Porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) are two of the most important pathogens affecting the swine industry worldwide. Co-infections are common on a global scale, resulting in pork production losses through reducing weight gain and causing respiratory disease in growing pigs. Our initial work demonstrated that the fecal microbiome was associated with clinical outcome of pigs 70days post-infection (dpi) with PRRSV and PCV2. However, it remained uncertain if microbiome characteristics could predispose response to viral infection. The purpose of this study was to determine if microbiome characteristics present at the time of virus exposure were associated with outcome after co-infection. Using the Lawrence Livermore Microbial Detection Array, we profiled the microbiome in feces prior to infection from pigs identified retrospectively as having high or low growth rates after co-infection. High growth rate pigs had less severe interstitial pneumonia, reduced virus replication, and a significant increase in average daily weight gain throughout the study. At the level of the fecal microbiome, high growth rate pigs had increased microbial diversity on both a family and species level. Shifts in the microbiome composition of high growth rate pigs included reduced Methanobacteriaceae species, increased Ruminococcaceae species, and increased Streptococcaceae species when compared to low growth rate pigs. The results indicate that both microbiome diversity and composition at the time of virus exposure may play a role in the subsequent response of pigs to PRRSV/PCV2 co-infection.

The dwarf cuttlefish, Sepia bandensis, a small cephalopod that exhibits dynamic camouflage, is an emerging model organism in neuroscience. Coleoid cephalopods (cuttlefish, octopus, and squid) evolved large, complex brains capable of learning, problem-solving, and memory. We used high resolution magnetic resonance imaging (MRI), deep learning, and fluorescent histology to generate a dwarf cuttlefish brain atlas and built an interactive web tool (cuttlebase.org) to host the data. Guided by observations in other cephalopods, we identified 38 brain lobes. The dwarf cuttlefish brain is partially encased in cartilage and includes two large optic lobes (74% the total volume of the brain), chromatophore lobes whose motor neurons directly innervate the skin, and a vertical lobe that has been implicated in learning and memory. Motor neurons emerging from the chromatophore lobe modulate the color, pattern, and texture of the skin to elicit camouflage. This brain atlas provides a valuable tool for exploring the neural basis of cuttlefish behavior.

Background Expression of receptor for advanced glycation end products (RAGE) plays an important role in diabetic peripheral artery disease. We proposed to show that treatment with an antibody blocking RAGE would improve hind limb perfusion and muscle viability in diabetic pig with femoral artery (FA) ligation. Methods and Results Purpose‐bred diabetic Yucatan minipigs with average fasting blood sugar of 357 mg/dL on insulin to maintain a glucose range of 300 to 500 mg/dL were treated with either a humanized monoclonal anti‐RAGE antibody (CR‐3) or nonimmune IgG. All pigs underwent intravascular occlusion of the anterior FA. Animals underwent ( 201 Tl) single‐photon emission computed tomography/x‐ray computed tomography imaging on days 1 and 28 after FA occlusion, angiogenesis imaging with [ 99m Tc]dodecane tetra‐acetic acid–polyethylene glycol–single chain vascular endothelial growth factor (scVEGF), muscle biopsies on day 7, and contrast angiogram day 28. Results showed greater increases in perfusion to the gastrocnemius from day 1 to day 28 in CR‐3 compared with IgG treated pigs ( P =0.0024), greater uptake of [99mTc]dodecane tetra‐acetic acid‐polyethylene glycol‐scVEGF (scV/Tc) in the proximal gastrocnemius at day 7, confirmed by tissue staining for capillaries and vascular endothelial growth factor A, and less muscle loss and fibrosis at day 28. Contrast angiograms showed better reconstitution of the distal FA from collaterals in the CR‐3 versus IgG treated diabetic pigs ( P =0.01). The gastrocnemius on nonoccluded limb at necropsy had higher 201 Tl uptake (percentage injected dose per gram) and reduced RAGE staining in arterioles in CR‐3 treated compared with IgG treated animals ( P =0.04). Conclusions A novel RAGE‐blocking antibody improved hind limb perfusion and angiogenesis in diabetic pigs with FA occlusion. Contributing factors are increased collaterals and reduced vascular RAGE expression. CR‐3 shows promise for clinical treatment in diabetic peripheral artery disease.

Myocardial Elastography (ME) is an ultrasound-based strain imaging method. A survival canine model (n=11) was employed to investigate ME's ability to image myocardial infarction (MI) formation and recovery. Infarcts were generated by ligation of the left anterior descending coronary artery. Canines were survived and imaged for four days (n=7) or four weeks (n=4), allowing sufficient time for recovery via collateral perfusion. A radial strain-based metric, percentage of healthy myocardium by strain (PHM ε ), was developed as a marker for healthy myocardial tissue. PHM ε was strongly linearly correlated with actual infarct size as determined by gross pathology (R 2 = 0.80). Mean PHM ε was reduced 1-3 days postinfarction (p<0.05) at the papillary and apical short-axis levels; full recovery was achieved by day 28, with mean PHM ε returning to baseline levels. ME was capable of diagnosing individual myocardial segments as non-infarcted or infarcted with high sensitivity (82%), specificity (92%), and precision (85%) (ROC AUC = 0.90).

