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Impaired healing wounds do not proceed through the normal healing processes in a timely and orderly manner, and while they do eventually heal, their healing is not optimal. Chronic wounds, on the other hand, remain unhealed for weeks or months. In the US alone, chronic wounds impact ~8.5 million people and cost ~USD 28-90 billion per year, not accounting for the psychological and physical pain and emotional suffering that patients endure. These numbers are only expected to rise in the future as the elderly populations and the incidence of comorbidities such as diabetes, hypertension, and obesity increase. Over the last few decades, scientists have used a variety of approaches to treat chronic wounds, but unfortunately, to date, there is no effective treatment. Indeed, while there are thousands of drugs to combat cancer, there is only one single drug approved for the treatment of chronic wounds. This is in part because wound healing is a very complex process involving many phases that must occur sequentially and in a timely manner. Furthermore, models that fully mimic human chronic wounds have not been developed. In this review, we assess various models currently being used to study the biology of impaired healing and chronic non-healing wounds. Among them, this paper also highlights one model which shows significant promise; this model uses aged and obese db/db-/- mice and the chronic wounds that develop show characteristics of human chronic wounds that include increased oxidative stress, chronic inflammation, damaged microvasculature, abnormal collagen matrix deposition, a lack of re-epithelialization, and the spontaneous development of multi-bacterial biofilm. We also discuss how important it is that we continue to develop chronic wound models that more closely mimic those of humans and that can be used to test potential treatments to heal chronic wounds.

Background Methods Results Conclusions Both global ischemia caused by circulatory arrest and extracorporeal circulation circuits have been shown to trigger cytokine release. We hypothesized that inserting a hemoadsorption device during thoraco‐abdominal normothermic regional perfusion (TA‐NRP) in the donation after circulatory death setting would mitigate the inflammatory response, potentially resulting in improved cardiac allograft function.In 15 pigs, circulatory arrest was induced by hypoxia. After a 15‐min no‐touch‐period, TA‐NRP was performed for 60 min. Eight pigs had a hemoadsorption device incorporated in the ECC, while seven did not. Plasma concentrations of IFN‐α, IFN‐γ, TNF‐α, IL‐1β, IL‐4, IL‐6, IL‐8, IL‐10, and IL‐12p40 were assessed by ELISA at baseline, immediately at start of TA‐NRP, 60 min after start of TA‐NRP (just before weaning from ECC), and at 30 and 60 min after weaning from ECC. Cardiac function was assessed with pressure–volume loop analysis.Hemoadsorption had no relevant effects on systemic cytokine levels post TA‐NRP. IL‐6 plasma levels gradually rose throughout the procedure for both groups. Hemoadsorption did not affect systolic or diastolic left ventricular function, nor were global hemodynamics improved by hemoadsorption.The insertion of a hemoadsorption device did not significantly affect plasma cytokine levels or cardiac function. Further research is necessary to assess the role of the inflammatory response in DCD heart transplantation and its modulation by TA‐NRP. [ABSTRACT FROM AUTHOR]

Increasing use of pigs as models in translational research, and growing focus on animal welfare are leading to better use of effective analgesics and anaesthetics when painful procedures are performed. However, there is a gap in basic knowledge such as pharmacokinetics of different anaesthetics in these species. The main objective of our study was to determine the pharmacokinetics of levobupivacaine in domestic pigs. Twelve female grower pigs weighing 31.17 ± 4.6 kg were subjected to general anaesthesia and experimental surgery, at the end of which they received 1 mg/kg levobupivacaine via peri-incisional subcutaneous infiltration. Plasma samples were collected before administration of levobupivacaine and at 0.5, 1, 2, 4, 8, 12, 24 and 48 h thereafter. Concentrations of levobupivacaine were determined by liquid chromatography coupled with tandem mass spectrometry. Following single dose of levobupivacaine, all animals had measurable plasma concentrations 0.5 h after drug administration, with most peak concentrations observed at the 1-h time point. In all 12 animals, levobupivacaine was below the limit of quantification 48 h after drug administration. The mean maximum plasma concentration, area under the curve and half-life were determined to be 809.98 μg/l, 6552.46 μg/l h and 6.25 h, respectively. Plasma clearance, volume of distribution and weight-normalized volume of distribution were 4.41 l/h, 35.57 l and 1.23 l/kg, respectively. Peak plasma concentrations in our study were well below concentrations that were found to produce toxicity in pigs. [ABSTRACT FROM AUTHOR]

Testosterone is secreted by Leydig cells (LCs), which play an important physiological role in preserving male secondary sex characteristics, protecting male reproductive function, and establishing the blood-testis barrier. Studies have shown that autophagy is particularly active in LCs; however, its involvement in testosterone synthesis in porcine LCs has not been fully explored. Therefore, this experiment aimed to investigate the influence of autophagy on testosterone secretion in porcine LCs and its potential regulatory mechanism. Our results demonstrated that both testicular autophagy and serum testosterone levels increased in piglets during postnatal development from 4 to 18 weeks. In addition, autophagy was found to degrade the Na+/H+ exchange regulatory factor 2 (NHERF2), leading to the up-regulation of scavenger receptor class B type 1 (SRB1). This process resulted in increased cholesterol intake and enhanced testosterone production. The observable level of sirtuin 1 (SIRT1) was directly proportional to the level of autophagy. In vitro investigations have shown that SIRT1 can affect the level of autophagy, cholesterol uptake as well as testosterone release. In conclusion, testosterone synthesis during pig development is regulated by SIRT1. SIRT1 mediates the degradation of NHERF2 through autophagy, thereby weakening its negative regulatory effect on the high-density lipoprotein receptor SRB1 in Leydig cells. This process increases cholesterol uptake and enhances testosterone synthesis. • Autophagy levels match testosterone secretion by Leydig cells in piglets from 4 to 18 weeks. • Autophagy facilitates cholesterol uptake in Leydig cells by degrading NHERF2, influencing testosterone production. • SIRT1 activates autophagy by promoting LC3 expression, thereby participating in testosterone synthesis. [ABSTRACT FROM AUTHOR]

Methionine adenosyltransferase 2A (MAT2A) is an essential enzyme in the methionine cycle that generates S-adenosylmethionine (SAM) by reacting with methionine and ATP. SAM acts as a methyl donors for histone and DNA methylation, which plays key roles in zygotic genome activation (ZGA). However, the effects of MAT2A on porcine ZGA remain unclear. To investigate the function of MAT2A and its underlying mechanism in porcine ZGA, MAT2A was knocked down by double-stranded RNA injection at the 1-cell stage. MAT2A is highly expressed at every stage of porcine embryo development. The percentages of four-cell-stage embryos and blastocysts were lower in the MAT2A-knockdown (KD) group than in the control group. Notably, depletion of MAT2A decreased the levels of H3K4me2, H3K9me2/3, and H3K27me3 at the four-cell stage, whereas MAT2A KD reduced the transcriptional activity of ZGA genes. MAT2A KD decreased embryonic ectoderm development (EED) and enhancer of zeste homolog 2 (EZH2) expression. Exogenous SAM supplementation rescued histone methylation levels and developmental arrest induced by MAT2A KD. Additionally, MAT2A KD significantly increased DNA damage and apoptosis. In conclusion, MAT2A is involved in regulating transcriptional activity and is essential for regulating histone methylation during porcine ZGA. • MAT2A affected porcine embryonic development. • MAT2A knockdown decreased histone methylation levels. • MAT2A knockdown decreased ZGA genes expression and induced apoptosis. • SAM supplementation rescued histone methylation levels and developmental arrest induced by MAT2A knockdown. [ABSTRACT FROM AUTHOR]

