The mTOR-regulated phosphoproteome reveals a mechanism of mTORC1-mediated inhibition of growth factor signaling

The mammalian target of rapamycin (mTOR) protein kinase is a master growth promoter that nucleates two complexes, mTORC1 and mTORC2. Despite the diverse processes controlled by mTOR, few substrates are known. We defined the mTOR-regulated phosphoproteome by quantitative mass spectrometry and characterized the primary sequence motif specificity of mTOR using positional scanning peptide libraries. We found that the phosphorylation response to insulin is largely mTOR dependent and that mTOR exhibits a unique preference for proline, hydrophobic, and aromatic residues at the +1 position. The adaptor protein Grb10 was identified as an mTORC1 substrate that mediates the inhibition of phosphoinositide 3-kinase typical of cells lacking tuberous sclerosis complex 2 (TSC2), a tumor suppressor and negative regulator of mTORC1. Our work clarifies how mTORC1 inhibits growth factor signaling and opens new areas of investigation in mTOR biology.     

Molecular mimicry regulates ABA signaling by SnRK2 kinases and PP2C phosphatases

Abscisic acid (ABA) is an essential hormone for plants to survive environmental stresses. At the center of the ABA signaling network is a subfamily of type 2C protein phosphatases (PP2Cs), which form exclusive interactions with ABA receptors and subfamily 2 Snfl-related kinase (SnRK2s). Here, we report a SnRK2-PP2C complex structure, which reveals marked similarity in PP2C recognition by SnRK2 and ABA receptors. In the complex, the kinase activation loop docks into the active site of PP2C, while the conserved ABA-sensing tryptophan of PP2C inserts into the kinase catalytic cleft, thus mimicking receptor-PP2C interactions. These structural results provide a simple mechanism that directly couples ABA binding to SnRK2 kinase activation and highlight a new paradigm of kinase-phosphatase regulation through mutual packing of their catalytic sites.     

Fear of predation slows plant-litter decomposition

Aboveground consumers are believed to affect ecosystem functioning by regulating the quantity and quality of plant litter entering the soil. We uncovered a pathway whereby terrestrial predators regulate ecosystem processes via indirect control over soil community function. Grasshopper herbivores stressed by spider predators have a higher body carbon-to-nitrogen ratio than do grasshoppers raised without spiders. This change in elemental content does not slow grasshopper decomposition but perturbs belowground community function, decelerating the subsequent decomposition of plant litter. This legacy effect of predation on soil community function appears to be regulated by the amount of herbivore protein entering the soil.     

Using an unmanned aerial vehicle to evaluate nitrogen variability and height effect with an active crop canopy sensor

Ground-based active sensors have been used in the past with success in detecting nitrogen (N) variability within maize production systems. The use of unmanned aerial vehicles (UAVs) presents an opportunity to evaluate N variability with unique advantages compared to ground-based systems. The objectives of this study were to: determine if a UAV was a suitable platform for use with an active crop canopy sensor to monitor in-season N status of maize, if UAV’s were a suitable platform, is the UAV and active sensor platform a suitable substitute for current handheld methods, and is there a height effect that may be confounding measurements of N status over crop canopies? In a 2013 study comparing aerial and ground-based sensor platforms, there was no difference in the ability of aerial and ground-based active sensors to detect N rate effects on a maize crop canopy. In a 2014 study, an active sensor mounted on a UAV was able to detect differences in crop canopy N status similarly to a handheld active sensor. The UAV/active sensor system (AerialActive) platform used in this study detected N rate differences in crop canopy N status within a range of 0.5–1.5 m above a relatively uniform turfgrass canopy. The height effect for an active sensor above a crop canopy is sensor- and crop-specific, which needs to be taken into account when implementing such a system. Unmanned aerial vehicles equipped with active crop canopy sensors provide potential for automated data collection to quantify crop stress in addition to passive sensors currently in use.     

