Clinical characterization of a hypersensitivity mixed bacterial and fungal dermatitis in a translational model of porcine NASH

IntroductionThe development of animal models of chronic liver disease via diet modification is a promising avenue for translational research but can lead to unexpected side effects that impact model adoption. While these side effects are well characterized in rodent models of nonalcoholic steatohepatitis (NASH), limited knowledge of these effects exists for novel porcine models of NASH. To close this gap, the present study investigates the side effects of diet-based NASH induction in pigs, with a systematic analysis of the pathologic mechanisms underlying dermatitis development and evaluation of treatment approaches.MethodTwelve pigs (10 large domestic pigs, 2 Goettingen minipigs) were fed a methionine- and choline-deficient, high-fat diet for 8 weeks to induce NASH. A retrospective review of each animal’s clinical record was performed to identify the side effects of the diet. Following the identification of diet-associated dermatitis, severity was judged by using a novel gradation system that characterized the individual lesions and body regions resulting in a cumulative evaluation. In addition to this clinical assessment, the etiology of the dermatitis was investigated via histopathologic and microbiologic testing. Furthermore, the success of prophylactic and therapeutic treatment approaches was evaluated by considering dermatitis development and clinical course.ResultsAll study animals demonstrated unexpected side effects of the methionine- and choline-deficient, high fat diet. In addition to marked dermatitis, study pigs showed impaired weight gain and developed steatorrhea and anemia. Based on the skin gradation system, five animals developed severe dermatitis, four animals moderate dermatitis, and three animals mild diet-associated dermatitis. Histological and microbiological evaluation of the affected skin showed signs of a hypersensitivity reaction with secondary infection by bacteria and fungi. The analysis showed that preemptive bathing extended the lesion-free duration by nearly 20 days. Furthermore, bathing in combination with a targeted antibiotic treatment represented a helpful treatment approach for diet-associated dermatitis.ConclusionThe provision of a methionine- and choline-deficient, high fat diet represents an effective approach for inducing NASH liver disease in pigs but predisposes study animals to multiple side effects. These side effects are universal to animals on study but can be adequately managed and do not represent a significant limitation of this model

Mars Phobos and Deimos Survey (M-PADS) – A Martian Moons Orbiter and Phobos Lander

We describe a Mars 'Micro Mission' for detailed study of the martian satellites Phobos and Deimos. The mission involves two ~330 kg spacecraft equipped with solar electric propulsion to reach Mars orbit. The two spacecraft are stacked for launch: an orbiter for remote investigation of the moons and in situ studies of their environment in Mars orbit, and another carrying a lander for in situ measurements on the surface of Phobos (or alternatively Deimos). Phobos and Deimos remain only partially studied, and Deimos less well than Phobos. Mars has almost always been the primary mission objective, while the more dedicated Phobos project (1988-89) failed to realise its full potential. Many questions remain concerning the moons' origins, evolution, physical nature and composition. Current missions, such as Mars Express, are extending our knowledge of Phobos in some areas but largely neglect Deimos. The objectives of M-PADS focus on: origins and evolution, interactions with Mars, volatiles and interiors, surface features, and differences. The consequent measurement requirements imply both landed and remote sensing payloads. M-PADS is expected to accommodate a 60 kg orbital payload and a 16 kg lander payload. M-PADS resulted from a BNSC-funded study carried out in 2003 to define candidate Mars Micro Mission concepts for ESA's Aurora programme

Diagnostic Survey of Biological Measurements Used to Determine Bone Mineralization in Pigs Across the US Swine Industry

