Swine models of Human Conditions

Comparative Medicine: Innovations for the Animal to Human Health CorridorSince the introduction of Dolly, the first cloned sheep, genetic engineering of livestock has advanced to the point that nearly any genetic modification appears feasible. Genes can be introduced, deleted or altered in cultured cells. These cells can be evaluated in culture and then used to regenerate live animals by somatic cell nuclear transfer (cloning). Since pigs share similar physiology with humans and have proven to be excellent models for many human conditions, genetic engineering and animal cloning are being used to produce models of human disease states and disorders. These models are proving to be particularly important for situations where laboratory rodents have proven to be inadequate. Current successes of genetically engineered swine biomedical models will be presented

Identification of H19 polymorphism for an assessment of biallelic expression

Abstract only availableAnimals produced from assisted reproductive technologies suffer from developmental abnormalities and early fetal death at a higher frequency than that observed in those produced by natural breeding. These symptoms are reminiscent of imprinting disruptions, suggesting the possibility of an alteration in the expression of imprinted genes such as biallelic expression or silencing. H19 is one of the imprinted genes first identified in mice and humans, but its imprinting status has not been determined in pigs. The objective of this study was to identify an H19 polymorphism and estimate its frequency in the commercial pig population. In this study a polymorphism in the H19 gene was identified. The PCR products contained a pooled genome with over 900 specimens to support this finding. From the positive PCR products, the DNA was cloned and transformed with a TOPO TA Cloning kit (Invitrogen). Positive colonies were identified and digested with an AciI enzyme, which cut the DNA in specific fragments that were identifiable in a gel. Analysis of the gel showed evidence that a polymorphism exists on the H19 gene.F.B. Miller Undergraduate Research Program in Animal Science

Oceanographic Structure and Light Levels Drive Patterns of Sound Scattering Layers in a Low-Latitude Oceanic System

Several factors have been reported to structure the spatial and temporal patterns of sound scattering layers, including temperature, oxygen, salinity, light, and physical oceanographic conditions. In this study, we examined the spatiotemporal variability of acoustically detected sound scattering layers in the northern Gulf of Mexico to investigate the drivers of this variability, including mesoscale oceanographic features [e.g., Loop Current-origin water (LCOW), frontal boundaries, and Gulf Common Water]. Results indicate correlations in the vertical position and acoustic backscatter intensity of sound scattering layers with oceanographic conditions and light intensity. LCOW regions displayed consistent decreases, by a factor of two and four, in acoustic backscatter intensity in the upper 200 m relative to frontal boundaries and Gulf Common Water, respectively. Sound scattering layers had greater backscatter intensity at night in comparison to daytime (25x for frontal boundaries, 17x for LCOW, and 12x for Gulf Common Water). The importance of biotic (primary productivity) and abiotic (sea surface temperature, salinity) factors varied across oceanographic conditions and depth intervals, suggesting that the patterns in distribution and behavior of mesopelagic assemblages in low-latitude, oligotrophic ecosystems can be highly dynamic

Abundance and Occurrence of Common Bottlenose Dolphins (Tursiops truncatus) in Three Estuaries of the Northwestern Gulf of Mexico

Current abundance estimates for populations of common bottlenose dolphins (Tursiops truncatus, Montagu, 1821) in bays, sounds, and estuaries are lacking throughout most of the northwestern Gulf of Mexico, including areas of Texas and western Louisiana. To address this issue, we conducted 92 small-boat photographic identification surveys covering ~2000 km2 and comprising ~11,000 km of track-line in winter and summer seasons in West Bay, TX (2014 – 2015, n = 25), the Galveston Bay, TX system (2016, n = 50), Sabine Lake, TX (2017, n = 17), and adjacent coastal waters. Individual dolphin encounter histories were constrained by spatiotemporal parameters to approximately represent 1) a “Bay” estimate of individuals limited to the interior of each embayment, and 2) a “Selective” estimate of the number of individuals in each survey area (including nearshore coastal waters), filtered for potential transient dolphins. Using the Selective dataset, estimated dolphins (95% CI) were (winter and summer, respectively) 38 (29 – 47) and 37 (33 – 40) for West Bay, 842 (694 – 990) and 1132 (846 – 1417) for Galveston Bay, and 122 (73 – 170) and 162 (114 – 210) for Sabine Lake. A range of 4-15% of marked individuals in each study area were identified as inter-bay matches. These results provide new insights on the potential spatial range of each population, update previous abundance estimates for West Bay and Galveston Bay, and contribute novel population information for Sabine Lake and adjacent coastal waters of the northwestern Gulf of Mexico

