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This study is concerned with the dynamics of a polydisperse ensemble of crystals in a single-component metastable solution/melt. A new theory based on the kinetic and balance equations is developed for the description of initial and intermediate stages of bulk crystallization. Such phenomena as unsteady growth rates of individual crystals with fluctuations, diffusion of the crystal-size distribution function in the space of particle radii, the Gibbs–Thomson and atomic kinetics effects, various crystal nucleation mechanisms are taken into account. The analytical solution is constructed in a parametric form with the modified time being the decision variable. Namely, the metastability degree, particle-radius distribution function, crystallization time, total number of crystals and their mean size are found as the functions of decision variable. The analytical solutions show that the metastability degree decreases with time as a result of liquid desupersaturation/desupercooling. As this takes place, the particle-radius distribution function moves to greater particle radii, becomes wider and lower with increasing the crystallization time. The theory is tested against experiments on the growth of such polypeptide hormones as porcine and bovine insulins. We show that the theory is in good agreement with the experimental data. [ABSTRACT FROM AUTHOR]

In pigs, stress factors such as pathogenic infections, environmental conditions, and various managemental practices can lead to a disease condition known as Porcine respiratory disease complex (PRDC). The main viral agents associated with this condition are Porcine circovirus 2 (PCV2), Porcine reproductive and respiratory syndrome virus (PRRSV), swine influenza virus (SIV), porcine respiratory coronavirus (PRCV), and pseudorabies virus. A study was undertaken to assess whether PCV2 and PRRSV, alone or in combination, are present in cases of respiratory ailments of pigs in North Kerala. The presence of PCV2 was detected by polymerase chain reaction (PCR) targeting the open reading frame 2 (ORF2) of the virus and PRRSV was detected by reverse transcriptase PCR (RT-PCR) targeting ORF6. A total of 54 tissue and lung samples were collected and screened for the presence of PCV2 and PRRSV. Of the samples tested, PCV2 could be detected in 22 (40.74 %) and PRRSV could be detected in 13 (24.07 %) samples. Two tissue samples showed mixed infections with PCV2 and PRRSV. The result shows that viral agents associated with PRDC are prevalent in pigs in North Kerala and that coinfections are also present. [ABSTRACT FROM AUTHOR]

The article discusses the potential of turning fat into muscle as an alternative to anti-obesity drugs like semaglutide. It highlights the effectiveness of GLP-1 receptor agonists in reducing body weight and introduces the concept of repartitioning drugs that stimulate muscle hypertrophy and reduce fat mass. The text suggests that interventions promoting global muscle hypertrophy may have anti-obesity and anti-diabetic effects, emphasizing the need for further research in this area to develop effective treatments for obesity and metabolic diseases. [Extracted from the article]

This study was aimed to assess the therapeutic potential (expressed by improvement of pelvic extremities motor functions) of a 5-hour local application of hypothermia with cold saline (4 °C), or saline at room temperature (≈ 24 °C) conveyed via perfusion chamber placed epidurally over the epicenter of spinal cord lesion in minipigs paraplegic due to acute spinal cord injuries (SCIs) inflicted through L3 laminectomy with the force of 8N, 15N, or 18N by a computer operated contusion apparatus. Eighteen 5–8-month-old minipigs (Göttingen-Minnesota-Liběchov crossbreed strains weighing 28–35 kg) were randomly divided into 6 subgroups (each containing three animals) another 3 minipigs were added as sham controls. To evaluate the pelvic extremities motor recovery was used the porcine 20-point neurological scale. Regular evaluations of motor scores showed gradual spontaneous recovery of this parameter in all experimental animals, however, the best results achieved minipigs after SCI inflicted by 8N impacts. The data achieved in the study suggest that local application of therapeutic hypothermia (TH) is well tolerated and may improve functional outcomes after SCI. Further experimental and preclinical studies in different SCI animal models are required before the introduction of the method in healthcare practice. [ABSTRACT FROM AUTHOR]

An experiment was conducted to assess the effects of porcine somatotropin (pST) on the responses to a near-ideal blend of AA on the AA composition of empty, whole-empty body (WEB) protein and WEB essential AA accretion rate in pigs from 22 to 60 kg BW. Forty Hampshire × Yorkshire gilts were individually penned and assigned to a 4 × 2 factorial arrangement of treatments consisting of four diets with and without pST injection. A fortified corn–soybean meal basal diet was formulated to contain 1.50% total Lys with Thr, Met, and Trp added to obtain a near-ideal blend of these AA relative to Lys. In three additional diets, Lys was reduced to 1.25%, 1.00%, and 0.75% by diluting the basal diet with cornstarch, cellulose, and sand such that the diets also contained the same ratios of AA. Pigs that received pST were administered a daily i.m. injection of 2 mg of pST. At 60 kg BW, the WEB (carcass, head, viscera, blood, nails, and hair) was ground and analyzed for proximate and AA composition. Administration of pST increased (P  < 0.001) accretion rates of WEB protein and essential AA. Increasing dietary essential AA increased (quadratic, P  < 0.03) accretion rate of WEB protein, His, Leu, Trp, and Val in pST-treated pigs, but not in untreated pigs. Lysine composition in the accreted WEB protein was not affected (P  > 0.05) by dietary Lys. The efficiency of Lys utilization for WEB Lys accretion was linearly affected (P  < 0.01) by dietary Lys. These results indicated that the dietary Lys needed to achieve maximum WEB Lys accretion is markedly increased by pST administration. [ABSTRACT FROM AUTHOR]

Since the emergence of 3D bioprinting technology, both synthetic and natural materials have been used to develop bioinks for producing cell-laden cardiac grafts. To this end, extracellular-matrix (ECM)-derived hydrogels can be used to develop scaffolds that closely mimic the complex 3D environments for cell culture. This study presents a novel cardiac bioink based on hydrogels exclusively derived from decellularized porcine myocardium loaded with human-bone-marrow-derived mesenchymal stromal cells. Hence, the hydrogel can be used to develop cell-laden cardiac patches without the need to add other biomaterials or use additional crosslinkers. The scaffold ultrastructure and mechanical properties of the bioink were characterized to optimize its production, specifically focusing on the matrix enzymatic digestion time. The cells were cultured in 3D within the developed hydrogels to assess their response. The results indicate that the hydrogels fostered inter-cell and cell-matrix crosstalk after 1 week of culture. In conclusion, the bioink developed and presented in this study holds great potential for developing cell-laden customized patches for cardiac repair. [ABSTRACT FROM AUTHOR]

Introduction The aim of this study was to evaluate outflow variation in different locations of the pyelocaliceal system with the use of different ureteral access sheath (UAS) sizes and different UAS positioning. Material and methods The experimental setup included an anaesthetised porcine model, a 7.5-Fr ureteroscope with a 200-µm laser fibre inserted in the working channel, a hand-held pumping irrigating system, and UAS of different sizes, namely: 9.5/11.5 Fr, 12/14 Fr, and 14/16 Fr. Each UAS was placed just below the ureteropelvic junction (UPJ) or in the mid-ureter. The ureteroscope was placed in the renal pelvis, upper and lower calyces, and outflow measurements were obtained with 3-second interval pumping for one minute in every experimental setup. Results The UAS positioning in the mid-ureter was associated with significantly higher outflow rates in the lower calyx (p = 0.041). While the UAS was below the UPJ, we observed a trend of lower outflow rate in the lower calyx, which was completely inverted when the UAS was in the mid-ureter. Increasing the UAS size from 9.5/11.5 Fr to 12/14 Fr led to a significant increase in outflow in the renal pelvis and upper calyx (p = 0.007), but not in the lower calyx. A further increase to 14/16 Fr did not produce increased flow. Conclusions Different locations of the pyelocaliceal system have different fluid mechanics during fURS. In the renal pelvis and upper calyx increasing the diameter of the UAS improved the outflow, whereas in the lower calyx the position of the UAS seems to be the most relevant factor. These variables should be considered when performing fURS, especially with high-power laser lithotripsy. [ABSTRACT FROM AUTHOR]

The article focuses on the use of a synthetic derivative of vitamin E, Trolox, as an antioxidant to improve the sperm functionality of refrigerated porcine semen. It discusses the challenges of preserving porcine sperm due to its sensitivity to cold storage and the impact of oxidative stress on sperm quality. It aims to find strategies to optimize sperm preservation and, consequently, enhance the reproductive efficiency of porcine production.

