RESUMEN

The use of direct nucleic acid amplification of pathogens from food matrices has the potential to reduce time to results over DNA extraction-based approaches as well as traditional culture-based approaches. Here we describe protocols for assay design and experiments for direct amplification of foodborne pathogens in food sample matrices using loop-mediated isothermal amplification (LAMP) and polymerase chain reaction (PCR). The examples provided include the detection of Escherichia coli in milk samples and Salmonella in pork meat samples. This protocol includes relevant reagents and methods including obtaining target sequences, assay design, sample processing, and amplification. These methods, though used for specific example matrices, could be applied to many other foodborne pathogens and sample types.

Asunto(s)

ADN Bacteriano , Microbiología de Alimentos , Leche , Técnicas de Amplificación de Ácido Nucleico , Reacción en Cadena de la Polimerasa , Salmonella , Técnicas de Amplificación de Ácido Nucleico/métodos , Microbiología de Alimentos/métodos , Animales , Leche/microbiología , Salmonella/genética , Salmonella/aislamiento & purificación , ADN Bacteriano/genética , ADN Bacteriano/aislamiento & purificación , Reacción en Cadena de la Polimerasa/métodos , Enfermedades Transmitidas por los Alimentos/microbiología , Escherichia coli/genética , Escherichia coli/aislamiento & purificación , Técnicas de Diagnóstico Molecular/métodos , Porcinos

RESUMEN

Fully restoring the lost population of cardiomyocytes and heart function remains the greatest challenge in cardiac repair post myocardial infarction. In this study, a pioneered highly ROS-eliminating hydrogel was designed to enhance miR-19a/b induced cardiomyocyte proliferation by lowering the oxidative stress and continuously releasing miR-19a/b in infarcted myocardium in situ. In vivo lineage tracing revealed that ∼20.47 % of adult cardiomyocytes at the injected sites underwent cell division in MI mice. In MI pig the infarcted size was significantly reduced from 40 % to 18 %, and thereby marked improvement of cardiac function and increased muscle mass. Most importantly, our treatment solved the challenge of animal death--all the treated pigs managed to live until their hearts were harvested at day 50. Therefore, our strategy provides clinical conversion advantages and safety for healing damaged hearts and restoring heart function post MI, which will be a powerful tool to battle cardiovascular diseases in patients.

Asunto(s)

Proliferación Celular , MicroARNs , Infarto del Miocardio , Miocitos Cardíacos , Estrés Oxidativo , Animales , MicroARNs/metabolismo , MicroARNs/genética , Miocitos Cardíacos/metabolismo , Infarto del Miocardio/metabolismo , Infarto del Miocardio/patología , Estrés Oxidativo/efectos de los fármacos , Ratones , Porcinos , Hidrogeles/química , Ratones Endogámicos C57BL , Especies Reactivas de Oxígeno/metabolismo

RESUMEN

Significance: ALA-PpIX and second-window indocyanine green (ICG) have been studied widely for guiding the resection of high-grade gliomas. These agents have different mechanisms of action and uptake characteristics, which can affect their performance as surgical guidance agents. Elucidating these differences in animal models that approach the size and anatomy of the human brain would help guide the use of these agents. Herein, we report on the use of a new pig glioma model and fluorescence cryotomography to evaluate the 3D distributions of both agents throughout the whole brain. Aim: We aim to assess and compare the 3D spatial distributions of ALA-PpIX and second-window ICG in a glioma-bearing pig brain using fluorescence cryotomography. Approach: A glioma was induced in the brain of a transgenic Oncopig via adeno-associated virus delivery of Cre-recombinase plasmids. After tumor induction, the pro-drug 5-ALA and ICG were administered to the animal 3 and 24 h prior to brain harvest, respectively. The harvested brain was imaged using fluorescence cryotomography. The fluorescence distributions of both agents were evaluated in 3D in the whole brain using various spatial distribution and contrast performance metrics. Results: Significant differences in the spatial distributions of both agents were observed. Indocyanine green accumulated within the tumor core, whereas ALA-PpIX appeared more toward the tumor periphery. Both ALA-PpIX and second-window ICG provided elevated tumor-to-background contrast (13 and 23, respectively). Conclusions: This study is the first to demonstrate the use of a new glioma model and large-specimen fluorescence cryotomography to evaluate and compare imaging agent distribution at high resolution in 3D.

