ABSTRACT
The mortality of birds resulting from collisions and electrocutions with overhead lines, such as power lines and phone lines, among others, has been implicated in the decline of various avian species globally. Specifically, overhead line collisions pose a significant threat to the conservation of the Canarian houbara bustard (Chlamydotis undulata fuertaventurae), an endangered subspecies endemic to the Canary Islands. This study centers on the postmortem findings of Canarian houbara bustards that have collided with overhead lines, providing insights into the post-collision outcomes for these birds. A complete standardized necropsy of nine Canarian houbara bustards revealed that trauma was the cause of death in all cases. The most notable gross lesions associated with trauma included bone fractures, soft tissue lacerations, hemorrhages, luxations, and hemocoelom. The inguinal area, chest, and wings were the body regions more frequently affected. A histopathology, immunohistochemistry, and entomology analysis confirmed that numerous birds survived the initial trauma. We concluded that when a houbara bustard collides with an overhead line, it frequently survives the initial trauma, with a survival time ranging from minutes to hours. The histopathology, immunohistochemistry, or entomologic analysis may be helpful to approximate the timing interval between trauma and death.
ABSTRACT
During cell fate transitions, cells remodel their transcriptome, chromatin, and epigenome; however, it has been difficult to determine the temporal dynamics and cause-effect relationship between these changes at the single-cell level. Here, we employ the heterokaryon-mediated reprogramming system as a single-cell model to dissect key temporal events during early stages of pluripotency conversion using super-resolution imaging. We reveal that, following heterokaryon formation, the somatic nucleus undergoes global chromatin decompaction and removal of repressive histone modifications H3K9me3 and H3K27me3 without acquisition of active modifications H3K4me3 and H3K9ac. The pluripotency gene OCT4 (POU5F1) shows nascent and mature RNA transcription within the first 24 h after cell fusion without requiring an initial open chromatin configuration at its locus. NANOG, conversely, has significant nascent RNA transcription only at 48 h after cell fusion but, strikingly, exhibits genomic reopening early on. These findings suggest that the temporal relationship between chromatin compaction and gene activation during cellular reprogramming is gene context dependent.
Subject(s)
Cellular Reprogramming , Chromatin Assembly and Disassembly , Histones , Humans , Cellular Reprogramming/genetics , Histones/metabolism , Single-Cell Analysis , Transcriptional Activation , Octamer Transcription Factor-3/metabolism , Octamer Transcription Factor-3/genetics , Chromatin/metabolism , Nanog Homeobox Protein/metabolism , Nanog Homeobox Protein/genetics , Induced Pluripotent Stem Cells/metabolism , Induced Pluripotent Stem Cells/cytologyABSTRACT
Slc4a genes encode various types of transporters, including Na+-HCO3- cotransporters, Cl-/HCO3- exchangers, or Na+-driven Cl-/HCO3- exchangers. Previous research has revealed that Slc4a9 (Ae4) functions as a Cl-/HCO3- exchanger, which can be driven by either Na+ or K+, prompting investigation into whether other Slc4a members facilitate cation-dependent anion transport. In the present study, we show that either Na+ or K+ drive Cl-/HCO3- exchanger activity in cells overexpressing Slc4a8 or Slc4a10. Further characterization of cation-driven Cl-/HCO3- exchange demonstrated that Slc4a8 and Slc4a10 also mediate Cl- and HCO3--dependent K+ transport. Full-atom molecular dynamics simulation on the recently solved structure of Slc4a8 supports the coordination of K+ at the Na+ binding site in S1. Sequence analysis shows that the critical residues coordinating monovalent cations are conserved among mouse Slc4a8 and Slc4a10 proteins. Together, our results suggest that Slc4a8 and Slc4a10 might transport K+ in the same direction as HCO3- ions in a similar fashion to that described for Na+ transport in the rat Slc4a8 structure.