Introduction Segmental bone defects (SBDs) are devastating injuries sustained by warfighters and are difficult to heal. Preclinical models that accurately simulate human conditions are necessary to investigate therapies to treat SBDs. We have developed two novel porcine SBD models that take advantage of similarities in bone healing and immunologic response to injury between pigs and humans. The purpose of this study was to investigate the efficacy of Bone Morphogenetic Protein-2 (BMP-2) to heal a critical sized defect (CSD) in two novel porcine SBD models. Materials and Methods Two CSDs were performed in Yucatan Minipigs including a 25.0-mm SBD treated with intramedullary nailing (IMN) and a 40.0-mm SBD treated with dual plating (ORIF). In control animals, the defect was filled with a custom spacer and a bovine collagen sponge impregnated with saline (IMN25 Cont, n  = 8; ORIF40 Cont, n  = 4). In experimental animals, the SBD was filled with a custom spacer and a bovine collage sponge impregnated with human recombinant BMP-2 (IMN25 BMP, n  = 8; ORIF40 BMP, n  = 4). Healing was quantified using monthly modified Radiographic Union Score for Tibia Fractures (mRUST) scores, postmortem CT scanning, and torsion testing. Results BMP-2 restored bone healing in all eight IMN25 BMP specimens and three of four ORIF40 BMP specimens. None of the IMN25 Cont or ORIF40 Cont specimens healed. mRUST scores at the time of sacrifice increased from 9.2 (±2.4) in IMN25 Cont to 15.1 (±1.0) in IMN25 BMP specimens (P  < .0001). mRUST scores increased from 8.2 (±1.1) in ORIF40 Cont to 14.3 (±1.0) in ORIF40 BMP specimens (P  < .01). CT scans confirmed all BMP-2 specimens had healed and none of the control specimens had healed in both IMN and ORIF groups. BMP-2 restored 114% and 93% of intact torsional stiffness in IMN25 BMP and ORIF40 BMP specimens. Conclusions We have developed two porcine CSD models, including fixation with IMN and with dual-plate fixation. Porcine models are particularly relevant for SBD research as the porcine immunologic response to injury closely mimics the human response. BMP-2 restored healing in both CSD models, and the effects were evident within the first month after injury. These findings support the use of both porcine CSD models to investigate new therapies to heal SBDs. [ABSTRACT FROM AUTHOR]

The growth and development of pigs is determined by their genotype and environmental conditions (primarily the level of feeding), however, the number of works aimed at studying the complex influence of genetic and non-genetic factors in their interaction is currently insufficient. The purpose of our work was to estimate the effect of the MC4R genotype, feeding level and interaction of these factors on growth and backfat thickness of crossbred pigs and to investigate the possibility of correcting the melanocortin-4-receptor gene polymorphism effect by adjusting the ration. Studies were conducted on 50 gilts obtained by crossing sows of the large white breed with landrace boars. Experimental pigs at the "Maxi 2010" farm were weighed at birth, then at the age of 28 days (at weaning) and at the age of 4, 6, 8 months. Fat thickness was measured at the age of 4, 6, 8months. Genetic studies were conducted in a certified laboratory of Institute of Pig Breeding and Agroindustrial Production. Analysis of 50 blood samples revealed that this group of pigs had a sufficient level of polymorphism for research (Polymorphism Information Content was equil 0.35). The frequency of genotype distribution at the MC4R / SNP c.1426 G>A locuswas 0.06 (AA) : 0.58 (GA) : 0.36 (GG). The type of feeding significantly influenced the live weight at the age of 4 months and the average daily gains of experimental pigs over the period of 28–120 days. Starting at the age of 6 months a significant effect of the interaction of organized factors (feeding + genotype) was recorded. At the age of 6 months, a significant influence of both the genotype and the level of feeding on the backfat thickness was established. Animals with the GG genotype receiving a restricted feed ration had significantly lower backfat thickness. At the age of 8 months, the difference in backfat thickness between the group with the GG genotype (restricted feed ration) and the AG genotype (high level feeding) reached a value of 12.9% (2.0 mm). Animals with the AG genotype had the lowest performance and the greatest fat thickness under feed limitation, which is important for raising young pigs for subsequent reproduction. Therefore, when selecting pigs to be used for further reproduction, the desired genotype is GG. In the future, it will be desirable to repeat the study on a larger number of pigs, so that the experiment involves a sufficient number of animals with the MC4R AA genotype for statistical processing. [ABSTRACT FROM AUTHOR]

Background: The vagus nerve affects innate immune responses by activating spleen-projecting sympathetic neurons, which modulate leukocyte function. Recent basic and clinical research investigating vagus nerve stimulation to engage the cholinergic anti-inflammatory pathway (CAP) has shown promising therapeutic results for a variety of inflammatory diseases. Abundant sympathetic innervation occurs in rodent spleens, and use of these species has dominated mechanistic research investigating the CAP. However, previous neuroanatomical studies of human spleen found a more restricted pattern of innervation compared to rodents. Therefore, our primary goal was to establish the full extent of sympathetic innervation of human spleens using donor tissue with the shortest procurement to fixation time. Parallel studies of porcine spleen, a large animal model, were performed as a positive control and for comparison. Methods: Human and porcine spleen tissue were fixed immediately after harvest and prepared for immunohistochemistry. Human heart and porcine spleen were stained in conjunction as positive controls. Several immunohistochemical protocols were compared for best results. Tissue was stained for tyrosine hydroxylase (TH), a noradrenergic marker, using VIP purple chromogen. Consecutive tissue slices were stained for neuropeptide Y (NPY), which often co-localizes with TH, or double-labelled for TH and CD3, a T cell marker. High-magnification images and full scans of the tissue were obtained and analyzed for qualitative differences between species. Results: TH had dominant perivascular localization in human spleen, with negligible innervation of parenchyma, but such nerves were abundant throughout ventricular myocardium. In marked contrast, noradrenergic innervation was abundant in all regions of porcine spleen, with red pulp having more nerves than white pulp. NPY stain results were consistent with this pattern. In human spleen, noradrenergic nerves only ran close to T cells at the boundary of the periarterial lymphatic sheath and arteries. In porcine spleen, noradrenergic nerves were closely associated with T cells in both white and red pulp as well as other leukocytes in red pulp. Conclusion: Sympathetic innervation of the spleen varies between species in both distribution and abundance, with humans and pigs being at opposite extremes. This has important implications for sympathetic regulation of neuroimmune interactions in the spleen of different species and focused targeting of the CAP in humans. [ABSTRACT FROM AUTHOR]

Nitrogen (N) is an essential element for life, but due to its inefficient utilization, N waste and emissions generated have exceeded the planetary boundaries and ability of the Earth to effectively overcome these environmentally detrimental effects. Global food animal production contributes 29 to 34% of total global N emissions, with pork supply chains contributing about 16% of these emissions. Life cycle N use efficiency (NUE) of U.S. pork production is only about 56%, which is similar to broiler (55%) and egg (50%) production, but greater than beef (10%) and milk (31%) production. However, only about 10 to 44% of dietary N is converted into edible pork products. While genetic improvement has increased NUE associated with improved lean growth rate, improvement in litter size has led to an increased proportion of low-birth-weight pigs that have reduced NUE. Although metabolic modifiers including porcine somatotropin, ractopamine, and immunocastration improve NUE, they are not feasible or acceptable to consumers in many countries. Separate sex feeding and reducing market weight can also improve NUE. Ultimately feeding program design and implementation have the greatest potential to improve NUE because feed is associated with 70% of N emissions in pork production systems. Multi-objective feed formulation using Life Cycle Assessment environmental impact data of feed ingredients, precision feed formulation and feeding practices to overcome variability in digestible nutrient content of feed ingredients and nutritional requirements of pigs fed in groups, feed processing to enhance energy and nutrient digestibility, and minimizing feed wastage offer the greatest opportunity to improve NUE and environmental sustainability. There are substantial differences in greenhouse gas emissions and embedded water and land use associated with growing-finishing feeding programs depending on the dietary proportions of corn, soybean meal, distillers dried grains with solubles (DDGS), and crystalline amino acids among U.S. geographic regions. Furthermore, addition of thermally processed supermarket food waste to corn-soybean meal (CSBM) diets can significantly reduce environmental impacts and diet cost without compromising growth performance and carcass composition compared with current commercial feeding programs. Although growing-finishing feeding programs using CSBM diets with minimal crystalline amino acids result in greater N intake, N retained, and urinary N excretion compared with feeding low protein, crystalline amino acid supplemented (LP) diets, and diets containing 30% DDGS, growth performance and carcass composition are optimized and impacts on climate change, marine and freshwater eutrophication, and fossil resource are reduced by feeding CSBM diets compared with feeding LP or DDGS diets. Sourcing and using feed ingredients with reduced environmental impact metrics along with precision diet formulation and feeding practices to ensure “getting the right amount of energy and nutrients in the right feed fed to the right pigs at the right time” can improve NUE and environmental sustainability of pork production systems. [ABSTRACT FROM AUTHOR]

Gonocytes in the neonatal testis have male germline stem cell properties and as such have important potential applications in fertility preservation and regenerative medicine. Such applications require further studies aimed at increasing gonocyte numbers and evaluating their pluripotency in vitro. The objective of the present study was to test the effects of basic fibroblast growth factor (bFGF), glial cell line-derived neurotrophic factor (GDNF), and leukemia inhibitory factor (LIF) on in vitro propagation, colony formation, and expression of pluripotency markers of neonatal porcine gonocytes. Testis cells from 1-week-old piglets were cultured in basic media (DMEM + 15% FBS), supplemented with various concentrations of bFGF, GDNF, and LIF, either individually or in combinations, in a stepwise experimental design. Gonocytes and/or their colonies were evaluated every 7 days and the gonocyte- (DBA) and pluripotency-specific markers (POU5F1, SSEA-1, E-cadherin, and NANOG) assessed on day 28. Greatest gonocyte numbers and largest colonies were found in media supplemented with 10 ng/mL bFGF and 10 ng/mL bFGF + 100 ng/mL GDNF + 1500 U/mL LIF, respectively. The resultant gonocytes and colonies expressed both germ cell- and pluripotency-specific markers. These results shed light on the growth hormone requirements of porcine gonocytes for in vitro proliferation and colony formation. [ABSTRACT FROM AUTHOR]