Asunto(s)

Neoplasias Encefálicas , Glioma , Imagenología Tridimensional , Verde de Indocianina , Animales , Verde de Indocianina/farmacocinética , Verde de Indocianina/química , Porcinos , Neoplasias Encefálicas/diagnóstico por imagen , Glioma/diagnóstico por imagen , Glioma/patología , Imagenología Tridimensional/métodos , Ácido Aminolevulínico/farmacocinética , Encéfalo/diagnóstico por imagen , Imagen Óptica/métodos , Modelos Animales de Enfermedad

RESUMEN

There are limited biosecurity measures directed at preventing airborne transmission of viruses in swine. The effectiveness of dust mitigation strategies such as oil sprinkling, to decrease risk of airborne virus transmission are unknown. Metagenomics and qPCR for common fecal viruses were used to hunt for a ubiquitous virus to serve as a proxy when evaluating the efficiency of mitigation strategies against airborne viral infectious agents. Air particles were collected from swine buildings using high-volume air samplers. Extracted DNA and RNA were used to perform specific RT-qPCR and qPCR and analyzed by high-throughput sequencing. Porcine astroviruses group 2 were common (from 102 to 105 genomic copies per cubic meter of air or gc/m3, 93% positivity) while no norovirus genogroup II was recovered from air samples. Porcine torque teno sus virus were detected by qPCR in low concentrations (from 101 to 102 gc/m3, 47% positivity). Among the identified viral families by metagenomics analysis, Herelleviridae, Microviridae, Myoviridae, Podoviridae, and Siphoviridae were dominant. The phage vB_AviM_AVP of Aerococcus was present in all air samples and a newly designed qPCR revealed between 101 and 105 gc/m3 among the samples taken for the present study (97% positivity) and banked samples from 5- and 15-year old studies (89% positivity). According to the present study, both the porcine astrovirus group 2 and the phage vB_AviM_AVP of Aerococcus could be proxy for airborne viruses of swine buildings.

Asunto(s)

Microbiología del Aire , Monitoreo del Ambiente , Metagenómica , Animales , Porcinos , Monitoreo del Ambiente/métodos , Aerosoles/análisis , Virus/aislamiento & purificación , Contaminación del Aire Interior/análisis , Vivienda para Animales

RESUMEN

The land application of livestock manure has been widely acknowledged as a beneficial approach for nutrient recycling and environmental protection. However, the impact of residual antibiotics, a common contaminant of manure, on the degradation of organic compounds and nutrient release in Eutric Regosol is not well understood. Here, we studied, how oxytetracycline (OTC) and ciprofloxacin (CIP) affect the decomposition, microbial community structure, extracellular enzyme activities and nutrient release from cattle and pig manure using litterbag incubation experiments. Results showed that OTC and CIP greatly inhibited livestock manure decomposition, causing a decreased rate of carbon (28%-87%), nitrogen (15%-44%) and phosphorus (26%-43%) release. The relative abundance of gram-negative (G-) bacteria was reduced by 4.0%-13% while fungi increased by 7.0%-71% during a 28-day incubation period. Co-occurrence network analysis showed that antibiotic exposure disrupted microbial interactions, particularly among G- bacteria, G+ bacteria, and actinomycetes. These changes in microbial community structure and function resulted in decreased activity of urease, ß-1,4-N-acetyl-glucosaminidase, alkaline protease, chitinase, and catalase, causing reduced decomposition and nutrient release in cattle and pig manures. These findings advance our understanding of decomposition and nutrient recycling from manure-contaminated antibiotics, which will help facilitate sustainable agricultural production and soil carbon sequestration.

Asunto(s)

Antibacterianos , Ganado , Estiércol , Microbiología del Suelo , Animales , Suelo/química , Secuestro de Carbono , Carbono/metabolismo , Fósforo , Reciclaje , Contaminantes del Suelo/metabolismo , Bovinos , Porcinos , Nitrógeno/análisis , Oxitetraciclina

RESUMEN

Oxidative stress is a major pathogenic factor in many intestinal diseases, such as inflammatory bowel disease (IBD) and colorectal cancer (CRC). The Nrf2 signaling pathway and mitophagy can reduce reactive oxygen species (ROS) and alleviate oxidative stress, but their relationship is unclear. Hydroxytyrosol (HT), a polyphenolic compound abundant in olive oil, has strong antioxidant activity and may help treat these diseases. We used pigs as a model to investigate HT's effect on intestinal oxidative damage and its mechanisms. Diquat (DQ) induced oxidative stress and impaired intestinal barrier function, which HT mitigated. Mechanistic studies in IPEC-J2 cells showed that HT protected against oxidative damage by activating the PI3K/Akt-Nrf2 signaling pathway and promoting mitophagy. Our study highlighted the synergistic relationship between Nrf2 and mitophagy in mediating HT's antioxidant effects. Inhibition studies confirmed that disrupting either pathway compromised HT's protective effects. Maintaining redox balance through Nrf2 and mitophagy is important for eliminating excess ROS. Nrf2 increases antioxidant enzymes to clear existing ROS, while mitophagy removes damaged mitochondria and reduces ROS generation. This study demonstrates that these pathways collaboratively modulate the antioxidant effects of HT, with neither being dispensable. Targeting Nrf2 and mitophagy could be a promising strategy for treating oxidative stress-related intestinal diseases, with HT as a potential treatment.