Subject(s)
Potassium , Sodium-Bicarbonate Symporters , Animals , Mice , Bicarbonates/metabolism , Binding Sites , Chloride-Bicarbonate Antiporters/metabolism , Chloride-Bicarbonate Antiporters/genetics , Chlorides/metabolism , Ion Transport , Molecular Dynamics Simulation , Potassium/metabolism , Sodium/metabolism , Sodium-Bicarbonate Symporters/metabolism , Sodium-Bicarbonate Symporters/geneticsABSTRACT
This study explores Neutrophil Extracellular Trap (NET) formation in equine neutrophils, which is crucial for eliminating infections and is implicated in various equine inflammatory diseases. We investigated the molecular pathways involved in NET release by equine neutrophils in response to stimuli. We use PMA, A23187, LPS, PAF, OZ, and cytokines, observing NET release in response to PMA, PAF, and A23187. In contrast, LPS, OZ, and the cytokines tested did not induce DNA release or did not consistently induce citrullination of histone 4. Peptidyl-arginine deiminase inhibition completely halted NET release, while NADPH oxidase and mitochondrial reactive oxygen species only played a role in PMA-induced NETs. Neutrophil elastase inhibition modestly affected PAF-induced NET liberation but not in PMA or A23187-induced NET, while myeloperoxidase did not contribute to NET release. We expect to provide a foundation for future investigations into the role of NETs in equine health and disease and the search for potential therapeutic targets.
Subject(s)
Extracellular Traps , Neutrophils , Animals , Horses , Neutrophils/metabolism , Extracellular Traps/metabolism , Calcimycin/metabolism , Lipopolysaccharides/metabolism , Cytokines/metabolism , Reactive Oxygen Species/metabolismABSTRACT
This study aimed to determine the performance of the averaged parasympathetic tone activity (PTAm) and its dynamic variation (ΔPTA) to assess intraoperative nociception in relation to heart rate (HR) and direct mean arterial pressure (MAP) in dogs undergoing laparoscopic ovariectomy. This prospective, observational, clinical study included 32 bitches. The PTAm, HR, MAP, and bispectral index (BIS) were assessed before (pre-stimulus), as well as 1 min and 2 min after, four surgical stimuli: insufflation, introduction of trocars, and removal of the left and right ovaries. A two-way ANOVA was performed to compare PTAm, HR, MAP, and BIS data across surgical stimuli. A ≥ 20% drop in PTAm or a ≥ 20% increase in HR and/or MAP regarding the pre-stimulus values was considered a PTAm-drop and/or a hemodynamic response, respectively. The performance of PTAm pre-stimulus, PTAm 1 min, and ΔPTA in predicting the hemodynamic response was assessed by calculation of the area under the receiver operating characteristic (ROC) curve. At insufflation, PTAm decreased after 1 (p = 0.010) and 2 (p = 0.045)min, and ΔPTA was different (p = 0.005) between dogs that presented hemodynamic response and dogs that did not. At PTAm-drop, MAP increased after 1 min (p = 0.001) and 2 min (p = 0.001) with respect to pre-stimulus value, whereas HR and BIS did not change. ROC curves showed a threshold value of PTAm pre-stimulus ≤51 to detect hemodynamic response (sensitivity 69%, specificity 52%). The PTAm and ΔPTA only assessed intraoperative nociception during insufflation. The PTAm pre-stimulus association to the hemodynamic response in anaesthetized dogs showed poor sensitivity and no specificity.
Subject(s)
Hemodynamics , Nociception , Female , Dogs , Animals , Remifentanil , Nociception/physiology , Prospective Studies , Heart Rate/physiologyABSTRACT
This study aimed to document the pathological findings observed in a common buzzard (Buteo buteo insularum) from Gran Canaria (Canary Islands, Atlantic Ocean), naturally infected with Buteo buteo herpesvirus (HV). Local authorities found the common buzzard alive, but it died after 10 days of specialized veterinary care. Postmortem investigation, including complete gross and histologic examination, immunohistochemistry, microbiology, and PCR, was performed. The animal presented necrotizing heterophilic and histiocytic bilateral conjunctivitis, stomatitis, pharyngitis, rhinitis, and sinusitis with secondary bacterial and fungal infections. Frequent eosinophilic intranuclear inclusion bodies were observed in the oral mucosa and esophagus epithelium. HV proteins and DNA were detected in tissues from this animal. The sequences obtained from the PCR product were identical to the reported sequences of Buteo buteo HV.
