Comparative and developmental functional morphology of the jaws of living and fossil gars (Actinopterygii: Lepisosteidae)

The feeding mechanism of gars (Ginglymodi : Lepisosteidae) is characterized by cranial elevation and lower jaw rotation but minimal cranial kinesis. Gar jaws have numerous, sharply pointed, elongate teeth for capture of evasive prey. Their mandibles range from relatively short to extremely long depending on the species. Jaw length and lever dimensions were hypothesized to affect the biomechanics of force and motion during feeding, according to simple mechanical models of muscles exerting force through first- or third-order levers. A morphometric protocol was used to measure the jaw structure of seven living and five fossil species of gar and these data were used to calculate the mechanical advantage (a measure of force transmission) for both opening and closing of the mandible. Gars were found to possess low mechanical advantage (MA) and high transmission of motion, although gars occupy a range of biomechanical states across the continuum of force vs. velocity transmission. The long-nose gar, Lepisosteus osseus, has one of the lowest jaw closing MAs (0.05) ever measured in fishes. Intraspecific lever mechanics were also calculated for a developmental series (from feeding larvae to adults) of L. osseus and Atractosteus spatula. A characteristic ontogenetic curve in MA of the lower jaw was obtained, with a large decrease in MA between larva and juvenile, followed by a steady increase during adult growth. This curve correlates with a change in prey type, with the small, robust-jawed individuals feeding mainly on crustaceans and insects and the large, long-jawed individuals of all species becoming mainly piscivorous. Principal components analysis of functionally important morphometrics shows that several gar species occupy different regions of functional morphospace. Some fossil gar species are also placed within functional morphospace using this approach.     

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.     

Depth of α-synuclein in a bilayer determined by fluorescence, neutron reflectometry, and computation

α-Synuclein (α-syn) membrane interactions are implicated in the pathogenesis of Parkinson's disease. Fluorescence and neutron reflectometry (NR) measurements reveal that α-syn penetrates ～9–14 Å into the outer leaflet of the bilayer, with a substantial portion of the membrane-bound polypeptide extending into the aqueous solvent. For the first time, to our knowledge, we used NR to obtain direct quantitative evidence of α-syn-induced membrane thinning. To examine the effect of specific residues on membrane penetration depths, we used a series of W4-containing N-terminal peptides. We identified that the first 15 residues (P15) nearly recapitulate the features of the full-length protein (i.e., partition constants, molecular mobility, and insertion of the W4 side chain into the bilayer), and found that as few as the first four N-terminal residues are sufficient for vesicle binding. Although at least one imperfect amphipathic repeat sequence (KAKEGV) is required for α-helical formation, secondary structural formation has little effect on membrane affinity. To develop an N-terminal α-syn model for bilayer interactions, we performed molecular-dynamics simulations of the P15 peptide submerged in a bilayer. The simulation results are highly consistent with experimental data indicating a broad low-energy region (8.5–14.5 Å) for W4 insertion.     

The extracellular loops of Smoothened play a regulatory role in control of Hedgehog pathway activation

The Hedgehog (Hh) signaling pathway plays an instructional role during development, and is frequently activated in cancer. Ligand-induced pathway activation requires signaling by the transmembrane protein Smoothened (Smo), a member of the G-protein-coupled receptor (GPCR) superfamily. The extracellular (EC) loops of canonical GPCRs harbor cysteine residues that engage in disulfide bonds, affecting active and inactive signaling states through regulating receptor conformation, dimerization and/or ligand binding. Although a functional importance for cysteines localized to the N-terminal extracellular cysteine-rich domain has been described, a functional role for a set of conserved cysteines in the EC loops of Smo has not yet been established. In this study, we mutated each of the conserved EC cysteines, and tested for effects on Hh signal transduction. Cysteine mutagenesis reveals that previously uncharacterized functional roles exist for Smo EC1 and EC2. We provide in vitro and in vivo evidence that EC1 cysteine mutation induces significant Hh-independent Smo signaling, triggering a level of pathway activation similar to that of a maximal Hh response in Drosophila and mammalian systems. Furthermore, we show that a single amino acid change in EC2 attenuates Hh-induced Smo signaling, whereas deletion of the central region of EC2 renders Smo fully active, suggesting that the conformation of EC2 is crucial for regulated Smo activity. Taken together, these findings are consistent with loop cysteines engaging in disulfide bonds that facilitate a Smo conformation that is silent in the absence of Hh, but can transition to a fully active state in response to ligand.     

Role of caspase 2 in apoptotic signaling in primate and murine germ cells

This study investigates the role of caspase 2 in apoptotic signaling of nonhuman primate male germ cells triggered by mild testicular hyperthermia, testosterone (T(e)) implants, or by combined interventions. Mean incidence of germ cell apoptosis increased significantly by Day 3 in the heat (H(e)) alone group and by Day 8 in the Te alone group but peaked at Day 3 in H(e) + T(e) group. We found activation of caspase 2 in both germ cells and Sertoli cells after induction of apoptosis. Most notably, active caspase 2 immunoreactivity was detected only in those germ cells susceptible to apoptosis compared with controls, where little or no such staining is detected. To further explore the role of caspase 2 in regulating male germ cell death, we next evaluated the efficacy of caspase 2 inhibition in preventing or attenuating heat-induced germ cell apoptosis in rats. Caspase 2 inhibition significantly (P < 0.05) prevented such heat-induced germ cell apoptosis. The protection offered by the caspase 2 inhibitor occurred upstream of mitochondria, involving suppression of mitogen-activated protein kinase (MAPK) 14 activation and inducible nitric oxide synthase (NOS2) induction and, in turn, suppression of cytochrome c-mediated death pathway. Together, our results show that caspase 2 is activated in male germ cells undergoing apoptosis in nonhuman primates after heat stress, hormonal deprivation, or after combined interventions. Blockade of caspase 2 activation prevents heat-induced germ cell apoptosis in rats by suppressing the MAPK14- and NO-mediated intrinsic pathway signaling.