Background: Juvenile panhypopituitarism is an endocrinopathy that affects young dogs and must be differentiated from other causes of height disorders, as one could have a deficiency of one or more adenohypophysis hormones, besides growth hormone. Diagnosis consists of excluding endocrine and non-endocrine causes. Treatment requires administration of heterologous porcine growth hormone or progestins, as well as supplementing other hormones that are deficient in the circulation. The prognosis for these patients tends to be unfavorable and they have a shorter life expectancy. The present case aimed to report the therapeutic challenges in a dog diagnosed with juvenile panhypopituitarism. Case: This study presents the case of a 6-month-old crossbred Labrador/Brazilian Fila dog, not neutered, 6.7 kg of body weight (body condition score 5/9), with a growth disorder and persistence of the puppy coat, when compared to other members of the same litter. During inspection, it was possible to observe an undersized dog and soft puppy coat, besides no bone irregularities or joint tenderness was noted. The other physical examination parameters were within the normal range for the species. No changes in complete blood count and only increases in urea, cholesterol and alkaline phosphatase activity were observed. Thyroid and abdominal ultrasound (US) examination did not reveal any remarkable changes. After serum dosage of insulin-like growth factor, thyroxine, thyrostimulating hormone, and cortisol, the patient was diagnosed with juvenile panhypopituitarism and underwent therapy with medroxyprogesterone and thyroid hormone supplementation. Monitoring was instituted at intervals of 3, 6, and 12 weeks, and currently every 3 or 4 months and the IGF-1 values normalized after 6 months with. After 8 months of therapy, the patient had good body growth and bone mineralization compared to the time of diagnosis. However, skeletal development was completed only 12 months after hormone replacement, accompanied by the presence of vertebral osteophytes and coxofemoral osteoarticular alterations. Considering chronic use of progestins, ovariosalpingohysterectomy (OSH) was recommended, but for personal reasons, the owners chose to do not submit her to surgery. After 18 months of treatment, the dog starts to exhibit prostration, selective appetite, and increased abdominal volume. After imaging exam, she was forwarded for OSH, due to consistent findings of pyometra. Subsequently, even the supervision of possible comorbidities involved in the chronic use of progestins was maintained at half-yearly intervals, the bitch returned to the service with skin thickening, increased limb volume, and macroglossia after 60 months of therapy. At this point IGF-1 values were higher when compared to the previous measurements and the application of medroxyprogesterone was suspended. Its supplementation was reintroduced only after 8 months when IGF-1 was significantly reduced. To date, the patient is close to 6 years of age and with a good quality of life. Discussion: The greatest therapeutic challenge for these patients involves dealing with the adverse effects of progestins, mainly related to reproductive disorders and alopecia at the application site, as well as maintaining adequate hormone replacement in order to avoid hypersomatotropism. Although longevity is lower in these individuals, the patient in this report has achieved 6 years of age and has had an excellent quality of life so far. [ABSTRACT FROM AUTHOR]

Quantitative estimation of soft tissue injuries due to penetration of sharp objects is a challenging task for forensic pathologists. The severity of injury depends on the force required to penetrate the tissue. This study focuses on investigating the amount of force required to penetrate porcine muscle tissue and gelatin simulants (10 % wt) to mimic human muscle tissue when subjected to sharp objects like nail at velocities below 5 m/s. A custom-made experimental setup was used to examine the influence of penetration velocity and nail diameter on penetration forces. Images captured by a high-speed camera were used to track the position and velocity of the nail. A finite element (FE) model was established to simulate the penetration behavior of the tissue and gelatin. The FE simulations of the nail penetration were validated through direct comparison with the experimental results. In tissues as well as in the simulant, penetration forces were seen to increase with increasing nail diameter and velocity. Porcine muscle tissue showed 23.9–46.5 % higher penetration forces than gelatin simulants (10 % wt) depending on nail diameter and velocity; the difference being higher for higher nail diameter and velocity. The ranges of maximum penetration forces measured were 8.6–59.1 N for porcine muscle tissue and 6.8–34.9 N for gelatin simulant. This study helps to quantify injuries caused by sharp nails at low velocities and offers insights with potential applications in injury management strategies and forensic studies. [Display omitted] • Injuries caused by a sharp object in soft tissues are a significant concern. • Developed a method to measure penetration forces in soft tissue by sharp nails. • Simulation of nail penetration into soft tissue is validated with experiments. • Penetration forces increase with both the nail size and penetration velocity. • Porcine muscle tissue showed higher penetration forces than the gelatin simulant. [ABSTRACT FROM AUTHOR]

Introduction: Retrobulbar hemorrhage (RBH) occurs in only 0.45% of ocular trauma, but failure to provide timely lateral canthotomy and cantholysis (LCC) risks permanent visual deficits. With ocular trauma rates as high as 8.5-10% amongst modern combat injuries, and more than 2,000 severe eye injuries documented over a 10 year span, this concern increases.12-15 However, given infrequent RBH occurrence in the non-combat environment, emergency medicine residents trained in stateside settings may not receive adequate LCC exposure prior to military deployment. Simulators should be evaluated for procedural confidence compared to expensive and cumbersome traditional live tissue training (LTT) options. We seek to compare procedural confidence and usability of emergency medicine military residents performing LCC on a novel simulator to those using LTT. Methods: This study randomized 32 emergency physician and physician assistant residents to perform LCC on a simulator or LTT model. All received a standardized brief on RBH recognition and LCC, then completed an 11-question survey using a 100-mm visual numerical rating scale about their ability to correctly identify RBH and perform LCC. The survey was repeated after LCC completion. All volunteers additionally completed a 10-question survey utilizing a 5-point Likert scale on the usability of the model to which they were randomized. Results: No significant difference in reported confidence changes between groups was found; however, significant increases were found across all reported confidence measures between pre- and post-trainer use in the overall sample population. LCC simulator users reported significantly higher usability in 7 of 10 ratings. Conclusion: The lack of a statistically significant difference between groups in procedural confidence suggests artificial LCC simulators may offer an attractive alternative to logistically-complicated porcine models. Further research is needed to evaluate non-inferiority and procedural performance. [ABSTRACT FROM AUTHOR]

The four genes coding for placental members of the human (h) growth hormone (GH) family include two that code independently for placental lactogen (PL), also known as chorionic somatomammotrophin hormone, one that codes for placental growth hormone (PGH) and a pseudogene for which RNA but no protein product is reported. These genes are expressed preferentially in the villus syncytiotrophoblast of the placenta in pregnancy. In higher primates, the placental members, including hPL and PGH, are the result of multiple duplication events of the GH gene. This contrasts with rodents and ruminants, where PLs result from duplication of the prolactin (PRL) gene. Thus, unlike their mouse counterparts, the hPL and PGH hormones bind both lactogenic and somatogenic receptors with varying affinity. Roles influenced by nutrient availability in both metabolic control in pregnancy and maternal behaviour are supported. However, the effect maternal obesity has on the activation of placental members of the hGH gene family, particularly the expression and function of those genes, is poorly understood. Evidence from partially humanised hGH/PL transgenic mice indicates that both the remote upstream hPL locus control region (LCR) and more gene‐related regulatory regions are required for placental expression in vivo. Furthermore, a specific pattern of interactions between the LCR and hPL gene promoter regions is detected in term placenta chromatin from women with a normal body mass index (BMI) in the range 18.5–25 kg m‐2 by chromosome conformation capture assay. This pattern is disrupted with maternal obesity (class II BMI > 35 kg m‐2) and associated with a > 40% decrease in term hPL RNA levels, as well as serum hPL but not PRL levels, during pregnancy. The relative importance of the chromosomal architecture and predicted properties for transcription factor participation in terms of hPL production and response to obesity are considered, based on comparison with components required for efficient human pituitary GH gene expression. [ABSTRACT FROM AUTHOR]

Porcine growth hormone (pGH) is most important hormone which is involved in the growth and development of pig. However, a series of studies have indicated that neonatal pig is insensitive to pGH; the reason for this phenomenon is still not fully understood. In this work, we try to investigate this issue from the angle of intracellular signaling induced by pGH. In the present study, porcine hepatocytes from neonatal pig were used as a model, and confocal laser scanning microscopy (CLSM), Western blot, co-immunoprecipitation and colocalization assay were used to study pGH's signaling properties in hepatocytes of neonatal pig and explore the possible mechanism(s) for why intracellular signaling is insensitive to pGH. The results indicated that Janus kinase 2 and signal transducers and activators of transcription 5/3/1 (JAK2-STATs) signaling are not activated. We further investigated the possible mechanism(s) by which JAK2-STATs' signaling is not activated by pGH and growth hormone receptor (GHR) and found that the negative regulatory molecules of JAK2-STATs signaling may be associated with this phenomenon in the hepatocytes of neonatal pig. In addition, we also explored pGH's biology in hepatocytes from neonatal pig, it can be found that pGH/GHR could translocate into the cell nucleus, which implies that pGH/GHR may exhibit physiological roles based on their nuclear localization. We found that pGH could not trigger intracellular signaling in the hepatocytes of neonatal pigs, but not young pigs, which provides an important explanation for why the growth of neonatal pig is GH independent. [ABSTRACT FROM AUTHOR]