Asunto(s)

Mitofagia , Factor 2 Relacionado con NF-E2 , Estrés Oxidativo , Alcohol Feniletílico , Fosfatidilinositol 3-Quinasas , Proteínas Proto-Oncogénicas c-akt , Especies Reactivas de Oxígeno , Transducción de Señal , Alcohol Feniletílico/análogos & derivados , Alcohol Feniletílico/farmacología , Factor 2 Relacionado con NF-E2/metabolismo , Animales , Estrés Oxidativo/efectos de los fármacos , Mitofagia/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Proteínas Proto-Oncogénicas c-akt/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Porcinos , Antioxidantes/farmacología , Intestinos/efectos de los fármacos , Línea Celular

RESUMEN

Introduction: Animal models that closely mimic human burn wound healing processes are essential for developing effective burn wound treatments. Pigs are useful animal models for studying burn wound healing. From their extensive literature review, Andrews and Cuttle (2017) reported mean temperature and exposure time values. This study was done to provide initial burn depth for another experiment comparing two burn wound treatments. The secondary goal was to validate a systematic review on porcine burn model standardization. Material and Methods: Six four-week-old Large White x Landrace gilts were housed in a closed structure for 10 days to acclimatize. The procedures were performed under general anesthesia. A round 2.5 cm copper plate welded to an aluminum rod with a wooden handle caused the injuries. The burning device was used to reach a contact temperature of 110 C on the pig's skin. The objective was to create a superficial partial thickness (SPT) burn for 10 seconds (Group 10s) and a deep partial thickness (DPT) burn for 20 seconds (Group 20s) using a plate heated at 110ºC. No stabilizer or pressure controller was used. Wounds were conclusively dressed and harvested 24 hours later. The usual hematoxylin-eosin protocol was used to cut and stain 4-micron sections. Results: A significant difference (p 0.01) was observed in dermis involvement, with a mean of 85.61 % (95% CI= 80.62 to 90.61) for group 10s and 123.71% (95% CI= 114.91 to 132.50) for group 20s. An exposure time of 20 seconds increased dermis depth-related total collagen denaturation by almost 50% compared to 10 seconds. Conclusions: In conclusion, our experiment produced DPT burns in 10 seconds and FT burns in 20 seconds without a pressure application device.

Asunto(s)

Quemaduras , Modelos Animales de Enfermedad , Cicatrización de Heridas , Animales , Quemaduras/terapia , Porcinos , Factores de Tiempo , Femenino , Humanos

RESUMEN

Although urine bladder transplantation is currently being conducted, the procedure is an incompletely resolved problem in clinical transplantology. A small number of en bloc bladder and kidney transplants from pediatric donors to adult recipients in humans have been reported. A small number of bladder transplants with and without combinations with kidneys have also been performed in experiments on different animal models. Here, we aimed to highlight the experiences of various scientists in bladder transplantation in humans and animals. We also presented our small experience in conducting transplant of 1 kidney, ureters, and a segment of the bladder in an experiment on pigs in 2023 (5 cases), which is a promising direction for further successful development of this technology in humans. In 2024, we plan to conduct another 10 transplants of a single block ofthe kidney and bladderin pigs, results of which will be published after the completion of the experimental work.

Asunto(s)

Trasplante de Riñón , Vejiga Urinaria , Trasplante de Riñón/efectos adversos , Animales , Humanos , Vejiga Urinaria/cirugía , Resultado del Tratamiento , Porcinos , Adulto , Niño , Donantes de Tejidos/provisión & distribución , Supervivencia de Injerto , Modelos Animales

RESUMEN

Incorporating 5-aminosalicylic acid (5-ASA) into a colon-specific carrier is crucial for treating inflammatory bowel diseases (IBD), as it enhances therapeutic efficacy, targets the affected regions directly, and minimizes side effects. This study evaluated the impact of incorporating cellulose nanofibers (CNF) on the in vitro and in vivo biological performance of retrograded starch/pectin (RS/P) microparticles (MPs) containing 5-ASA. Using Fourier Transform Infrared (FTIR) Spectroscopy, shifts in the spectra of retrograded samples containing CNF were observed with increasing CNF proportions, suggesting the establishment of new supramolecular interactions. Liquid absorption exhibited pH-dependent behaviors, with reduced absorption in simulated gastric fluid (∼269 %) and increased absorption in simulated colonic fluid (∼662 %). Increasing CNF concentrations enhanced mucoadhesion in porcine colonic sections, with a maximum force of 3.4 N at 50 % CNF. Caco-2 cell viability tests showed biocompatibility across all tested concentrations (0.0625-2.0000 mg/mL). Evaluation of intestinal permeability in Caco-2 cell monolayers demonstrated up to a tenfold increase in 5-ASA permeation, ranging from 29 % to 48 %. An in vivo study using Galleria mellonella larvae, with inflammation induced by LPS, showed reduction of inflammation. Given the scalability of spray-drying, these findings suggest the potential of CNF-incorporated RS/P microparticles for targeted 5-ASA delivery in IBD.

