Search Results (48)

Cynops orientalis, a member of the Salamandridae family, can regenerate severely injured limbs throughout its lifespan. The molecular mechanisms regulating limb regeneration in C. orientalis remain largely unknown. SERCA1 (sarcoplasmic/endoplasmic reticulum calcium ATPase 1) is a major sarco/endoplasmic reticulum calcium pump in skeletal muscle, and our previous transcriptomic sequencing results showed increased SERCA1 mRNA expression during the early stages of newt limb regeneration. In the present study, we investigated the role and regulatory mechanism of SERCA1 during newt limb regeneration by using a limb-amputation model, shRNA-mediated knockdown, Thapsigargin-mediated SERCA1 inhibition, miRNAomics analysis, dual-luciferase reporter assays and Agomir therapy. Knockdown of SERCA1 expression or suppression of SERCA1 activity impaired skeletal muscle cell dedifferentiation, disrupted AEC and blastema formation, reduced cell proliferation, and significantly delayed limb regeneration compared with control animals. Through miRNAomics analysis and in vitro dual-luciferase reporter assays, we identified miR-20487-5p as a negative regulator of SERCA1 expression in C. orientalis. In vivo treatment with miR-20487-5p Agomir down-regulated SERCA1 expression resulted in limb-regenerative defects. Mechanistically, shRNA-SERCA1 or miR-20487-5p Agomir treatment reduced ERK phosphorylation in regenerating limb tissue, suggesting an association between the miR-20487-5p/SERCA1 axis and MAPK/ERK activation. Taken together, our study supports the significant role of the miR-20487-5p/SERCA1/MAPK/ERK axis in early newt limb regeneration. This study expands the current understanding of molecular mechanisms initiating newt limb regeneration. Full article

Background: Diabetic peripheral neuropathy (DPN) is a major complication of type 2 diabetes mellitus (T2D) that reduces quality of life and increases the risk of foot ulcers and amputations. Early detection is essential, and blood-based biomarkers may support improved screening and timely intervention. This study aimed to identify novel circulating biomarkers for the identification of DPN in patients with T2D. Methods: In the screening phase, plasma samples from 43 participants (10 healthy volunteers [HV], 20 T2D without complications, and 13 T2D with DPN) were analyzed using an antibody array targeting 310 proteins. Thirteen differentially expressed proteins were identified, and six hub proteins were selected through bioinformatic analysis. In the validation phase, plasma concentrations of the six proteins were measured by ELISA in 252 subjects (100 HV, 97 T2D without complications, and 55 T2D with DPN). Mucin-1 expression in sciatic nerves was further evaluated in db/db mice. Results: Of the six hub proteins (TGFB1, MUC1, PF4, IL2RA, SELL, B2M), only mucin-1 showed a significant increase in the DPN group. Plasma mucin-1 positively correlated with MNSI scores and negatively with motor and sensory nerve conduction velocities. In db/db mice, sciatic nerve mucin-1 expression was elevated, while CD31 expression was reduced. Conclusions: Plasma mucin-1 is strongly associated with DPN in both humans and animals and may serve as a promising biomarker for the screening and early identification of DPN. Full article

Background/Objectives: This article brings forward a novel methodology for the intra-op approach of forearm amputation stumps to facilitate their subsequent wireless connection to a neural prosthesis. A neural prosthesis offers the amputee more motor functions compared to myoelectric prostheses, but the neural prosthesis must be connected to the patient’s stump nerves. Methods: An experimental animal study was conducted on 15 Wistar rats. Under anesthesia, the sciatic nerve was carefully dissected and preserved using a folding technique to maintain maximum length without tension. Nerves were repositioned with consideration for future use with biocompatible conduits. Morphometric measurements (nerve length, external diameter, fascicle count) were performed, followed by statistical analysis of length–diameter correlations. Results: The techniques show that the length of the nerves in the amputation stump can be preserved and integrated into the muscle masses with appropriate methods and biomaterials, which ensures the transmission of motor impulses to control the movements of a prosthesis. Fibrosis and mechanical injury have a lower risk of occurring with the nerves protected in the muscle mass. Through statistical analysis we find that sciatic nerve length and diameter have a positive correlation (r = 0.71, p = 0.003), supporting anatomic plausibility for human extrapolation of results. Conclusions: The amputation technique preserves much of the nerve length and viability and is simple to perform. Neural electrode implantation can be facilitated by folding the nerve within a large muscle mass and using biomaterial conduits. Better rehabilitation of the patient may occur with the use of a prosthesis equipped with more functions and superior control. Full article