Extracellular vesicles, also known as exosomes, influence numerous cellular functions by regulating different signaling pathways. However, their role in animal reproduction remains understudied. This study aimed to evaluate the effects of porcine follicular fluid-derived exosomes (pff-Exos) on porcine oocyte in vitro maturation and parthenogenetic embryo development. We obtained pff-Exos through mixed-method ultracentrifugation and size-exclusion chromatography. Transmission electron microscopy revealed an increase in the expression of exosome markers in the first four of thirteen fractions. The number of pff-Exo was 2.2 × 106 particles per microliter. The highest maturation rate of porcine oocytes treated with pff-Exo was observed with 1.1 × 107 particles of pff-Exo in the absence of porcine follicular fluid (pFF) culture conditions. Moreover, increased expression of Gdf9 and Bmp15 was observed. The developmental rate was the highest upon treatment with 1.1 × 107 particles of pff-Exo, which increased the total cell number in blastocysts. Embryonic development to the 2-cell stage was similar between the control and pff-Exo groups; however, development to the 4-cell stage and blastocyst was significantly increased in the pff-Exo group (61.6 ± 6.08 % and 29.72 ± 1.41 %, respectively; P < 0.05) compared with that in the control group (42.0 ± 5.19 % and 18.14 ± 1.78 %, respectively). The expression levels of Oct4 , Sox2 , Bcl2 , Elf4 , and Gcn5 significantly increased at the pff-Exo 2-cell stage, whereas those of Bax, Hdac1, Hdac6 , and Sirt6 decreased. Specifically, the Oct4, Sox2, Elf4, Gcn5, and Hdac6 levels remained stable in pff-Exo 4-cell embryos, whereas those of p53 and Hat1 were reduced and increased, respectively. Treatment with pffExos significantly increased H3K9 and H3K14 acetylation levels. These results demonstrate that pff-Exo affects the in vitro maturation of porcine oocytes and early embryonic development by regulating gene expression. • The developmental rate was highest upon treatment with 1.1 × 107 pff-Exo particles. • Development to the 2-cell stage was similar between control and pff-Exo groups. • Development to the 4-cell and blastocyst stages was high in the pff-Exo group. • Treatment with pffExos significantly increased H3K9 and H3K14 acetylation levels. [ABSTRACT FROM AUTHOR]

Background: Extracellular vesicles (EVs) regulate cell metabolism and various biological processes by delivering specific proteins and nucleic acids to surrounding cells. We aimed to investigate the effects of the cargo contained in EVs derived from adipose-derived stem cells (ASCs) on the porcine embryonic development. Methods: ASCs were isolated from porcine adipose tissue and characterized using ASC-specific markers via flow cytometry. EVs were subsequently extracted from the conditioned media of the established ASCs. These EVs were added to the in vitro culture environment of porcine embryos to observe qualitative improvements in embryonic development. Furthermore, the proteins within the EVs were analyzed to investigate the underlying mechanisms. Results: We observed a higher blastocyst development rate and increased mitochondrial activity in early stage embryos in the ASC-EVs-supplemented group than in the controls (24.8% ± 0.8% vs. 28.6% ± 1.1%, respectively). The terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay of blastocysts also revealed significantly reduced apoptotic cells in the ASC-EVs-supplemented group. Furthermore, through proteomics, we detected the proteins in ASC-EVs and blastocysts from each treatment group. This analysis revealed a higher fraction of proteins in the ASC-EVs-supplemented group than in the controls (1,547 vs. 1,495, respectively). Gene analysis confirmed that ASC-EVs showed a high expression of tyrosine-protein kinase (SRC), whereas ASC-EVs supplemented blastocysts showed a higher expression of Cyclin-dependent kinase 1 (CDK1). SRC is postulated to activate protein kinase B (AKT), which inhibits the forkhead box O signaling pathway and activates CDK1. Subsequently, CDK1 activation influences the cell cycle, thereby affecting in vitro embryonic development. Conclusion: ASC-EVs promote mitochondrial activity, which is crucial for the early development of blastocysts and vital in the downregulation of apoptosis. Additionally, ASC-EVs supply SRC to porcine blastocysts, thereby elongating the cell cycle. [ABSTRACT FROM AUTHOR]

Collagen-based membrane is the most commonly used biomaterial for guided bone and tissue regeneration; however, its barrier function can be threatened by its rapid degradation pattern, affecting the success of the regeneration process. Differences in the origin and functionalization of the membrane to obtain better properties can alter the degradation rate. The objective of this study was to examine the biodegradation pattern of two commercially available collagen membranes (Jason® and Collprotect®) manufactured using porcine pericardium or dermis, doped or not with zinc-ions or doxycycline, in a period up to 21 days. The membrane specimens were subjected to hydrolytic and bacterial degradation tests. The different immersion times were carried out from 12 h up to 21 days. At each time point, quantitative measurements of thickness and weight were made using a digital caliper and an analytic microbalance, respectively. ANOVA and Student–Newman–Keuls tests were carried out for comparison purposes (p < 0.05). The differences between time-points within the same membranes and solutions were assessed by pairwise comparisons (p < 0.001). Unfunctionalized Jason membrane made of porcine pericardium attained the highest resistance to both degradation tests. The functionalization of the membranes did not alter the biodegradation patterns. All the membranes completely degraded before 48 h in the bacterial collagenase solution, which was the most aggressive test. [ABSTRACT FROM AUTHOR]

The coalescence of nanocrystals that occurs during low-temperature thermal treatments during the purification of hydroxyapatite from pigs has not yet been studied in the context of low-temperature environmentally friendly purification. To preserve the nature of hydroxyapatite, two extraction methods were investigated: Route 1 (autoclave and Soxhlet) and Route 2 (calcination at low temperatures with different times). The focus is on observing the effectiveness of removing fats and proteins while maintaining crystallite size and morphology. DSC showed that coalescence of crystallites occurred at 452, 566, 648, and 704 °C. Raw has a higher IR FWHM (phosphate band) due to the organic material; R1 had a lower FWHM (partial removal of lipids and proteins). Increasing the calcination temperature decreases the FWHM, indicating a shift in vibrational states from surface to bulk, leading to an increase in apparent crystallite size due to coalescence. For 20–80 h at quasi-stable temperatures of 400 and 500 °C, no significant changes occur in the IR spectrum. X-ray diffraction indicates the removal of organic material during calcination, as the peaks become clearer with increasing temperature. The crystallite size is not affected and remains in similar ranges. Purification with hydroxide shows an increase in both lattice parameters, in addition to the fluctuations that occur due to the ionic substitutions that naturally occur in biogenic HAp. And the TEM study confirms that the changes in apparent size from 2D to 3D are the result of the phenomenon of interplanar coalescence at low temperatures that occurs in sintering processes. [ABSTRACT FROM AUTHOR]

The optimal repair of rigid mineralized tissues, such as bone, in cases of fracture, surgical resection, or prosthetic placement, is a complex process often necessitating the use of bone graft materials. Autogenous bone from the patient is generally the gold standard in terms of outcomes but also has disadvantages, which have resulted in extensive research in the field of tissue engineering to develop better and more convenient alternatives. In the dental field, several initiatives have demonstrated that the dentin material derived from extracted teeth produces excellent results in terms of repairing bone defects and supporting dental implants. Dentin is acellular and thus, in contrast to autogenous bone, cannot provide osteoblasts or other cellular elements to the grafted region, but it does contain growth and differentiation factors, and has other properties that make it an impressive material for bone repair. In this review, the beneficial properties of dentin and the ways it interacts with the host bone are described in the context of bone graft materials. Autogenous tooth material has limitations, particularly in terms of the need for tooth extraction and the limited amount available, which currently restrict its use to particular dental procedures. The development of a xenograft dentin-derived material, which retains the properties of autogenous dentin, is described. Such a material could potentially enable the use of dentin-derived material more widely, particularly in orthopedic indications where its properties may be advantageous. [ABSTRACT FROM AUTHOR]

Background/Objectives: Sports-related concussions are a main cause of cognitive dysfunction and somatic complaints, particularly in youth. While the majority of concussion symptoms resolve within one week, cognitive effects may persist. In this study, we sought to study changes to cognition within this acute time frame. Methods: In this current study, we use an established swine model of traumatic brain injury (TBI) to study the effects of single and repeated head rotations on resting-state electroencephalography (rs-EEG) in awake piglets in the acute (within 7 days) time period after injury. We studied both healthy and experimental groups to (1) establish healthy reference ranges (RRs; N = 23) for one-minute rs-EEG in awake piglets, (2) compare the effects of single (N = 12) and repeated head rotations (N = 13) on rs-EEG, and (3) examine the acute time course (pre-injury and days 1, 4, and 7 post-injury) in animals administered single and repeated head rotations. EEG data were Fourier transformed, and total (1–30 Hz) and relative power in the alpha (8–12 Hz), beta (16.5–25 Hz), delta (1–4 Hz), and theta (4–7.5 Hz) bands were analyzed. Results: Total power and relative alpha, beta, delta, and theta power were consistent measures across days in healthy animals. We found a significant and transient increase in relative alpha power after repeated injury on day 1 in all regions and a rise above the healthy RR in the frontal and left temporal regions. Conclusions: Future studies will expand the study duration to investigate and inform clinical prognoses from acute measurements of rs-EEG. [ABSTRACT FROM AUTHOR]

