Innovative models for teaching reproduction in small animals: The experience at DIMEVET of Bologna University

For a long time, the main way to acquire the skills necessary for good veterinary practice has been the traditional apprenticeship model (observe, assist, and perform under supervision). However, in the last years, more creative and innovative teaching models have been adopted by academic institutions and, parallelly, the opportunities to gain hands-on experience for clinical and surgical procedures are becoming more limited. For that reason, the introduction of the models can provide a potential solution to the ethical and legal implications related to the use of live animals for educational purposes and the biosafety risks deriving from the manipulation of human/animal cadavers. The activity on this topic at DIMEVET and, in particular, the experience about teaching reproduction includes in-house production and use of odels for learning skills both for degree (fifth and third years of course) and post-graduate courses. Our models are designed on the basis of two fundamental aspect: the student level and the teacher target. The aim of this study was to evaluate, for the first time in literature, how much the use of simulators impacts on practical teaching in degrees and post graduate courses at DIMEVET, considering different learners with different levels of knowledge and skills. Namely, effectiveness, adequacy and quality of simulators have been checked assessing students’ learning experience and teacher’s opinion by specific satisfaction questionnaires and considering the attendance at the training sessions and the results of a final examination (PHY group) and an in vivo test (POST group). The rate of satisfaction was high among the three groups considered and the participants that had used the models had a higher success, both in the final examination and in vivo test. Data collected have been useful in order to show an improvement in teaching at DIMEVET in the field of reproduction, despite the relatively short years of experience using simulation. With this study we demonstrate that the adequacy of the models is not correlated to how simulators is built, whether with basic materials or not faithfully in terms of anatomical appearance, but it is important that it is realistic in terms of psychomotor procedure. In conclusion, the use of models permits the student to gain the hand-eye coordination and dexterity necessary to perform certain skills and the models proposed are meeting the desired educational goals

Diagnosis and Treatment of Foetal Anasarca in Two English bulldog Puppies

This case report provides the first description of foetal anasarca, from prenatal diagnosis to neonatal treatment, in two English Bulldog puppies. Anasarca affected 2 out of 3 puppies of the same litter. The dam was a 3 year-old primipara and was ultrasonographically monitored during pregnancy. The diagnosis of foetal anasarca was performed on d53. The most severely affected puppy was a female, which showed subcutaneous oedema and fluid in the thoracic and abdominal cavities. The other affected puppy was a male, which showed subcutaneous oedema but no fluid in thoracic and abdominal cavities. At birth, the female puppy did not respond to resuscitation, whereas the male puppy was successfully treated with furosemide after resuscitation and completely recovered within 36 hours. The third puppy was a healthy female. Two weeks after birth, the male puppy was healthy, although it was lighter compared to its sister puppy. In conclusion, ultrasound examination during routine pregnancy monitoring permits an early diagnosis of foetal anasarca, at least in the most severe cases. Prenatal diagnosis can be useful for the planned c-section, in order to apply an adequate treatment as soon as possible at delivery

Additive Functions in Boolean Models of Gene Regulatory Network Modules

Gene-on-gene regulations are key components of every living organism. Dynamical abstract models of genetic regulatory networks help explain the genome’s evolvability and robustness. These properties can be attributed to the structural topology of the graph formed by genes, as vertices, and regulatory interactions, as edges. Moreover, the actual gene interaction of each gene is believed to play a key role in the stability of the structure. With advances in biology, some effort was deployed to develop update functions in Boolean models that include recent knowledge. We combine real-life gene interaction networks with novel update functions in a Boolean model. We use two sub-networks of biological organisms, the yeast cell-cycle and the mouse embryonic stem cell, as topological support for our system. On these structures, we substitute the original random update functions by a novel threshold-based dynamic function in which the promoting and repressing effect of each interaction is considered. We use a third real-life regulatory network, along with its inferred Boolean update functions to validate the proposed update function. Results of this validation hint to increased biological plausibility of the threshold-based function. To investigate the dynamical behavior of this new model, we visualized the phase transition between order and chaos into the critical regime using Derrida plots. We complement the qualitative nature of Derrida plots with an alternative measure, the criticality distance, that also allows to discriminate between regimes in a quantitative way. Simulation on both real-life genetic regulatory networks show that there exists a set of parameters that allows the systems to operate in the critical region. This new model includes experimentally derived biological information and recent discoveries, which makes it potentially useful to guide experimental research. The update function confers additional realism to the model, while reducing the complexity and solution space, thus making it easier to investigate

Plasma Concentration Rise after the Intramuscular Administration of High Dose Medetomidine (0.13 mg/kg) for Semen Collection in Cats

High dose medetomidine 0.13 mg/kg can be used for semen collection in cats with variable results in terms of quantity and quality. Therefore, a variation in terms of distribution and elimination among patients has been hypothesised. The aim of the study was to characterise the pharmacokinetics of medetomidine (0.13 mg/kg) administered intramuscularly (IM) in healthy male cats. Eighteen male cats undergoing castration were included, and medetomidine (0.13 mg/kg) was administered IM. Venous blood samples were collected at 20, 30, 40, 50, 60, 75 and 90 minutes after medetomidine administration. Before orchiectomy, at T20, sperm collection was attempted. Plasma medetomidine concentrations were determined by liquid chromatography/mass spectrometry analysis. Semen collection was successful in 15/18 cats. The medetomidine plasma concentration following the IM administration of a bolus was best described using a non-compartment model. Time of maximum concentration was observed at 40 minutes (range 20-90); maximum concentration was 32.8 ng/mL (range 26.8-51.2). The median apparent clearance was 11.9 mL/kg/minute (range 0.7-43.8). In conclusion, medetomidine administered IM at 0.13 mg/kg reached its peak plasma concentration slowly and with variability among patients. In addition, it was characterised by low total body clearance probably due to the cardiovascular alterations associated with medetomidine administration

