The cell and molecular basis of mechanical, cold, and inflammatory pain

Peripheral pain pathways are activated by a range of stimuli. We used diphtheria toxin to kill all mouse postmitotic sensory neurons expressing the sodium channel Nav1.8. Mice showed normal motor activity and low-threshold mechanical and acute noxious heat responses but did not respond to noxious mechanical pressure or cold. They also showed a loss of enhanced pain responses and spontaneous pain behavior upon treatment with inflammatory insults. In contrast, nerve injury led to heightened pain sensitivity to thermal and mechanical stimuli indistinguishable from that seen with normal littermates. Pain behavior correlates well with central input from sensory neurons measured electrophysiologically in vivo. These data demonstrate that Na(v)1.8-expressing neurons are essential for mechanical, cold, and inflammatory pain but not for neuropathic pain or heat sensing.     

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.     

C and N dynamics of a range of biogas slurries as a function of application rate and soil texture: a laboratory experiment

Digestates vary in composition and studies regarding their impact on C and N dynamics in soils are scarce. The objective was to analyse the C and N dynamics of digestates originating from various substrates applied to a sandy Cambisol and a silty Anthrosol. In three laboratory experiments (4–6 weeks), the effects of digestate properties, N rate and water content were tested. Averaged over both soils, 21% of the C supplied was emitted as CO₂. Potential NH₃ emissions during the first week ranged between 6% and 12% of NH₄⁺ present in the digestates. The emission factors in the sandy Cambisol were on average 1.2 and 2 times higher for CO₂ and potential NH₃, respectively, compared to the silty Anthrosol. Similarly, net nitrogen mineralization in the sandy Cambisol was approximately twice the N mineralized in the silty Anthrosol. Net nitrification was not influenced by soil texture or different digestates, but increased with increasing application rates and had highest values at 75% of water holding capacity. Our results indicate that the type of substrate input for anaerobic digestion influences the properties of the digestate and therefore the dynamics of C and N. However, soil texture can affect these dynamics markedly.     

TRANS-SPLENIC PORTAL SCINTIGRAPHY IN NORMAL DOGS

The purpose of this study was to (1) establish a technique for ultrasound-guided trans-splenic portal scintigraphy (TSPS) using 99mTcO4(-), (2) evaluate portal vein morphology, (3) compare the radiation exposures for TSPS vs. per-rectal portal scintigraphy (PRPS), and (4) compare the quality of numerical data from the TSPS vs. PRPS. Eight juvenile dogs underwent PRPS and TSPS (minimum of 48h between studies) after initial screening tests. PRPS was done according to established protocol using 425 +/- 36MBq (mean +/- SD) of 99mTcO4(-). TSPS was done with the dogs in right lateral recumbency over the gamma camera. 99mTcO4(-) (57 +/- 13.9 MBq) was injected into the spleen 1-2s following initiation of the dynamic acquisition. The frame rate was 4 frames/s for 5 min. There was significantly lower radioactivity of 99mTcO4(-) given and significantly higher total counts recorded in the liver and heart during the TSPS compared with PRPS. The total counts for the TSPS and PRPS were 7120 +/- 4386 and 830 +/- 523, respectively. Percent absorption from the spleen was 52.5 +/- 19.1% compared with 9.2 +/- 5.7% for the colon. Calculated transit time for the TSPS studies was 7 +/- 2.3s. In TSPS studies, the splenic and portal veins were clearly identified. Radiation exposure levels of the dogs were significantly lower following TSPS than after PRPS. TSPS appears superior to PRPS as a method to image the portal venous system representing a valid alternative diagnostic test for animals with suspected portosystemic shunts.     

USE OF COMPUTED TOMOGRAPHY AND RADIOLABELED LEUKOCYTES IN A CAT WITH PANCREATITIS

The normal feline pancreas has been evaluated using radiolabeled leukocytes (99mTc-HMPAO) and computed tomography. The purpose of this report is to describe a clinical case where both modalities were utilized to assess the inflamed feline pancreas. A nine year old female cat presented with anorexia, depression and some vomiting. Blood values were unremarkable. Radiographs and ultrasound were suggestive of pancreatitis. The cat's leukocytes were separated and labeled according to an established protocol. Whole body images were acquired immediately, at 5 and 30 min, and at 1, 2, 4, and 17 hours post injection. Approximately 48 h later, the animal was anesthetized and computed tomography of the abdomen was preformed both pre and post contrast. Surgical biopsies were taken. The distribution of the WBCs was similar to that documented in normal animals, however, at 2 h there was faint uptake seen in the region of the pancreas. This uptake became more intense at 4 h and persisted at 17 h. Computed tomography showed irregular margination of the pancreas, it was larger than normal and inhomogeneous. Contrast enhancement was inhomogeneous and its peak enhancement was not reached until 10 min post injection; normal feline pancreas enhances homogeneously and peaks immediately. Histopathology confirmed pancreatitis with lymphocytic, plasmacytic, neutrophilic and eosinophilic inflammation and fibrosis. Radiolabeled leukocytes can be used to document pancreatic inflammation and this is best seen 4 h after injection. Computed tomography allows superior visualization of the pancreas. Both the appearance and contrast enhancement pattern of the inflamed pancreas differ from normal.     