The zinc oxide nanoparticles (ZnO NPs) is one of the most extensively utilized metal oxide nanoparticles in biomedicine, human food, cosmetics and livestock farming. However, growing evidence suggests that there is a potential risk for humans and animals because of the accumulation of ZnO NPs in cells, which leads to cell death through several different pathways. Nevertheless, the effects of ZnO NPs on porcine granulosa cells (PGCs) and how ZnO NPs regulate the follicular cells are unknown. In this study, we aimed to elucidate the role of ZnO NPs in the porcine ovary by using PGCs. Firstly, we identified the characterization of ZnO NPs used in this study and the results showed that the size of ZnO NPs was 29.0 nm. The results also demonstrated that ZnO NPs impaired cell viability and decreased steroid hormone secretion in PGCs. In addition, ZnO NPs induced reactive oxygen species (ROS) production, leading to oxidative stress of PGCs. Meanwhile, ZnO NPs also triggered autophagy in PGCs by increasing the ratio of LC3-II/LC3-I, along with the expression of SQSTM1 and ATG7. Finally, the results from N-acetylcysteine (NAC) addition suggested that ZnO NPs promoted autophagy through the enhancement of ROS production. In summary, this study demonstrates that ZnO NPs impair the viability and function of PGCs through autophagy, which is regulated by ROS production. [ABSTRACT FROM AUTHOR]

The interactions between myogenic cells and adipocytes play an important role in improving carcass traits and the efficiency of energy utilization. However, there are few reports about the interaction between them mediated by small extracellular vesicles (sEV). In this study, sEV derived from porcine primary skeletal muscle stem cells (MuSCs) was found to be involved in the inhibition of porcine primary adipocyte viability, triglyceride content, Oil Red O enrichment and the expression of adipogenic genes. When the MuSCs were treated with insulin (INS) and oleic acid (OA), the effects of their secreted sEVs on adipose precursor cells were reversed, suggesting that the signaling effects of sEV are related to their own heterogeneity. Further by component heterogeneity analysis, miR-146a-5p was found to be enriched in sEVs of MuSCs and to regulate and suppress adipogenesis through its heterogeneity. This study provides an important mechanism and molecular target for small extracellular vesicles to regulate the interaction between muscle and adipose tissue and improve carcass traits at the intercellular level. [ABSTRACT FROM AUTHOR]

Background: The primary objective of this study was to advance our understanding of active drug uptake at brain barriers in higher species than rodents, by examining oxycodone brain concentrations in pigs. Methods: This was investigated by a microdialysis study in healthy and endotoxemic conditions to increase the understanding of inter-species translation of putative proton-coupled organic cation (H+/OC) antiporter-mediated central nervous system (CNS) drug delivery in health and pathology, and facilitate the extrapolation to humans for improved CNS drug treatment in patients. Additionally, we sought to evaluate the efficacy of lumbar cerebrospinal fluid (CSF) exposure readout as a proxy for brain unbound interstitial fluid (ISF) concentrations. By simultaneously monitoring unbound concentrations in blood, the frontal cortical area, the lateral ventricle (LV), and the lumbar intrathecal space in healthy and lipopolysaccharide (LPS)-induced inflammation states within the same animal, we achieved exceptional spatiotemporal resolution in mapping oxycodone transport across CNS barriers. Results: Our findings provide novel evidence of higher unbound oxycodone concentrations in brain ISF compared to blood, yielding an unbound brain-to-plasma concentration ratio (Kp,uu,brain) of 2.5. This supports the hypothesis of the presence of the H+/OC antiporter system at the blood–brain barrier (BBB) in pigs. Despite significant physiological changes, reflected in pig Sequential Organ Failure Assessment, pSOFA scores, oxycodone blood concentrations and its active net uptake across the BBB remained nearly unchanged during three hours of i.v. infusion of 4 µg/kg/h LPS from Escherichia coli (O111:B4). Mean Kp,uu,LV values indicated active uptake also at the blood-CSF barrier in healthy and endotoxemic pigs. Lumbar CSF concentrations showed minimal inter-individual variability during the experiment, with a mean Kp,uu,lumbarCSF of 1.5. LPS challenge caused a slight decrease in Kp,uu,LV, while Kp,uu,lumbarCSF remained unaffected. Conclusions: This study enhances our understanding of oxycodone pharmacokinetics and CNS drug delivery in both healthy and inflamed conditions, providing crucial insights for translating these findings to clinical settings. [ABSTRACT FROM AUTHOR]

Objectives of the current study were to examine the effects of exogenous expression of PGC-1α, which is a transcription factor responsive for controlling mitochondrial DNA (mtDNA) replication, mitochondria quantity control, mitochondrial biogenesis, and reactive oxygen species (ROS) maintenance, in porcine oocytes during in-vitro maturation (IVM) on the developmental competence, as well as mitochondrial quantity and function. Exogenous over-expression of PGC-1α by injection of the mRNA construct into oocytes 20 h after the start of IVM culture significantly increased the copy number of mtDNA in the oocytes, but reduced the incidences of oocytes matured to the metaphase-II stage after the IVM culture for totally 44 h and completely suppressed the early development in vitro to the blastocyst stage following parthenogenetic activation. The exogenous expression of PGC-1α also significantly induced spindle defects and chromosome misalignments. Furthermore, markedly higher ROS levels were observed in the PGC-1α-overexpressed mature oocytes, whereas mRNA level of SOD1, encoded for a ROS scavenging enzyme, was decreased. These results conclude that forced expression of PGC-1α successfully increase mtDNA copy number but led to increased ROS production, evidently by downregulation of SOD1 gene expression, inducement of spindle aberration/chromosomal misalignment, and consequently reduction in the meiotic and developmental competences of porcine oocytes. • Microinjection of PGC-1α mRNA into porcine oocytes achieved the successful exogenous expression and increased mtDNA copy number. • Microinjection of PGC-1α mRNA made aberrant spindles and misaligned chromosomes and reduced the meiotic and developmental competences. • In the oocytes overexpressed PGC-1α, significantly higher ROS and lower SOD1 expression levels were also observed. [ABSTRACT FROM AUTHOR]

Introduction: Although Campylobacter spp., including Campylobacter coli, have emerged as important zoonotic foodborne pathogens globally, the understanding of the genomic epidemiology of C. coli of porcine origin is limited. Methods: As pigs are an important reservoir of C. coli, we analyzed C. coli genomes that were isolated (n = 3) from pigs and sequenced (this study) them along with all other C. coli genomes for which pig intestines, pig feces, and pigs were mentioned as sources in the NCBI database up to January 6, 2023. In this paper, we report the pan-genomic features, the multi-locus sequence types, the resistome, virulome, and mobilome, and the phylogenomic analysis of these organisms that were obtained from pigs. Results and discussion: Our analysis revealed that, in addition to having an open pan-genome, majority (63%) of the typeable isolates of C. coli of pig origin belonged to a single clonal complex, ST-828. The resistome of these C. coli isolates was predominated by the genes tetO (53%), blaOXA-193 (49%), and APH (3')-IIIa (21%); however, the virulome analysis revealed a core set of 37 virulence genes. Analysis of the mobile genetic elements in the genomes revealed wide diversity of the plasmids and bacteriophages, while 30 transposons were common to all genomes of C. coli of porcine origin. Phylogenomic analysis showed two discernible clusters comprising isolates originating from Japan and another set of isolates comprising mostly copies of a type strain stored in three different culture collections. [ABSTRACT FROM AUTHOR]

This study aimed to compare the effectiveness, in terms of viability and quality post‐warming, when vitrifying in vitro‐produced (IVP) pig blastocysts with either a modified Cryotop (n = 161; 20 blastocysts/device) or the conventional Superfine Open Pulled Straw (SOPS; n=160; 5‐6 blastocysts/device systems. Blastocyst viability, morphology, and apoptosis parameters were evaluated after 24 h of in vitro culture. The Cryotop system yields better results in terms of higher embryo viability and total cell numbers (p <.05) and lower apoptosis (p <.05) parameters than the SOPS procedure, defining a high effectiveness to simultaneously vitrify 20 pig IVP blastocysts at one time, thus increasing the yield of IVP blastocysts readily available for embryo transfer. [ABSTRACT FROM AUTHOR]