Porcine growth hormone (pGH) is a class of peptide hormones secreted from the pituitary gland, which can significantly improve growth and feed utilization of pigs. However, it is unstable and volatile in vitro. It needs to be encapsulated in liposomes when feeding livestock, whose high cost greatly limits its application in pig industry. Therefore we attempted to express pGH as intracellular soluble protein in Pichia pastoris and feed these yeasts with partial wall-breaking for swine, which could release directly pGH in intestine tract in case of being degraded in intestinal tract with low cost. In order to improve the intracellular soluble expression of pGH protein in Pichia pastoris and stability in vitro, we optimized the pGH gene, and screened molecular chaperones from E. coli and Pichia pastoris respectively for co-expressing with pGH. In addition, we had also explored conditions of mechanical crushing and fermentation. The results showed that the expression of intracellular soluble pGH protein was significantly increased after gene optimized and co-expressed with Ssa1–Sis1 chaperone from Pichia pastoris. Meanwhile, the optimal conditions of partial wall-breaking and fermentation of Pichia pastoris were confirmed, the data showed that the intracellular expression of the optimized pGH protein co-expressed with Ssa1–Sis1 could reach 340 mg/L with optimal conditions of partial wall-breaking and fermentation. Animal experiments verified that the optimized pGH protein co-expression with Ssa1–Sis1 had the best promoting effects on the growth of piglets. Our study demonstrated that Ssa1–Sis1 could enhance the intracellular soluble expression of pGH protein in Pichia pastoris and that partial wall-breaking of yeast could prevent pGH from degradation in vitro, release targetedly in the intestine and play its biological function effectively. Our study could provide a new idea to cut the cost effectively, establishing a theoretical basis for the clinic application of unstable substances in vitro. [ABSTRACT FROM AUTHOR]

It is well known that zinc ion (Zn2+) can regulate the biological activity of growth hormone (GH). However, until now, the mechanism by which Zn2+ regulates GH biological activity remains unclear. In the current study, we first performed molecular docking between Zn2+ and porcine GH (pGH) using computational biology. We then explored the effect of Zn2+ on the GH signaling ability in the cell model expressing porcine growth hormone receptor (GHR). It was found that the phosphorylation levels of Janus kinase 2, signal transducers and activators of transcription 5/3/1, and GHR increased significantly under Zn2+ treatment, indicating that Zn2+ can enhance the signaling ability of GH/GHR. On this basis, we further explored how Zn2+ regulates the biological activity of GH/GHR. The results showed that downregulation and turnover of GHR changed under Zn2+/pGH treatment. Zn2+ enhanced the membrane residence time of pGH/GHR and delayed GHR downregulation. Further investigation showed that the internalization dynamic of pGH/GHR was changed by Zn2+, which prolonged the residence time of pGH/GHR in the cell membrane. These factors acted together to upregulate the signaling of GH/GHR. This study lays a foundation for further exploration of the biological effects of Zn2+ on GH. [ABSTRACT FROM AUTHOR]

Present study explored the quality changes in meat emulsion during storage at refrigerated temperature prepared with inclusion of three different levels of liver protein hydrolysate (LPH) in meat emulsion (LPH‐1: 0.03, LPH‐2: 0.06, and LPH‐3: 0.09%) and compared with control (LPH: 0.00%) and positive control (butylated hydroxytoluene: 0.02% w/w meat emulsion). Physico‐chemicals, antioxidant activities, lipid oxidation, color profile, microbial quality, and microbial challenge test (MCT) were assessed for all groups. Results indicated that all evaluated attributes were considerably improved with increase in LPH concentration. Among treated groups, LPH‐3 was maintained comparatively better for every attribute assessed during storage studying. Regarding microbial quality, LPH‐3 showed (P ≤ 0.05) lower aerobic plate count, coliforms, and yeast mold counts than others. Likewise for MCT, significantly (P ≤ 0.05) lower microbial counts were recorded in LPH‐3 during storage. Results concluded that LPH can be a good alternative substance for the preservation of meat for lower oxidation activity and bacterial growth. Practical Application: Meat emulsion is more prone to lipid oxidation and microbial contamination than fresh meat. However, for the preparation of convenient, value added meat products, better utilization of freezed meat and reduced cooking time have enhanced the demand of emulsion‐based meat products. Liver protein hydrolysate can be a good alternative substance for the preservation of emulsion‐based meat products. Treated groups better retained their physico‐chemical properties, color indices, and showed lower oxidation and microbial counts than control. It can be exploited commercially for the preparation of functional foods, nutraceuticals, and cosmeceuticals. [ABSTRACT FROM AUTHOR]

The goal of this project was to determine if increasing insulin-like growth factor-1 (IGF-1) concentrations in late pregnancy can stimulate mammogenesis in gilts. Yorkshire × Landrace gilts of a similar body weight (BW ; 196.2 ± 6.2 kg) on day 89 of gestation were separated in 2 groups, namely, controls (CTL, n = 17) that were injected with sterile water, and porcine somatotropin-treated (pST, n = 20) that received injections of 5 mg of pST (Reporcin). Injections were given daily from days 90 to 109 of gestation and gilts were slaughtered on day 110 to collect mammary glands for compositional analyses. Blood samples were obtained on days 89, 96, 103, and 109 of gestation to measure IGF-1, free fatty acids (FFA), urea, glucose, and insulin concentrations. Treated gilts gained more BW (22.7 vs. 18.2 kg, P < 0.05) and lost more backfat (P < 0.05) than CTL gilts during the treatment period. There was a treatment × day effect (P < 0.01) on IGF-1, glucose, and urea concentrations. Concentrations of IGF-1 increased 4-fold (P < 0.01) in pST compared with CTL gilts on days 96, 103, and 109 of gestation. Insulin values were also greater on days 96 (P < 0.01) and 103 (P = 0.01), and tended to be greater (P < 0.10) on day 109 of gestation in pST gilts. Glucose was greater in pST than CTL gilts on days 96 (P < 0.01), 103 (P < 0.01), and 109 (P = 0.01). Concentrations of urea were lower (P < 0.01) on days 96, 103, and 109 of gestation in gilts receiving pST injections, and FFA was not altered by treatment on any sampling day (P > 0.10). Injections of pST did not affect mammary extraparenchymal tissue weight (P > 0.10) but increased mammary parenchymal mass (1922 vs. 1576 ± 124 g, P < 0.05). The composition of parenchymal tissue was also altered by treatment. Mammary parenchyma from pST gilts contained more (P < 0.05) protein, DNA and RNA and less fat (P < 0.05) and dry matter (P < 0.01) than that from CTL gilts. These findings provide a clear demonstration that increasing circulating IGF-1 in late-pregnant gilts can stimulate mammary development both in terms of total parenchymal mass and of parenchymal tissue composition. [ABSTRACT FROM AUTHOR]

Abstract The analysis of the uterine contraction have become a general practice in an effort to improve the clinical management of uterine contractions during pregnancy and labour in human beings. The fluctuations in uterine activity may occur without affecting progress of gestation, however the painful and fashion contractions may be the first threat of miscarriage. While pigs were considered as an referential preclinical model, the computational modelling of spontaneous myoelectrical activity of complex systems of porcine myometrium in peri-fertilization period has been proposed. The higher order statistic, multivariate cumulants and Joint Skewness Band Selection method, have been applied to study the dependence structure of electromyographic (EMG) signal with an effective EMG feature. Than the model of recognition of multivariate, myoelectricaly changes according to crucial stages for successful fertilization and early pregnancy maintenance has been estimated. We found that considering together time and frequency features of EMG signal was extremely non-Gaussian distributed and the higher order multivariate statistics such as cumulants, have to be used to determine the pattern of myoelectrical activity in reproductive tract. We confirmed the expectance that the probabilistic model changes on a daily base. We demonstrated the changes in proposed model at the crucial time points of in peri-fertilization period. We speculate the activity of the middle of uterine horn and the power (minimum and maximum) and pauses between myoelectrical burst features are essential for the functional role of uterine contractility in peri-fertilization period. [ABSTRACT FROM AUTHOR]

Primary porcine bronchial epithelial cells (PBECs) are an ideal model to study the molecular and pathogenic mechanisms of various porcine respiratory pathogens. However, the short lifespan of primary PBECs greatly limit their application. Here, we isolated and cultured primary PBECs and established immortalized PBECs by transfecting primary PBECs with the pEGFP‐hTERT recombinant plasmid containing human telomerase reverse transcriptase (hTERT). Immortalized PBECs (hTERT‐PBECs) retained the morphological and functional features of primary PBECs as indicated by cytokeratin 18 expression, telomerase activity assay, proliferation assays, karyotype analysis, and quantitative reverse‐transcriptase polymerase chain reaction. Compared to primary PBECs, hTERT‐PBECs had higher telomerase activity, extended replicative lifespan, and displayed enhanced proliferative activity. Moreover, this cell line is not transformed in vitro and does not exhibit a malignant phenotype in vivo, suggesting that it can be safely used in further studies. Besides, hTERT‐PBECs were susceptible to swine influenza virus of H3N2 subtype and porcine circovirus type 2. In conclusion, the immortalized hTERT‐PBECs represent a valuable in vitro model, which can be widely used in the study of porcine respiratory pathogenic infections. In summary, we provided detailed protocols for bronchus isolation from piglets and in vitro expansion of primary PBECs. Furthermore, we established the immortalized hTERT‐PBEC line, which retains the morphological and functional features of primary PBECs. The establishment of immortal hTERT‐PBECs is of great importance as an in vitro model for pathogenicity studies of porcine respiratory pathogens. [ABSTRACT FROM AUTHOR]