Zootechnical practices such as tail docking are still in use in dairy sheep farming, performed in the first week of life, mainly by rubber ring and only rarely by surgical methods. In this study, we evaluated the impact of caudectomy on ovine stress levels, inflammation, and health status by comparing tail docking carried out using rubber rings or surgical amputation. Twenty-one lambs were randomly selected and equally allocated into three groups: controls (n = 7), lambs with tail cut by rubber rings (n = 7), and lambs with caudectomy performed by surgical practice (n = 7). Several biochemical parameters and inflammatory markers were monitored at different times post-caudectomy, as well as wool levels of the stress marker cortisol. Our data revealed that lambs that underwent tail docking by rubber rings, but not by surgical procedure, presented inflammation and stress, as well as a moderate increase in muscular damage markers. These results are useful for the evaluation of animal welfare in dairy sheep that underwent caudectomy, highlighting the need to re-evaluate this procedure, as well as the ways in which it is performed. Full article

Bycatch is the most common cause of death of small delphinids worldwide, including the Mediterranean Sea. The diagnosis of bycatch as cause of death in stranded cetaceans depends on the cumulative presence of multiple findings, termed bycatch criteria. In this study, we retrospectively evaluated the presence of bycatch criteria in 138 necropsied cetaceans, 136 stranded and 2 confirmed bycaught, in the Catalan Mediterranean Sea across a 13-year period. With the aim of identifying the most specific and reliable bycatch criteria, the animals’ cause of death was classified as either bycaught or other causes. Animals were necropsied according to standard procedures with complete histopathological examination and ancillary diagnostic techniques. We reviewed the necropsy reports and photographs of 138 cetaceans of seven species. Bycatch had been determined as the cause of death/stranding in 40 (29%) necropsied cetaceans. Both sexes were equally represented in the bycatch group. Bycatch was diagnosed in the Mediterranean common bottlenose dolphin (10/14; 71.4%), striped dolphin (29/108; 26.9%), and Risso’s dolphin (1/11; 9.1%). Sixty-seven out of 98 (68.3%) cetaceans that had been classified as non-bycatch had one or two bycatch criteria. Cetaceans with two and three major criteria had an overlap of causes of death, as some animals were diagnosed with bycatch and others with other causes of mortality. Animals with four criteria were invariably diagnosed as being bycaught. Recent feeding, absence of disease, good nutritional status, marks of fishing gear, multiorgan intravascular gas bubbles, hyphema and amputations or sharp incisions presumably inflicted by humans were significantly more likely to result in a diagnosis of bycatch, while loss of teeth and cranial fractures were not. None of the dolphins diagnosed as bycatch had ingested fishing gear. Our results highlight the relevance of bycatch as the cause of death of dolphins in the Mediterranean and suggest that some criteria traditionally linked to bycatch are not specific for bycatch in our region. Full article

Introduction and objective: This literature review aims to provide an overview of the progress in metabolomic assessment in animal and cell models and in humans with diabetic neuropathy (DN). Methods: Metabolomics has emerged as an important approach for investigating, identifying, and describing biomarkers related to DN. None has yet been validated for use in clinical practice. Results: DN induced significant alterations in energy metabolism and carbohydrates, lipids, amino acids, peptides, and proteins. Several treatments for DN, evaluated using metabolomics, were proved to have promising results. Conclusions: The ideal metabolite or set of metabolites that could be used as biomarkers should identify patients with diabetes prone to develop DN or those prone to progress to severe forms of sensory loss, associated with risk of ulcerations and amputation. Another potential use of a metabolite might be as an indicator of treatment response in clinical trials using agents with potential disease-modifying properties. Full article

Deep-sea mineral water (DSW) consists of different compositions and properties. The composition can be varied in different seas all around the world. However, previous studies have investigated that DSW is a potential candidate that can be used to prevent different inflammatory diseases. Even though inflammation is an important protective mechanism in an animal, excessive inflammation causes organ failure and ultimate death. Therefore, the present study was carried out to investigate the anti-inflammatory effect of DSW extracted from the South Korean Sea to unveil its potential as an anti-inflammatory drug. To perform this, first, we have compared the cytotoxic effect of DSW on RAW 264.7 murine macrophage cells with NaCl and normal-sea water (NSW). Results reveal that DSW enhances cell survival while other treatments negatively affect cell survival. Furthermore, we have investigated that DSW reduces the LPS-induced cell apoptosis compared to the NaCl- and NSW-treated cells. Moreover, DSW has the ability to suppress the pro-inflammatory cytokine transcription (TNF-α, IL-1β, and IL-6) and NO production upon LPS treatment. In-vivo survival assay in zebrafish larvae shows a more than 50% survival rate in 10, 20, 30, 40, or 50% concentrations of DSW-treated larvae compared to NaCl- or NSW-treated larvae. Further investigations unveiled that DSW can negatively regulate the neutrophil and macrophage recruitment to the inflammatory site, which was induced by fin-fold amputation in zebrafish larvae and pro-inflammatory cytokine (tnf-α, il-1β, and il-6) secretion. Taken together, the present study concluded that DSW may have the ability to act as an anti-inflammatory drug to suppress excessive inflammation and subsequent consequences. Full article