The effect of breed,sex, and oral meloxicam administration on pain biomarkers following hot-iron branding in Hereford and Angus calves

Hot-iron branding uses thermal injury to permanently identify cattle causing painful tissue damage. The primary objective was to examine the physiological and behavioral effects of oral meloxicam (MEL), compared to a control, administered at the time of hot-iron branding in Angus and Hereford steers and heifers. The secondary objectives were to investigate breed and sex effects on pain biomarkers. A total of 70 yearlings, consisting of 35 heifers and 35 steers (Angus, Hereford, or Angus × Hereford), were enrolled in the study. Animals were blocked by sex, randomized across weight, and assigned to receive MEL (1 mg/kg) or a placebo (CON). Biomarkers were assessed for 48 h after branding and included infrared thermography (IRT), mechanical nociceptive threshold (MNT), accelerometry and a visual analog scale (VAS), and serum cortisol and prostaglandin E₂ metabolites (PGEM). Wound healing was assessed for 12 wk. Hair samples to quantify cortisol levels were taken prior to and 30 d post-branding. Responses were analyzed using repeated measures with calf nested in treatment as a random effect, and treatment, time, treatment by time interaction, breed, and sex as fixed effects. There was a treatment by time interaction for PGEM (P < 0.01) with MEL having lower values than CON at 6, 24, and 48 h (MEL: 18.34 ± 3.52, 19.61 ± 3.48, and 22.24 ± 3.48 pg/mL, respectively; CON: 32.57 ± 3.58, 37.00 ± 3.52, and 33.07 ± 3.48 pg/mL; P < 0.01). MEL showed less of a difference in maximum IRT values between the branded (2.27 ± 0.29 °C) and control site (3.15 ± 0.29 °C; P < 0.01). MEL took fewer lying bouts at 0–12 h (4.91 bouts ± 0.56) compared with CON (6.87 bouts ± 0.55; P < 0.01). Compared with Hereford calves, Angus calves exhibited greater serum but lower hair cortisol, greater PGEM, more lying bouts, and less healed wound scores at 3, 4, and 5 wk. Compared with heifers, steers exhibited lower PGEM, lower branding site and ocular IRT, higher MNT, and lower plasma meloxicam levels. Steers spent more time lying, took more lying bouts and had greater VAS pain, and more healed wound scores at 5 wk than heifers. Meloxicam administration at branding reduced branding and control site temperature differences and reduced lying bouts for the first 12 h. Breed and sex effects were observed across many biomarkers. Changes from baseline values for IRT, MNT, lying time, step count, VAS pain, and wound scoring all support that branding cattle is painful.     

Diagnostic and public health investigation of Mycobacterium tuberculosis infection in a dog in Ontario,Canada

A 4-y-old, female mixed-breed dog was presented to the Ontario Veterinary College for further evaluation of multiple pulmonary and hepatic masses, intrathoracic lymphadenitis, and recent development of a pyogranulomatous pleural effusion. Along with other comprehensive tests, a thoracic lymph node biopsy was performed, and Mycobacterium tuberculosis complex infection was confirmed by real-time PCR. The dog’s condition declined post-operatively, and euthanasia was elected. Postmortem examination confirmed severe granulomatous pneumonia, hepatitis, intrathoracic and intraabdominal lymphadenitis, omentitis, and nephritis. Line-probe assays performed on samples collected postmortem confirmed the species as M. tuberculosis. 24-loci MIRU-VNTR genotyping, spoligotyping, and whole-genome sequencing revealed relations to known human isolates, but no epidemiologic link to these cases was investigated. Given the concern for potential human exposure during this animal’s disease course, a public health investigation was initiated; 45 individuals were tested for M. tuberculosis exposure, and no subsequent human infections related to this animal were identified. Our case highlights the need for more readily available, minimally invasive testing for the diagnosis of canine mycobacteriosis, and highlights the ability of canid species to act as potential contributors to the epidemiology of M. tuberculosis infections.     

