Relationship between Mycobacterium avium subspecies paratuberculosis, IL-1alpha, and TRAF1 in primary bovine monocyte-derived macrophages

Mycobacterium avium subspecies paratuberculosis (MAP) is a facultative intracellular pathogen that resides in host macrophage cells. Presently, little is known about how MAP is able to subvert the normal bacteriocidal functions of infected macrophages. Previously, we reported that ileal tissues from MAP infected cattle contained high levels of interleukin-1 alpha (IL-1alpha) and tumor necrosis factor receptor-associated factor 1 (TRAF1), relative to ileal tissues from uninfected cattle. High-level expression of these two proteins could have profound effects on macrophage function, intracellular signaling, and apoptosis. We now demonstrate that high levels of TRAF1 protein are located primarily within macrophages infiltrating areas of MAP infection. We have also utilized cultured bovine monocyte-derived macrophage cells (MDM) either infected with live MAP or stimulated with recombinant IL-1alpha (rIL-1alpha) to determine if there is a relationship between IL-1alpha and TRAF1 expression. These studies have identified a dose dependent increase in TRAF1 protein levels in bovine MDM in response to infection with live MAP or following treatment with rIL-1alpha. Sustained TRAF1 protein expression was dependent upon interaction of rIL-1alpha with it's receptor and rIL-1beta was also able to enhance TRAF1 gene expression. Our results suggest that MAP may use the IL-1-TRAF1 system to enhance TRAF1 protein expression in infected bovine MDM. These novel results provide evidence for a new avenue of research on the effect of MAP and other intracellular pathogens on macrophage signaling and apoptosis.     

Modeling the impacts of contrasting nutrient and residue management practices on grain yield of sorghum (Sorghum bicolor (L.) Moench) in a semi-arid region of Ghana using APSIM

The cropping systems model APSIM (Agricultural Production Systems sIMulator) was applied to assess the response of sorghum grain yield to inorganic fertilizers applications and residue retention in diverse farmers’ management systems (homestead fields and bush farms). The model was parameterized using data collected from experiments under optimum growth conditions (limited water or nutrient stress). Independent data from field experiments with three levels of P and four levels of N fertilizers conducted at two different locations and soils were used to evaluate the model. Soil water and fertility parameters measured were used for simulations while same starting conditions were assumed for unmeasured parameters for all trials. APSIM predicted the grain yield response of sorghum to both N and P applications with an overall modified internal coefficient of efficiency of 0.64. Following model parameterization, a long-term simulation study was conducted using a stochastic weather data derived from historical weather data to assess the effects of crop residue management on grain production. A gradual decline in sorghum grain yield was simulated over the 30-year simulation period in both the homestead fields and the bush farms, with yields being much lower in the latter under farmers’ management practices. Half the amount of mineral N fertilizer used in the bush farms was needed in the homestead fields to produce the average grain yields produced on the bush farm with full fertilization, if crop residues were returned to the fields in the homestead. Year-to-year variability in grain yield was consistently higher with the removal of crop residues, irrespective of management systems. APSIM was responsive to both organic and inorganic fertilizer applications in the study area and also highlighted the essential role of crop residues and inorganic fertilizer in influencing the temporal sorghum grain production and hence the impact of farmers’ management practices on food security. This was evident in the rapid decline in soil organic carbon (SOC) accompanied by a decline in grain yield over the 30 years of cropping. The use of inorganic fertilizer and retention of crop residues (SOC) are critical for attaining food security in the study area.     

Silicon pools and fluxes in soils and landscapes—a review

Silicon (Si) is the second‐most abundant element in the earth's crust. In the pedosphere, however, huge spans of Si contents occur mainly caused by Si redistribution in soil profiles and landscapes. Here, we summarize the current knowledge on the different pools and fluxes of Si in soils and terrestrial biogeosystems. Weathering and subsequent release of soluble Si may lead to (1) secondarily bound Si in newly formed Al silicates, (2) amorphous silica precipitation on surfaces of other minerals, (3) plant uptake, formation of phytogenic Si, and subsequent retranslocation to soils, (4) translocation within soil profiles and formation of new horizons, or (5) translocation out of soils (desilication). The research carried out hitherto focused on the participation of Si in weathering processes, especially in clay neoformation, buffering mechanisms for acids in soils or chemical denudation of landscapes. There are, however, only few investigations on the characteristics and controls of the low‐crystalline, almost pure silica compounds formed during pedogenesis. Further, there is strong demand to improve the knowledge of (micro)biological and rhizosphere processes contributing to Si mobilization, plant uptake, and formation of phytogenic Si in plants, and release due to microbial decomposition. The contribution of the biogenic Si sources to Si redistribution within soil profiles and desilication remains unknown concerning the pools, rates, processes, and driving forces. Comprehensive studies considering soil hydrological, chemical, and biological processes as well as their interactions at the scale of pedons and landscapes are necessary to make up and model the Si balance and to couple terrestrial processes with Si cycle of limnic, fluvial, or marine biogeosystems.     

