Kidney-derived proteins in urine as biomarkers of induced acute kidney injury in sheep

Acute kidney injury (AKI) is a life-threatening condition for which an early diagnosis is problematic. The aim of the present study was to identify kidney-derived urinary proteins specific to AKI in sheep. AKI was induced in six sheep by an overdose of ketoprofen. Six untreated sheep served as controls. Urine samples were collected for up to 24 h after drug administration and pooled according to time and treatment. Tissue samples from kidney were taken immediately after euthanasia. Urinary proteins were separated by two-dimensional gel electrophoresis (2DE) and the proteins of interest were identified by mass spectrometry. Calbindin-D28k, retinol-binding protein 4 and CD1d were identified in ketoprofen-treated sheep, but not in controls. In addition, calbindin-D28k and CD1d were localized in kidney tissues by immunohistochemical staining. These preliminary results suggest that urinary calbindin-D28k and CD1d represent potential useful biomarkers of AKI, at least in sheep.     

Release of Carbon in Different Molecule Size Fractions from Decomposing Boreal Mor and Peat as Affected by Enchytraeid Worms

Terrestrial export of dissolved organic carbon (DOC) to watercourses has increased in boreal zone. Effect of decomposing material and soil food webs on the release rate and quality of DOC are poorly known. We quantified carbon (C) release in CO₂, and DOC in different molecular weights from the most common organic soils in boreal zone; and explored the effect of soil type and enchytraeid worms on the release rates. Two types of mor and four types of peat were incubated in laboratory with and without enchytraeid worms for 154 days at +?15 °C. Carbon was mostly released as CO₂; DOC contributed to 2–9% of C release. The share of DOC was higher in peat than in mor. The release rate of CO₂ was three times higher in mor than in highly decomposed peat. Enchytraeids enhanced the release of CO₂ by 31–43% and of DOC by 46–77% in mor. High molecular weight fraction dominated the DOC release. Upscaling the laboratory results into catchment level allowed us to conclude that peatlands are the main source of DOC, low molecular weight DOC originates close to watercourse, and that enchytraeids substantially influence DOC leaching to watercourse and ultimately to aquatic CO₂ emissions.     

The genetic diversity of Icelandic populations of two barley leaf pathogens, Rhynchosporium commune and Pyrenophora teres

Twelve species of fungi have been found growing on barley leaves in Icelandic fields. The study presented here on the population structure of two of these species, the pathogens Rhynchosporium commune and Pyrenophora teres f. teres, reveals high levels of genetic diversity, low levels of migration, and a significant differentiation from other European populations, despite the short history of continuous barley cultivation in Iceland. The gene diversity for R. commune in Iceland was 0.55 compared to a range of 0.43?C0.73 for six European populations. The gene diversity for P. teres was higher in Iceland than in populations from Russia and Finland. The two mating types were found to overlap in distribution for both fungi making sexual reproduction a possibility, supported by the few clones and the gametic equilibrium within the Icelandic populations. When the high levels of diversity, low levels of migration, and the genetic differentiation observed between Icelandic and Scandinavian populations are put into context with the short history of barley cultivation in Iceland, it raises questions regarding the origin of the Icelandic fungal populations. It also underlines the importance of proper analysis of pathogens prior to starting resistance breeding projects. The findings are an addition to the ongoing analysis of the global diversity of barley fungal pathogens in general and an important input into future barley breeding projects in Iceland in particular.     

Phenotypic and genetic characterization of Salmonella enterica subsp. enterica serovar Typhimurium isolated from pigs in Rio Grande do Sul, Brazil

This study aimed to investigate the relatedness of porcine Salmonella enterica subsp. enterica (S.) serovar Typhimurium strains isolated in Southern Brazil. Sixty-six isolates from pigs belonging to three commercial companies were submitted to phage typing, XbaI-macrorestriction (PFGE), IS200 hybridization, rep-PCR, antimicrobial susceptibility testing, and PCR assay targeting the spvR region. All strains presented a unique rep-PCR pattern and 63 strains had a common IS200 profile. One pulse-type (XA) was the most prevalent (39/66 strains) and included strains of phage types DT177, DT192, DT194 and RDNC. The spvR region was detected in three strains, which harboured plasmids of 90 kb. High rates of tetracycline, sulfonamide and streptomycin resistance were found. Isolates from farms located in different geographic regions but associated to the same commercial companies clustered together and presented a common resistance profile. Results suggested that clonal groups of S. Typhimurium are present in pig commercial companies in Southern Brazil.     