Asunto(s)

Enfermedades Inflamatorias del Intestino , Mesalamina , Nanofibras , Pectinas , Almidón , Mesalamina/química , Mesalamina/farmacología , Mesalamina/administración & dosificación , Animales , Células CACO-2 , Humanos , Enfermedades Inflamatorias del Intestino/tratamiento farmacológico , Nanofibras/química , Nanofibras/toxicidad , Porcinos , Pectinas/química , Almidón/química , Portadores de Fármacos/química , Sistemas de Liberación de Medicamentos , Supervivencia Celular/efectos de los fármacos , Colon/efectos de los fármacos , Colon/metabolismo , Colon/patología , Antiinflamatorios no Esteroideos/química , Antiinflamatorios no Esteroideos/farmacología , Antiinflamatorios no Esteroideos/administración & dosificación

RESUMEN

Domestic pigs serve as significant hosts and reservoirs for multiple parasite species, some specific to pigs and many others of zoonotic importance. This study aimed to assess the prevalence of intestinal parasites in pigs within a rural area in northeastern Argentina. We also examined demographic information, breeding conditions, and exposure factors associated with parasite presence. Pig feces were subjected to coprological examination through flotation and sedimentation techniques. Modified Ziehl-Neelsen technique was employed to examine oocysts of Cryptosporidium spp. In total, 29 family farms with pig pens were analyzed, and 42 stool samples were collected from pigs on these farms. At the farm level, the presence of at least one parasite species was recorded in 27 rural houses (93.1%). We found that 90.4% of pigs were parasitized, with a specific parasitoses of 10 species, with a maximum of six species in a single host. The most prevalent protozoa were Entamoeba spp. (57.1%) and Blastocystis sp. (45.2%), followed by Iodamoeba butschlii (33.3%), Neobalantidium coli (21.4%), coccidia (14.2%), Cryptosporidium spp. (9.5%) and Giardia spp. (2.3%). The most prevalent helminths were Strongylidae eggs (52.3%), Ascaris spp. (14.2%) and Trichuris spp. (2.3%). We advocate for an urgent need to implement a comprehensive prophylaxis program prioritizing general hygiene practices such as regular cleaning, removal of fecal material and renewal of drinking water. Additionally, vaccination and deworming protocols should be implemented. Furthermore, this study highlights the necessity for molecular-level evaluations to detect potential zoonotic genotypes of the identified protozoa.

Asunto(s)

Granjas , Heces , Parasitosis Intestinales , Enfermedades de los Porcinos , Animales , Argentina/epidemiología , Enfermedades de los Porcinos/epidemiología , Enfermedades de los Porcinos/parasitología , Prevalencia , Porcinos , Parasitosis Intestinales/epidemiología , Parasitosis Intestinales/veterinaria , Parasitosis Intestinales/parasitología , Heces/parasitología , Femenino , Sus scrofa/parasitología , Masculino

RESUMEN

The lung comprises multiple components including the parenchyma, airways, and visceral pleura, where each constituent displays specific material properties that together govern the whole organ's properties. The structural and mechanical complexity of the lung has historically undermined its comprehensive characterization, especially compared to other biological organs, such as the heart or bones. This knowledge void is particularly remarkable when considering that pulmonary disease is one of the leading causes of morbidity and mortality across the globe. Establishing the mechanical properties of the lung is central to formulating a baseline understanding of its operation, which can facilitate investigations of diseased states and how the lung will potentially respond to clinical interventions. Here, we present established and widely accepted experimental protocols for pulmonary material quantification, specifying how to extract, prepare, and test each type of lung constituent under planar biaxial tensile loading to investigate the mechanical properties, such as physiological stress-strain profiles, anisotropy, and viscoelasticity. These methods are presented across an array of commonly studied species (murine, rat, and porcine). Additionally, we highlight how such material properties may inform the construction of an inverse finite element model, which is central to implementing predictive computational tools for accurate disease diagnostics and optimized medical treatments. These presented methodologies are aimed at supporting research advancements in the field of pulmonary biomechanics and to help inaugurate future novel studies. © 2024 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: General procedures in lung biaxial testing Alternate Protocol 1: Parenchymal-specific preparation and loading procedures Alternate Protocol 2: Airway-specific preparation and loading procedures Alternate Protocol 3: Visceral pleura-specific preparation and loading procedures Basic Protocol 2: Computational analysis.