Porcine circovirus 2 (PCV2) is a circular single-stranded DNA virus responsible for a group of diseases collectively known as PCV2 Associated Diseases (PCVAD). Variation in the incidence and severity of PCVAD exists between pigs suggesting a host genetic component involved in pathogenesis. A large-scale genome-wide association study of experimentally infected pigs (n = 974), provided evidence of a host genetic role in PCV2 viremia, immune response and growth during challenge. Host genotype explained 64% of the phenotypic variation for overall viral load, with two major Quantitative Trait Loci (QTL) identified on chromosome 7 (SSC7) near the swine leukocyte antigen complex class II locus and on the proximal end of chromosome 12 (SSC12). The SNP having the strongest association, ALGA0110477 (SSC12), explained 9.3% of the genetic and 6.2% of the phenotypic variance for viral load. Dissection of the SSC12 QTL based on gene annotation, genomic and RNA-sequencing, suggested that a missense mutation in the SYNGR2 (SYNGR2 p.Arg63Cys) gene is potentially responsible for the variation in viremia. This polymorphism, located within a protein domain conserved across mammals, results in an amino acid variant SYNGR2 p.63Cys only observed in swine. PCV2 titer in PK15 cells decreased when the expression of SYNGR2 was silenced by specific-siRNA, indicating a role of SYNGR2 in viral replication. Additionally, a PK15 edited clone generated by CRISPR-Cas9, carrying a partial deletion of the second exon that harbors a key domain and the SYNGR2 p.Arg63Cys, was associated with a lower viral titer compared to wildtype PK15 cells (>24 hpi) and supernatant (>48hpi)(P < 0.05). Identification of a non-conservative substitution in this key domain of SYNGR2 suggests that the SYNGR2 p.Arg63Cys variant may underlie the observed genetic effect on viral load. [ABSTRACT FROM AUTHOR]

TP53 (which encodes p53) is one of the most frequently mutated genes in cancers. In this study, we generated TP53-mutant pigs by gene editing via electroporation of the Cas9 protein (GEEP), a process that involves introducing the Cas9 protein and single-guide RNA (sgRNA) targeting exon 3 and intron 4 of TP53 into in vitro-fertilized zygotes. Zygotes modified by the sgRNAs were transferred to recipients, two of which gave birth to a total of 11 piglets. Of those 11 piglets, 9 survived. Molecular genetic analysis confirmed that 6 of 9 live piglets carried mutations in TP53, including 2 piglets with no wild-type (WT) sequences and 4 genetically mosaic piglets with WT sequences. One mosaic piglet had 142 and 151 bp deletions caused by a combination of the two sgRNAs. These piglets were continually monitored for 16 months and three of the genome-edited pigs (50%) exhibited various tumor phenotypes that we presumed were caused by TP53 mutations. Two mutant pigs with no WT sequences developed mandibular osteosarcoma and nephroblastoma. The mosaic pig with a deletion between targeting sites of two sgRNAs exhibited malignant fibrous histiocytoma. Tumor phenotypes of TP53 mosaic mutant pigs have not been previously reported. Our results indicated that the mutations caused by gene editing successfully induced tumor phenotypes in both TP53 mosaic- and bi-allelic mutant pigs. [ABSTRACT FROM AUTHOR]

Background: Transmissible gastroenteritis virus (TGEV) infection can activate NF-κB pathway in porcine intestinal epithelial cells and result in severe inflammation. Non-coding RNAs (ncRNAs) are not translated into proteins and play an important role in many biological and pathological processes such as inflammation, viral infection, and mitochondrial damage. However, whether ncRNAs participate in TGEV-induced inflammation in porcine intestinal epithelial cells is largely unknown. Results: In this study, the next-generation sequencing (NGS) technology was used to analyze the profiles of mRNAs, miRNAs, and circRNAs in Mock- and TGEV-infected intestinal porcine epithelial cell-jejunum 2 (IPEC-J2) cell line. A total of 523 mRNAs, 65 microRNAs (miRNAs), and 123 circular RNAs (circRNAs) were differentially expressed. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed differentially expressed mRNAs were linked to inflammation-related pathways, including NF-κB, Toll-like receptor, NOD-like receptor, Jak-STAT, TNF, and RIG-I-like receptor pathways. The interactions among mRNA, miRNA, and circRNA were analyzed. The data showed that ssc_circ_009380 and miR-22 might have interaction relationship. Dual-luciferase reporter assay confirmed that miR-22 directly bound to ssc_circ_009380. We also observed that overexpression of miR-22 led to a reduction of p-IκB-α and accumulation of p65 in nucleus in TGEV-infected IPEC-J2 cells. In contrast, inhibition of miR-22 had the opposite effects. Moreover, silencing of ssc_circ_009380 inhibited accumulation of p65 in nucleus and phosphorylation of IκB-α. Conclusions: The data revealed that differentially expressed mRNAs and ncRNAs were primarily enriched in inflammation-related pathways and ssc_circ_009380 promoted activation of NF-κB pathway by binding miR-22 during TGEV-induced inflammation. [ABSTRACT FROM AUTHOR]

Highlights • Feed intake of the lactating sow is a critical issue determining lactation performance and sow longevity. • Spray-dried porcine plasma is well known to improve feed intake, growth rate, and feed efficiency in nursery pigs. • Spray-dried porcine plasma addition to lactation diets of older, mature sows tended to increase measures of sow lactation performance. • However, when evaluating the response across all parities, spray-dried porcine plasma addition to lactation diets did not affect sow or litter performance. Abstract A cooperative study, utilizing 404 primi- and multi-parous crossbred sows from five experiment stations (AL, KY, OK, VA, and Brazil), was conducted to determine the effects of dietary inclusion of 0.50% spray-dried porcine plasma (SDPP ; Appetein, APC, Inc., Ankeny, IA) on sow and litter performance. Crossbred sows were fed corn and soybean meal-based diets containing a minimum of 0.50% SID lysine during gestation. Sows were allotted at farrowing, based on body wt, to two dietary treatments. The control (C) diet was a fortified corn, soybean meal based diet. Spray-dried porcine plasma was added to the control diet at 0.50% at the expense of soybean meal to formulate the test diet. Both diets were formulated to 0.90% SID lysine and 3414 kcal/kg ME. A common trace mineral and vitamin premix was used at all stations with the exception of Brazil. Sows were allowed ad libitum access to feed. Sows were weighed within 24 h after farrowing, and all litters were adjusted to ≥10 pigs/litter by d 2 following farrowing. Average sow parity, number of pigs on d 2, and lactation length for the C and SDPP treatments were, respectively: 2.5 and 2.3; 11.0 and 10.7; and 23 and 23 d The ADFI during lactation for C and SDPP diets was similar (5.65 kg/d; P > 0.10). The number of pigs weaned (9.9 vs. 9.7), litter weaning weights (68.4 vs. 69.1 kg), litter weight gains (51.6 vs. 52.2), sow weight change during lactation (−6.0 vs. −8.3 kg), and wean-to-estrus interval (7.5 vs. 7.2 d) were not affected (P > 0.10) by dietary treatment. There were no station x treatment interactions (P > 0.10). After evaluating the entire data set, the data were sorted by parity (≤3 vs. ≥4) to evaluate the effects of SDPP in young vs. older sows. In older, mature sows, SDPP tended to increase (P ≤ 0.10) the no. of pigs weaned (9.6 vs. 10.3), percentage survival (86 vs. 91), litter wt at weaning (64.5 vs. 70.3 kg), and daily litter wt change (2.20 vs. 2.47 kg). Overall, these data suggest no benefit of inclusion of spray-dried porcine plasma in lactation diets of younger sows; however, SDPP increased litter performance in older, mature sows. [ABSTRACT FROM AUTHOR]

Few effective porcine models of myocardial infarction (MI) related to platelet thrombus formation are available. In this study, we established a novel porcine MI model and examined the effect of dual antiplatelet therapy (DAPT) with aspirin and prasugrel, a P2Y 12 antagonist, using this MI model. Thrombotic MI was photochemically induced using rose bengal. Male miniature pigs were divided into 3 treatment groups: Sham, MI, and DAPT. In the DAPT group, aspirin (10 mg/kg, p.o.) and prasugrel (1 mg/kg, p.o.) were administered 4 h before photo-irradiation. Platelet aggregation, MI volume, and cardiac function were evaluated 24 h after photo-irradiation. Inhibition of ADP-induced platelet aggregation in the DAPT group was about 45%, similar to the effects of DAPT in a clinical setting. No MI was observed in the Sham group, and MI volume was 12.9 ± 2.9% in the left ventricle ( P  = 0.0016) in the MI group. Additionally, an increase in end-systolic volume ( P  = 0.0006), and a decrease in stroke volume ( P  = 0.0001) and ejection fraction ( P  < 0.0001) were observed in the MI group compared to the Sham group without any changes in end-diastolic volume. DAPT significantly decreased MI volume ( P  = 0.0006) and ameliorated cardiac dysfunction compared to the MI group. In conclusion, a novel porcine model of thrombotic MI with cardiac dysfunction was established. In this model, DAPT decreased MI volume and ameliorated of cardiac dysfunction, suggesting that this porcine MI model could be useful for future research on MI and antithrombotic agents. [ABSTRACT FROM AUTHOR]