Activation of complement system in kidney after ketoprofen-induced kidney injury in sheep

Background

Non-steroidal anti-inflammatory drugs (NSAIDs) are widely used to treat inflammatory pain in humans and animals. An overdose of an NSAID is nephrotoxic and can lead to acute kidney injury (AKI). Complement activation occurs in several types of renal disorders with proteinuria. The aim of this study was to investigate whether complement system becomes activated in kidneys after a high dose of NSAID. Kidney tissue and urine samples were collected from six sheep with ketoprofen-induced AKI and from six healthy control sheep. The localization of complement proteins in kidney tissue was carried out using immunohistochemical stainings, and excretion of C3 was tested by immunoblotting.

Results

The complement system was found to become activated in the kidney tissue as demonstrated by positive immunostaining for C1q, C3c, C4c, C5, C9 and factor H and by Western blotting analysis of C3 activation products in urine samples in sheep with AKI.

Conclusions

Our results thus suggest that the alternative complement pathway is activated, and it may contribute to the acute tubular injury seen in the kidneys of NSAID-induced AKI sheep. Inhibition of complement activation may serve as potential therapeutic target for intervention in drug-induced AKI.     

Carbon and nitrogen release from decomposing Scots pine,Norway spruce and silver birch stumps

Stumps are the largest coarse woody debris component in managed forests, but their role in nutrient cycling is poorly understood. We studied carbon (C) and nitrogen (N) dynamics in Scots pine (Pinus sylvestris), Norway spruce (Picea abies), and silver birch (Betula pendula) stumps, which had decomposed for 0, 5, 10, 20, 30 and 40 years after clear-cutting in southern Finland. Carbon and N were released significantly faster from birch stumps than from conifer stumps. In 40 years, conifer stumps lost 78% and birch stumps 90% of their initial C. In contrast, the amount of N in stumps increased, indicating that external N accumulated in the stumps. After 40 years of decomposition, the amount of N was 1.7 and 2.7 times higher than the initial amount in pine and spruce stumps, respectively. Nitrogen was released from birch stumps, but only after they had decomposed for 20 or more years. On average, 59% of N stored in birch stumps was released during 40 years. The results indicate that the stumps of the major tree species in Fennoscandian forests are long-term C and, especially, N pools which serve as N sinks, thus potentially diminishing N leaching into ground water and watercourses after harvesting. This suggests that the removal of stumps for bioenergy production may markedly affect the nutrient status and nutrient cycling of boreal forests.     

Circadian variation in ghrelin and certain stress hormones in crib-biting horses

Crib-biting is classified as an oral stereotypy, which may be initiated by stress susceptibility, management factors, genetic factors and gastrointestinal irritation. Ghrelin has been identified in the gastric mucosa and is involved in the control of food intake and reward, but its relationship to crib-biting is not yet known. The aim of this study was to examine the concentration and circadian variation of plasma ghrelin, cortisol, adrenocorticotropic hormone (ACTH) and β-endorphin in crib-biting horses and non-crib-biting controls. Plasma samples were collected every second hour for 24h in the daily environment of eight horses with stereotypic crib-biting and eight non-crib-biting controls. The crib-biting horses had significantly higher mean plasma ghrelin concentrations than the control horses. The circadian rhythm of cortisol was evident, indicating that the sampling protocol did not inhibit the circadian regulation in these horses. Crib-biting had no statistically significant effect on cortisol, ACTH or β-endorphin concentrations. The inter-individual variations in β-endorphin and ACTH were higher than the intra-individual differences, which made inter-individual comparisons difficult and complicated the interpretation of results. Further research is therefore needed to determine the relationship between crib-biting and ghrelin concentration.     

Upregulation of CRISP-3 and kallikrein in stallion seminal plasma is associated with poor tolerance of cooled storage

For unknown reasons, stallion fertility and sperm longevity during cooled storage of semen vary markedly between individuals. Spermatozoa from individual stallions react differently to the presence, or the removal, of seminal plasma (SP). The aim was to evaluate differences in protein content in stallion seminal plasma with either a positive or a negative effect on sperm chromatin integrity during storage. Stallion semen samples from different ejaculate fractions were stored at 5°C for 24 hr. Sperm survival was assessed after storage using a sperm chromatin structure assay. Protein expression in SP with either positive or negative effects on sperm survival during storage was studied using two-dimensional differential gel electrophoresis and liquid chromatography–mass spectrometry. Lower sperm chromatin integrity was associated with upregulation of the proteins kallikrein, CRISP-3 and HSP-1, while higher chromatin integrity was associated with upregulation of TIMP-2. In the sperm-rich fractions, kallikrein and CRISP-3 differed significantly between SP samples with differing effects on sperm chromatin integrity. In the sperm-poor fractions, TIMP-2 and HSP-1 differed significantly between the two SP groups. Differences in the seminal plasma proteome are associated with sperm longevity during cooled storage.