Asunto(s)

Pulmón , Animales , Pulmón/fisiología , Ratas , Fenómenos Biomecánicos , Porcinos , Ratones , Análisis de Elementos Finitos , Estrés Mecánico

RESUMEN

Polyphenolic compounds are common constituents of human and animal diets and undergo extensive metabolism by the gut microbiota before entering circulation. In order to compare the transformations of polyphenols from yerba mate, rosemary, and green tea extracts in the gastrointestinal tract, simulated gastrointestinal digestion coupled with colonic fermentation were used. For enhancing the comparative character of the investigation, colonic fermentation was performed with human, pig and rat intestinal microbiota. Chemical analysis was performed using a HPLC system coupled to a diode-array detector and mass spectrometer. Gastrointestinal digestion diminished the total amount of phenolics in the rosemary and green tea extracts by 27.5 and 59.2 %, respectively. These reductions occurred mainly at the expense of the major constituents of these extracts, namely rosmarinic acid (-45.7 %) and epigalocatechin gallate (-60.6 %). The yerba mate extract was practically not affected in terms of total phenolics, but several conversions and isomerizations occurred (e.g., 30 % of trans-3-O-caffeoylquinic acid was converted into the cis form). The polyphenolics of the yerba mate extract were also the least decomposed by the microbiota of all three species, especially in the case of the human one (-10.8 %). In contrast, the human microbiota transformed the polyphenolics of the rosemary and green extracts by 95.9 and 88.2 %, respectively. The yerba mate-extract had its contents in cis 3-O-caffeoylquinic acid diminished by 78 % by the human microbiota relative to the gastrointestinal digestion, but the content of 5-O-caffeoylquinic acid (also a chlorogenic acid), was increased by 22.2 %. The latter phenomenon did not occur with the rat and pig microbiota. The pronounced interspecies differences indicate the need for considerable caution when translating the results of experiments on the effects of polyphenolics performed in rats, or even pigs, to humans.

Asunto(s)

Colon , Depsidos , Digestión , Fermentación , Ilex paraguariensis , Extractos Vegetales , Polifenoles , Ácido Rosmarínico , Rosmarinus , Animales , Humanos , Extractos Vegetales/metabolismo , Rosmarinus/química , Ratas , Ilex paraguariensis/química , Porcinos , Depsidos/metabolismo , Depsidos/análisis , Polifenoles/metabolismo , Polifenoles/análisis , Colon/metabolismo , Colon/microbiología , Masculino , Cinamatos/metabolismo , Cinamatos/análisis , Microbioma Gastrointestinal , Té/química , Ácido Quínico/análogos & derivados , Ácido Quínico/metabolismo , Ácido Quínico/análisis , Catequina/análogos & derivados , Catequina/metabolismo , Catequina/análisis , Cromatografía Líquida de Alta Presión , Camellia sinensis/química

RESUMEN

A novel gradient-temperature heating regime was proposed to improve the texture of braised pork. Compared with one-stage pressure heat treatment of around 107 °C, the gradient-temperature heat regime of preheating at 60 °C, followed by a slow increase of temperature to 107 °C and simmering at 97 °C increased the retention of immobilized water and reduced the shear force of meat. In this cooking regime, preheating treatment at 50-60 °C could promote the dissociation of thin and thick myofilaments, which contributed to a weakened shrinkage of myofibrils during the subsequent high temperature heating process. Pressure-heating treatment with a slow increasing temperature and the medium-temperature simmering significantly reduced (p < 0.05) the oxidation of sulfhydryl groups and the loss of α-helical, which weakened the excessive aggregation of protein and promoted the formation of myofibril network. Both the weakened shrinkage and the formation of myofibril network during gradient-temperature heating contributed to the decreased shear force and an increased immobilized water. Hence, the reduction of the oxidation and aggregation of the proteins is the key to improve the tenderness of the braised meat.

Asunto(s)

Culinaria , Calor , Animales , Culinaria/métodos , Porcinos , Miofibrillas/química , Oxidación-Reducción , Agua/química , Carne de Cerdo/análisis , Resistencia al Corte , Manipulación de Alimentos/métodos

RESUMEN

Boar semen production plays a pivotal role in modern swine breeding programmes, influencing the genetic progress and overall efficiency of the pork industry. This review explores the current challenges and emerging trends in liquid-preserved boar semen production, addressing key issues that impact the quality and quantity of boar semen. Advances in new reproductive technologies, boar selection, housing, semen processing, storage and transport, and the need for sustainable practices including the use of artificial intelligence are discussed to provide a comprehensive overview of the field.