Abstract: In vitro expansion of undifferentiated porcine primed embryonic stem (ES) cells is facilitated by use of non‐cellular niches that mimic three‐dimensional (3D) microenvironments enclosing an inner cell mass of porcine blastocysts. Therefore, we investigated the integrin heterodimers on the surface of undifferentiated porcine primed ES cells for the purpose of developing a non‐cellular niche to support in vitro maintenance of the self‐renewal ability of porcine primed ES cells. Immunocytochemistry and a fluorescence immunoassay were performed to assess integrin α and β subunit levels, and attachment and antibody inhibition assays were used to evaluate the function of integrin heterodimers. The integrin α3, α5, α6, α9, αV, and β1 subunits, but not the α1, α2, α4, α7, and α8 subunits, were identified on the surface of undifferentiated porcine primed ES cells. Subsequently, significant increase of their adhesion to fibronectin, tenascin C, and vitronectin were observed and functional blocking of integrin heterodimer α5β1, α9β1, or αVβ1 showed significantly inhibited adhesion to fibronectin, tenascin C, or vitronectin. No integrin α6β1 heterodimer‐mediated adhesion to laminin was detected. These results demonstrate that active α5β1, α9β1, and αVβ1 integrin heterodimers are present on the surface of undifferentiated porcine primed ES cells, together with inactive integrin α3 (presumed) and α6 subunits. [ABSTRACT FROM AUTHOR]

Abstract: Discovery and development of enzymes for the synthesis of chiral amines have been a hot topic for basic and applied aspects of biocatalysts. Based on our X‐ray crystallographic analyses of porcine kidney D‐amino acid oxidase (pkDAO) and its variants, we rationally designed a new variant that catalyzed the oxidation of (S)‐4‐Cl‐benzhydrylamine (CBHA) from pkDAO and obtained it by functional high‐throughput screening with colorimetric assay. The variant I230A/R283G was constructed from the variant R283G which had completely lost the activity for D‐amino acids, further gaining new activity toward (S)‐chiral amines with the bulky substituents. The variant enzyme (I230A/R283G) was characterized to have a catalytic efficiency of 1.85 s−1 for (S)‐CBHA, while that for (R)‐1‐phenylethylamine was diminished 10‐fold as compared with the Y228L/R283G variant. The variant was efficiently used for the synthesis of (R)‐CBHA in 96 % ee from racemic CBHA by the deracemization reaction in the presence of reducing agent such as NaBH4 in water. Furthermore, X‐ray crystallographic analysis of the new variant complexed with (S)‐CBHA, together with modelling study clearly showed the basis of understanding the structure‐activity relationship of pkDAO. [ABSTRACT FROM AUTHOR]

Pigs are commonly used as an animal model to evaluate the toxic effects of exogenous compounds. Cytochrome P450 1A1 (CYP1A1) metabolizes numerous exogenous compounds and is abundantly expressed in the liver, kidneys, and intestines. The high amino acid similarity between human and porcine CYP1A1 indicates that they probably have the same metabolic characteristics. Therefore, understanding the regulatory mechanism of CYP1A1 expression in pigs is particularly important for predicting the toxicology and metabolic kinetics of exogenous chemicals. Currently, the transcriptional regulation of porcine CYP1A1 has rarely been studied, especially regarding basal transcription. In this study, we first confirmed that the key regulatory elements of porcine CYP1A1 basal transactivation are in the proximal promoter region using promoter truncation analysis via a dual luciferase assay in a porcine kidney cell line LLC-PK1. Two overlapping cis -elements, the xenobiotic response element (XRE) and GC box, in this proximal region potentially play key roles in the basal transactivation of porcine CYP1A1. Furthermore, using electrophoretic mobility shift assay and chromatin immunoprecipitation, the GC box binding protein Sp1 was confirmed to bind to the proximal promoter of porcine CYP1A1 , instead of AhR, the XRE binding protein. In LLC-PK1 cells, by knocking down either Sp1 or AhR, the expression of porcine CYP1A1 at the mRNA level and protein level was significantly downregulated, suggesting both proteins are important for porcine CYP1A1 expression. However, promoter activity analysis in LLC-PK1 cells treated with an AhR agonist and antagonist confirmed that AhR does not participate in the basal regulation of porcine CYP1A1 at the proximal promoter. In conclusion, our study revealed that the proximal promoter is the key regulatory region for porcine CYP1A1 basal expression. Although AhR plays an important role in the transactivation of porcine CYP1A1 expression, the key determinant transcription factor for its basal transactivation is Sp1 at the proximal promoter of porcine CYP1A1. [ABSTRACT FROM AUTHOR]

Inhibins regulate folliculogenesis, gametogenesis and hormone secretion via endocrine, paracrine and autocrine manners, and play roles in encouraging or suppressing proliferation of cells. In order to investigate the effects of inhibin A on proliferation and apoptosis of porcine granulosa cells (GCs), GCs were isolated from ovarian follicles (3–6 mm), treated with inhibin A at different concentrations. The cell viability, mitochondrial membrane potential (MMP), expression of proliferation-related genes and cell cycle were detected. Inhibin α subunit ( INHA ) gene was silenced to detect the effect of down-regulation of inhibin on expression of proliferation-related genes in GCs. The results showed that cell viability was associated with inhibin A concentration and significantly enhanced by inhibin A at high doses (50 and 100 ng/mL, P  < 0.05) compared to control group (0 ng/mL). Meanwhile, the MMP boosted after treated with 100 ng/mL inhibin A for 48 h. Expression of proliferating cell nuclear antigen ( PCNA ) (200 ng/mL) and CyclinB1 (100 and 200 ng/mL) was promoted while Caspase-3 (100 and 200 ng/mL) and BAX (200 ng/mL) was inhibited in dose dependent manner after the cells were incubated with inhibin A for 24 h ( P  < 0.05) compared to control group, thereby the transition from G1 phase to S phase was promoted and the number of S phase cells was increased. After silencing the INHA gene expression, expression of Caspase-3 was enhanced and CyclinB1 was inhibited ( P  < 0.05) compared to Ri-negative group. All the results pointed to the conclusion that inhibin A promotes the proliferation of GCs while inhibits apoptosis. [ABSTRACT FROM AUTHOR]

Abstract: The use of adhesives for fracture fixation can revolutionize the surgical procedures toward more personalized bone repairs. However, there are still no commercially available adhesive solutions mainly due to the lack of biocompatibility, poor adhesive strength, or inadequate fixation protocols. Here, a surgically realizable adhesive system capitalizing on visible light thiol–ene coupling chemistry is presented. The adhesives are carefully designed and formulated from a novel class of chemical constituents influenced by dental resin composites and self‐etch primers. Validation of the adhesive strength is conducted on wet bone substrates and accomplished via fiber‐reinforced adhesive patch (FRAP) methodology. The results unravel, for the first time, on the promise of a thiol–ene adhesive with an unprecedented shear bond strength of 9.0 MPa and that surpasses, by 55%, the commercially available acrylate dental adhesive system Clearfil SE Bond of 5.8 MPa. Preclinical validation of FRAPs on rat femur fracture models details good adhesion to the bone throughout the healing process, and are found biocompatible not giving rise to any inflammatory response. Remarkably, the FRAPs are found to withstand loads up to 70 N for 1000 cycles on porcine metacarpal fractures outperforming clinically used K‐wires and match metal plates and screw implants. [ABSTRACT FROM AUTHOR]

A thorough understanding of the well-functioning, native aortic root is pivotal in an era, where valve sparing surgical techniques are developed and used with increasing frequency. The objective of this study was to characterize the local structural stiffness of the native aortic root, to create a baseline for understanding how different surgical interventions affect the dynamics of the aortic root. In this acute porcine study (N = 10), two dedicated force transducers were implanted to quantify the forces acting on both the annular plane and on the sinotubular junction (STJ). To assess the changes in geometry, eleven sonomicrometry crystals were implanted within the aortic root. The combination of force and length measurements yields the radial structural stiffness for each segment of the aortic root. The least compliant segment at the annular plane was the right-left interleaflet triangle with a stiffness modulus of 1.1 N mm −1 (SD0.4). At the sinotubular junction the same segment (right-left) was most compliant, compared with the two other segments, however not statistically significant different. The elastic energy storage was derived from the aortic root pressure volume relationship; the mean elastic energy storage was 826 µJ (SD529). In conclusion, the aortic root has been characterized in terms of both segmental forces, segmental change in length and elastic energy storage. This study is the first to assess the radial structural stiffness of different segments of the aortic root. The presented data is reference for further studies regarding the impact of surgical interventions on the aortic root. [ABSTRACT FROM AUTHOR]