Although supplementation with docosahexaenoic acid (DHA) during porcine oocyte IVM is well-established, the available data are limited due to the lack of consistency. Moreover, to our knowledge, the anti-oxidant effects of DHA on porcine oocytes have not been reported. Hence, this study aimed to examine the effects of DHA supplementation on the regulation of energy metabolism during porcine oocyte maturation to improve oocyte maturation and embryonic development. By supplementing the IVM medium with various DHA concentrations, 25 μM DHA was identified as the optimal concentration which improved intraoocyte glutathione content and enhanced embryonic development after parthenogenesis. Compared to embryos derived from the control group, those derived from SCNT or IVF showed significantly improved blastocyst formation upon DHA supplementation during IVM. In addition, various transcription factors associated with oocyte development and apoptosis in mature oocytes were beneficially regulated in the DHA-treated oocytes. Moreover, DHA improved the AMP-activated protein kinase (AMPK)-regulatory ability of porcine oocytes and ameliorated nuclear maturation and embryonic development, which were decreased by artificially downregulating AMPK. To our knowledge, this is the first study to examine the effects of DHA as an AMPK regulator on oocyte maturation and embryo development in pigs. Furthermore, DHA addition to the IVM medium upregulated the relative expression of genes associated with mitochondrial potential and lipid metabolism. Therefore, the membrane potential of mitochondria (evaluated based on the JC-1 aggregate/JC-1 monomer ratio) and the levels of fatty acids and lipid droplets in matured oocytes increased, resulting in increased ATP synthesis. In conclusion, the DHA treatment of porcine oocytes with 25 μM DHA during IVM enhances the homeostasis of energy metabolism by improving mitochondrial function and lipid metabolism, leading to improved quality of matured oocytes and enhanced embryonic developmental potential of in vitro produced (IVP) embryos. Thus, 25 μM DHA supplementation could serve as a tool for improving the quality of IVP embryos. The study findings provide a basis for further research on improving the production efficiency of cloned animals by securing high-quality matured oocytes and enhancing energy metabolism in mammalian oocytes, including those of pigs. • DHA (25 μM) optimally improves porcine oocyte quality and embryonic development. • DHA maintains the homeostasis of energy metabolism of porcine oocytes. • DHA improves pivotal functions of mitochondria in matured porcine oocytes. • DHA supplementation during porcine oocyte maturation improves lipid metabolism. [ABSTRACT FROM AUTHOR]

Background: The main goal of the study was to find the magnetic resonance imaging (MRI) parameters that optimize contrast between tissue and thermal lesions produced by focused ultrasound (FUS) using T1-weighted (T1-W) and T2-weighted (T2-W) fast spin echo (FSE) sequences. Methods: FUS sonications were performed in ex vivo porcine tissue using a single-element FUS transducer of 2.6 MHz in 1.5 and 3 T MRI scanners. The difference in relaxation times as well as the impact of critical MRI parameters on the resultant contrast-to-noise ratio (CNR) between coagulated and normal tissues were assessed. Discrete and overlapping lesions were inflicted in tissue with simultaneous acquisition of T2-W FSE images. Results: FUS lesions are characterized by lower relaxation times than intact porcine tissue. CNR values above 80 were sufficient for proper lesion visualization. For T1-W imaging, repetition time values close to 1500 ms were considered optimum for obtaining sufficiently high CNR at the minimum time cost. Echo time values close to 50 ms offered the maximum lesion contrast in T2-W FSE imaging. Monitoring of acute FUS lesions during grid sonications was performed successfully. Lesions appeared as hypointense spots with excellent contrast from surrounding tissue. Conclusion: MRI monitoring of signal intensity changes during FUS sonication in grid patterns using optimized sequence parameters can provide useful information about lesion progression and the success of ablation. This preliminary study demonstrated the feasibility of the proposed monitoring method in ex vivo porcine tissue and should be supported by in vivo studies to assess its clinical potential. [ABSTRACT FROM AUTHOR]

Parabens (PBs) are widely used in the cosmetic, pharmaceutical, and food industries as preservatives of products. Because of its great use, humans and other organisms are highly exposed daily. However, little is known about the effect of PBs on male infertility. Therefore, the aim of the present study was to evaluate the effect of methylparaben (MePB) and propylparaben (PrPB), alone or in combination, on the physiological characteristics of pig in vitro exposed sperm to different concentrations (0, 200, 500, and 700 μM) for viability, motility, and acrosome integrity evaluation and (0, 200, 500, 700, 1000, and 2000 μM) for DNA fragmentation index evaluation, after 4 h of exposure. The results showed that sperm viability decreased after exposure to MePB from the concentration of 500 μM. In the PrPB and mixture groups, viability decreased at all concentrations except for the control. The decrease in viability of sperm exposed to PrPB was greater than that of the mixture and MePB groups. Sperm motility decreased in all the experimental groups exposed to PBs, at all concentrations, except for the control group. Acrosome integrity was not decreased in the MePB group; however, in the PrPB group, it decreased at a concentration of 200 μM and in the mixture at 500 μM. All groups exhibited DNA damage at different concentrations, except for the control group. Additionally, the effect of PBs on sperm quality was concentration‐dependent. The results demonstrated that MePB and PrPB alone or in combination can have adverse effects on sperm quality parameters. MePB had lower toxicity than did both PrPB and the mixture. The mixture did not have an additive effect on any of the parameters evaluated. This could partially explain the link between PB exposure and infertility. Sperm viability decreased after exposure to MePB from the concentration of 500 μM. In the PrPB and mixture groups, viability decreased at all concentrations. Sperm motility decreased in all the experimental groups exposed to PBs at all concentrations. Acrosome integrity was not decreased in the MePB group; however, in the PrPB group, it decreased at a concentration of 200 μM and in the mixture at 500 μM. All groups exhibited DNA damage at different concentrations. [ABSTRACT FROM AUTHOR]

Background: The main goal of the study was to find the magnetic resonance imaging (MRI) parameters that optimize contrast between tissue and thermal lesions produced by focused ultrasound (FUS) using T1-weighted (T1-W) and T2-weighted (T2-W) fast spin echo (FSE) sequences. Methods: FUS sonications were performed in ex vivo porcine tissue using a single-element FUS transducer of 2.6 MHz in 1.5 and 3 T MRI scanners. The difference in relaxation times as well as the impact of critical MRI parameters on the resultant contrast-to-noise ratio (CNR) between coagulated and normal tissues were assessed. Discrete and overlapping lesions were inflicted in tissue with simultaneous acquisition of T2-W FSE images. Results: FUS lesions are characterized by lower relaxation times than intact porcine tissue. CNR values above 80 were sufficient for proper lesion visualization. For T1-W imaging, repetition time values close to 1500 ms were considered optimum for obtaining sufficiently high CNR at the minimum time cost. Echo time values close to 50 ms offered the maximum lesion contrast in T2-W FSE imaging. Monitoring of acute FUS lesions during grid sonications was performed successfully. Lesions appeared as hypointense spots with excellent contrast from surrounding tissue. Conclusion: MRI monitoring of signal intensity changes during FUS sonication in grid patterns using optimized sequence parameters can provide useful information about lesion progression and the success of ablation. This preliminary study demonstrated the feasibility of the proposed monitoring method in ex vivo porcine tissue and should be supported by in vivo studies to assess its clinical potential.

Abstract Background Extracellular vesicles (EVs) regulate cell metabolism and various biological processes by delivering specific proteins and nucleic acids to surrounding cells. We aimed to investigate the effects of the cargo contained in EVs derived from adipose-derived stem cells (ASCs) on the porcine embryonic development. Methods ASCs were isolated from porcine adipose tissue and characterized using ASC-specific markers via flow cytometry. EVs were subsequently extracted from the conditioned media of the established ASCs. These EVs were added to the in vitro culture environment of porcine embryos to observe qualitative improvements in embryonic development. Furthermore, the proteins within the EVs were analyzed to investigate the underlying mechanisms. Results We observed a higher blastocyst development rate and increased mitochondrial activity in early stage embryos in the ASC-EVs-supplemented group than in the controls (24.8% ± 0.8% vs. 28.6% ± 1.1%, respectively). The terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay of blastocysts also revealed significantly reduced apoptotic cells in the ASC-EVs-supplemented group. Furthermore, through proteomics, we detected the proteins in ASC-EVs and blastocysts from each treatment group. This analysis revealed a higher fraction of proteins in the ASC-EVs-supplemented group than in the controls (1,547 vs. 1,495, respectively). Gene analysis confirmed that ASC-EVs showed a high expression of tyrosine-protein kinase (SRC), whereas ASC-EVs supplemented blastocysts showed a higher expression of Cyclin-dependent kinase 1 (CDK1). SRC is postulated to activate protein kinase B (AKT), which inhibits the forkhead box O signaling pathway and activates CDK1. Subsequently, CDK1 activation influences the cell cycle, thereby affecting in vitro embryonic development. Conclusion ASC-EVs promote mitochondrial activity, which is crucial for the early development of blastocysts and vital in the downregulation of apoptosis. Additionally, ASC-EVs supply SRC to porcine blastocysts, thereby elongating the cell cycle.