Asunto(s)

Preservación de Semen , Semen , Animales , Masculino , Preservación de Semen/veterinaria , Preservación de Semen/métodos , Porcinos , Semen/fisiología , Cruzamiento/métodos , Inseminación Artificial/veterinaria , Criopreservación/veterinaria , Criopreservación/métodos , Análisis de Semen/veterinaria , Sus scrofa/fisiología

RESUMEN

The chronic airway infections with Pseudomonas aeruginosa are the major co-morbidity in people with cystic fibrosis (CF). Within CF lungs, P. aeruginosa persists in the conducting airways together with human mucins as the most abundant structural component of its microenvironment. We investigated the adhesion of 41 serial CF airway P. aeruginosa isolates to airway mucin preparations from CF sputa. Mucins and bacteria were retrieved from five modulator-naïve patients with advanced CF lung disease. The P. aeruginosa isolates from CF airways and non-CF reference strains showed a strain-specific signature in their adhesion to ovine, porcine and bovine submaxillary mucins and CF airway mucins ranging from no or low to moderate and strong binding. Serial CF clonal isolates and colony morphotypes from the same sputum sample were as heterogeneous in their affinity to mucin as representatives of other clones thus making 'mucin binding' one of the most variable intraclonal phenotypic traits of P. aeruginosa known to date. Most P. aeruginosa CF airway isolates did not adhere more strongly to CF airway mucins than to plastic surfaces. The strong binders, however, exhibited a strain-specific affinity gradient to O-glycans, CF airway and mammalian submaxillary mucins.

Asunto(s)

Adhesión Bacteriana , Fibrosis Quística , Mucinas , Infecciones por Pseudomonas , Pseudomonas aeruginosa , Esputo , Fibrosis Quística/microbiología , Pseudomonas aeruginosa/metabolismo , Pseudomonas aeruginosa/aislamiento & purificación , Mucinas/metabolismo , Humanos , Animales , Esputo/microbiología , Infecciones por Pseudomonas/microbiología , Porcinos , Bovinos , Ovinos

RESUMEN

BACKGROUND: Prosthetic joint infections (PJI) are recalcitrant, hard-to-treat infections and severe complications of joint arthroplasty. Therefore, there is a need to develop new effective treatment strategies, and animal models of high clinical relevance are needed. This study aimed to develop a detailed surgical protocol for hip hemiarthroplasty in Göttingen minipigs and a thorough post-mortem sampling protocol to pave the way for creating a minipig PJI model. METHODS: Three adult female Göttingen minipigs underwent surgery with insertion of a hip hemiarthroplasty, using the anterior approach to the hip joint. After surgery the minipigs were followed closely with daily clinical evaluation and gait scoring. Comprehensive post-mortem analyses were performed with evaluation of macroscopic lesions, microbiology, synovial fluid analysis and histology. RESULTS: The study resulted in the first Göttingen minipig with hip hemiarthroplasty and identified several points of awareness when inserting a hip prosthesis in minipigs, especially the high risk of joint dislocation. A spontaneous PJI occurred in one of the minipigs, revealing an impaired ability of the immune cells to reach the bacteria at the bone-prosthesis interface. CONCLUSION: The present study provides a detailed description of surgical technique and post-mortem sampling and validates the suitability of the hip hemiarthroplasty minipig model for future experimental modeling of PJI.

Asunto(s)

Artroplastia de Reemplazo de Cadera , Hemiartroplastia , Infecciones Relacionadas con Prótesis , Porcinos Enanos , Animales , Porcinos , Hemiartroplastia/métodos , Hemiartroplastia/efectos adversos , Femenino , Artroplastia de Reemplazo de Cadera/métodos , Artroplastia de Reemplazo de Cadera/efectos adversos , Infecciones Relacionadas con Prótesis/etiología , Modelos Animales de Enfermedad , Prótesis de Cadera/efectos adversos

