Students Satisfaction with the Use of PlayDoh® as a Tool to Actively Learn 3D Veterinary Anatomy More Accurately.

Understanding veterinary anatomy is an essential skill for the study of veterinary medicine as well as for diagnostic imaging and therapy. Dissection facilities are increasingly limited in some schools and its alternatives have often focussed on using two-dimensional images. However, the study of veterinary anatomy is mainly concerned with identifying structures and spatial relationships between them within a 3D space, and the use of 2D teaching approaches does not provide accurate information. We tested whether PlayDoh® student-built models could be an inexpensive potential tool beneficial to veterinary students learning anatomy in three distinct scenarios: (1) during a lecture, introducing a new concept; (2) during a flipped classroom approach where a video-podcast lecture was to be watched by the students prior to the lecture and (3) as a revision session where students brought their own questions and created, under supervision, their own models to respond to them. PlayDoh® sessions benefitted 172 first-year Veterinary Medicine and Animal Science students. The most accurate visualisation of anatomical structures in 3D was the principal benefit mentioned by the learners (35%). In addition, the learners noted that the technique would help with 'retention' (18%). According to the students' preferences, it was possible to create four groups: A, B, C and D. Group A encompassed the methodologies most liked by students and consisted of lectures, dissection and demonstrations. Group B included demonstrations and 3D modelling using PlayDoh®. Group C consisted of 3D modelling using PlayDoh®, books and online and, finally, group D included the methodologies least preferred by students, i.e. online and PBL. Our findings suggest that using 3D PlayDoh® modelling has potential as a method to enhance the learning of veterinary anatomy and may be most valuable to those students learning more complex subject areas that require a 3D teaching approach in practice. Supplementary Information: The online version contains supplementary material available at 10.1007/s40670-023-01892-y.

Comparison of volume of the forebrain, subarachnoid space and lateral ventricles between dogs with idiopathic epilepsy and controls using a stereological approach: Cavalieri's principle.

BACKGROUND: Canine idiopathic epilepsy (IE) is the most common chronic neurological brain disease in dogs, yet it can only be diagnosed by exclusion of all other potential causes. In people, epilepsy has been associated with a reduction in brain volume. The objective was to estimate the volume of the forebrain (FB), subarachnoid space (SAS) and lateral ventricles (LV) in dogs with IE compared to controls using Cavalieri's principle. MRI scans of case and control dogs were identified from two neurology referral hospital databases. Eight breeds with increased odds of having IE were included: Golden Retriever, Labrador Retriever, Cocker Spaniel, Border terrier, German Shepherd dog, Parson Jack Russell terrier, Boxer, and Border Collie. Five dogs of each breed with IE and up to five controls were systematically and uniformly randomly sampled (SURS). The volume of the FB, SAS and LV were estimated from MRI scans by one blinded observer using Cavalieri's principle. RESULTS: One hundred-two dogs were identified; 56 were diagnosed with IE and 46 were controls. There was no statistically significant difference in FB, SAS and LV volume between dogs with IE and controls. Dogs with a history of status epilepticus had significantly larger FB than those without (p = 0.05). There was a border-line trend for LV volume to increase with increasing length of seizure history in the IE group (p = 0.055). CONCLUSION: The volumes of the FB, SAS and LV are not different between dogs with IE and controls, so IE remains a diagnosis of exclusion with no specific neuroanatomical biomarkers identified. This is the first time FB and SAS volume has been compared in dogs with IE. Unfortunately, we have shown that the results reporting significantly larger FBs in dogs with status epilepticus and LV volume increase with length of seizure history were likely confounded by breed and should be interpreted cautiously. Whilst these associations are interesting and clinically relevant, further investigation with breed-specific or larger, breed-diverse populations are required to permit strong conclusions. The Cavalieri principle provided an effective estimation of FB, SAS and LV volumes on MRI, but may be too time-intensive for use in clinical practice.

Stereology shows that damaged liver recovers after protein refeeding.

OBJECTIVE: The aim of the present study was to investigate the putative effects of a low-protein diet on the three-dimensional structure of hepatocytes and determine whether this scenario could be reversed by restoring the adequate levels of protein to the diet. METHODS: Using design-based stereology, the total number and volume of hepatocytes were estimated in the liver of mice in healthy and altered (by protein malnutrition) conditions and after protein renutrition. RESULTS: This study demonstrated a 65% decrease in the liver volume (3302 mm3 for the control for undernourished versus 1141 mm3 for the undernourished group) accompanied by a 46% reduction in the hepatocyte volume (8223 µm3 for the control for undernourished versus 4475 µm3 for the undernourished group) and a 90% increase in the total number of binucleate hepatocytes (1 549 393 for the control for undernourished versus 2 941 353 for the undernourished group). Reinstating a normoproteinic diet (12% casein) proved to be effective in restoring the size of hepatocytes, leading to an 85% increase in the total number of uninucleate hepatocytes (15 988 560 for the undernourished versus 29 600 520 for the renourished group), and partially reversed the liver atrophy. CONCLUSIONS: Awareness of these data will add to a better morphologic understanding of malnutrition-induced hepatopathies and will help clinicians improve the diagnosis and treatment of this condition in humans and in veterinary practice.