Pigs from 64 commercial sites across 14 production systems in the Midwest US were used to evaluate the baseline biological measurements used to determine bone mineralization. Three pigs were selected from each commercial site representing: 1) a clinically normal pig (healthy); 2) a pig with evidence of clinical lameness (lame); and 3) a pig from a hospital pen that is assumed to have recent low feed intake (unhealthy). Pigs ranged in age from nursery to market weight, with the three pigs sampled from each site representing the same age or phase of production. Blood, urine, metacarpal, fibula, 2nd rib, and 10th rib were collected and analyzed. Serum was analyzed for Ca, P, and 25(OH)D3, and urine was collected and analyzed for Ca, P, and creatinine. Each bone was measured for density, ash (defatted and non-defatted technique), and breaking strength. A bone × pig type interaction (P \u3c 0.001) was observed for defatted and non-defatted bone ash, density, and breaking strength. For defatted bone ash, there were no differences (P \u3e 0.10) between pig types for the fibulas, 2nd rib, and 10th rib, but metacarpals from healthy pigs had greater (P \u3c 0.05) percentage bone ash compared to unhealthy pigs, with the lame pigs intermediate. For non-defatted bone ash, there were no differences (P \u3e 0.10) between pig types for metacarpals and fibulas, but unhealthy pigs had greater (P \u3c 0.05) non-defatted percentage bone ash for 2nd and 10th ribs compared to healthy pigs, with lame pigs intermediate. Healthy and lame pigs had greater (P \u3c 0.05) bone density than unhealthy pigs for metacarpals and fibulas, with no difference (P \u3e 0.10) observed for ribs. Healthy pigs had bones with increased breaking strength compared to lame and unhealthy pigs for metacarpals and 10th ribs (P \u3c 0.05) with no differences (P \u3e 0.05) between pig types for fibula and 2nd rib. Healthy pigs had greater (P \u3c 0.05) serum Ca and 25(OH)D3 compared to unhealthy pigs, with lame pigs intermediate. Healthy pigs had greater (P \u3e 0.05) serum P compared to unhealthy and lame pigs, with no differences (P \u3e 0.05) between the unhealthy and lame pigs. Unhealthy pigs excreted more (P \u3c 0.05) P and creatinine in the urine compared to healthy pigs, with lame pigs intermediate. In summary, there are differences in serum Ca, P, and vitamin D between healthy, lame, and unhealthy pigs. Differences in bone mineralization between the pig types varied depending on the analytical procedure and bone. There was a considerable range in values within pig type across the 14 production systems sampled

One-pot synthesis and AFM imaging of a triangular aramide macrocycle

Macrocyclizations in exceptionally good yields were observed during the self-condensation of N-benzylated phenyl p-aminobenzoates in the presence of LiHMDS to yield three-membered cyclic aramides that adopt a triangular shape. An ortho-alkyloxy side chain on the N-benzyl protecting group is necessary for the macrocyclization to occur. Linear polymers are formed exclusively in the absence of this Li-chelating group. A model that explains the lack of formation of other cyclic congeners and the demand for an N-(o-alkoxybenzyl) protecting group is provided on the basis of DFT calculations. High-resolution AFM imaging of the prepared molecular triangles on a calcite(10.4) surface shows individual molecules arranged in groups of four due to strong surface templating effects and hydrogen bonding between the molecular triangles

The Effect of Bone and Analytical Methods on the Assessment of Bone Mineralization Response to Dietary Phosphorus, Phytase, and Vitamin D in Nursery Pigs

Three hundred-fifty pigs (initially 26.2 ± 1.23 lb) were used to evaluate the effects of bone and analytical methods on the assessment of bone mineralization response to dietary P and vitamin D in nursery pigs. Pens of pigs (5 or 6 pigs/pen) were randomized to 6 dietary treatments in a randomized complete block design with 10 pens per treatment. Treatments were formulated to have varying levels of P, phytase, and vitamin D to provide differences in bone characteristics. After feeding diets for 28 d, eight pigs per treatment were euthanized for bone, blood, and urine analysis. The response to treatment for bone density and ash was dependent upon the bone analyzed (density × bone interaction, P = 0.044; non-defatted bone ash × bone interaction, P = 0.060; defatted bone ash × bone interaction, P = 0.068). Pigs fed 0.19% STTD P had decreased (P \u3c 0.05) bone density and ash (non-defatted and defatted) for all bones compared to 0.44% STTD P, with 0.33% STTD P generally intermediate or similar to 0.44% STTD P. Pigs fed 0.44% STTD P with no vitamin D had greater (P \u3c 0.05) non-defatted fibula ash compared to all treatments other than 0.44% STTD P with added HyD. Pigs fed the three diets with 0.44% STTD P had greater (P \u3c 0.05) defatted 2nd rib ash compared to pigs fed 0.19% STTD P or 0.33% STTD P with no phytase. In summary, bone density and ash responses varied depending on the bone analyzed. Differences in bone density and ash in response to P and vitamin D were most apparent with fibulas and 2nd ribs. The difference between bone ash procedures was more apparent than the differences between treatments. For histopathology, 10th ribs were more sensitive than 2nd ribs or fibulas for detection of lesions