ABSTRACT
Introduction: Liquid biopsy based on the analysis of circulating cell-free DNA (cfDNA), as well as on detection of point mutations by digital droplet PCR (ddPCR), has revolutionized the research in oncology. In recent years, this technique has been pioneering in veterinary medicine since it is a minimally invasive approach with very promising results for characterization of tumors. Methods: The aim of this study was, firstly, to analyze the concentration and the fragmentation pattern of cfDNA of dogs with mammary tumors (n = 36) and healthy dogs (n = 5) and its correlation with clinicopathological data. Secondly, analysis of TP53 gene expression and the point mutation in the codon 245 were performed in cfDNA and in tumor tissues to assess their potential as plasma biomarkers. Results and discussion: Our results highlighted that those dogs with worse clinicopathological characteristics (simple or undifferentiated carcinomas, higher histological grade and presence of peritumoral inflammation) shown higher cfDNA concentration and higher concentrations of short-fragments (<190 bp) than healthy dogs. In addition, although no detection of the point mutation in codon 245 of TP53 gene could be detected neither in plasma nor tumor tissue, an increased TP53 expression was detected in animals with tumors bearing malignant characteristics. Finally, a high concordance with TP53 gene expression in plasma and tumor tissue and cfDNA concentration was also found. The results derived from this work confirm the valuable potential of cfDNA and its fragments, as well as the analysis of TP53 expression in plasma as useful liquid biomarkers for clinical application in veterinary oncology.
ABSTRACT
In this paper we present a new way to compute the odometry of a 3D lidar in real-time. Due to the significant relation between these sensors and the rapidly increasing sector of autonomous vehicles, 3D lidars have improved in recent years, with modern models producing data in the form of range images. We take advantage of this ordered format to efficiently estimate the trajectory of the sensor as it moves in 3D space. The proposed method creates and leverages a flatness image in order to exploit the information found in flat surfaces of the scene. This allows for an efficient selection of planar patches from a first range image. Then, from a second image, keypoints related to said patches are extracted. This way, our proposal computes the ego-motion by imposing a coplanarity constraint between pairs
ABSTRACT
This paper proposes LTC-Mapping, a method for building object-oriented semantic maps that remain consistent in the long-term operation of mobile robots. Among the different challenges that compromise this aim, LTC-Mapping focuses on two of the more relevant ones: preventing duplicate instances of objects (instance duplication) and handling dynamic scenes. The former refers to creating multiple instances of the same physical object in the map, usually as a consequence of partial views or occlusions. The latter deals with the typical assumption made by object-oriented mapping methods that the world is static, resulting in outdated representations when the objects change their positions. To face these issues, we model the detected objects with 3D bounding boxes, and analyze the visibility of their vertices to detect occlusions and partial views. Besides this geometric modeling, the boxes are augmented with semantic information regarding the categories of the objects they represent. Both the geometric entities (bounding boxes) and their semantic content are propagated over time through data association and a fusion technique. In addition, in order to keep the map curated, the non-detection of objects in the areas where they should appear is also considered, proposing a mechanism that removes them from the map once there is evidence that they have been moved (i.e., multiple non-detections occur). To validate our proposal, a number of experiments have been carried out using the Robot@VirtualHome ecosystem, comparing its performance with a state-of-the-art alternative. The results report a superior performance of LTC-Mapping when modeling both geometric and semantic information of objects, and also support its online execution.
Subject(s)
Robotics , Semantics , EcosystemABSTRACT
STUDY DESIGN: Retrospective review of prospectively collected data. OBJECTIVE: To analyze the postoperative factors that led delayed discharge in patients who would have been eligible for ambulatory lumbar fusion (ALF). SUMMARY OF BACKGROUND DATA: Assessing postoperative inefficiencies is vital to increase the feasibility of ALF. MATERIALS AND METHODS: Patients who underwent single-level minimally invasive transforaminal lumbar interbody fusion and would have met the eligibility criteria for ALF were included. Length of stay (LOS); time in postanesthesia recovery unit (PACU); alertness and neurological examination, and pain scores at three and six hours; type of analgesia; time to physical therapy (PT) visit; reasons for PT nonclearance; time to per-oral (PO) intake; time to voiding; time to readiness for discharge were assessed. Time taken to meet each discharge criterion was calculated. Multiple regression analyses were performed to study the effect of variables on postoperative parameters influencing discharge. RESULTS: Of 71 patients, 4% were discharged on the same day and 69% on postoperative day 1. PT clearance was the last-met discharge criterion in 93%. Sixty-six percent did not get PT evaluation on the day of surgery. Seventy-six percent required intravenous opioids and <60% had adequate pain control. Twenty-six percent had orthostatic intolerance. The median postoperative LOS was 26.9 hours, time in PACU was 4.2 hours, time to PO intake was 6.5 hours, time to first void was 6.3 hours, time to first PT visit was 17.7 hours, time to PT clearance was 21.8 hours, and time to discharge readiness was 21.9 hours. Regression analysis showed that time to PT clearance, time to PO intake, time to voiding, time in PACU, and pain score at three hours had a significant effect on LOS. CONCLUSIONS: Unavailability of PT, surgery after 1 pm , orthostatic intolerance, inadequate pain control, prolonged PACU stay, and long feeding and voiding times were identified as modifiable factors preventing same-day discharge. LEVEL OF EVIDENCE: 4.