Abstracts The interactions between myogenic cells and adipocytes play an important role in improving carcass traits and the efficiency of energy utilization. However, there are few reports about the interaction between them mediated by small extracellular vesicles (sEV). In this study, sEV derived from porcine primary skeletal muscle stem cells (MuSCs) was found to be involved in the inhibition of porcine primary adipocyte viability, triglyceride content, Oil Red O enrichment and the expression of adipogenic genes. When the MuSCs were treated with insulin (INS) and oleic acid (OA), the effects of their secreted sEVs on adipose precursor cells were reversed, suggesting that the signaling effects of sEV are related to their own heterogeneity. Further by component heterogeneity analysis, miR-146a-5p was found to be enriched in sEVs of MuSCs and to regulate and suppress adipogenesis through its heterogeneity. This study provides an important mechanism and molecular target for small extracellular vesicles to regulate the interaction between muscle and adipose tissue and improve carcass traits at the intercellular level.

Abstract Endothelial cell dysfunction can lead to various vascular diseases. Blood flow disorder is a common symptom of vascular diseases. Regenerative angiogenesis, which involves transplanting vascular cells or stem cells into the body to shape new vasculature, can be a good therapeutic strategy. However, there are several limitations to using autologous cells from the patients themselves. We sought to investigate the new vascular cells that can play a role in the formation of angiogenesis in vivo using stem cells from alternative animals suitable for cellular therapy. Porcine is an optimal animal model for xenotransplantation owing to its physiological similarity to humans. We used differentiated porcine endothelial cells (pECs) as a therapeutic strategy to restore vessel function. Differentiated pECs formed vessel-like structures in mice, distinguishing them from stem cells. MMPs activity and migration assays indicated that differentiated pECs possessed angiogenic potential. Tube formation and 3D spheroid sprouting assays further confirmed the angiogenic phenotype of the differentiated pECs. Immunofluorescence and immunoprecipitation analyses revealed claudin-mediated tight junctions and connexin 43-mediated gap junctions between human ECs and differentiated pECs. Additionally, the movement of small RNA from human ECs to differentiated pECs was observed under co-culture conditions. Our findings demonstrated the in vivo viability and angiogenetic potential of differentiated pECs and highlighted the potential for intercellular communication between human and porcine ECs. These results suggest that transplanted cells in vascular regeneration completed after cell therapy have the potential to achieve intercellular communication within the body.

In vitro maturation (IVM) plays a critical role in embryo production. However, the quality of IVM oocytes often suffers from oxidative stress due to the excessive accumulation of ROS. Equol, a metabolite of soybean flavonoids, exhibits potent antioxidant activity. This study investigated the effects of equol on porcine oocyte IVM. Our findings showed that treatment with 5 μM equol significantly enhanced cumulus cell expansion and the first polar body extrusion in porcine oocytes. Moreover, equol also improved the subsequent embryonic development capacity of the oocytes after parthenogenetic activation. Additionally, equol improved mitochondrial function by increasing mitochondrial content, membrane potential, and ATP levels, while promoting lipid droplet accumulation in oocytes. Equol also reduced DNA damage and early apoptosis, with an associated upregulation of BCL2 and downregulation of BAX expression. Notably, equol decreased ROS levels, likely through activation of the NRF2/KEAP1 antioxidant pathway, leading to increased expression of HO-1 , CAT , GPX1 , and SOD. In conclusion, equol improves porcine oocyte IVM by mitigating oxidative stress via activation of the NRF2/KEAP1 pathway, offering a potential strategy for optimizing the IVM system in porcine oocytes. [Display omitted] [ABSTRACT FROM AUTHOR]

During fetal development, primordial oocytes maintain their developmental potential through a ROS-minimizing metabolic mechanism. Maturation increases ROS levels, causing stress and damage, which are countered by in vivo antioxidants. In vitro maturation (IVM) worsens this due to fewer antioxidant presence and medium factors. To address this, we evaluated the effects of incorporating various natural antioxidants in the porcine oocyte IVM media. Our findings revealed that 10 μM Dendrobine (DEN), 1 μM Polydatin (PD), 20 μM Limonin (LIM), and 25 μM Nobiletin (NOB) significantly improved the first polar body extrusion rates (p < 0.05), reduced ROS, and increased GSH levels. Individual addition of 100 μg/mL Lycium barbarum polysaccharides (LBP), 0.1 μM Kaempferol (KAE), 250 μM Salidroside (SAL), 10 μM Curcumin (CUR), DEN, PD, LIM, and NOB to the porcine IVM system showed that KAE, LIM, NOB, and LBP treatments yielded the most favorable results. At the gene level, LIM, LBP, and NOB were found to upregulate the expression levels of GPX1, SIRT1, and TFAM, while downregulating Caspase3 and increasing the BCL2/BAX ratio. The inclusion of LIM, NOB, and LBP, either alone or in combination, into the IVM media effectively alleviated oxidative stress in porcine oocytes, decreased cell apoptosis, preserved mitochondrial membrane potential, and enhanced the blastocyst rate. These results offer valuable insights for optimizing the porcine oocyte IVM culture system. • Dendrobine (10 μM), Polydetin (1 μM), Limonin (20 μM), and Nobiletin (25 μM) enhance porcine oocyte maturation. • Limonin (LIM), Nobiletin (NOB), and Lycium barbarum polysaccharides (LBP) enhance blastocyst formation. • LIM, NOB, and LBP reduce oxidative stress, decrease apoptosis, preserve mitochondrial membrane potential in porcine oocytes. [ABSTRACT FROM AUTHOR]

Background: Typical difficulties encountered during in vitro fertilization (IVF) to produce embryos in pigs include poor pronucleus formation and poor-quality fertilized embryos because of high polysperm invasion. In this study, we evaluated the effects of supplementation with apple seed extract (ASE) and coculture systems on porcine in vitro-fertilized embryo culture. Methods: Slaughterhouse-derived ovaries were used to obtain cumulus-oocyte complexes (COCs). COCs were conventionally used to perform IVF. We examined the differences in apoptosis and metabolism during development following addition of ASE to normal culture and coculture systems. Matrix metalloproteinases (MMPs), cell development-related factors, and apoptotic proteins were compared in porcine embryos produced under different conditions. Results: The expression of genes related to insulin-like growth factor (IGF) signaling was increased in the coculture system. In the ASE group, early apoptosis and necrosis were reduced in fertilized embryos and the late survival rate increased. Supplementation of the coculture system with ASE led to increased expression of BCL-2 and decreased expression of Casp-3 in the cytoplasm, thereby lowering the apoptosis rate and inducing MMP expression. In addition, compared with the extract-supplemented group in normal culture, the activity of MMP-2 decreased in the coculture system supplemented with ASE, activity of MMP-9 increased, and the expression of dynactin p62 and BrdU in the cytoplasm was higher than that in the other groups. Conclusions: The coculture system increased the activity of the embryonic cytoplasm compared with the non-coculture system. Supplementation with ASE may induce cell activity and inhibit the expression of apoptotic factors.

Abstract Background Cortisone is derived from cortisol through the action of the enzyme 11β-hydroxysteroid dehydrogenase type II, and it has gained importance in recent years as a biomarker of stress. This study aimed to develop and validate an assay for the measurement of cortisone in pig saliva and evaluate whether its concentration varies in stressful situations. For this purpose, a specific immunoassay was developed and validated analytically, and a study was performed to evaluate whether cortisone concentrations in saliva can vary under heat stress conditions. Results The assay proved to be accurate, reliable, and sensitive for the measurement of cortisone in pig saliva. The limit of detection of the assay was set at 0.006 ng/ml, and the lower limit of quantification was 0.023 ng/ml. It also correlated significantly with the results obtained by LC‒MS/MS (P = 0.003; r = 0.64). In addition, the cortisone concentration in animals subjected to prolonged heat stress decreased significantly 15 days after treatment (P

Abstract Background Developmental dysplasia of the hip causes secondary osteoarthritis. Finite element analysis suggests high hip joint contact pressure in patients with hip dysplasia and a reduction in contact pressure after periacetabular osteotomy. However, few biomechanical studies have examined the load distribution in the hip joint. This study aimed to investigate the biomechanical properties of load distribution in porcine hip joints at different acetabular coverages. Methods Six porcine hip joints were analyzed using three models: 1) neutral coverage, 2) 15° under-coverage (defined as dysplasia model), and 3) 15° over-coverage created by varying the acetabular coverage. The load distribution was assessed using a pressure-mapping sensor system after applying a loading force of 100 N to the hip joint. Results In the dysplasia model, the load was concentrated at the acetabular rim; in the neutral and over-coverage models, it was dispersed. The average contact pressure was significantly higher in the dysplasia model than in the neutral coverage model ([0.42 vs. 0.3 MPa]; p = 0.004). The contact area was significantly smaller in the dysplasia model than in the neutral coverage model ([250.7 vs. 345.0 mm2]; p = 0.004). No significant differences were observed in contact pressure or area between the neutral and over-coverage models. Conclusions Insufficient acetabular coverage in the dysplasia model demonstrated higher contact pressure and smaller contact area than the neutral model. Conversely, the contact pressure and area in the over-coverage model did not differ significantly from those in the normal model. Therefore, surgeons should note that acetabular coverage overcorrection has limited effect; normalization is crucial during periacetabular osteotomy.