Studies on environmental enrichment for captive bearded dragons (Pogona vitticeps) are still very scarce in the literature, but they are of the utmost importance in the search for quality of life for these animals when they are under human care. Environmental enrichment items should be inclusive, allowing disabled animals to explore them and providing good experiences. Therefore, this study aimed to evaluate food enrichment strategies to increase the foraging behavior of captive non-disabled and disabled bearded dragons. This work was carried out with six adult bearded dragons, three of which had one limb amputated and three that had all limbs, kept at the São Paulo Aquarium. Live kingworm larvae (Zophobas morio) were offered as food in three different levels of enrichment challenge (low, medium, high). Behavioral data were collected using focal sampling with instantaneous recording, with a time interval of every 30 s. Results showed that the more complex the food enrichment challenge, the greater the frequency of bearded dragon foraging. In addition, both amputees and non-amputees responded similarly to the enrichment items, differing primarily in their activity levels, where amputee individuals showed more tendencies toward ‘stationary’ and ‘lying down’ behaviors. More studies with reptiles should be encouraged so that good management practices become daily protocols for a better quality of life and for rescuing the natural behaviors of animals kept under human care, even those with anatomical disabilities. Full article

The amputation of a limb in quadrupeds can overload the remaining limbs, especially the contralateral one. The compensatory effort is especially high if it is a forelimb. It is, therefore, important to objectively know the changes in weight redistribution that occur in the animal while walking and standing still. With this objective, static (postural) and dynamic kinetic examinations were carried out on five French bulldogs with an amputated forelimb and five intact French bulldogs. For this examination, force and pressure platforms were used. The results were statistically compared using the student t-test. The parameters derived from the ground reaction forces were significantly higher in the amputee group. Surprisingly, postural examination showed that amputated dogs reached the same stability as healthy ones. Tripedal support in dogs does not objectively imply a loss of balance in quantitative terms; although the increase in force used by the remaining limb, as well as its altered cranial disposition during the support phase, may potentially predispose the animal to additional injuries in the future due to an overuse of different musculoskeletal units. Full article

The development of realistic dummies for training the distal phalanx amputation (DPA) technique in mouse pups is a promising alternative to reduce and replace animals in training for research and teaching. To test this, we obtained micro-CT data from postnatal day-five mouse pups, meticulously segmented them, and converted them into a 3D mesh format suitable for 3D printing. Once the dummy was printed, it was evaluated during actual training courses in two different groups: in the first group, users received no dummies to train the DPA, and in the second group, users were trained with three dummies. To assess the effectiveness of the dummy, we conducted a survey followed by an expert veterinarian evaluation. Our results showed that DPA is a complex procedure, and it is commonly poorly performed. When implementing the dummies, users who were not provided with dummies to practice only had an 8.3% success rate in DPA, while users provided with three dummies had a 45.5% success rate, respectively. Despite additional research being needed, our dummy offered improved practical training by providing a safe and effective alternative in line with ethical considerations while demonstrating the feasibility of using 3D printing technology to promote the 3Rs in experimental research. Full article

Animal personality, which describes inter-individual differences and intra-individual consistency in behaviors across time and contexts, has been widely observed and has significance for both ecology and evolution. Morphological modifications, particularly during early life stages, may highly influence animal behavior in adulthood; thus, exploring this relationship can elucidate personality development throughout ontogeny. In this study, we reared juvenile crayfish (Procambarus clarkii) with different degrees of cheliped mutilation and explored their personality patterns, including exploration and aggression, when they reached sexual maturity. Male crayfish showed repeatability in exploration, and both sexes showed repeatability in aggression. We observed no significant correlation between the two behavioral traits, indicating the absence of behavioral syndromes. Moreover, exploration did not differ according to the type of mutilation, but crayfish with more intact chelipeds were more aggressive, and males were more aggressive than females. These results indicate that cheliped mutilation may modify the average levels of personality traits associated with competition or self-defense. Our study provides insights into how morphological modifications may shape animal personalities in adulthood. Full article