Subject(s)
Orthostatic Intolerance , Spinal Fusion , Humans , Length of Stay , Lumbar Vertebrae/surgery , Pain , Patient Discharge , Retrospective Studies , Spinal Fusion/adverse effects , Treatment OutcomeABSTRACT
Burhinus oedicnemus distinctus is an endemic subspecies of Eurasian Stone-curlew present in the Canary Islands. Their populations are rapidly declining, mainly because of anthropogenic impacts. This report describes valvular endocarditis and septicemia in a Eurasian Stone-Curlew with left foot loss and severe contralateral bumblefoot.
Subject(s)
Bacteremia , Charadriiformes , Endocarditis , Streptococcal Infections , Animals , Bacteremia/veterinary , Endocarditis/veterinary , Streptococcal Infections/veterinary , StreptococcusABSTRACT
Ion channels are potentially exploitable as pharmacological targets to treat asthma. This study evaluated the role of KCa3.1 channels, encoded by Kcnn4, in regulating the gene expression of mouse airway epithelium and the development of asthma traits. We used the ovalbumin (OVA) challenge as an asthma model in wild-type and Kcnn4-/- mice, performed histological analysis, and measured serum IgE to evaluate asthma traits. We analyzed gene expression of isolated epithelial cells of trachea or bronchi using mRNA sequencing and gene ontology and performed Ussing chamber experiments in mouse trachea to evaluate anion secretion. Gene expression of epithelial cells from mouse airways differed between trachea and bronchi, indicating regional differences in the inflammatory and transepithelial transport properties of proximal and distal airways. We found that Kcnn4 silencing reduced mast cell numbers, mucus, and collagen in the airways, and reduced the amount of epithelial anion secretion in the OVA-challenged animals. In addition, gene expression was differentially modified in the trachea and bronchi, with Kcnn4 genetic silencing significantly altering the expression of genes involved in the TNF pathway, supporting the potential of KCa3.1 as a therapeutic target for asthma.
Subject(s)
Asthma , Trachea , Animals , Asthma/genetics , Asthma/metabolism , Asthma/pathology , Bronchi/metabolism , Disease Models, Animal , Gene Expression , Mice , Mice, Inbred BALB C , Ovalbumin/metabolism , Trachea/metabolism , Trachea/pathologyABSTRACT
In mammals, the daily variation in the ecology of the intestinal microbiota is tightly coupled to the circadian rhythm of the host. On the other hand, a close correlation between increased body weight and light pollution at night has been reported in humans and animal models. However, the mechanisms underlying such weight gain in response to light contamination at night remain elusive. In the present study, we tested the hypothesis that dim light pollution at night alters the colonic microbiota of mice, which could correlate with weight gain in the animals. By developing an experimental protocol using a mouse model that mimics light contamination at night in urban residences (dLAN, dim light at night), we found that mice exposed to dLAN showed a significant weight gain compared with mice exposed to control standard light/dark (LD) photoperiod. To identify possible changes in the microbiota, we sampled two stages from the resting period of the circadian cycle of mice (ZT0 and ZT10) and evaluated them by high-throughput sequencing technology. Our results indicated that microbial diversity significantly differed between ZT0 and ZT10 in both LD and dLAN samples and that dLAN treatment impacted the taxonomic composition, functions, and interactions of mouse colonic microbiota. Together, these results show that bacterial taxa and microbial metabolic pathways might be involved with the mechanisms underlying weight gain in mice subjected to light contamination at night.