Placing the pieces: Reconstructing the original property mosaic in a warrant and patent watershed

Recent research shows that land use history is an important determinant of current ecosystem function. In the United States, characterization of land use change following European settlement requires reconstruction of the original property mosaic. However, this task is difficult in unsystematically surveyed areas east of the Appalachian Mountains. The Gwynns Falls watershed (Baltimore, MD) was originally surveyed in the 1600-1700s under a system of warrants and patents (commonly known as ‘metes and bounds’). A method for the reconstruction and mapping of warrant and patent properties is presented and used to map the original property mosaic in the Gwynns Falls watershed. Using the mapped mosaic, the persistence of properties and property lines in the current Gwynns Falls landscape is considered. The results of this research indicate that as in agricultural areas, the original property lines in the Gwynns Falls watershed are persistent. At the same time, the results suggest that the property mosaic in heavily urbanized/suburbanized areas is generally ‘reset.’ Further, trends in surveying technique, parcel size, and settlement patterns cause property line density and property shape complexity to increase in the less urbanized upper watershed. The persistence of original patterns may be damping expression of heterogeneity gradients in this urban landscape. This spatial pattern of complexity in the original mosaic is directly opposite of hypothesized patterns of landscape heterogeneity arising from urbanization. The technique reported here and the resulting observations are important for landscape pattern studies in areas settled under unsystematic survey systems, especially the heavily urbanized areas of the eastern United States. This revised version was published online in July 2006 with corrections to the Cover Date.     

Nutritional Enhancement of Biocontrol Activity of Candida sake (CPA-1) against Penicillium expansum on Apples and Pears

Pome fruits are poor in nitrogenous compounds and the addition of nitrogen can improve colonisation of the fruits by antagonists. Twenty-two nitrogenous compounds were evaluated for their effect on Candida sake (CPA-1) growth in vitro. Ten compounds that induced greater growth were applied with the antagonist to wounded fruits to evaluate their effect on enhancing control of Penicillium expansum. Calcium chloride and 2-deoxy-D-glucose were also tested. L-serine and L-aspartic acid enhanced biocontrol by C. sake against P. expansum on apples. On apples and pears, ammonium molybdate, calcium chloride and 2-deoxy-D-glucose improved the capacity of the antagonist to control P. expansum. The addition of ammonium molybdate at 1 mM allowed C. sake to be used on apples and pears at a lower concentration without diminishing control. Similar results were observed with the addition of calcium chloride to the antagonist. 2-deoxy-D-glucose at 6 and 18 mM enhanced biocontrol on pears by over 81%, but on apples the improvement of biocontrol was observed only at 6 mM. In cold storage, the combination of ammonium molybdate and C. sake completely eliminated the incidence of blue mould on pears, and reduced its severity and incidence by more than 80% on apples.     

Treatment of equine glaucoma by transscleral neodymium:yttrium aluminum garnet laser cyclophotocoagulation: a retrospective study of 23 eyes of 16 horses

Contact neodymium:yttrium aluminum garnet (Nd:YAG) laser transscleral cyclophotocoagulation (TSCP) was performed on 23 eyes of 16 horses for treatment of glaucoma. The mean highest preoperative IOP was 51 ± 17 mmHg. Follow-up evaluation was available for 19 eyes 1 day after surgery, 14 eyes from 1 to 2 weeks, 16 eyes from 4 to 6 weeks, 9 eyes from 12 to 16 weeks, and 10 eyes greater than 20 weeks after laser treatment. The mean intraocular pressure (IOP) the day following surgery was 34 ± 13 mmHg. The mean IOP for each follow-up period was: one to two weeks postoperative, 23 ± 9 mmHg; four to six weeks, 24 ± 7 mmHg; 12–16 weeks, 28 ± 10 mmHg; and  20 weeks, 22 ± 9 mmHg. IOP measurements were significantly different from pretreatment values for all follow-up intervals except for weeks 12–16 ( P < 0.05). Treatment success was defined as maintenance of IOP < 30 mmHg. Treatment success was achieved in 93%, 88%, 78%, and 70% of the treated eyes at the 1–2 weeks, 4–6 weeks, 12–16 weeks, and the  20 weeks re-evaluation, respectively. No significant difference was found between the number of eyes visual at presentation (52.2%) and visual at 20 weeks (60%). The most common laser complications were conjunctival hyperemia (21.7%) and corneal ulcers (13.0%). Results of this study indicate that Nd:YAG TSCP is an effective method of controlling IOP and preserving vision in horses with glaucoma. An effective Nd:YAG laser protocol for TSCP in the equine glaucomatous eye is a power setting of 11 W, duration of 0.4 s, applied 5 mm posterior to the limbus at 60 sites, resulting in a total energy dose of 264 J.