Abstract Seminal plasma (SP) is rich in extracellular vesicles (EVs), which are still poorly studied, especially in livestock species. To better understand their functional role in both spermatozoa and endometrial epithelial cells, proper characterization of EVs is an essential step. The objective was to phenotypically characterize porcine seminal EVs (sEVs) using cryogenic electron microscopy (cryo-EM), which allows visualization of EVs in their native state. Porcine ejaculates are released in fractions, each containing SP from different source. This allows characterization sEVs released from various male reproductive tissues. Two experiments were performed, the first with SP from the entire ejaculate (n:6) and the second with SP from three ejaculate fractions (n:15): the first 10 mL of the sperm-rich ejaculate fraction (SRF-P1) with SP mainly from the epididymis, the remainder of the SRF (SRF-P2) with SP mainly from the prostate, and the post-SRF with SP mainly from the seminal vesicles. The sEVs were isolated by size exclusion chromatography and 1840 cryo-EM sEV images were acquired using a Jeol-JEM-2200FS/CR-EM. The size, electron density, complexity, and peripheral corona layer were measured in each sEV using the ImageJ software. The first experiment showed that sEVs were structurally and morphologically heterogeneous, although most (83.1%) were small (less than 200 nm), rounded, and poorly electrodense, and some have a peripheral coronal layer. There were also larger sEVs (16.9%) that were irregularly shaped, more electrodense, and few with a peripheral coronal layer. The second experiment showed that small sEVs were more common in SRF-P1 and SRF-P2, indicating that they originated mainly from the epididymis and prostate. Large sEVs were more abundant in post-SRF, indicating that they originated mainly from seminal vesicles. Porcine sEVs are structurally and morphologically heterogeneous. This would be explained by the diversity of reproductive organs of origin.

The emergence of porcine deltacoronavirus (PDCoV) presents a significant threat to both human and animal health due to its ability to cause highly contagious enteric diseases. This underscores the crucial need for timely and accurate diagnosis to facilitate effective epidemiological investigation and clinical management. This research aimed to establish a visual detection method based on reverse transcription loop-mediated isothermal amplification (RT-LAMP) for PDCoV testing. In this study, six pairs of primers were designed according to the conserved sequences of PDCoV ORF1a/b genes. The primer sets and parameters that affect LAMP reaction were optimized. The visual RT-LAMP method was developed by incorporating methyl red into the optimized reaction system, it exclusively detected PDCoV without cross-reactivity with other viruses and the detection limits for PDCoV could reach 10 copies/µL. In comparison with RT-PCR for testing 132 clinical samples, the relative specificity and sensitivity of the visual RT-LAMP were found to be 99.2 and 100%, respectively, with a concordance rate of 99.2% and a kappa value of 0.959, indicating that the visual RT-LAMP is a reliable method for the application of PDCoV detection in clinical samples. [ABSTRACT FROM AUTHOR]

Introduction: Since boar spermatozoa show a marked deterioration in sperm quality when cooled, insemination doses are usually stored at 16-18 °C. However, maintaining this temperature during transport of semen doses is challenging, particularly during the summer months. An alternative could be to store the doses at 4 °C if cold-shock to the sperm could be prevented. The objective of this study was to evaluate boar sperm quality and fertility in in vitro fertilization after storage in AndroStar Premium at 4 °C for 1 week. Methods: Insemination doses (n =9) in AndroStar Premium from a commercial boar semen collection station were transported to the laboratory at approximately 20 °C. At the laboratory, sperm quality evaluation and was preformed and each dose was split; half of each ejaculate was stored in a climate-controlled box at 16-18 °C, the other was slowly cooled to 4 °C. Both samples were stored for 1 week before further sperm quality evaluation and in vitro fertilization (IVF) were performed. Mean values were tested using generalized linear regression, with treatment and boar as fixed factors; p ≤ 0.05 was considered significant. Results: Sperm membrane integrity (mean±sem: 91±0.05 and 83±0.09% for 16 and 4 °C, respectively) and superoxide production (6.79±2.37 and 13.54±6.23% for 16 and 4 °C, respectively), were different between treatments. The DNA fragmentation index was lower in cold-stored samples than in conventionally stored samples (3.74±2.25 and 7.40±3.36% for 4 and 16 °C, respectively). The numbers of oocytes developing to blastocyst on Day 6 (mean±sd: 9.0±8.0 and 6.0±5.0%, for storage at 16 and 4 °C, respectively) were not different between treatments. Discussion: Therefore, storage of boar semen doses in AndroStar Premium at 4 °C for up to 7days would be a viable alternative to current praxis. [ABSTRACT FROM AUTHOR]

Short-chain fatty acids (SCFAs), a subset of organic fatty acids with carbon chains ranging from one to six atoms in length, encompass acetate, propionate, and butyrate. These compounds are the endproducts of dietary fiber fermentation, primarily catalyzed by the glycolysis and pentose phosphate pathways within the gut microbiota. SCFAs act as pivotal energy substrates and signaling molecules in the realm of animal nutrition, exerting a profound influence on the intestinal, immune system, and intestinal barrier functions. Specifically, they contibute to 60-70% of the total energy requirements in ruminants and 10-25% in monogastric animals. SCFAs have demonstrated the capability to effectively modulate intestinal pH, optimize the absorption of mineral elements, and impede pathogen invasion. Moreover, they enhance the expression of proteins associated with intestinal tight junctions and stimulate mucus production, thereby refining intestinal tissue morphology and preserving the integrity of the intestinal structure. Notably, SCFAs also exert anti-inflammatory properties, mitigating inflammation within the intestinal epithelium and strengthening the intestinal barrier's defensive capabilities. The present review endeavors to synthesize recent findings regarding the role of SCFAs as crucial signaling intermediaries between the metabolic activities of gut microbiota and the status of porcine cells. It also provides a comprehensive overview of the current literature on SCFAs' impact on immune responses within the porcine intestinal mucosa. [ABSTRACT FROM AUTHOR]

Circular RNAs (circRNAs) are a class of circular non-coding RNAs that play essential roles in the intricate and dynamic networks governing cell growth, development, and apoptosis. Resveratrol (RSV), a non-flavonoid polyphenol, is known to participate in follicular development and ovulation. In our previous research, we established a model using porcine ovarian granulosa cells (POGCs) treated with resveratrol, which confirmed its regulatory effects on long non-coding RNAs (lncRNAs) and microRNAs (miRNAs) within these cells. However, the influence of resveratrol on circRNA expression has not been thoroughly investigated. To explore how resveratrol affects circRNA levels in POGCs, we designed an experiment with three groups: a control group (CON, n = 3, 0 μM RSV), a low-dose RSV group (LOW, n = 3, 50 μM RSV), and a high-dose RSV group (HIGH, n = 3, 100 μM RSV) for circRNA sequencing. We identified a total of 10,045 candidate circRNAs from POGCs treated with different concentrations of resveratrol (0, 50, and 100 μM). Differential expression analysis indicated that 96 circRNAs were significantly altered in the LOW vs. CON group, while 109 circRNAs showed significant changes in the HIGH vs. CON group. These circRNAs were notably enriched in biological processes associated with cell metabolism, apoptosis, and oxidative stress. Functional enrichment analysis of the host genes revealed their involvement in critical signaling pathways, including mTOR, AMPK, and apoptosis pathways. Additionally, we identified potential miRNA sponge candidates among the differentially expressed circRNAs, particularly novel_circ_0012954 and novel_circ_0004762, which exhibited strong connectivity within miRNA-target networks. Our findings provide valuable insights into the regulatory mechanisms of circRNAs in the context of resveratrol-induced apoptosis in POGCs, highlighting their potential as innovative therapeutic targets in reproductive biology. [ABSTRACT FROM AUTHOR]