Subject(s)
Colon/microbiology , Gastrointestinal Microbiome , Light Pollution/adverse effects , Weight Gain , Animals , MiceABSTRACT
Neutrophils display an array of biological functions including the formation of neutrophil extracellular traps (NETs), web-like structures specialized in trapping, neutralizing, killing and preventing microbial dissemination within the host. However, NETs contribute to a number of inflammatory pathologies, including severe equine asthma. Tamoxifen (TX) is a selective estrogen receptor modulator which belongs to the triphenylethyllenes group of molecules, and which is used as a treatment in all stages of estrogen-positive human breast cancer. Our previous results suggest that tamoxifen can modulate neutrophil functionality and promote resolution of inflammation; this would partly explain the clinical beneficial effect of this drug in horses with airway inflammation. Enhanced NETs production has been reported with tamoxifen use in humans, but minimal data exists regarding the drug's effect on NETs in horses. The aim of this study is to assess the in vitro effect of TX on NETs formation from peripheral blood of healthy horses. Five clinically healthy mixed-breed adult horses were enrolled in the study. For this, cellular free DNA quantification, immunofluorescence for the visualization of NETs, assessment of different types of NETs, and detection of mitochondrial superoxide. TX induced NETs formation at a concentration of 10 uM. Our results show that only two types of NETs were induced by TX: 95% spread NETs (sprNETs) and 5% aggregated NETs (aggNETs). Furthermore, induction of these NETs could be influenced by mitochondrial ROS. Future research should involve an In vivo study of horses with severe asthma and TX treatment, to evaluate BALF neutrophil NET formation. In conclusion, this in vitro study suggests that the resolution of inflammation by TX in horses with airway inflammation is due to inhibition of other neutrophilic functions but not to NET formation.
ABSTRACT
Nitric oxide (NO) is a key factor in inflammation. Endothelial nitric oxide synthase (eNOS), whose activity increases after stimulation with proinflammatory cytokines, produces NO in endothelium. NO activates two pathways: 1) soluble guanylate cyclase-protein kinase G and 2) S-nitrosylation (NO-induced modification of free-thiol cysteines in proteins). S-nitrosylation affects phosphorylation, localization, and protein interactions. NO is classically described as a negative regulator of leukocyte adhesion to endothelial cells. However, agonists activating NO production induce a fast leukocyte adhesion, which suggests that NO might positively regulate leukocyte adhesion. We tested the hypothesis that eNOS-induced NO promotes leukocyte adhesion through the S-nitrosylation pathway. We stimulated leukocyte adhesion to endothelium in vitro and in vivo using tumor necrosis factor-α (TNF-α) as proinflammatory agonist. ICAM-1 changes were evaluated by immunofluorescence, subcellular fractionation, immunoprecipitation, and fluorescence recovery after photobleaching (FRAP). Protein kinase Cζ (PKCζ) activity and S-nitrosylation were evaluated by Western blot analysis and biotin switch method, respectively. TNF-α, at short times of stimulation, activated the eNOS S-nitrosylation pathway and caused leukocyte adhesion to endothelial cells in vivo and in vitro. TNF-α-induced NO led to changes in ICAM-1 at the cell surface, which are characteristic of clustering. TNF-α-induced NO also produced S-nitrosylation and phosphorylation of PKCζ, association of PKCζ with ICAM-1, and ICAM-1 phosphorylation. The inhibition of PKCζ blocked leukocyte adhesion induced by TNF-α. Mass spectrometry analysis of purified PKCζ identified cysteine 503 as the only S-nitrosylated residue in the kinase domain of the protein. Our results reveal a new eNOS S-nitrosylation-dependent mechanism that induces leukocyte adhesion and suggests that S-nitrosylation of PKCζ may be an important regulatory step in early leukocyte adhesion in inflammation.NEW & NOTEWORTHY Contrary to the well-established inhibitory role of NO in leukocyte adhesion, we demonstrate a positive role of nitric oxide in this process. We demonstrate that NO induced by eNOS after TNF-α treatment induces early leukocyte adhesion activating the S-nitrosylation pathway. Our data suggest that PKCζ S-nitrosylation may be a key step in this process.