Reflection imaging of China ink-perfused brain vasculature using confocal laser-scanning microscopy after clarification of brain tissue by the Spalteholz method.

Confocal laser-scanning microscopy is a useful tool for visualizing neurons and glia in transparent preparations of brain tissue from laboratory animals. Currently, imaging capillaries and venules in transparent brain tissues requires the use of fluorescent proteins. Here, we show that vessels can be imaged by confocal laser-scanning microscopy in transparent cortical, hippocampal and cerebellar preparations after clarification of China ink-injected specimens by the Spalteholz method. This method may be suitable for global, three-dimensional, quantitative analyses of vessels, including stereological estimations of total volume and length and of surface area of vessels, which constitute indirect approaches to investigate angiogenesis.

Sensitivity of primary fibroblasts in culture to atmospheric oxygen does not correlate with species lifespan.

Differences in the way human and mouse fibroblasts experience senescence in culture had long puzzled researchers. While senescence of human cells is mediated by telomere shortening, Parrinello et al. demonstrated that senescence of mouse cells is caused by extreme oxygen sensitivity. It was hypothesized that the striking difference in oxygen sensitivity between mouse and human cells explains their different rates of aging. To test if this hypothesis is broadly applicable, we cultured cells from 16 rodent species with diverse lifespans in 3% and 21% oxygen and compared their growth rates. Unexpectedly, fibroblasts derived from laboratory mouse strains were the only cells demonstrating extreme sensitivity to oxygen. Cells from hamster, muskrat, woodchuck, capybara, blind mole rat, paca, squirrel, beaver, naked mole rat and wild-caught mice were mildly sensitive to oxygen, while cells from rat, gerbil, deer mouse, chipmunk, guinea pig and chinchilla showed no difference in the growth rate between 3% and 21% oxygen. We conclude that, although the growth of primary fibroblasts is generally improved by maintaining cells in 3% oxygen, the extreme oxygen sensitivity is a peculiarity of laboratory mouse strains, possibly related to their very long telomeres, and fibroblast oxygen sensitivity does not directly correlate with species' lifespan.

Stereological and allometric studies on neurons and axo-dendritic synapses in superior cervical ganglia.

The superior cervical ganglion (SCG) plays an important role in neuropathies including Horner's syndrome, stroke, and epilepsy. While mammalian SCGs seem to share certain organizational features, they display natural differences related to the animal size and side and the complexity and synaptic coverage of their dendritic arborizations. However, apart from the rat SCG, there is little information concerning the number of SCG neurons and synapses, and the nature of relationships between body weight and the numbers and sizes of neurons and synapses remain uncertain. In the recognition of this gap in the literature, in this chapter, we reviewed the current knowledge on the SCG structure and its remodeling during postnatal development across a plethora of large mammalian species, focusing on exotic rodents and domestic animals. Instrumentally, we present stereology as a state-of-the-art 3D technology to assess the SCG 3D structure unbiasedly and suggest future research directions on this topic.

New thymidylate synthase inhibitors induce apoptosis in melanoma cell lines.

Malignant melanoma is particularly resistant to conventional chemotherapy and radiotherapy. For this reason in the past years a huge variety of new compounds has been developed with potential chemotherapeutic activity which needs to be tested in vitro and in vivo. We investigated the in vitro action of three new experimental antifolate substances (MR7, MR21 and MR36) with a critical target for thymidylate synthase (TS), an essential enzyme for DNA synthesis. The response of two melanoma cell lines (SK-MEL-2 derived from malignant melanoma metastasis and SK-MEL-28 derived from primary malignant melanoma) was examined after treatment with these substances. The antifolate agents induced apoptosis in SK-MEL-2 and SK-MEL-28 cells as confirmed by the TUNEL technique and Comet Assay. Western-blot analysis showed a down-regulation of Bcl-2 protein level and PARP cleavage, otherwise p53 and Bax expressions were not modulated. Moreover, these antifolate-induced apoptosis was accompanied by both pro-caspase-9 and -8 activations. These results were supported by the use of the pan-caspases inhibitor Z-VAD-FMK that almost completely decreased the amount of apoptosis in both the melanoma cell lines treated with antifolate. In conclusion our results show that TS inhibitors are able to induce apoptosis through a caspase-mediated pathway, but without the involvement of the p53/Bax signalling.

Regulation of Entamoeba invadens encystation and gene expression with galactose and N-acetylglucosamine.

Encystation of Entamoeba invadens parasites is prevented by the presence of free galactose or N-acetylglucosamine in the encystation medium. Galactose prevents the formation of amoeba cellular aggregates which develop during the early phase of encystation, suggesting the presence of functional cell surface galactose-binding molecules, whereas N-acetylglucosamine allows aggregation to occur and prevents cyst formation at a later point. While studying sugar inhibition of amoeba encystation, it was found that high efficiency encystation required the inclusion in encystation medium of precise amounts of polyvalent galactose-terminated molecules, and these molecules could be supplied by serum or by defined glycoconjugates, including mucin. Addition of free galactose to encystation medium prevented the accumulation of three transcripts which are normally upregulated during encystation, and N-acetylglucosamine prevented accumulation of one of the transcripts. These results suggest the presence of distinct sugar-sensitive pathways that regulate differentiation of the amoeba trophozoite into infectious cysts.