Potential Role of mRNA in Estimating Postmortem Interval: A Systematic Review.

Although the postmortem interval estimation still represents one of the main goals of forensic medicine, there are still several limitations that weigh on the methods most used for its determination: for this reason, even today, precisely estimating the postmortem interval remains one of the most important challenges in the forensic pathology field. To try to overcome these limitations, in recent years, numerous studies have been conducted on the potential use of the mRNA degradation time for reaching a more precise post mortem interval (PMI) estimation. An evidence-based systematic review of the literature has been conducted to evaluate the state of the art of the knowledge focusing on the potential correlation between mRNA degradation and PMI estimation. The research has been performed using the electronic databases PubMed and Scopus. The analysis conducted made it possible to confirm the potential applicability of mRNA for reaching a more precise PMI estimation. The analysis of the results highlighted the usefulness of some mRNAs, such as ß-actin and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA, especially in short time frames, within a few hours or days of death. The matrices on which these analyses were conducted were also analyzed, resulting in less exposure to the external environment, including the heart, brain, and dental pulp. The major limitations were also reported, including the short time intervals analyzed in most of the articles, the lack of mathematical models, and the failure to report the error rate between the mRNA degradation time and PMI. Given the still small number of published articles, the lack of globally recognized standardized methods, and the numerous techniques used to evaluate the mRNA degradation times, numerous and larger studies are still necessary to reach more solid and shared evidence.

Digestive enzymatic activity during ontogenetic development in zebrafish (Danio rerio).

Despite the growing importance of zebrafish (Danio rerio) as an experimental model in biomedical research, some aspect of physiological and related morphological age dependent changes in digestive system during larval development are still unknown. In this paper, a biochemical and morphological study of the digestive tract of zebrafish was undertaken to record the functional changes occurring in this species during its ontogenetic development, particularly from 24 hr to 47 days post fertilization (dpf). Endo- and exo-proteases, as well as α-amylase enzymes, were quantified in zebrafish larvae before first feeding (7 dpf). The most morphologically significant events during the ontogenesis of the gut occurred between 3 dpf (mouth opening) and 7 dpf (end of exocrine pancreas differentiation). The presence of a wide range of digestive enzymes, already active at earlier zebrafish larval stages, closely related with the omnivorous diet of this species. Increasing enzyme activities were found with increasing age, probably in relation with intestinal mucosa folding and consequent absorption surface increase. J. Exp. Zool. (Mol. Dev. Evol.) 324B: 699-706, 2015. © 2015 Wiley Periodicals, Inc.

The odd couple: Caffeine and microplastics. Morphological and physiological changes in Mytilus galloprovincialis.

In recent years, the presence of pharmaceuticals and microplastics (MPs) in aquatic ecosystems has raised concerns about their environmental impact. This study explores the combined effects of caffeine, a common pharmaceutical pollutant, and MPs on the marine mussel Mytilus galloprovincialis. Caffeine, at concentrations of 20.0 µg L-1, and MPs (1 mg L-1, 35-50 µm size range), was used to mimic real-world exposure scenarios. Two hundred M. galloprovincialis specimens were divided into four groups: caffeine, MPs, Mix (caffeine + MPs), and Control. After a two-week acclimation period, the mollusks were subjected to these pollutants in oxygen-aerated aquariums under controlled conditions for 14 days. Histopathological assessments were performed to evaluate gill morphology. Cellular volume regulation and digestive gland cell viability were also analyzed. Exposure to caffeine and MPs induced significant morphological changes in M. galloprovincialis gills, including cilia loss, ciliary disk damage, and cellular alterations. The chitinous rod supporting filaments also suffered damage, potentially due to MP interactions, leading to hemocyte infiltration and filament integrity compromise. Hemocytic aggregation suggested an inflammatory response to caffeine. In addition, viability assessments of digestive gland cells revealed potential damage to cell membranes and function, with impaired cell volume regulation, particularly in the Mix group, raising concerns about nutrient metabolism disruption and organ function compromise. These findings underscore the vulnerability of M. galloprovincialis to environmental pollutants and emphasize the need for monitoring and mitigation efforts. RESEARCH HIGHLIGHTS: The synergy of caffeine and microplastics (MPs) in aquatic ecosystems warrants investigation. MPs and caffeine could affect gill morphology of Mytilus galloprovincialis. Caffeine-exposed cells had lower viability than the control group in the NR retention test. MPs and mix-exposed cells struggled to recover their volume.

Morphological characteristics of the blackspot seabream (Pagellus bogaraveo) tongue: A structural and immunohistochemical study.