RESUMEN

Intratumoral injections have the potential for enhanced cancer treatment efficacy while reducing costs and systemic exposure. However, intratumoral drug injections can result in substantial off-target leakage and are invisible under standard imaging modalities like ultrasound (US) and x-ray. A thermosensitive poloxamer-based gel for drug delivery was developed that is visible using x-ray imaging (computed tomography (CT), cone beam CT, fluoroscopy), as well as using US by means of integrating perfluorobutane-filled microbubbles (MBs). MBs content was optimized using tissue mimicking phantoms and ex vivo bovine livers. Gel formulations less than 1% MBs provided gel depositions that were clearly identifiable on US and distinguishable from tissue background and with minimal acoustic artifacts. The cross-sectional areas of gel depositions obtained with US and CT imaging were similar in studies using ex vivo bovine liver and postmortem in situ swine liver. The gel formulation enhanced multimodal image-guided navigation, enabling fusion of ultrasound and x-ray/CT imaging, which may enhance targeting, definition of spatial delivery, and overlap of tumor and gel. Although speculative, such a paradigm for intratumoral drug delivery might streamline clinical workflows, reduce radiation exposure by reliance on US, and boost the precision and accuracy of drug delivery targeting during procedures. Imageable gels may also provide enhanced temporal and spatial control of intratumoral conformal drug delivery.

Asunto(s)

Sistemas de Liberación de Medicamentos , Hidrogeles , Hígado , Poloxámero , Ultrasonografía , Poloxámero/química , Animales , Hidrogeles/química , Hígado/diagnóstico por imagen , Hígado/metabolismo , Bovinos , Ultrasonografía/métodos , Sistemas de Liberación de Medicamentos/métodos , Microburbujas , Porcinos , Fantasmas de Imagen , Tomografía Computarizada por Rayos X/métodos , Tomografía Computarizada de Haz Cónico/métodos

RESUMEN

BACKGROUND: In June 2019, a patient presented with persistent fever and multiple organ dysfunction after a tick bite at a wetland park in Inner Mongolia. Next-generation sequencing in this patient revealed an infection with a previously unknown orthonairovirus, which we designated Wetland virus (WELV). METHODS: We conducted active hospital-based surveillance to determine the prevalence of WELV infection among febrile patients with a history of tick bites. Epidemiologic investigation was performed. The virus was isolated, and its infectivity and pathogenicity were investigated in animal models. RESULTS: WELV is a member of the orthonairovirus genus in the Nairoviridae family and is most closely related to the tickborne Hazara orthonairovirus genogroup. Acute WELV infection was identified in 17 patients from Inner Mongolia, Heilongjiang, Jilin, and Liaoning, China, by means of reverse-transcriptase-polymerase-chain-reaction assay. These patients presented with nonspecific symptoms, including fever, dizziness, headache, malaise, myalgia, arthritis, and back pain and less frequently with petechiae and localized lymphadenopathy. One patient had neurologic symptoms. Common laboratory findings were leukopenia, thrombocytopenia, and elevated d-dimer and lactate dehydrogenase levels. Serologic assessment of convalescent-stage samples obtained from 8 patients showed WELV-specific antibody titers that were 4 times as high as those in acute-phase samples. WELV RNA was detected in five tick species and in sheep, horses, pigs, and Transbaikal zokors (Myospalax psilurus) sampled in northeastern China. The virus that was isolated from the index patient and ticks showed cytopathic effects in human umbilical-vein endothelial cells. Intraperitoneal injection of the virus resulted in lethal infections in BALB/c, C57BL/6, and Kunming mice. The Haemaphysalis concinna tick is a possible vector that can transovarially transmit WELV. CONCLUSIONS: A newly discovered orthonairovirus was identified and shown to be associated with human febrile illnesses in northeastern China. (Funded by the National Natural Science Foundation of China and the Chinese Academy of Medical Sciences Innovation Fund for Medical Sciences.).

Asunto(s)

Fiebre , Nairovirus , Mordeduras de Garrapatas , Adulto , Anciano , Animales , Femenino , Humanos , Masculino , Ratones , Persona de Mediana Edad , Adulto Joven , Anticuerpos Antivirales/sangre , China/epidemiología , Fiebre/diagnóstico , Fiebre/epidemiología , Fiebre/virología , Nairovirus/genética , Nairovirus/aislamiento & purificación , Nairovirus/patogenicidad , Filogenia , Mordeduras de Garrapatas/complicaciones , Mordeduras de Garrapatas/virología , Prevalencia , Modelos Animales de Enfermedad , Ovinos , Caballos , Porcinos , Lactante , Preescolar , Niño , Adolescente , Anciano de 80 o más Años