Subject(s)
Abdominal Muscles/blood supply , Cell Adhesion , Endothelial Cells/drug effects , Leukocytes/metabolism , Nitric Oxide Synthase Type III/metabolism , Nitric Oxide/metabolism , Tumor Necrosis Factor-alpha/pharmacology , Animals , Cell Line , Coculture Techniques , Endothelial Cells/enzymology , Enzyme Activation , Humans , Intercellular Adhesion Molecule-1/metabolism , Male , Mice, Inbred C57BL , Phosphorylation , Protein Kinase C/metabolism , Protein Processing, Post-Translational , Signal Transduction , Time FactorsABSTRACT
Ae4 transporters are critical for Cl- uptake across the basolateral membrane of acinar cells in the submandibular gland (SMG). Although required for fluid secretion, little is known about the physiological regulation of Ae4. To investigate whether Ae4 is regulated by the cAMP-dependent signaling pathway, we measured Cl-/HCO3- exchanger activity in SMG acinar cells from Ae2-/- mice, which only express Ae4, and found that the Ae4-mediated activity was increased in response to ß-adrenergic receptor stimulation. Moreover, pretreatment with H89, an inhibitor of the cAMP-activated kinase (PKA), prevented the stimulation of Ae4 exchangers. We then expressed Ae4 in CHO-K1 cells and found that the Ae4-mediated activity was increased when Ae4 is coexpressed with the catalytic subunit of PKA (PKAc), which is constitutively active. Ae4 sequence analysis showed two potential PKA phosphorylation serine residues located at the intracellular NH2-terminal domain according to a homology model of Ae4. NH2-terminal domain Ser residues were mutated to alanine (S173A and S273A, respectively), where the Cl-/HCO3- exchanger activity displayed by the mutant S173A was not activated by PKA. Conversely, S273A mutant kept the PKA dependency. Together, we conclude that Ae4 is stimulated by PKA in SMG acinar cells by a mechanism that probably depends on the phosphorylation of S173.NEW & NOTEWORTHY We found that Ae4 exchanger activity in secretory salivary gland acinar cells is increased upon ß-adrenergic receptor stimulation. The activation of Ae4 was prevented by H89, a nonselective PKA inhibitor. Protein sequence analysis revealed two residues (S173 and S273) that are potential targets of cAMP-dependent protein kinase (PKA). Experiments in CHO-K1 cells expressing S173A and S273A mutants showed that S173A, but not S273A, is not activated by PKA.
Subject(s)
Acinar Cells/enzymology , Chloride-Bicarbonate Antiporters/metabolism , Cyclic AMP-Dependent Protein Kinase Catalytic Subunits/metabolism , Salivary Glands/enzymology , Animals , CHO Cells , Chloride-Bicarbonate Antiporters/chemistry , Chloride-Bicarbonate Antiporters/genetics , Cricetulus , Cyclic AMP-Dependent Protein Kinase Catalytic Subunits/genetics , Female , Mice, Inbred C57BL , Mice, Knockout , Models, Molecular , Mutation , Phosphorylation , Protein Conformation , Salivary Glands/cytology , Structure-Activity RelationshipABSTRACT
Face recognition is a technology with great potential in the field of robotics, due to its prominent role in human-robot interaction (HRI). This interaction is a keystone for the successful deployment of robots in areas requiring a customized assistance like education and healthcare, or assisting humans in everyday tasks. These unconstrained environments present additional difficulties for face recognition, extreme head pose variability being one of the most challenging. In this paper, we address this issue and make a fourfold contribution. First, it has been designed a tool for gathering an uniform distribution of head pose images from a person, which has been used to collect a new dataset of faces, both presented in this work. Then, the dataset has served as a testbed for analyzing the detrimental effects this problem has on a number of state-of-the-art methods, showing their decreased effectiveness outside a limited range of poses. Finally, we propose an optimization method to mitigate said negative effects by considering key pose samples in the recognition system's set of known faces. The conducted experiments demonstrate that this optimized set of poses significantly improves the performance of a state-of-the-art, cutting-edge system based on Multitask Cascaded Convolutional Neural Networks (MTCNNs) and ArcFace.
Subject(s)
Facial Recognition , Robotics , Face , Head , Humans , Neural Networks, ComputerABSTRACT
Bone lengthening and bone transport are regeneration processes that commonly rely on distraction osteogenesis, a widely accepted surgical procedure to deal with numerous bony pathologies. Despite the extensive study in the literature of the influence of biomechanical factors, a lack of knowledge about their mechanobiological differences prevents a clinical particularization. Bone lengthening treatments were performed on sheep metatarsus by reproducing the surgical and biomechanical protocol of previous bone transport experiments. Several in vivo monitoring techniques were employed to build an exhaustive comparison: gait analysis, radiographic and CT assessment, force measures through the fixation, or mechanical characterization of the new tissue. A significant initial loss of the bearing capacity, quantified by the ground reaction forces and the limb contact time with the ground, is suffered by the bone lengthening specimens. The potential effects of this anomaly on the musculoskeletal force distribution and the evolution of the bone callus elastic modulus over time are also analyzed. Imaging techniques also seem to reveal lower bone volume in the bone lengthening callus than in the bone transport one, but an equivalent mineralization rate. The simultaneous quantification of biological and mechanical parameters provides valuable information for the daily clinical routine and numerical tools development.