International validation of Enhanced Recovery After Surgery Society guidelines on enhanced recovery for gynecologic surgery

Background: Enhanced Recovery After Surgery Society publishes guidelines on perioperative care, but these guidelines should be validated prospectively. Objective: To evaluate the association between compliance with Enhanced Recovery After Surgery Gynecologic/Oncology guideline elements and postoperative outcomes in an international cohort. Study Design: The study comprised 2101 patients undergoing elective gynecologic/oncology surgery between January 2011 and November 2017 in 10 hospitals across Canada, the United States, and Europe. Patient demographics, surgical/anesthesia details, and Enhanced Recovery After Surgery protocol compliance elements (pre-, intra-, and postoperative phases) were entered into the Enhanced Recovery After Surgery Interactive Audit System. Surgical complexity was stratified according to the Aletti scoring system (low vs medium/high). The following covariates were accounted for in the analysis: age, body mass index, smoking status, presence of diabetes, American Society of Anesthesiologists class, International Federation of Gynecology and Obstetrics stage, preoperative chemotherapy, radiotherapy, operating time, surgical approach (open vs minimally invasive), intraoperative blood loss, hospital, and Enhanced Recovery After Surgery implementation status. The primary end points were primary hospital length of stay and complications. Negative binomial regression was used to model length of stay, and logistic regression to model complications, as a function of compliance score and covariates. Results: Patient demographics included a median age 56 years, 35.5% obese, 15% smokers, and 26.7% American Society of Anesthesiologists Class III-IV. Final diagnosis was malignant in 49% of patients. Laparotomy was used in 75.9% of cases, and the remainder minimally invasive surgery. The majority of cases (86%) were of low complexity (Aletti score ≤3). In patients with ovarian cancer, 69.5% had a medium/high complexity surgery (Aletti score 4–11). Median length of stay was 2 days in the low- and 5 days in the medium/high-complexity group. Every unit increase in Enhanced Recovery After Surgery guideline score was associated with 8% (IRR, 0.92; 95% confidence interval, 0.90–0.95; P\u3c.001) decrease in days in hospital among low-complexity, and 12% (IRR, 0.88; 95% confidence interval, 0.82–0.93; P\u3c.001) decrease among patients with medium/high-complexity scores. For every unit increase in Enhanced Recovery After Surgery guideline score, the odds of total complications were estimated to be 12% lower (P\u3c.05) among low-complexity patients. Conclusion: Audit of surgical practices demonstrates that improved compliance with Enhanced Recovery After Surgery Gynecologic/Oncology guidelines is associated with an improvement in clinical outcomes, including length of stay, highlighting the importance of Enhanced Recovery After Surgery implementation

Short-Haul Revitalization Study Final Report

A feasibility study was performed for an advanced commercial short-haul aircraft to evaluate the potential for increased service for short-haul flights that operate out of regional and community airports. An analysis of potential origin-destination markets and trip distances resulted in a seat capacity selection of 48 passengers and a design range of 600 NM. A down-select of advanced technologies resulted in a hybrid-electric propulsion system being chosen as the primary enabling technology. A conceptual design of the advanced aircraft was developed, and a mission and sizing analysis was performed, comparing variants of the advanced aircraft with different levels of electrification. Fairly aggressive levels of electrification and battery specific energy are needed for the hybridelectric architecture to realize any benefit in terms of total energy cost for the 600 NM design mission. The development and operational costs were estimated for the advanced aircraft and compared to the baseline. This analysis demonstrated the negative effect of the cost to develop the hybrid-electric technology on the eventual operating cost. A market analysis was performed to determine possible passenger demand for the advanced shorthaul aircraft. According to the market analysis, there is potential demand for such an aircraft, but not necessarily in many of the smaller regional and community airports that were the intended beneficiaries of this new aircraft concept