Background: New therapies to treat diabetic peripheral artery disease (PAD) require target-specific non-invasive imaging modalities to follow efficacy. As a translational study, we performed targeted imaging of receptors for vascular endothelial growth factor (VEGF) in response to anterior femoral artery occlusion (FAO) in Yucatan minipigs and compare the normal response to response in diabetic Yucatan minipigs. Methods: Eleven Yucatan minipigs, 6 non-diabetic (non-D) and 5 purpose bred diabetic (D) (Sinclair, Auxvasse MO), underwent intravascular total occlusion of the anterior femoral artery (FA). At days 1 and 28, pigs underwent SPECT/ CT 201 Tl hindlimb perfusion imaging and at day 7 were injected with [ 99m Tc]DOTA-PEG-scVEGF (scV/Tc) tracer targeting VEGF receptor, and underwent biopsies of the hindlimb muscles for gamma counting and histology, followed by imaging. One day after the final scan, pigs underwent contrast angiography of the lower extremities. Counts from scans were converted to percentage injected activity (%IA). Results: Perfusion was lower in the occluded hindlimb compared to non-occluded on day 1 in both the D and non-D pigs. At day 7, scV/Tc count ratio of counts from ROIs drawn in proximal gastrocnemius muscle for the occluded over non-occluded limb was significantly higher in non-D vs. D pigs (1.32 ± 0.06 vs. 1.04 ± 0.13, P = 0.02) reflecting higher level of angiogenesis. Perfusion increased between days 1 and 28 in the muscles in the occluded limb for the non-diabetic pigs while the diabetic pig showed no increase (+ 0.13 ± 0.08 %IA vs. − 0.13 ± 0.11, P = 0.003). The anterior FA showed poor contrast filling beyond occluder and qualitatively fewer bridging collaterals compared to non-D pigs at 28 days. Conclusion: VEGF receptor targeted imaging showed the effects of diabetes to suppress angiogenesis in response to occlusion of the anterior femoral artery of purpose bred diabetic Yucatan minipigs and indicates potential applicability as a marker to follow efficacy of novel therapies to improve blood flow by stimulating angiogenesis in diabetic PAD.

Models of type-I diabetes are well-characterized and commonly used in the preclinical evaluation of drugs and medical devices. The diabetic minipig is an excellent example of a translational model. However, chronic glucose monitoring in this species can be challenging; frequent blood sampling can be technically difficult and poorly tolerated in conscious swine. Skin-patch continuous blood glucose monitors are FDA-approved for human use and offer a potential refinement to cageside blood collection. However, this modality has not been evaluated in pigs. In this study, young adult male STZ-induced diabetic Yucatan minipigs (n = 4) and healthy York pigs (n = 4) were implanted with a 14-d skin-patch continuous glucose monitor. Readings from continuous glucose monitors were time-matched to whole blood samples, with glucose measurements performed using point-of-care blood glucose monitors, serum chemistry or both. The aims of the study were to assess if a continuous glucose monitoring system could accurately detect glucose levels in swine, and to compare the readings toboth point-of-care glucometers and serum chemistry results. We hypothesized that a continuous glucose monitoring system would accurately detect glucose levels in swine in comparison with a validated analyzer and could serve as an animal welfarerefinement for studies of diabetes. We found that the continuous glucose monitor used in this study provided an adequateadjunct for clinical management in the stable diabetic pig and a minimally invasive and inexpensive option for colony maintenanceof chronically diabetic swine.

BACKGROUND: MG53, a member of the tripartite motif (TRIM) protein family, plays an essential role in cell membrane repair and promotes cell survival.Recent studies show that systemic delivery of recombinant human MG53 (rhMG53) protein markedly attenuates tissue injury/ inflammation, and facilitates healing. This study was performed to test whether intravenous administration of rhMG53 protein would decrease the lesion size in a clinically relevant large animal model of traumatic brain injury (TBI). METHOD:Yorkshire swine (40-45 kg; n = 5/group) were subjected to controlled cortical impact TBI and randomized to either: (1) rhMG53 protein (2 mg/kg, intravenous) or (2) normal saline control. Hemodynamics, intracranial pressure, and brain oxygenation were monitored for 7 hours. Brains were then harvested and sectioned into 5-mm slices and stained with 2,3,5-triphenyltetrazolium chloride to quantify the lesion size. Blood-brain barrier permeability of MG53 in the brain was determined by Western blot and immunohistochemistry. Bcl-2 and phospho-GSK β levels were measured as makers of prosurvival pathway activation. RESULTS:Hemodynamic parameters were similar in both groups, but the lesion size in the rhMG53-treated group (2,517 ± 525.4 mm 3 ) was significantly ( p < 0.05) smaller than the control group (3,646 ± 740.1 mm 3 ). In the treated animals, rhMG53 was detected in the regions surrounding the TBI, but it was absent in the saline-treated control animals. Bcl-2 and phospho-GSK β levels in the brains were upregulated in the rhMG53-treated animals. CONCLUSION: Intravenously administered rhMG53 localizes to the injured areas of the brain, with the treated animals demonstrating a significant attenuation in the brain lesion size following TBI.

No abstract