Crystal structure of a lipid G protein-coupled receptor

The lyso-phospholipid sphingosine 1-phosphate modulates lymphocyte trafficking, endothelial development and integrity, heart rate, and vascular tone and maturation by activating G protein-coupled sphingosine 1-phosphate receptors. Here, we present the crystal structure of the sphingosine 1-phosphate receptor 1 fused to T4-lysozyme (S1P(1)-T4L) in complex with an antagonist sphingolipid mimic. Extracellular access to the binding pocket is occluded by the amino terminus and extracellular loops of the receptor. Access is gained by ligands entering laterally between helices I and VII within the transmembrane region of the receptor. This structure, along with mutagenesis, agonist structure-activity relationship data, and modeling, provides a detailed view of the molecular recognition and requirement for hydrophobic volume that activates S1P(1), resulting in the modulation of immune and stromal cell responses.     

An investigation of Salmonella Fluntern illnesses linked to leopard geckos—United States, 2018

Reptile contact can result in zoonotic non‐typhoidal salmonellosis. In April 2018, Oregon Public Health Division contacted CDC about a cluster of four Salmonella serovar Fluntern (SF) illnesses in four states (OR, CA, IA, NY); patients reported contact with geckos, a popular reptile pet. PulseNet, the national molecular subtyping network of food‐borne disease surveillance, subsequently identified additional SF clinical isolates. Twelve cases in 11 states were identified; median age was 5 years (range: <1–58 years). Three patients were hospitalized; no deaths were reported. Of those with exposure information (n = 10), all reported reptile exposure; 9 (90%) specified contact with leopard geckos. No common source of geckos was identified from reported purchase locations. Los Angeles County (LAC) health officials isolated SF from one patient's leopard gecko. Five reptile/gecko isolates were identified from the USDA National Veterinary Services Laboratories (NVSL) from 2015 to 2018. Five countries responded to an Epidemic Intelligence Information System post by PulseNet; reptile isolate sequence data were received from Czech Republic. A clinical case from England was identified through the National Center for Biotechnology Information pathogen detection pipeline; the patient did not report contact with leopard geckos. Whole genome sequencing analysis revealed substantial genetic diversity between clinical and animal isolates; however, gecko and clinical isolates from LAC were highly related (1 allele difference). This investigation linking SF illnesses to leopard geckos highlights an important public health risk from pets. A better understanding of how geckos are distributed by the pet industry in the United States could improve traceability to points of origin and mitigate Salmonella transmission at gecko breeders. Earlier NVSL reports of SF isolates from geckos suggest the risk of human SF infection from geckos is not new. This investigation demonstrates a need to educate gecko breeders, retailers and gecko owners about the continued Salmonella infection risk from pet geckos.     

Effect of forage conservation method on microbial flora and fermentation pattern in forage and in equine colon and faeces

The influence of forage conservation method on microbial and chemical composition in forage and on colon content and faeces in horses fed with the forages was investigated. Hay (815 g DM/kg), haylage (548 g DM/kg) and silage (343 g DM/kg) was produced from the same grass crop and fed to four fistulated horses in a crossover study. Horses were sampled in right ventral colon and of faeces after a feeding period of 21 days (for each forage). Fermentation kinetics of right ventral colon was also studied on two subsequent days in each period, with sampling times immediately before and 2, 4, 8 and 12 h after the morning meal. Forage conservation method influenced microbial and chemical composition of the forages. Silage differed from haylage and hay in more variables than haylage differed from hay. However, the different forage types had similar influence on microbial and chemical composition of right ventral colon content and faeces in horses at Day 21, with the exception of counts of Streptococci which were highest when horses were fed with hay. However, interaction among forage types and sampling locations (right ventral colon and faeces) was also present for Streptococci as well as for pH. In general, pH was lower and counts of lactate utilizing and lactate producing bacteria were higher in faeces compared to right ventral colon. Fermentation kinetics in colon varied little among forage types and sampling times, and no interaction between forage types and sampling times were found, indicating that silage, haylage and hay had very similar fermentation kinetics in right ventral colon of horses. In conclusion, silage, haylage and hay produced from the same grass crop tended to produce similar responses in microbial and chemical composition in right ventral colon and faeces in horses fed with the forages.     