Subject(s)
Bone Regeneration/physiology , Bony Callus/physiology , Gait/physiology , Metatarsal Bones/physiology , Osteogenesis, Distraction , Animals , Biomechanical Phenomena , Bony Callus/diagnostic imaging , Bony Callus/surgery , Elastic Modulus , Female , Metatarsal Bones/diagnostic imaging , Metatarsal Bones/surgery , Sheep , Tomography, X-Ray ComputedABSTRACT
The amount of proteins of the regulatory pluripotency network can be determinant for somatic cell reprogramming into induced pluripotent stem cells (iPSCs) as well as for the maintenance of pluripotent stem cells (PSCs). Here, we report a transposon-based reprogramming system (PB-Booster) that allowed high expression levels of a polycistronic transgene containing Myc, Klf4, Oct4 and Sox2 (MKOS) and showed increased reprogramming efficiency of fresh mouse embryonic fibroblasts (MEFs) into iPSCs under low, but not under high, MKOS expression levels. In contrast, MEFs after 2 passages derived into a similar number of iPSC colonies as fresh MEFs at a high MKOS dose, but this number was reduced at a low MKOS dose. Timing of reprogramming was not affected by MKOS expression levels but, importantly, exogenous MKOS expression in established PSCs caused a significant cell loss. At high but not at low MKOS expression levels, MEFs of the CD1 strain produced more initial cell clusters than iPSCs and, although reprogrammed at a similar efficiency as MEFs of the 129/Sv strain, iPSCs could not be maintained in the absence of exogenous MKOS. In CD1-iPSCs, Oct4, Nanog, Rex1 and Esrrb expression levels were reduced when compared with the levels in PSCs derived from the 129/Sv strain. Culture of CD1-iPSCs in medium with MEK and GSK3ß inhibitors allowed their self-renewal in the absence of exogenous MKOS, but the expression levels of Oct4, Nanog, Rex1 and Esrrb were only partially increased. Despite the reduced levels of those pluripotency factors, CD1-iPSC kept high capacity for contribution to chimeric mouse embryos. Therefore, levels of regulatory pluripotency factors influence reprogramming initiation and PSC maintenance in vitro without affecting their differentiation potential in vivo.
Subject(s)
Cellular Reprogramming , Induced Pluripotent Stem Cells , Animals , Cell Differentiation , Cells, Cultured , Cellular Reprogramming/genetics , Fibroblasts , Mice , Mice, 129 Strain , TransgenesABSTRACT
Photoplethysmography is an extensively-used, portable, and noninvasive technique for measuring vital parameters such as heart rate, respiration rate, and blood pressure. The deployment of this technology in veterinary medicine has been hindered by the challenges in effective transmission of light presented by the thick layer of skin and fur of the animal. We propose an injectable capsule system to circumvent these limitations by accessing the subcutaneous tissue to enable reliable signal acquisition even with lower light brightness. In addition to the reduction of power usage, the injection of the capsule offers a less invasive alternative to surgical implantation. Our current prototype combines two application-specific integrated circuits (ASICs) with a microcontroller and interfaces with a commercial light emitting diode (LED) and photodetector pair. These ASICs implement a signal-conditioning analog front end circuit and a frequency-shift keying (FSK) transmitter respectively. The small footprint of the ASICs is the key in the integration of the complete system inside a 40-mm long glass tube with an inner diameter of 4 mm, which enables its injection using a custom syringe similar to the ones used with microchip implants for animal identification. The recorded data is transferred wirelessly to a computer for post-processing by means of the integrated FSK transmitter and a software-defined radio. Our optimized LED duty cycle of 0.4% at a sampling rate of 200 Hz minimizes the contribution of the LED driver (only 0.8 mW including the front-end circuitry) to the total power consumption of the system. This will allow longer recording periods between the charging cycles of the batteries, which is critical given the very limited space inside the capsule. In this work, we demonstrate the wireless operation of the injectable system with a human subject holding the sensor between the fingers and the in vivo functionality of the subcutaneous sensing on a pilot study performed on anesthetized rat subjects.