The Effect of Different Bone and Analytical Methods on the Assessment of Bone Mineralization to Dietary Phosphorus, Phytase, and Vitamin D in Finishing Pigs

Eight hundred eighty-two pigs (initially 73.2 ± 0.7 lb) were used to evaluate the effects of different bones and analytical methods on the assessment of bone mineralization response to dietary P and vitamin D in growing-finishing pigs. Pens of pigs (20 pigs per pen) were randomized to 1 of 5 dietary treatments in a completely randomized design with 9 pens per treatment. Treatments were formulated to have varying levels of P, phytase, and vitamin D to potentially provide wide differences in bone characteristics. After feeding diets for 112 d, nine pigs per treatment were euthanized for bone, blood, and urine analysis. There were no significant differences for final BW, ADG, ADFI, F/G (P \u3e 0.10), or bone ash (bone ash × bone interaction, P \u3e 0.10) regardless of the ashing method. The response to treatment for bone density and bone mineral content was dependent upon the bone (density interaction, P = 0.053; mineral interaction, P = 0.078). There were no treatment differences for bone density and bone mineral content for metacarpals, fibulas, and 2nd rib (P \u3e 0.05). For 10th rib bone density, pigs fed industry levels of P and vitamin D had increased (P \u3c 0.05) bone density compared to pigs fed NRC levels with phytase, with pigs fed deficient P, NRC levels of P with no phytase, and extra 25(OH)D3 vitamin D (HyD) intermediate. Pigs fed extra vitamin D from HyD had increased (P \u3c 0.05) 10th rib bone mineral content compared to pigs fed deficient P and NRC levels of P with phytase, with pigs fed industry P and vitamin D, and NRC P with monocalcium intermediate. In summary, bone density and bone mineral content responses varied depending on the bone. The difference between bone ash procedures was more apparent than the differences between diets. Differences in bone density and mineral content in response to P and vitamin D were most apparent with the 10th ribs

Interleukin-21 Drives Proliferation and Differentiation of Porcine Memory B Cells into Antibody Secreting Cells.

Immunological prevention of infectious disease, especially viral, is based on antigen-specific long-lived memory B cells. To test for cellular proliferation and differentiation factors in swine, an outbred model for humans, CD21+ B cells were activated in vitro with CD40L and stimulated with purported stimulatory cytokines to characterize functional responses. IL-21 induced a 3-fold expansion in total cell numbers with roughly 15% of all B cells differentiating to IgM or IgG antibody secreting cells (ASCs.) However, even with robust proliferation, cellular viability rapidly deteriorated. Therefore, a proliferation inducing ligand (APRIL) and B cell activating factor (BAFF) were evaluated as survival and maintenance factors. BAFF was effective at enhancing the viability of mature B cells as well as ASCs, while APRIL was only effective for ASCs. Both cytokines increased approximately two-fold the amount of IgM and IgG which was secreted by IL-21 differentiated ASCs. Mature B cells from porcine reproductive and respiratory virus (PRRSV) immune and naïve age-matched pigs were activated and treated with IL-21 and then tested for memory cell differentiation using a PRRSV non-structural protein 7 ELISPOT and ELISA. PRRSV immune pigs were positive on both ELISPOT and ELISA while naïve animals were negative on both assays. These results highlight the IL-21-driven expansion and differentiation of memory B cells in vitro without stimulation of the surface immunoglobulin receptor complex, as well as the establishment of a defined memory B cell culture system for characterization of vaccine responses in outbred animals