Hierarchical patterning modes orchestrate hair follicle morphogenesis

Two theories address the origin of repeating patterns, such as hair follicles, limb digits, and intestinal villi, during development. The Turing reaction–diffusion system posits that interacting diffusible signals produced by static cells first define a prepattern that then induces cell rearrangements to produce an anatomical structure. The second theory, that of mesenchymal self-organisation, proposes that mobile cells can form periodic patterns of cell aggregates directly, without reference to any prepattern. Early hair follicle development is characterised by the rapid appearance of periodic arrangements of altered gene expression in the epidermis and prominent clustering of the adjacent dermal mesenchymal cells. We assess the contributions and interplay between reaction–diffusion and mesenchymal self-organisation processes in hair follicle patterning, identifying a network of fibroblast growth factor (FGF), wingless-related integration site (WNT), and bone morphogenetic protein (BMP) signalling interactions capable of spontaneously producing a periodic pattern. Using time-lapse imaging, we find that mesenchymal cell condensation at hair follicles is locally directed by an epidermal prepattern. However, imposing this prepattern’s condition of high FGF and low BMP activity across the entire skin reveals a latent dermal capacity to undergo spatially patterned self-organisation in the absence of epithelial direction. This mesenchymal self-organisation relies on restricted transforming growth factor (TGF) β signalling, which serves to drive chemotactic mesenchymal patterning when reaction–diffusion patterning is suppressed, but, in normal conditions, facilitates cell movement to locally prepatterned sources of FGF. This work illustrates a hierarchy of periodic patterning modes operating in organogenesis

Use of Accelerometers in a Large Field-Based Study of Children: Protocols, Design Issues, and Effects on Precision

Background: Objective methods can improve accuracy of physical activity measurement in field studies but uncertainties remain about their use. Methods: Children age 11 years from the Avon Longitudinal Study of Parents and Children (ALSPAC), were asked to wear a uni-axial accelerometer (MTI Actigraph) for 7 days. Results: Of 7159 children who attended for assessment, 5595 (78%) provided valid measures. The reliability coefficient for 3 days of recording was .7 and the power to detect a difference of 0.07 SDs (P ≤ .05) was \u3e 90%. Measures tended to be higher on the first day of recording (17 counts/min; 95% CI, 10-24) and if children wore the monitor for fewer days, but these differences were small. The children who provided valid measures of activity were different from those who did not, but the differences were modest. Conclusion: Objective measures of physical activity can be incorporated into large longitudinal studies of children

A Genetic Porcine Model of Cancer

The large size of the pig and its similarity in anatomy, physiology, metabolism, and genetics to humans make it an ideal platform to develop a genetically defined, large animal model of cancer. To this end, we created a transgenic oncopig line encoding Cre recombinase inducible porcine transgenes encoding KRASG12D and TP53R167H, which represent a commonly mutated oncogene and tumor suppressor in human cancers, respectively. Treatment of cells derived from these oncopigs with the adenovirus encoding Cre (AdCre) led to KRASG12D and TP53R167H expression, which rendered the cells transformed in culture and tumorigenic when engrafted into immunocompromised mice. Finally, injection of AdCre directly into these oncopigs led to the rapid and reproducible tumor development of mesenchymal origin. Transgenic animals receiving AdGFP (green fluorescent protein) did not have any tumor mass formation or altered histopathology. This oncopig line could thus serve as a genetically malleable model for potentially a wide spectrum of cancers, while controlling for temporal or spatial genesis, which should prove invaluable to studies previously hampered by the lack of a large animal model of cancer