GPIHBP1 is expressed restrictively in capillary endothelial cells, but the transcriptional regulation mechanisms of GPIHBP1 in capillaries are still unknown. Here, we analyzed the promoter activities of porcine GPIHBP1 using dual luciferase reporter assays and detected single nucleotide polymorphisms (SNPs) in the promoter region. Activity analysis discovered that the sequence from-187 to +174 is a basal promoter region, the sequence from-187 to-582 is a positive regulation region, whereas the sequence from-582 to-1321 is a negative regulation region. 11 SNPs were found in the promoter region of porcine GPIHBP1, and the g.-796G>A SNP within the NF-l-binding site is associated with pig back fat thickness. The work performed here will serve as a valuable resource for discovering the expression regulation mechanism of GPIHBP1 and promoting further development of pig as a model organism for human cardiovascular disease research. [ABSTRACT FROM AUTHOR]

Glycine, a component of glutathione (GSH), plays an important role in protection from reactive oxygen species (ROS) and inhibition of apoptosis. The aim of this study was to determine the effect of glycine on in vitro maturation (IVM) of porcine oocytes and their developmental competence after parthenogenetic activation (PA). We examined nuclear maturation, ROS levels, apoptosis, mitochondrial membrane potential (ΔΨm), and ATP concentration, as well as the expression of several genes related to oocyte maturation and development. Our studies found that treatment with glycine in IVM culture medium increased nuclear maturation rate, but varying the concentrations of glycine (0.6, 6, or 12 mM) had no significant effect. Furthermore, 6 mM glycine supported greater blastocyst formation rates and lesser apoptosis after PA than the other concentrations (P < 0.05). All the glycine treatment groups had decreased levels of ROS in both matured oocytes and at the 2-cell stage (P < 0.05). At the 2-cell stage, the 6 mM glycine group had ROS levels that were lesser than the other 2 glycine treatment groups (0.6 and 12 mM). From this, we deemed 6 mM to be the optimal condition, and we then investigated the effects of 6 mM glycine on gene expression. The expression of both FGFR2 and Hsf1 were greater than the control group in mature oocytes. The glycine treatment group had greater levels of expression of an antiapoptotic gene (Bcl2) in mature oocytes and cumulus cells and lesser levels of expression of a proapoptotic gene (Bax) in PA blastocysts (P < 0.05). In addition, mitochondrial ΔΨm and ATP concentration were increased in 6 mM glycine group compared with the control group. In conclusion, our results suggest that glycine plays an important role in oocyte maturation and later development by reducing ROS levels and increasing mitochondrial function to reduce apoptosis. [ABSTRACT FROM AUTHOR]

Glycine, a component of glutathione (GSH), plays an important role in protection from reactive oxygen species (ROS) and inhibition of apoptosis. The aim of this study was to determine the effect of glycine on in vitro maturation (IVM) of porcine oocytes and their developmental competence after parthenogenetic activation (PA). We examined nuclear maturation, ROS levels, apoptosis, mitochondrial membrane potential (ΔΨm), and ATP concentration, as well as the expression of several genes related to oocyte maturation and development. Our studies found that treatment with glycine in IVM culture medium increased nuclear maturation rate, but varying the concentrations of glycine (0.6, 6, or 12 mM) had no significant effect. Furthermore, 6 mM glycine supported greater blastocyst formation rates and lesser apoptosis after PA than the other concentrations (P < 0.05). All the glycine treatment groups had decreased levels of ROS in both matured oocytes and at the 2-cell stage (P < 0.05). At the 2-cell stage, the 6 mM glycine group had ROS levels that were lesser than the other 2 glycine treatment groups (0.6 and 12 mM). From this, we deemed 6 mM to be the optimal condition, and we then investigated the effects of 6 mM glycine on gene expression. The expression of both FGFR2 and Hsf1 were greater than the control group in mature oocytes. The glycine treatment group had greater levels of expression of an antiapoptotic gene (Bcl2) in mature oocytes and cumulus cells and lesser levels of expression of a proapoptotic gene (Bax) in PA blastocysts (P < 0.05). In addition, mitochondrial ΔΨm and ATP concentration were increased in 6 mM glycine group compared with the control group. In conclusion, our results suggest that glycine plays an important role in oocyte maturation and later development by reducing ROS levels and increasing mitochondrial function to reduce apoptosis. [ABSTRACT FROM AUTHOR]

Abstract: BACKGROUND: The use of protein hydrolyzates in food and feed ingredients is a growing area of interest. The present study was conducted aiming to determine the standardized ileal digestibility (SID) and to evaluate protein quality of porcine and bovine hydrolyzates. The SID values were determined in pigs and these values were used to calculate digestible indispensable amino acid score (DIAAS) values. RESULTS: The SID of crude protein of hydrolyzates of porcine plasma, bovine muscle, bovine collagen, porcine heart and porcine muscle was 0.81, 0.84, 0.79, 0.79 and 0.91, respectively. Based on the DIAAS, hydrolyzates of porcine plasma and porcine heart (DIAAS = 87 and 76) are considered as good quality protein sources for younger children. For older children, adolescents and adults, the hydrolyzates of bovine muscle and porcine heart (DIAAS = 81 and 87) are considered as good quality protein sources and the hydrolyzate of porcine plasma (DIAAS = 102) is considered as a high quality protein source. CONCLUSION: DIAAS values indicate that the porcine and bovine hydrolyzates tested in the present study have low and medium to high protein values and also that the protein products can be included in human nutrition. © 2017 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Objective: Recent studies have challenged the traditional paradigm that growth hormone receptor (GHR) displays physiological functions only in the cell membrane. It has been demonstrated that GHR localizes to the cell nucleus and still exhibits important physiological roles. The phenomenon of nuclear localization of growth hormone (GH)-induced GHR has previously been described in vitro. However, until recently, whether GH could induce nuclear localization of GHR in vivo was unclear. Methods: In the present study, we used pig as an animal model, and porcine growth hormone (pGH) or saline was injected into the inferior vena cava. We subsequently observed the localization of porcine growth hormone receptor (pGHR) using multiple techniques, including, immunoprecipitation and Western-blotting, indirect immunofluorescence assay and electronmicroscopy. Results: The results showed that pGH could induce nuclear localization of pGHR. Taken together, the results of the present study provided the first demonstration that pGHR was translocated to cell nuclei under pGH stimulation in vivo. Conclusion: Nuclear localization of pGHR induced by the in vivo pGH treatment suggests new functions and/or novel roles of nuclear pGHR, which deserve further study. [ABSTRACT FROM AUTHOR]

Autophagy is a homeostatic process that has been shown to be vital in the innate immune defense against pathogens. However, little is known about the regulatory role of autophagy in porcine teschovirus 2 (PTV-2) replication. In this study, we found that PTV-2 infection induces a strong increase in GFP-LC3 punctae and endogenous LC3 lipidation. However, PTV-2 infection did not enhance autophagic protein degradation. When cellular autophagy was pharmacologically inhibited by wortmannin or 3-methyladenine, PTV-2 replication increased. The increase in virus yield via autophagy inhibition was further confirmed by silencing atg5, which is required for autophagy. Furthermore, PTV-2 replication was suppressed when autophagy was activated by rapamycin. Together, the results suggest that PTV-2 infection activates incomplete autophagy and that autophagy then inhibits further PTV-2 replication. [ABSTRACT FROM AUTHOR]

1. Humans and animals are commonly exposed to indole-3-carbinol (I3C) and resveratrol (RES) via food or beverages. Moreover, these compounds have been demonstrated to potentially cause food–drug interactions. However, information about their combined effects is limited. Therefore, we investigated the effects of I3C and RES, both as single compounds and in combination, on cytochrome P450 1A and 3A activity and gene expression. 2. Using porcine microsomes, we demonstrated that RES caused non-competitive inhibition of CYP1A activity and un-competitive inhibition of CYP3A activity. Compared to the effect of single compounds, co-treatment with I3C and RES increased a degree of inhibition of CYP1A activity. 3. In porcine primary hepatocytes, treatment with I3C and RES resulted in induction of CYP1A1, CYP1A2 and CYP3A29 mRNA expression. 4. In conclusion, we demonstrated that both RES and I3C could cause food–drug interactions and that the combined effect could be more potent in doing so. [ABSTRACT FROM AUTHOR]

The Sine oculis homeobox 1 (Six1) gene is important for skeletal muscle growth and fiber specification; therefore, it is considered as a promising candidate gene that may influence porcine growth and meat quality traits. Nevertheless, the association of Six1 with these processes and the mechanisms regulating its expression remain unclear. The objectives of this study were to identify variant sites of Six1 in different pig breeds, conduct association analysis to evaluate the relationship between polymorphisms of these variants and porcine production traits in Pietrain × Duroc × Landrace × Yorkshire commercial pigs, and explore the potential regulatory mechanisms of Six1 affecting production traits. A total of 12 variants were identified, including 10 single-nucleotide variations (SNVs), 1 insertion-deletion (Indel), and 1 (AC)n microsatellite. Association analysis demonstrated that the SNV, g.1595A>G, was significantly associated with meat color (redness, a*); individuals with the G allele had greater a* values (P < 0.05). Moreover, our results demonstrated that the (AC)n polymorphism in the Six1 promoter was significantly associated with weaning weight (P < 0.05), carcass weight (P < 0.05), and thoracic and lumbar back fat (P < 0.01). In addition, we found that the (AC)n variant was closely related with Six1 expression levels and demonstrated this polymorphism on promoter activity by in vitro experiments. Overall, this study provides novel evidence for elucidating the effects of Six1 on porcine production traits as promising candidate and describes two variants with these traits, which are potential reference markers for pig molecular breeding. In addition, our data on the relationship between porcine Six1 expression and the polymorphic (AC)n microsatellite in its promoter may facilitate similar studies in other species. [ABSTRACT FROM AUTHOR]