A method for astral microtubule tracking in fluorescence images of cells doped with taxol and nocodazole

In this paper we describe an algorithm that performs automatic detection and tracking of astral microtubules in fluorescence confocal images. This sub-population of microtubules only exists during and immediately before mitosis and aids in the spindle orientation by connecting it to the cell cortex. Anomalies in their dynamic behaviour play a causal role in many diseases, such as development disorders and cancer. The main novelty of the proposed algorithm lies in the fact it provides a fully automated estimation of parameters related to microtubule dynamic instability (growth velocity, track length and track lifetime), and helps in understanding the effects of intermediate drug concentrations. Its performance has been objectively assessed using publicly available synthetic data and largely employed metrics. Moreover, we present experiments addressing cell cultures doped with different concentrations of taxol and nocodazole. Such drugs are known to suppress the microtubule dynamic instability, but their effects at intermediate concentrations are not completely assessed. The algorithm been compared with other stateof- the-art approaches, tested on consistent real datasets. The results are encouraging in terms of performance, robustness and simplicity of use, and the algorithm is now routinely employed in our Department of Molecular Biotechnology

Retrospective evaluation of the induction of anaesthesia with alfaxalone or propofol in cats undergoing caesarean section

Objectives: Several anaesthetic protocols for caesarean section (c-section) have been described in dogs; however, anaesthesiological studies in cats undergoing c-section are lacking. The aim of this study was to assess the viability and outcome of kittens born from queens undergoing emergency c-section and receiving propofol or alfaxalone for anaesthetic induction and isoflurane for maintenance of general anaesthesia. Methods: All cats admitted to the Veterinary Teaching Hospital of the University of Bologna between January 2014 and December 2022 for dystocia that had undergone an emergency c-section were analysed in this retrospective study. The queens received propofol 2-6 mg/kg IV (group P) or alfaxalone 1-3 mg/kg IV (group A) administered slowly; general anaesthesia was maintained with isoflurane. Survival of all the kittens was evaluated at birth and after 24 h; in addition, heart rate, respiratory rate and mucous membrane colour were evaluated in the kittens at birth. Results: Of the 14 cats included in the study, eight received alfaxalone (group A) and six received propofol (group P) for induction of the general anaesthesia. A total of 50 kittens were born by c-section: 30 kittens in group A and 20 in group P. The overall survival of the kittens was 90% at birth: 96.7% (29/30) for group A and 80% (16/20) for group P. The 24 h survival rate was 93.1% for group A and 87.5% for group P. Conclusions and relevance: The results of the present study demonstrated that in cats undergoing c-section, both alfaxalone and propofol are feasible for the induction of general anaesthesia

Sex-sorted canine sperm cryopreservation: Limits and procedural considerations

The aim of this study was to define a protocol to store dog sperm before and after sorting to obtain an insemination dose sufficient to allow the conception by artificial insemination. Experiment 1 and 2 were performed to evaluate the more appropriate extender for preserving at room temperature dog sperm before and after sorting. Four extenders were tested: (1) Tris-fructose-citrate (TFC), (2) Tris-glucose-citrate (TGC), (3) modified Tyrode\u2019s albumin lactate pyruvate medium (mTALP), and (4) third fraction of the ejaculate (after centrifugation at 5000 g for 10 minutes; III FRAC). Experiment 3 and 4 were performed to evaluate the ability of dog semen to withstand sex sorting and freezing/thawing. Modified Tyrode\u2019s albumin lactate pyruvate medium was the best extender for canine sperm storage at room temperature (20 C\u201325 C) before (total motility: TFC, 8.3 1.7; TGC, 50.0 11.5; mTALP, 70.0 0.1; III FRAC, 25.0 1 0.4; P < 0.05) and after sorting (total motility: TFC, 7.3 1.5; TGC, 10.3 1.5; mTALP, 33.3 6.7; III FRAC, 8.7 5.8; P < 0.05), even if at 24- hour sorted sperm quality was impaired in all extenders tested herein. Sperm quality decreased after sorting (total motility: control, 92.5 0.9; sorted, 52.9 6.0; P < 0.05) and, especially, after freezing/thawing (total motility: frozen control, 25.7 4.1; frozen sorted, 2.4 1.2; P < 0.05). In conclusion, mTALP is an appropriate medium for canine sperm storage before and soon after sorting (hours), but a long storage period of sexed sperm at room temperature is not adequate. Cryopreservation greatly impaired sperm quality, and further studies are needed to optimize the freezing protocol for sexed dog sperm

ZIKA virus elicits P53 activation and genotoxic stress in human neural progenitors similar to mutations involved in severe forms of genetic microcephaly

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