Effects of Water Hardness and Calcium: Magnesium Ratios on Reproductive Performance and Offspring Quality of Macrobrachium rosenbergii

This study evaluated the effect of specific calcium and magnesium ratios at two hardness values on reproductive performance and offspring quality of Macrobrachium rosenbergii brood‐females. Hatchery water containing 110.5 mg/L CaCO3 hardness was considered baseline as it has been used successfully to hatch and rear larvae and was used as the control treatment. At each hardness value of 150 and 190 mg/L CaCO₃, four experimental water treatments were made in triplicate. Each treatment was adjusted using soluble salts (CaCl₂·H₂O, MgSO₄·7H₂O, and MgCl₂·6H₂O) to provide the hardness and calcium to magnesium ratios of 0:20, 20:80, 50:50, and 80:20 needed. Each of the experimental tanks (140 L) were stocked with six females and one male. The results indicated that both hardness and Ca:Mg ratios affect brood‐female reproductive qualitative parameters such as intermolt period, egg hatchability, egg dry weight, and egg‐clutch somatic index (ESI) parameters (P < 0.05) but not fecundity and eggs per spawn. The results revealed that brood‐females at 150 mg/L hardness showed greater reproductive performance than at 190 and 110.5 mg/L hardness. The findings also demonstrated that the treatment 50Ca50Mg at 150 mg/L hardness with 38.8 mg/L calcium and 12.9 mg/L magnesium had optimum reproductive performance and offspring quality for M. rosenbergii brood‐females.     

Dwarf shrub litter as a food source for macro-decomposers in alpine pastureland

Dwarf shrub litter is thought to be of poor quality and palatability for macro-decomposers. In laboratory feeding experiments, however, it was found that this litter type represents a potential food source for earthworms and millipedes. Here we tested whether this is true under natural conditions by conducting a 1 year field experiment on an abandoned alpine pasture in Tyrol (Austria). As the natural isotopic signatures of plant litter do not allow discriminating between different plant species, dwarf shrub litter, artificially enriched in ¹⁵N, was offered in experimental plots to the macro-decomposer community. Both the earthworm Lumbricus rubellus and the millipede Cylindrojulus fulviceps fed on dwarf shrub litter as indicated by their increased δ¹⁵N signatures, but IsoError analysis indicated a clear preference for grass litter for all macro-decomposer species investigated.Soil δ¹⁵N signatures were only marginally affected by the experimental provision of ¹⁵N-enriched dwarf shrub litter, whereas litter from other, unlabelled, plants became enriched by the isotopic tracer to some extent. Except for one grass species, plants harvested at the end of the experiment were not enriched in ¹⁵N, suggesting that the N-uptake from decomposing litter material by plants on abandoned alpine pastureland is slow.     

Invasive black spiny‐tailed iguanas (Ctenosaura similis) on Gasparilla Island,Florida, USA

The native range of Ctenosaura similis extends from southern Mexico through Panama. From an initial introduction of 3 animals in 1979, the species now numbers in the thousands on Gasparilla Island in southwest Florida. In response to complaints of property damage from residents and threats to native species, local officials and the US Department of Agriculture Wildlife Services began a removal program in 2008. Through 2011, trappers removed 9467 ctenosaurs. The number removed declined from 32 iguanas/day in 2008 to 1.9 iguanas/day in 2011 despite no easing of the control effort. We necropsied 2757 ctenosaurs to document aspects of their natural history. Females outnumbered males overall, although the largest size class (>300 mm snout–vent length) included 32 males and just 2 females. Reproduction was seasonal. We found oviducal eggs in females from early Apr to early Jun, approximately 2 months later than C. similis in its native range. We trapped hatchlings from late Jul to early Oct coincident with the summer rainy season. Clutch size increased with female body size, with 62 being the largest clutch size recorded. In general, the biology of the invasive population on Gasparilla Island resembles native C. similis populations in Central America, except for the lack of large individuals. We suggest that shorter day length and colder temperatures create environmental conditions that are suboptimal for individual growth compared to those in the native range.