Simple Summary: Prolonged stress negatively affects pig health, welfare, and productivity. Herein, we used a porcine model of stress during the growing period, divided into stressed and control groups. Stressed pigs experienced reduced space and were mixed twice, leading to decreased body weight and feed efficiency. Differences in feeding behavior were also observed; stressed pigs visited feeders less frequently and spent more time per meal. The microbiota of stressed pigs showed an increase in opportunistic bacteria, while control pigs had a higher abundance of beneficial butyrate- and propionate-producing bacteria. This study highlights the potential of using specific fecal microorganisms as non-invasive biomarkers to assess stress and well-being in pigs, with implications for improving both animal welfare and research applied to the human gut-brain axis. Stress significantly affects the health, welfare, and productivity of farm animals. We performed a longitudinal study to evaluate stress's effects on pig performance, feeding behavior, and fecal microbiota composition. This study involved 64 Duroc pigs during the fattening period, divided into two experimental groups: a stress group (n = 32) and a control group (n = 32). Stressed groups had less space and were mixed twice during the experiment. We monitored body weight, feed efficiency, feeding behavior, and fecal microbiota composition. Compared to the control group, the stressed pigs exhibited reduced body weight, feed efficiency, fewer feeder visits, and longer meal durations. In the fecal microbiota, resilience was observed, with greater differences between groups when sampling was closer to the stressful stimulus. Stressed pigs showed an increase in opportunistic bacteria, such as Streptococcus, Treponema and members of the Erysipelotrichaceae family, while control pigs had more butyrate- and propionate-producing genera like Anaerobutyricum, Coprococcus and HUN007. Our findings confirm that prolonged stress negatively impacts porcine welfare, behavior, and performance, and alters their gut microbiota. Specific microorganisms identified could serve as non-invasive biomarkers for stress, potentially informing both animal welfare and similar gut-brain axis mechanisms relevant to human research. [ABSTRACT FROM AUTHOR]

Simple Summary: Maintaining a sustainable supply of animal protein is the mission of the global swine production industry. The entry of infectious pathogens to swine populations can cause significant animal welfare issues, increase the use of antibiotics, challenge environmental stability, and interrupt/reduce the supply of pork; therefore, preventing pathogen entry is critical to achieve its mission using science-based biosecurity programs. Biosecurity is the application of science-based protocols to minimize the risk of pathogen entry. The objective of this review is to summarize basic biosecurity terms and concepts, review the transmission of porcine reproductive and respiratory syndrome virus (PRRSV) and the biosecurity protocols designed to mitigate these risk factors, and discuss how the swine industry is applying Next Generation Biosecurity to prevent PRRSV infection of the breeding herd. The prevention of disease introduction into swine herds requires the practice of science-based protocols of biosecurity that have been validated to reduce the risk of the entry of targeted pathogens. The fundamental pillars of biosecurity include bio-exclusion, biocontainment, and bio-management. Biosecurity protocols must be science-based, a way of life, continuously validated, cost-effective, and benchmarked over time. This paper will review these concepts, the direct and indirect routes of transmission of porcine reproductive and respiratory syndrome virus (PRRSV), and the interventions that have been designed and validated to prevent infection of the breeding herd. It will close with a review of Next Generation Biosecurity, describing how a science-based approach is being used to prevent PRRSV infection in breeding herds from a large commercial pork production system in the US. [ABSTRACT FROM AUTHOR]

HT-2 toxin is a type of mycotoxin which is shown to affect gastric and intestinal lesions, hematopoietic and immunosuppressive effects, anorexia, lethargy, nausea. Recently, emerging evidences indicate that HT-2 also disturbs the reproductive system. In this study, we investigated the impact of HT-2 toxin exposure on the organelles of porcine oocytes. Our results found that the abnormal distribution of endoplasmic reticulum increased after HT-2 treatment, with the perturbation of ribosome protein RPS3 and GRP78 expression; Golgi apparatus showed diffused localization pattern and GM130 localization was also impaired, thereby affecting the Rab10-based vesicular transport; Due to the impairment of ribosomes, ER, and Golgi apparatus, the protein supply to lysosomes was hindered, resulting in lysosomal damage, which further disrupted the LC3-based autophagy. Moreover, the results indicated that the function and distribution of mitochondria were also affected by HT-2 toxin, showing with fragments of mitochondria, decreased TMRE and ATP level. Taken together, our study suggested that HT-2 toxin exposure induces damage to the organelles for endomembrane system, which further inhibited the meiotic maturation of porcine oocytes. • HT-2 caused abnormal distribution of ER and GRP78-based ER stress in oocytes. • HT-2 disrupted GM130-based Golgi apparatus and Rab10-based vesicular transport. • HT-2 induced lysosome damage and disrupted LC3-based autophagy in oocytes. • HT-2 disturbed mitochondria distribution, MMP and ATP production in oocytes. [ABSTRACT FROM AUTHOR]

There are few existing publications that describe transvaginal ultrasound-guided Ovum Pick-Up (OPU) in sows, and the impacts of the procedure for the welfare of the animals are unknown. In this study, we evaluated the effects of OPU, performed following restraint in a claw-trimming chute, on the animal welfare and reproductive health of second parity hybrid sows. The study utilized a generalized randomized block design at a commercial sow pool. We assessed salivary cortisol levels before, during, and after the procedure to compare the physiological stress response between OPU and restraint chute procedures (control group). We found a significant increase in salivary cortisol caused by the physical restraint procedure, and that the salivary cortisol level at the end of the procedure did not differ between OPU and control groups (p = 0.51). Furthermore, we conducted a novel approach-aversion test for sows, designed to assess if a feed reward would motivate the animals to willingly participate in the OPU-procedure. The animals were trained daily to enter the chute to access a feed reward. Ten animals in each group failed to complete the training period and did not voluntarily enter the restraint chute on the experimental day. This indicates that even the short daily restraint procedure during the four-day long training period was aversive to some animals. There was no difference in aversion towards the restraint chute between OPU and control groups one day after the procedure. The reproductive performance of the animals was subsequently evaluated through oestrus synchronization and insemination of the sows after the experiment. There was no observed difference in the farrowing rate (p = 0.72) and total number of born piglets (p = 0.84) between OPU and control sows. On average, we retrieved 9.0 ± 5.9 oocytes during the OPU-sessions (N = 26). Our results show that a majority of the sows prioritize the motivation for feed over their aversion to the OPU procedure. However, the physical restraint procedure is unpleasant for the animals and elicits a temporary stress response. We suggest that transvaginal OPU may be used for the recovery of oocytes from live sows, but refinements are needed to avoid stress during the lifting procedure. Such modifications could also potentially reduce the observed inter-individual variations in oocyte recovery outcomes. • Physical restraint and OPU causes significant increase in cortisol levels. • Average oocyte retrieval during OPU was 9.0 ± 5.9. • Sows prioritized feed reward over aversion to OPU. • No difference in subsequent reproductive performance in OPU vs control groups. • Refinements are needed to reduce stress and increase efficiency of OPU in sows. [ABSTRACT FROM AUTHOR]

Clonazepam causes sedation and psychomotor impairment in people. Due to similarities between people and swine in response to benzodiazepines, clonazepam may represent a viable option to produce mild‐to‐moderate tranquillization in pigs. The objective of this study was to determine the pharmacokinetic profile of a single oral dose (0.5 mg/kg) of clonazepam in eight healthy, growing commercial cross pigs. Serial plasma samples were collected at baseline and up to 96 h after administration. Plasma concentrations were quantified using reverse‐phase high‐performance liquid chromatography, and compartment models were fit to time–concentration data. A one‐compartment first‐order model best fits the data. Maximum plasma concentration was 99.5 ng/mL, and time to maximum concentration was 3.4 h. Elimination half‐life was 7.3 h, mean residence time 7.4 h, and apparent volume of distribution 5.7 L/kg. Achieved plasma concentrations exceeded those associated with psychomotor impairment in people although pharmacodynamic effects have not been investigated in pigs. A simulated oral regimen consisting of 0.35 mg/kg administered every 8 h to pigs would achieve plasma concentrations above 32 ng/mL which are shown to produce psychomotor impairment in people. Further studies to test the clinical efficacy of these dosages in commercial and miniature pigs are warranted. [ABSTRACT FROM AUTHOR]

Gelatin and collagen are two animal-derived ingredients that are widely used in various industries. Both have distinctive physico-chemical characteristic that made them ingredients of interest for many industrial players to be applied as there are vast arrays of usage in the food, cosmetic and biomedical fields. However, the origin of gelatin and collagen poses ethical and religious concerns, especially for Muslims and Jews who have restrictions on food consumption. Porcine by-products are of concern for religious and health reasons, and there is a demand for precise and reliable detection techniques. The limitation of DNA detection is due to extreme environment in food processing which results in low extractability of DNA. Therefore, peptide-based detection using mass spectrometry is required. However, identify the suitable marker is like searching needle in haystacks. Hence, combination of bioinformatics and mass spectrometry is proposed. This study aims to identify the specific peptide biomarkers by employing bioinformatics technique which can be applied to identify gelatin and collagen sources with the aid of mass spectrometry. In these approach, combination of Petunia Trans-Proteomic Pipeline (TPP, version 5.2.0) and sequence alignment ClustalW were applied to facilitate the MS data (LC-QTOF-MS) and peptide identification. As a result, 69 fasta file of protein sequence from both UniProtKB and NCBInr have been collected, 81 collagen peptides sequence and 118 gelatine peptides has been attainable that have the potential to distinguish different species. In conclusion, in silico protein sequence approaches helps to enable rapid screening of proteotypic peptides that can serve as species biomarkers proficiently. [ABSTRACT FROM AUTHOR]