Non-iterative variance component estimation in QTL analysis

In variance component quantitative trait loci (QTL) analysis, a mixed model is used to detect the most likely chromosome position of a QTL. The putative QTL is included as a random effect and a method is needed to estimate the QTL variance. The standard estimation method used is an iterative method based on the restricted maximum likelihood (REML). In this paper, we present a novel non-iterative variance component estimation method. This method is based on Henderson's method 3, but relaxes the condition of unbiasedness. Two similar estimators were compared, which were developed from two different partitions of the sum of squares in Henderson's method 3. The approach was compared with REML on data from a European wild boar × domestic pig intercross. A meat quality trait was studied on chromosome 6 where a functional gene was known to be located. Both partitions resulted in estimated QTL variances close to the REML estimates. From the non-iterative estimates, we could also compute good approximations of the likelihood ratio curve on the studied chromosome.     

Cytopathological and histopathological diagnosis of canine splenic disorders

Objectives To determine (1) the common types of canine splenic disorders, and the breeds affected, that are diagnosed by cytopathological and histopathological examination in Sydney, Australia and (2) the accuracy of cytopathological examination compared with histopathological examination for the diagnosis of canine splenic disorders. Design 69 cytopathological and 51 histopathological diagnoses of canine splenic disorders presented to the Veterinary Pathology Diagnostic Services, The University of Sydney during 2006 and 2007 were tabulated and analysed; 17 cases examined both cytopathologically and histopathologically during 2001–07 were also analysed. Results The most common cytopathological diagnoses were benign disorders of growth, vascular disturbances and necrosis (29%), followed by no abnormalities detectable (28%), malignant neoplasms (20%), equivocal diagnoses (20%) and inflammatory disorders (3%). The most common breeds were Kelpie crosses and mixed breeds. The most common histopathological diagnoses were benign disorders of growth, vascular disturbances and necrosis (49%), followed by malignant neoplasms (43%) and inflammatory disorders (8%). The most common breeds were German Shepherd Dogs, Boxers and Maltese Terriers. Cytopathological and histopathological diagnoses were in complete agreement in 59% of cases, partial agreement in 29% and disagreement in 12%. Conclusion Benign disorders of growth, vascular disturbances and necrosis were the most commonly diagnosed canine splenic disorders, both cytopathologically and histopathologically. Kelpie crosses presented most frequently for cytopathological examination. German Shepherd Dogs were the most common breed diagnosed histopathologically with haemangiosarcoma. Although cytopathological and histopathological splenic examinations are complementary for diagnosis, this study has shown a high correlation for complete and partial agreement between the two.     

Topical anaesthesia alleviates short-term pain of castration and tail docking in lambs

Objective To investigate the effect of a topical anaesthetic formulation on pain alleviation, wound healing and systemic levels of local anaesthetic actives in lambs undergoing castration and tail docking. Design Three placebo-controlled and/or randomised experiments were conducted using three groups of Merino lambs (n = 62, 68 and 19) undergoing routine castration and tail docking. Procedure Surgical castration, with either surgical or hot-iron tail docking, was performed with and without the application of topical anaesthetic (Tri-Solfen®) or placebo. The effects of this procedure were compared with those of rubber ring castration and tail docking, and of the handled but unmarked controls. Wound pain was assessed using calibrated Von-Frey monofilaments over a 4-h period, pain-related behaviour was assessed over 5 h, wound healing was assessed at 14 and 28 days, and the plasma levels of lignocaine and bupivacaine were determined. Results Rapid and up to 4 h primary hyperalgesia developed following surgical castration and tail docking in the untreated and placebo-treated lambs. It was absent in the castration wounds, and significantly reduced in the tail-docking wounds, of the treated lambs. Hot-iron docking was associated with mild and transient secondary hyperalgesia, which was abolished by the topical anaesthesia. There was a significant reduction in pain-related behaviours in treated lambs, which were not significantly different in their behaviour to the sham-operation handled controls. Plasma lignocaine and bupivacaine levels were below the toxic thresholds in all tested lambs. Conclusion Topical anaesthesia alleviates wound pain and significantly reduces pain-related behaviours in lambs undergoing surgical castration plus surgical or hot-iron tail docking, without a negative effect on wound healing or a risk of systemic toxicity.     

The influence of compost addition on heavy metal distribution between operationally defined geochemical fractions and on metal accumulation in plant

Purpose

Metal distribution patterns among geochemical fractions are informative for metal phytoavailability. Compost added to polluted soils may adsorb metals on the less phytoavailable fractions. A bioassay experiment was conducted to establish possible correlations between metal concentrations in different soil fractions and metal contents in edible plant parts and to investigate the influence of different compost loads on heavy metal availability to plants.

