct role in subunit assembly by binding to monomeric subunits within the endoplasmic reticulum and promoting tetrameric channel assembly. Zn(2+)-less Kv4.2 channels expressed with KChIP3 demonstrate several distinct kinetic changes in channel gating, including a reduced time to peak and faster entry into the inactivated state as well as extending the time to recover from inactivation by 3-4 fold.     

Solar furnaces or swamp coolers: costs and benefits of water use by solar-tracking flowers of the alpine snow buttercup, Ranunculus adoneus

Solar tracking or heliotropism simultaneously raises organ temperature and light interception. For leaves and flowers carbon gain is maximized at the expense of water loss. In this study I explore how costs and benefits associated with water use by solar-tracking flowers of the alpine snow buttercup, Ranunculus adoneus change with ambient temperature. First, I test whether heliotropism increases the water cost of reproduction in the snow buttercup under extant alpine conditions. I then explore whether water use for evaporative cooling in solar-tracking flowers reduces the risk of over-heating as temperatures increase. Solar tracking, by elevating floral temperature and irradiance causes a 29% increase in water uptake by flowers. Gas exchange measurements suggest that the extra water taken up by solar-tracking flowers is released through transpiration. Transpirational cooling in turn allows solar-tracking flowers to gain advantages of enhanced light interception and warmth while reducing the risk of over-heating. Transpiration reduces excess temperature in solar-tracking flowers, but at a water cost. Results show that even in cool alpine habitats, flower heliotropism has water costs to balance its reproductive advantages. Plants with solar-tracking flowers may tolerate hotter conditions if soil moisture is plentiful, but not under drought.     

Characterization of Expressed Sequence Tag–Derived Simple Sequence Repeat Markers for Aspergillus flavus: Emphasis on Variability of Isolates from the Southern United States

Simple sequence repeat (SSR) markers were developed from Aspergillus flavus expressed sequence tag (EST) database to conduct an analysis of genetic relationships of Aspergillus isolates from numerous host species and geographical regions, but primarily from the United States. Twenty-nine primers were designed from 362 tri-nucleotide EST-SSR sequences. Eighteen polymorphic loci were used to genotype 96 Aspergillus species isolates. The number of alleles detected per locus ranged from 2 to 24 with a mean of 8.2 alleles. Haploid diversity ranged from 0.28 to 0.91. Genetic distance matrix was used to perform principal coordinates analysis (PCA) and to generate dendrograms using unweighted pair group method with arithmetic mean (UPGMA). Two principal coordinates explained more than 75?% of the total variation among the isolates. One clade was identified for A. flavus isolates (n?=?87) with the other Aspergillus species (n?=?7) using PCA, but five distinct clusters were present when the others taxa were excluded from the analysis. Six groups were noted when the EST-SSR data were compared using UPGMA. However, the latter PCA or UPGMA comparison resulted in no direct associations with host species, geographical region or aflatoxin production. Furthermore, there was no direct correlation to visible morphological features such as sclerotial types. The isolates from Mississippi Delta region, which contained the largest percentage of isolates, did not show any unusual clustering except for isolates K32, K55, and 199. Further studies of these three isolates are warranted to evaluate their pathogenicity, aflatoxin production potential, additional gene sequences (e.g., RPB2), and morphological comparisons.     

Effects of grazing and invasive grasses on desert vertebrates in California

Much of California's San Joaquin Valley is a desert and, like portions of other North American deserts, is experiencing an ecological shift from being dominated by ephemeral native forbs, with widely spaced shrubs, to fire-prone non-native annual grasses. Small terrestrial vertebrates, many of which are adapted to open desert habitats, are declining. One hypothesis is that the invasive plants contribute to the decline by altering vegetative structure. Although cattle may have originally been a factor in the establishment of these non-native plants, their grazing may benefit the terrestrial vertebrates by maintaining an open structure, especially during average or wet winters when the exotic grasses grow especially dense. We experimentally tested the effect of cattle grazing on invasive plants and a community of small vertebrates at a site in the southwestern San Joaquin Desert. We established and monitored 4 treatment (grazed) and 4 control (ungrazed) plots from 1997 to 2006, and assessed the abundances of blunt-nosed leopard lizards (Gambelia sila), giant kangaroo rats (Dipodomys ingens), short-nosed kangaroo rats (Dipodomys nitratoides nitratoides), and San Joaquin antelope squirrels (Ammospermophilus nelsoni), all of which are listed as threatened or endangered by state or federal agencies. We also recorded abundances of the non-protected western whiptail lizards (Aspidoscelis tigris), side-blotched lizards (Uta stansburiana), San Joaquin pocket mice (Perognathus inornatus inornatus), and Heermann's kangaroo rats (Dipdomys heermanni). Based on repeated measures analysis of variance (ANOVA) and a 0.05 alpha level, only Heermann's kangaroo rats showed a treatment effect; they were more abundant on the control plots. However, this effect could be accounted for by the natural re-establishment of saltbush (Atriplex spp.) on part of the study site. Saltbush return also favored western whiptail lizards and San Joaquin antelope squirrels. A regression analysis indicated that populations of blunt-nosed leopard lizard and giant kangaroo rat increased significantly faster in grazed plots than the ungrazed controls, and abundances of 6 of 8 species were negatively correlated with increased residual dry matter. With relaxed alpha values to decrease Type II error, populations of blunt-nosed leopard lizards (500% greater), San Joaquin antelope squirrels (85% greater), and short-nosed kangaroo rats (73% greater) increased significantly on grazed plots over the course of the study compared to ungrazed plots. We did not find grazing to negatively affect abundance of any species we studied. When herbaceous cover is low during years of below average rainfall in deserts and other arid areas, grazing may not be necessary to maintain populations of small vertebrates. However, if cattle grazing is closely monitored in space and time to minimize adverse effects on the habitat, it could be an effective tool to control dense stands of non-native grasses and benefit native wildlife. © 2011 The Wildlife Society.     

Exogenous fluorescent tracer agents based on pegylated pyrazine dyes for real-time point-of-care measurement of glomerular filtration rate

Novel pyrazine carboxamides bearing hydrophilic poly(ethylene glycol) (PEG) moieties were designed, synthesized, and evaluated for use as fluorescent glomerular filtration rate (GFR) tracer agents. Among these, compounds 4d and 5c that contain about 48 ethylene oxide units in the PEG chain exhibited the most favorable physicochemical and renal clearance properties. In vitro studies show that these two compounds have low plasma protein binding, a necessary condition for renal excretion. In vivo animal model results show that 4d and 5c have a higher urine recovery of the injected dose than iothalamate (a commonly considered gold standard GFR agent). Pharmacokinetic studies show that these two compounds exhibit a plasma clearance equivalent to iothalamate, but with a faster (i.e. lower) terminal half-life than iothalamate (possibly from restricted distribution into the extracellular space due to large molecular size and hydrodynamic volume). Furthermore, the plasma clearance of 4d and 5c remained unchanged upon blockage of the tubular secretion pathway with probenecid, a necessary condition for establishment of clearance via glomerular filtration exclusively. Finally, noninvasive real-time monitoring of this class of compounds was demonstrated by pharmacokinetic clearance of 5c by optical measurements in rat model, which correlates strongly with plasma concentration of the tracer. Hence, 4d and 5c are promising candidates for translation to the clinic as exogenous fluorescent tracer agents in real-time point-of-care monitoring of GFR.