[Objectives] Porcine CD205, as a receptor specific to porcine dendritic cells (DC), played a key role in the antigen presentation. In this study, nanobodies against porcine CD205 CysR-FN Ⅱ protein were screened by phage display technology, which laid a foundation for the research of porcine CD205-targeted antigen presentation. [Methods] CysR-FN Ⅱ protein of porcine CD205 molecule prepared in the previous research was used for the immunization of alpacas. After subcutaneous immunization for 6 times, peripheral blood lymphocytes of alpaca were collected, their total RNA was extracted and cDNA was obtained by reverse transcription. The gene fragments encoding nanobodies were obtained by PCR amplification, and the porcine CD205 phage display nanobody library was constructed. Bio-spannings were performed by phage display technology and monoclonal colonies were randomly selected to screen specific nanobodies against porcine CD205 CysR-FN Ⅱ protein by Phage-ELISA. [Results] After 4 rounds of bio-spanning, a total of 12 porcine CD205 CysR-FN Ⅱ protein specific nanobodies were selected by Phage-ELISA and sequence analysis with their molecular weight about 17×10³. [Conclusions] This study successfully selected and obtained porcine CD205 CysR-FN Ⅱ protein specific nanobodies. [ABSTRACT FROM AUTHOR]

Recently, neurological diseases associated with astroviruses (AstVs) have been reported in pigs, ruminants, minks, and humans. In 2017, neuro-invasive porcine astrovirus (Ni-PAstV) 3 was detected in the central nervous system (CNS) of pigs with encephalomyelitis in Hungary and the USA. In the process of diagnosing domestic pigs exhibiting neurological signs, histopathologic lesions of non-suppurative encephalomyelitis with meningitis, neuronal vacuolation, and gliosis were detected, and PAstV was identified using reverse transcriptase PCR in CNS samples of four pigs in three farms from August to September in 2020, South Korea. Subsequently, the ORF2 region was successfully acquired from three brain samples, facilitating subsequent analysis. Four genotypes of PAstV (PAstV1, 3, 4, and 5) were detected, and coinfection of PAstV with multiple genotypes was observed in brain samples. This is the first study to report Ni-PAstV infection in pigs in South Korea. [ABSTRACT FROM AUTHOR]

Normothermic machine perfusion (NMP) of donor kidneys provides the opportunity to assess and improve organ viability prior to transplantation. This study explored the necessity of an oxygen carrier during NMP and whether the hemoglobin-based oxygen carrier (HBOC-201) is a suitable alternative to red blood cells (RBCs). Porcine kidneys were perfused with a perfusion solution containing either no-oxygen carrier, RBCs, or HBOC-201 for 360 min at 37°C. Renal flow and resistance did not differ significantly between groups. NMP without an oxygen carrier showed lower oxygen consumption with higher lactate and aspartate aminotransferase levels, indicating that the use of an oxygen carrier is necessary for NMP. Cumulative urine production and creatinine clearance in the RBC group were significantly higher than in the HBOC-201 group. Oxygen consumption, injury markers, and histology did not differ significantly between these two groups. However, methemoglobin levels increased to 45% after 360 min in the HBOC-201 group. We conclude that HBOC-201 could be used as an alternative for RBCs, but accumulating methemoglobin levels during our perfusions indicated that HBOC-201 is probably less suitable for prolonged NMP. Perfusion with RBCs, compared to HBOC-201, resulted in more favorable renal function during NMP. [ABSTRACT FROM AUTHOR]

Porcine Teschoviruses (PTVs) are ubiquitous enteric viral pathogens that infect pigs and wild boars worldwide. PTVs have been responsible for causing the severe clinical disease (Teschen disease) to asymptomatic infections. However, to date, limited information is available on large-scale epidemiological data and molecular characterization of PTVs in several countries. In this study, we report epidemiological data on PTVs based on screening of 534 porcine fecal samples from different states of India and a RT-PCR based detection of PTVs shows a percent positivity of 8.24% (44/534). The PTV prevalence varied among different regions of the country with the highest detection rates observed in the state of Karnataka (38.1%). Phylogenetic analysis based on VP1 gene reveals the presence of PTV genotype 6 and 13 along with some unassigned novel genotypes which did not cluster with any of the established PTV genotypes (PTV 1–PTV 13). Indian PTV 6 strains are genetically closest to the Spanish strains (85.7–94.4%) whereas PTV 13 and novel genotype strains were found to be more similar to the Chinese strains (88.1–99.1%). Using recombination detection software, no Indian PTVs found to be recombinant on VP1 gene and selection pressure analysis revealed the purifying selection in the several sites of the VP1 gene of PTVs. The Bayesian analysis of Indian PTVs shows 1.16 × 10–4 substitution/site/year as the mean evolutionary rate. Further, isolation of the novel PTV strains from India and more detailed investigation much needed to know the evolutionary history of PTV strains circulating in porcine populations in India. [ABSTRACT FROM AUTHOR]

Burn injury remains a health problem, specifically in Indonesia. In major burns, xenograft has been proved to be useful as temporary wound coverage. However, some xenografts are not widely available due to cultural, financial and religious backgrounds, or have an unesthetic appearance, such as the scaly appearance of tilapia fish xenograft.Striped catfish (Pangasius hypophthalmus) is a scaleless fish that has abundant type 1 collagen. This study aimed to compare striped catfish skin to commonly used xenograft (Nile tilapia and porcine skin) as xenograft material for burn wound. In this experimental study, nine different skin samples of striped catfish, Nile tilapia and porcine were prepared and histologically examined using hematoxylin-eosin stained samples. Macroscopic and microscopic features of each sample were documented and analysed. The macroscopic skin appearances of striped catfish were hairless and scaleless with black-silver color and moderate thickness. As for microscopic features, the epidermal thickness of striped catfish skin (8.49±1.60 μm) was significantly different to both Nile tilapia (2.18±0.37 μm; p<0.001) and porcine skin (42.22±14.85 μm; p=0.002). The dermal thickness of striped catfish skin (288.46±119.04 μm) was similar to Nile tilapia (210.68±46.62 μm; p=0.783) but differs significantly to porcine skin (1708.44±505.12 μm; p<0.001). The integrity and collagen organization of striped catfish was also similar to tilapia based on semi-quantitative histology scoring system (p>0.05). Striped catfish had potential macroscopic appearance and comparable microscopic features to Nile tilapia; smoother macroscopic appearance, thicker epidermis, and similar dermis thickness. Therefore, we believe it can be potentially used as a xenograft material. Further studies are required to evaluate the effectiveness and feasibility of striped catfish xenograft in burn wound management. [ABSTRACT FROM AUTHOR]

Background: Currently, high polyspermy remains a significant obstacle to achieving optimal efficiency in in vitro fertilization (IVF) and in vitro embryo production (IVP) systems in pigs. Developing strategies that would prevent polyspermy is essential in overcoming this challenge and maximizing the potential of this reproductive biotechnology. Previous results have demonstrated that using boar spermatozoa subjected to a high‐extension and reconcentration procedure and then cryopreserved resulted in significant improvements in IVF/IVP systems with high rates of monospermy and penetration. Objective: The aim of the present study was to unveil the molecular mechanisms that may underlie the changes in fertilization patterns exhibited by highly extended and cryopreserved boar spermatozoa. Materials and methods: To achieve this goal, we used quantitative proteomic analysis (LC‒ESI‒MS/MS SWATH) to identify differentially abundant proteins (DAPs) between highly extended (HE) and conventionally (control; CT) cryopreserved boar spermatozoa. Prior to the analysis, we evaluated the in vitro post‐thawing fertilizing ability of the sperm samples. The results demonstrated a remarkable improvement in monospermy and IVF efficiency when using HE spermatozoa in IVF compared with CT spermatozoa. Results: At the proteomic level, the combination of high‐extension and cryopreservation had a significant impact on the frozen–thawed sperm proteome. A total of 45 proteins (24 downregulated and 21 upregulated) were identified as DAPs (FC > 1 or ≤1; p < 0.05) when compared with CT spermatozoa. Some of these proteins were primarily linked to metabolic processes and the structural composition of sperm cells. The dysregulation of these proteins may have a direct or indirect effect on essential sperm functions and significantly affect spermatozoa–oocyte interaction and, therefore, the sperm fertilization profile under in vitro conditions. While these findings are promising, further research is necessary to comprehend how the disturbance of specific proteins affects sperm fertilization ability. [ABSTRACT FROM AUTHOR]