Materials and methods

Chinese cabbage plants were grown in pots with sandy and clayey soils and soils mixed with different doses of biosolid compost spiked with soluble heavy metal salts (Cd, Cu, and Pb). The metals’ distribution pattern in the soil and mixed samples was determined by sequential extraction procedure (modified BCR protocol). The studied fractions, from most to least bioavailable, were water-extractable (WE), exchangeable-adsorbed (EXC), associated with carbonates and acetic acid-soluble forms (CARB), occluded by reducible (hydro)oxides of Fe and Mn (RO), and associated with organic matter (OM) and a residual fraction (RES). Metal concentrations in soil extracts and in the digested plant tissue were measured by ICP-AES.

Results and discussion

The highest compost doses (72 and 115 Mg ha^?1) enhanced cabbage yield significantly. No excessive phytoaccumulation of metals was observed in plants grown in the clayey soil or its mixtures with compost. The compost dose of 72 Mg ha^?1 was optimal in decreasing Cu accumulation by plants grown in sandy soil, and 28.8 Mg ha^?1 was found to be effective in reducing Cd and Pb uptake. Metals were accumulated in plants primarily from the WE, EXC, and CARB fractions, whereas other fractions decreased phytoaccumulation. Compost addition suppressed heavy metal mobility, but different fractions were active in pollutant sorption, depending on soil type and metal.

Conclusions

Compost addition increased metal proportions in the RO and OM fractions, reducing metal phytoavailability. This is especially important for sandy soils with low adsorption ability and higher vulnerability to metal pollution than clayey soils. A compost dose of 20% v/v (or 28.8 Mg ha^?1) effectively reduced plant accumulation of Cd and Pb. We propose using the first three steps of the modified BCR protocol as a three-step sequential-extraction procedure for the most phytoavailable fractions of heavy metal: WE, EXC, and CARB.     

Potassium uptake characteristics of prunus rootstocks: Influence of solution Ca/Mg ratios and solution nickel

Abstract

Nutrient solution experiments were conducted in the growth chamber to study the influence of rootstock, solution Ca/Mg ratios and solution nickel on K uptake. The experimental plants were one‐year‐old prune trees: ‘French’ prune (Prunus domestica L.) scions grafted on Myrobalan 29C (P. cerasifera Ehrh.), Marianna 2624 (P. cerasifera x P. munsoniana?) or Nemaguard (P. persica x P. davidiana) rootstocks. Ion uptake parameters Imax, Km, and Cmin were calculated from ion depletion measurements over a 6 to 10‐hr period.

With K solution concentrations initially adjusted to 100 μM, K uptake rates of Prunus rootstocks were constant down to approximately 20–30 μM, then declined. Rootstocks were able to deplete solution K to concentrations less than 1 μM. There were no significant differences in K uptake parameters among the rootstocks tested.

Varying solution Ca/Mg ratio from 2.75/1 to 1/4 (Ca + Mg = 3.75 mM) had no effect on K uptake. Potassium uptake rates of Myrobalan 29C rootstocks in the presence of 100 μM nickel were not significantly different from those in the absence of nickel. Rates of nickel uptake were significantly lower than those of K. After eight days of pretreatment in solutions adjusted daily to 100 μM Ni(NO₃)₂, prune leaves began to show signs of interveinal chlorosis. Potassium uptake by nickel pretreated trees was not significantly different from that by control trees. Results are discussed in relation to field observations of K deficiency in prune orchards.     

The Effect of Calcium Sprays and Fruit Thinning on Bitter Pit Incidence and Calcium Content in ‘Honeycrisp’ Apple

‘Honeycrisp’ is a recently released cultivar of apple (Malus x domestica) developed by the University of Minnesota, now in high demand by growers and consumers. This cultivar is susceptible to bitter pit, especially when the trees are young and the fruit load is low. The effects of calcium (Ca) sprays, fruit thinning, vegetative growth, fruit and leaf nutrient concentration on bitter pit incidence, and two sampling methods for nutrient analysis were studied. Bitter pit incidence was associated with several variables, including Ca and phosphorus (P) concentrations in leaves and fruits, magnesium (Mg) concentration in leaves, and yield parameters. Fruit Ca concentration and content for the growing season were modeled, which allowed the determinations of Ca levels associated with bitter pit incidence. Our data suggest that plug analysis was less sensitive than fruit-segment analysis for finding differences in Ca concentrations.