Correlation of enzyme-linked immunosorbent assay titers with protection against infectious bursal disease virus.

We examined the utility of baculovirus-expressed infectious bursal disease virus (IBDV) proteins to act as antigens in the enzyme-linked immunosorbent assay (ELISA). The three IBDV protein antigens tested included 1) a truncated VP2, 2) whole VP2, and 3) the polyprotein products VP2, VP3, and VP4. Serum samples from 2-wk-old commercially reared broilers were collected and tested in the three ELISAs. Serum samples were obtained from 34 different commercial broiler flocks. An average of 14 serum samples (range = 11-17) were tested for each flock. The ELISA results were compared with the percentage of protection of these birds following challenge with IBDV. Fifty 2-wk-old chicks from each of the 34 broiler flocks were challenged with STC classic virus or Del-E variant virus. At 7 days postchallenge, the bursa from each of the birds was removed and bursa/body weights were recorded. Percentage of protection was determined by the number of birds in each challenge group that had normal relative bursal weights compared with unchallenged controls. No evidence was found of a relationship between ELISA data generated with the polyprotein antigen (VP2, VP3, VP4) and percentage of protection observed in the STC and Del-E challenged birds. A significant relationship was found between ELISA data and percentage of protection to STC and Del-E when the truncated VP2 or whole VP2 antigens were used in the ELISA. The results of this study indicate that predicting the percentage of protection against classic or variant IBDV strains in broilers from vaccinated breeder flocks can be improved when VP2 is used as the only antigen in the ELISA.     

Oxygenation and plasma endothelin‐1 concentrations in healthy horses recovering from isoflurane anaesthesia administered with or without pulse‐delivered inhaled nitric oxide

ObjectiveTo assess oxygenation, ventilation‐perfusion (V/Q) matching and plasma endothelin (ET‐1) concentrations in healthy horses recovering from isoflurane anaesthesia administered with or without pulse‐delivered inhaled nitric oxide (iNO).Study DesignProspective experimental trial.AnimalsHealthy adult Standardbred horses.MethodsHorses were anaesthetized with isoflurane in oxygen and placed in lateral recumbency. Six control (C group) horses were anaesthetized without iNO delivery and six horses received pulse‐delivered iNO (NO group). After 2.5 hours of anaesthesia isoflurane and iNO were abruptly discontinued, inhaled oxygen was reduced from 100% to approximately 30%, and the horses were moved to the recovery stall. At intervals during a 30‐minute period following the discontinuation of anaesthesia, arterial and mixed venous blood gas values, shunt fraction (Qs/Qt), plasma ET‐1 concentration, pulse rate and respiratory rate were measured or calculated. Repeated measures anova and a Bonferroni post hoc test was used to analyze data with significance set at p <0.05.ResultsAt all time points in the recovery period, NO horses maintained better arterial oxygenation (oxygen partial pressure: NO 13.2 ± 2.7–11.1 ± 2.7 versus C 6.7 ± 1.1–7.1 ± 1.1 kPa) and better V/Q matching (Qs/Qt NO 0.23 ± 0.05–0.14 ± 0.06 versus C 0.48 ± 0.03–0.32 ± 0.08%) than C horses. Mixed venous oxygenation was higher in NO for 25 minutes following the discontinuation of anaesthesia (NO 6.3 ± 0.2–4.5 ± 0.07 versus C 4.7 ± 0.6–3.7 ± 0.3 kPa). In both groups of horses arterial oxygenation remained fairly stable; venous oxygenation declined over this time period in the NO group but still remained higher than venous oxygen in the C group. ET‐1 concentrations were higher at most time points in C than NO. Changes in other parameters were either minor or absent.Conclusions and Clinical RelevanceDelivery of iNO to healthy horses during anaesthesia results in better arterial and venous oxygenation and V/Q matching (as determined by lower Qs/Qt) and lower ET‐1 concentrations throughout a 30‐minute anaesthetic recovery period.     