RESUMEN

Pro-inflammatory cytokines in muscle play a pivotal role in physiological responses and in the pathophysiology of inflammatory disease and muscle atrophy. Lactobacillus delbrueckii (LD), as a kind of probiotics, has inhibitory effects on pro-inflammatory cytokines associated with various inflammatory diseases. This study was conducted to explore the effect of dietary LD on the lipopolysaccharide (LPS)-induced muscle inflammation and atrophy in piglets and to elucidate the underlying mechanism. A total of 36 weaned piglets (Duroc × Landrace × Large Yorkshire) were allotted into three groups with six replicates (pens) of two piglets: (1) Nonchallenged control; (2) LPS-challenged (LPS); (3) 0.2% LD diet and LPS-challenged (LD+LPS). On d 29, the piglets were injected intraperitoneally with LPS or sterilized saline, respectively. All piglets were slaughtered at 4 h after LPS or saline injection, the blood and muscle samples were collected for further analysis. Our results showed that dietary supplementation of LD significantly attenuated LPS-induced production of pro-inflammatory cytokines IL-6 and TNF-α in both serum and muscle of the piglets. Concomitantly, pretreating the piglets with LD also clearly inhibited LPS-induced nuclear translocation of NF-κB p65 subunits in the muscle, which correlated with the anti-inflammatory effects of LD on the muscle of piglets. Meanwhile, LPS-induced muscle atrophy, indicated by a higher expression of muscle atrophy F-box, muscle RING finger protein (MuRF1), forkhead box O 1, and autophagy-related protein 5 (ATG5) at the transcriptional level, whereas pretreatment with LD led to inhibition of these upregulations, particularly genes for MuRF1 and ATG5. Moreover, LPS-induced mRNA expression of endoplasmic reticulum stress markers, such as eukaryotic translational initiation factor 2α (eIF-2α) was suppressed by pretreatment with LD, which was accompanied by a decrease in the protein expression levels of IRE1α and GRP78. Additionally, LD significantly prevented muscle cell apoptotic death induced by LPS. Taken together, our data indicate that the anti-inflammatory effect of LD supply on muscle atrophy of piglets could be likely regulated by inhibiting the secretion of pro-inflammatory cytokines through the inactivation of the ER stress/NF-κB singling pathway, along with the reduction in protein degradation.

Asunto(s)

Estrés del Retículo Endoplásmico , Lactobacillus delbrueckii , Lipopolisacáridos , Atrofia Muscular , Animales , Lipopolisacáridos/toxicidad , Porcinos , Estrés del Retículo Endoplásmico/efectos de los fármacos , Atrofia Muscular/inducido químicamente , Atrofia Muscular/metabolismo , Atrofia Muscular/prevención & control , Atrofia Muscular/patología , Destete , Proteolisis , Probióticos/farmacología , Inflamación/metabolismo , Miositis/inducido químicamente , Miositis/metabolismo , Miositis/patología , Citocinas/metabolismo , Músculo Esquelético/metabolismo , Músculo Esquelético/patología , Músculo Esquelético/efectos de los fármacos

RESUMEN

Toxoplasma gondii, a common protozoan parasite, poses significant public health risks due to its potential to cause toxoplasmosis in humans and can be contracted from pigs, which are considered its critical intermediate host. The aim of this study is to evaluate the prevalence of T. gondii in slaughtered pigs for human consumption, emphasizing the zoonotic implications and the need for improved biosecurity and monitoring practices in pig farming. A total of 1,526 pig samples (1,051 whole blood samples and 384 lung tissue samples from the local slaughterhouse and 91 aborted fetus samples from local farms) were collected throughout the whole country of Korea in 2020. Among them, 6 (0.4%) were found to be infected with T. gondii by nested PCR. When compared by sample type, the prevalence of T. gondii was significantly higher in the aborted fetus samples (2.2%, 2/91) than in the blood (0.3%, 3/1,051) and lung tissue samples (0.3%, 1/384). The B1 gene sequence of T. gondii was similar (97.9-99.8%) to that of the other T. gondii isolates. This study represents the first molecular genotyping survey of T. gondii in the lung tissue of fattening pigs and aborted fetuses in Korea. Our findings indicated the importance of adopting preventive measures including the implementation of rigorous farm hygiene protocols and the promotion of public awareness about the risks of consuming undercooked pork. By addressing the gaps in current control strategies and encouraging the One Health approach, this study contributes to the development of more effective strategies to mitigate the transmission of T. gondii from pigs to humans, ultimately safeguarding public health.

Asunto(s)

Genotipo , Enfermedades de los Porcinos , Toxoplasma , Toxoplasmosis Animal , Animales , Toxoplasma/genética , Toxoplasma/aislamiento & purificación , República de Corea/epidemiología , Porcinos , Toxoplasmosis Animal/epidemiología , Toxoplasmosis Animal/parasitología , Enfermedades de los Porcinos/parasitología , Enfermedades de los Porcinos/epidemiología , Enfermedades de los Porcinos/transmisión , Prevalencia , Mataderos , Pulmón/parasitología , Reacción en Cadena de la Polimerasa , Humanos , ADN Protozoario/genética , Feto Abortado/parasitología