Conditional gene targeting at porcine IκBα may be a solution to delayed xenograft rejection, the main barrier to xenotransplantation. An oligonucleotide-based method was applied to construct the vector for conditional targeting of porcine IκBα. This method was free from PCR amplification during the assembling of the different vector elements, avoiding introduction of unwanted mutations. With the help of short double-stranded DNA fragments produced by annealing oligonucleotides, nondirectional cloning has also been avoided. By making the best of directional cloning, a highly complex targeting vector was built within 3 weeks. The present study also explained why the two recombination-based methods (recombineering and gateway recombination), although having demonstrated to be highly efficient in constructing ordinary targeting vectors, were not appropriate in this context. The description in the present study of an additional method to efficiently construct targeting vectors is suggested to introduce more flexibility in the field therefore helping to meet the different needs of the researchers. [ABSTRACT FROM AUTHOR]

Green gemini surfactants (m-E2-m) containing cleavable ester functionalities were allowed to interact with porcine serum albumin (PSA) and the binding mechanism along with concomitant physicochemical changes were analyzed using a multi-technique approach. Intrinsic fluorescence study reveals that the concerned gemini surfactants have a substantial affinity for PSA at a pH well above its isoelectric point. Static quenching is indicated by the relevant value of bimolecular quenching constant (k q ). Synchronous, three-dimensional and pyrene fluorescence data depict the PSA−m-E2-m complex formation together with significant conformational changes induced in PSA. UV–vis studies are also indicative of ground state complexation involving the biomolecule and the amphiphile. Far-UV CD spectra indicate the stabilization of protein structure as deciphered by the increased α-helical content, whereas near-UV CD spectra signify a tertiary structure close to that of the native state. In order to validate the results obtained by fluorescence, cyclic voltammetry is employed, which unveils the formation of electrochemically inactive PSA−m-E2-m complex. Moreover, it is revealed that gemini with longer chain interacts more efficiently than the one with shorter chain owing to stronger hydrophobic forces. All these findings suggest that in future, the m-E2-m geminis may be potentially utilized as excipients in drug, skincare and immunoassay reagent formulations. Furthermore, results of this research work will also contribute in understanding the physicochemical effect of surfactant architecture on its interaction with biomacromolecules. [ABSTRACT FROM AUTHOR]

Ferulic Acid (FA) is a metabolite of phenylalanine and tyrosine, a phenolic compound commonly found in fruits and vegetables. Several studies have shown that FA has various functions such as antioxidant effect, prevention of cell damage from irradiation, protection from cell damage caused by oxygen deficiency, anti-inflammatory action, anti-aging action, liver protective effect and anti-cancer action. In this study, we investigated the maturation rate, intracellular glutathione (GSH) and reactive oxygen species (ROS) of porcine oocytes by adding FA to the in vitro maturation (IVM) medium and examined subsequent embryonic developmental competence at 5% oxygen through parthenogenesis. There is no significant difference between the control group (0μM) and treatment groups (5μM, 10μM, 20μM) on maturation rates. Intracellular GSH levels in oocyte treated with 5μM of FA significantly increased (P < 0.05), and 20μM of FA revealed significant decrease (P < 0.05) in intracellular ROS levels compared with the control group. Oocytes treated with FA exhibited significantly higher cleavage rates (79.01% vs 89.19%, 92.20%, 90.89%, respectively) than the control group. Oocytes treated with 10μM showed significantly higher blastocyst formation rates (28.3% vs 40.3%, respectively) after PA than the control group. Total cell numbers in blastocyst of 10μM FA displayed significantly higher (39.4 vs 51.9, respectively) than the control group. In conclusion, these results suggested that treatment with FA during IVM improved the developmental potential of porcine embryos by increasing intracellular GSH synthesis and reducing ROS levels. Also, there was an improvement of cleavage rate, blastocyst formation and total cell numbers in blastocysts. It might be associated with Keap1-Nrf2 pathway as an antioxidant regulate pathway that plays a crucial role in determining the sensitivity of cells to oxidative damages by regulating the basal and inducible expression of enzymes which is related to detoxification and anti-oxidative effects, stress response enzymes and/or proteins and ABC transporters. [ABSTRACT FROM AUTHOR]

CD1d-restricted invariant natural killer T (iNKT) cells are innate-like T cells that share phenotypic characteristics of both NK and conventional T cells (Tconv). Although iNKT cells have been well characterized in mice and humans, functional CD1d and CD1d-restricted iNKT cells are not universally expressed in mammals. Swine express iNKT cells that can be detected using α-galactosylceramide (α-GalCer)-loaded CD1d tetramers. In the present study, we characterized iNKT cells from the blood, spleen, lymph node, lung and liver of commercial mixed-breed pigs, and compared their phenotype to NK cells and Tconv. The principal findings are that pig iNKT cells are CD8α and CD44 positive and CD11b and Nkp46 negative. Most are also negative for the CD4 co-receptor, which is used to distinguish functionally distinct mouse and human iNKT cells subsets. The frequency of IFN-γ-producing CD8α bright iNKT cells was 3–4-fold higher than CD8α dull iNKT cells, suggesting that CD8α expression identifies iNKT cells with a unique functional role in immune responses. Finally, large variability was detected among pigs in interactions between iNKT cells and monocytes when iNKT cells were activated with α-GalCer, which raises a cautionary note about manipulating iNKT cells for immunotherapy. Collectively, our study provides important phenotypic and functional information about porcine iNKT cells that will be useful for understanding how iNKT cells contribute to immune responses in swine, with potential implications for human health. [ABSTRACT FROM AUTHOR]

Background There might be much benefit in xenotransplantation, however, the risk of infections across species barriers remains, especially porcine endogenous retrovirus (PERV). To date, many attempts have been made to knock down active PERVs by inhibitory RNA (RNAi) and micro RNA (miRNA), which target different genes of PERV. There are a few studies that have explored whether targeting promoter regions of PERV could exert an inhibition effect. Methods miRNAs were automatically selected based on an online program BLOCK-iT RNAi Designer. The inhibition efficiency between miRNAs was compared based on their inhibition of different PERV genes: long terminal repeats (LTR), gag, and pol. Both relative quantitative real-time polymerase chain reaction (PCR) and C-type reverse transcriptase activity were performed. Results The results demonstrated that miRNA targeting the LTR region degraded the target sequence, and simultaneously inhibited the mRNA expression of both gag and pol genes of PERV. The LTR1, LTR2, and dual LTR1 + LTR2 miRNA inhibited 76.2%, 22%, and 76.8% of gag gene expression, respectively. Similarly, the miRNA was found to knock down the pol gene expression of 69.8%, 25.5%, and 77.7% for single targeting miRNA (LTR1 and LTR2) and multi-targeting miRNA (LTR1 + LTR2), respectively. A stable PK15 clone constitutively expressed dual LTR1 + LTR2 miRNA and exhibited higher inhibitory up to 82.8% and 92.7% of the expressions of the gag and pol genes, respectively. Also, the result of co-cultivation of dual LTR1 + LTR2 miRNA transfected PK15 cell with a human cell line inhibited expression of LTR, gag, and pol genes of PERV. Conclusions In conclusion, this study suggested that the LTR might be an alternative target for gene silencing of PERV, and that multi-targeting miRNA had better inhibitory effect than single- targeting miRNA. In an in vitro model, the presence of miRNA was able to reduce PERV infectivity in a human cell line. [ABSTRACT FROM AUTHOR]

Background Xenotransplantations of pig corneas have become an attractive alternative to human corneas. Such xenotransplantations carry the danger, however, of transmission of porcine endogenous retroviruses (PERVs). Here, we investigated whether porcine corneas harbor viral DNA and RNA and whether transplantation to a nonhuman primate would lead to host PERV infection. Methods Monkey vein endothelial cells (MVECs) were inoculated with porcine aortic endothelial cell (PAEC) supernatants, and DNA and total RNA of MVECs were tested for PERV by polymerase chain reaction (PCR) and reverse transcription PCR (RT-PCR) assays. Corneas were harvested from Wuzhishan miniature pigs, and the presence of PERV proviral DNA and RNA was analyzed by PCR and RT-PCR, respectively. Fresh or dehydrated corneas were then transplanted to rhesus monkeys, and PERV proviral DNA and RNA were analyzed in host peripheral blood lymphocytes at 6 and 24 months. Furthermore, the presence of PERV sequences was analyzed in the transplant at 24 months. Results PCR analysis showed PERV transfection from PAECs to MVECs in vitro. PCR and RT-PCR gave positive signals for PERV subtypes A and B, but not PERV-C, regardless of how the corneas were prepared. No evidence was found for PERV transmission to the host, and the transplant had lost its viral signal at the end of the 24-month period. Conclusions Regardless of cornea preparation and storage, PERV transmission from pig to host could not be detected, despite that the transplant was initially PERV-positive. The use of the Wuzhishan miniature pig as the donor may be advantageous because it lacks PERV-C and hence potentially infectious A/C recombinants. [ABSTRACT FROM AUTHOR]