A Stable-Isotope Mass Spectrometry-Based Metabolic Footprinting Approach to Analyze Exudates from Phytoplankton

Phytoplankton exudates play an important role in pelagic ecology and biogeochemical cycles of elements. Exuded compounds fuel the microbial food web and often encompass bioactive secondary metabolites like sex pheromones, allelochemicals, antibiotics, or feeding attractants that mediate biological interactions. Despite this importance, little is known about the bioactive compounds present in phytoplankton exudates. We report a stable-isotope metabolic footprinting method to characterise exudates from aquatic autotrophs. Exudates from ¹³C-enriched alga were concentrated by solid phase extraction and analysed by high-resolution Fourier transform ion cyclotron resonance mass spectrometry. We used the harmful algal bloom forming dinoflagellate Alexandrium tamarense to prove the method. An algorithm was developed to automatically pinpoint just those metabolites with highly ¹³C-enriched isotope signatures, allowing us to discover algal exudates from the complex seawater background. The stable-isotope pattern (SIP) of the detected metabolites then allowed for more accurate assignment to an empirical formula, a critical first step in their identification. This automated workflow provides an effective way to explore the chemical nature of the solutes exuded from phytoplankton cells and will facilitate the discovery of novel dissolved bioactive compounds.     

Characterization of Transgenic Pigs That Express Human Decay Accelerating Factor and Cell Membrane‐tethered Human Tissue Factor Pathway Inhibitor

The expression of human complement regulatory proteins (hCRP; hDAF, hCD59, and hMCP) in pig tissues has been suggested as one of strategies to overcome the hyperacute rejection (HAR) in pig‐to‐human transplantation. Expression of human tissue factor pathway inhibitor (hTFPI) in porcine endothelial cells has been suggested as a remedy to overcome microvascular thrombosis. To investigate the effects of these combined transgenes, we established transformed pig cells expressing human decay accelerating factor (hDAF) under the control of enhancer promoter (5′LTR‐PCMVIE), and the fusion protein (hTFPI/hCD4) consisting of the functional domains (K1 and K2) of hTFPI and membrane‐tethering domains (D3 and D4) of hCD4 under the control of PCMVIE. Transgenic pigs were generated with the transformed porcine cells through somatic cell nuclear transfer (SCNT) technology. Analysis of quantitative PCR and real‐time quantitative RT‐PCR showed that four copies of hDAF were integrated and 391 copies of hDAF mRNA expressed in the cells of the transgenic pig. The enhancing activity of 5′LTR was approximately 2 fold compared to CMVIE promoter only. The cell viability test showed that more than 80% of ear cells were viable in the presence of 50% human serum. The chromogenic substrate assay and immunocytochemical staining with tail cells showed that the TFPI activity of fusion protein was observed on the cell membrane. The membrane localization of hDAF and hTFPI proteins was observed by immunocytochemical staining, and the expression of transgenes in heart and liver tissues was also confirmed by immunohistochemistry.     

Effects of barefoot trimming on hoof morphology

Objective To monitor changes in hoof morphology in response to barefoot trimming. Methods Seven horses were trimmed every 6 weeks according to barefoot trimming principles, which involved levelling the hoof to live sole, lowering the heels, bevelling the toe and rounding the peripheral wall, while leaving the sole, frog and bars intact. A 4‐month period was allowed to lower the heels sufficiently to achieve a hoof shape representative of the barefoot trim. This was regarded as the starting point for morphological adaptations in response to maintenance of the trim. Hoof morphology was measured from lateral, dorsal and solar view photographs and lateromedial radiographs taken at 0, 4 and 16 months. Changes from 0 to 4 months represented differences between a natural hoof shape and the trim, while changes from 4 to 16 months represented adaptive effects during hoof growth. Results Establishment of the barefoot trim involved significant shortening of the toe, heel and medial and lateral walls, with increases in angulation at the toe, medial and lateral walls, but not at the heel. Maintenance of the trim resulted in a palmar/plantar migration of the heels, with increases in support length, heel angle and solar angle of the distal phalanx (P3). Conclusions Bevelling the toe and engaging the frog and bars in the weight‐bearing function of the foot resulted in elevation of the heel angle and solar angle of P3. These changes may be beneficial in treating under‐run heels and negative solar plane angulation of P3.     

Coupled 186Os and 187Os evidence for core-mantle interaction

Osmium isotopic analyses of picritic lavas from Hawaii show enrichments in the osmium-186/osmium-188 ratio (186Os/188Os) of 0. 008 to 0.018%, relative to a chondritic upper mantle, that are positively correlated with enrichments in 187Os/188Os of 5.4 to 9.0%. The most viable mechanism to produce these coupled 186Os and 187Os enrichments is by addition of 0.5 to 1 weight percent of outer core metal to a portion of the D" layer and subsequent upwelling of the mixture. These data suggest that some plumes originate at the core-mantle boundary and that Os isotopes may be used to distinguish plumes derived from shallow versus deep mantle sources.