Traumatic nerve injuries are common lesions that affect several hundred thousand humans, as well as dogs and cats. The assessment of nerve regeneration through animal models may provide information for translational research and future therapeutic options that can be applied mutually in veterinary and human medicine, from a One Health perspective. This review offers a hands-on vision of the non-invasive and conservative approaches to peripheral nerve injury, focusing on the role of neurorehabilitation in nerve repair and regeneration. The peripheral nerve injury may lead to hypersensitivity, allodynia and hyperalgesia, with the possibility of joint contractures, decreasing functionality and impairing the quality of life. The question remains regarding how to improve nerve repair with surgical possibilities, but also considering electrical stimulation modalities by modulating sensory feedback, upregulation of BDNF, GFNF, TrKB and adenosine monophosphate, maintaining muscle mass and modulating fatigue. This could be improved by the positive synergetic effect of exercises and physical activity with locomotor training, and other physical modalities (low-level laser therapy, ultrasounds, pulsed electromagnetic fields, electroacupuncture and others). In addition, the use of cell-based therapies is an innovative treatment tool in this field. These strategies may help avoid situations of permanent monoplegic limbs that could lead to amputation. Full article

We studied the regeneration of orthotopic and heterotopic tails in larval axolotls. First, we analyzed tail regeneration following reciprocal exchange of cuffs of tail integument between dark-colored (wild-type) and yellow-colored (hybrid) larval animals. Second, we studied tail regeneration in larval axolotls following transplantation of cuffs of tail integument from metamorphosed dark-colored conspecifics and from an adult fire salamander. In all cases, the amputation planes involved the transplanted integumental cuffs. In the first experiment, the regenerated tails showed the color of the host animals, not that of the transplanted cuffs. This suggests that the melanocytes of the regenerated tails were derived from the host hypodermis. Following transplantation of metamorphosed skin from axolotls and a fire salamander onto larval axolotls, the metamorphosed epidermis reverted to a larval condition. This indicates that the state of differentiation of the metamorphosed epidermis was not permanent. Rather, in order to maintain the metamorphosed epidermal structure, a continuous exposure of the animals to sufficient levels of thyroid hormones was required. Transplantation of tail buds from yellow-colored onto dark-colored axolotl embryos caused the formation of yellow-colored tails both in the head and the anterior limb region of the hosts. Incomplete resection of these heterotopic tails was followed by tail regeneration, while no tail regeneration occurred following complete resection of the heterotopic tails. Successful tail regeneration depended on the presence of neural tissue along the resection plane. Full article

The study aims to construct an inertial measuring system for the application of amputee subjects wearing a prosthesis. A new computation scheme to process inertial data by installing seven wireless inertial sensors on the lower limbs was implemented and validated by comparing it with an optical motion capture system. We applied this system to amputees to verify its performance for gait analysis. The gait parameters are evaluated to objectively assess the amputees’ prosthesis-wearing status. The Madgwick algorithm was used in the study to correct the angular velocity deviation using acceleration data and convert it to quaternion. Further, the zero-velocity update method was applied to reconstruct patients’ walking trajectories. The combination of computed walking trajectory with pelvic and lower limb joint motion enables sketching the details of motion via a stickman that helps visualize and animate the walk and gait of a test subject. Five participants with above-knee (n = 2) and below-knee (n = 3) amputations were recruited for gait analysis. Kinematic parameters were evaluated during a walking test to assess joint alignment and overall gait characteristics. Our findings support the feasibility of employing simple algorithms to achieve accurate and precise joint angle estimation and gait parameters based on wireless inertial sensor data. Full article

Approximately 6% of adults worldwide suffer from peripheral artery disease (PAD), primarily caused by atherosclerosis of lower limb arteries. Despite optimal medical care and revascularization, many PAD patients remain symptomatic and progress to critical limb ischemia (CLI) and risk major amputation. Delivery of pro-angiogenic factors as proteins or DNA, stem, or progenitor cells confers vascular regeneration and functional recovery in animal models of CLI, but the effects are not well replicated in patients and no pro-angiogenic biopharmacological procedures are approved in the US, EU, or China. The reasons are unclear, but animal models that do not represent clinical PAD/CLI are implicated. Consequently, it is unclear whether the obstacles to clinical success lie in the toxic biochemical milieu of human CLI, or in procedures that were optimized on inappropriate models. The question is significant because the former case requires abandonment of current strategies, while the latter encourages continued optimization. These issues are discussed in the context of relevant preclinical and clinical data, and it is concluded that preclinical mouse models that include age and atherosclerosis as the only comorbidities that are consistently present and active in clinical trial patients are necessary to predict clinical success. Of the reviewed materials, no biopharmacological procedure that failed in clinical trials had been tested in animal models that included advanced age and atherosclerosis relevant to PAD/CLI. Full article