The blackspot seabream (Pagellus bogaraveo, Brünnich, 1768) is an omnivorous, predominantly carnivorous fish. In aquaculture, it is fed with pellets rich in proteins and fat. The morphological and functional aspects of the fish tongue, the feeding modality and the tasting capacity are strictly related. Therefore, the aim of this study was to describe by scanning electron, light and confocal laser microscopy, the morphological characteristics of the tongue in this species. It showed an apex, a body and a root. There were rows of teeth on the edges of the mouth and taste pores on all the tongue dorsal surface with folds and furrows. In addition, body and root showed several fungiform-like papillae in the mucosa of the folds, covered by a weakly keratinized stratified squamous epithelium, can be observed. The papillae were innervated by S100 positive fibres. In the apex, a mesenchymal tissue with vimentin positive star-shaped stem cells was evident. The results could give a support for a wider use of the blackspot seabream as a farmed species, considering the morphological data as correlated with the potentiality of food discrimination. This provides a basis for possible applications in feeding strategies. The presence, localization and characteristics of the mesenchymal stem cells, as seen also in previous studies, could represent a further basis for future applications in clinical trials.

The Alimentary Tract of African Bony-Tongue, Heterotis niloticus (Cuvier, 1829): Morphology Study.

A morphological study of the alimentary tract, from the oropharyngeal cavity to the rectum, including the attached glands, of African bony-tongue, Heterotis niloticus (Cuvier, 1829) was carried out by gross anatomy, and light microscope analysis. This study aimed to give a deeper knowledge of the alimentary tract morphological features of this species of commercial interest. H. niloticus is distinguished by individual morphological characteristics showing a digestive tract similar to that of reptiles and birds. Within the oropharyngeal cavity, two tubular structures with digitiform ends are arranged on both lateral sides of the triangular tongue. The oropharyngeal cavity connects the stomach by a short esophagus. This latter is adapted to mechanical trituration, and it is divided into a pars glandularis and a thick-walled pars muscularis. The gizzard flows into the anterior intestine and two blind pyloric appendages, which exhibit specific functions, including immune defense for the presence of secondary lymphoid organs. The anterior intestine continues with the middle and posterior tracts up into the rectum. According to the histological observations, all regions of the alimentary tract have common structural features, typical of hollow organs, with differences in the mucosa structure that reflects the different functions of the apparatus, from mouth to anus. Within this study, we provided the first basis for future studies on optimizing rearing conditions, feed conversion ratio, and the digestive capacity, improving the growth performance of this species, and ensuring its conservation.

Morphology of the Atlantic salmon (Salmo salar) tongue.

The Atlantic salmon (Salmo salar) is a freshwater and marine fish of the family Salmonidae, widely farmed in aquaculture facilities in several countries. The salmon are carnivorous, but in aquaculture, alternative foods have been experienced. It is well known that feeding in captivity should cause adaptation and modifications of the morphological characteristics of the oral cavity, especially of tongue; therefore, the aim of this study was to investigate, by light, laser confocal and scanning electron microscopy, the morphological characteristics of the tongue dorsal surface, considering the importance of the correlations between feeding habits and the anatomy of the tongue. Scanning electron microscopy demonstrates the presence of caniniform teeth with oro-aboral orientation surrounded by numerous filiform papillae, single, fused or arranged in row. Oro-aborally, the papillae show an appearance like a rosette and they disappear at level of the root. Light and laser confocal microscopy demonstrates that the mucosa is covered by a non-keratinized stratified pavement epithelium with, in the deepest layer, the presence of a triangular structure whose apex is cranially directed and base facing aborally. In this structure, spindle-shaped cells are present, with a vimentin immunoreactivity, that for their characteristics could be adult mesenchymal stem cells. The obtained data could be useful not only for further studies on the nutrition, but it is interesting the detection of tissues typical of the embryo-fetal phase in the adult specimens tongue, thus giving a basis for studies of potential applications, if any, regarding cell therapies for different clinical indications.

Anatomical, histological and immunohistochemical study of the tongue in the rainbow trout (Oncorhynchus mykiss).

The rainbow trout (Oncorhynchus mykiss Walbaum, 1792) is a fish commercially farmed all over the world. These fish are usually fed, in aquaculture, with pellets rich in proteins and fat. It is well known that there are close relationships among the adaptation of vertebrates to their environment, the capacity and the modality of feeding and the oral cavity morphology, especially the tongue one. No data are so far available about the morphology of the rainbow trout tongue, and therefore, the aim of this study was to investigate by light, scanning electron and confocal laser microscopy, the morphological characteristics of the tongue. An apex, a body and a root can be distinguished in the tongue, and the presence of teeth, taste buds and fungiform-like papillae was demonstrated. Light microscopy shows the presence of an adipose tissue pad in the deeper layer of the apex and in the most superficial layer of the root. In the deeper layer of the body, a triangular-shaped pad consisting of fusiform cells immersed in abundant extracellular matrix of the mesenchymal tissue was observed. The confocal laser microscopy shows the presence of cells with a fibroblast-like morphology positive for vimentin. In the deepest layer of the tongue root, a large area of osteo-cartilaginous tissue was observed. The results, besides the description of the morphological characteristics of the tongue, related to studies regarding the feeding, could be considered for the eventual applications of the use of mesenchymal cells, observed in adult fish, in cell therapies in different pathologies.

LM and SEM study on the swordfish (Xiphias gladius) tongue.

Swordfish (Xiphias gladius L. 1758) is a predatory and migratory fish. Its characteristic feature is a flat and sharp upper jaw forming a "sword". The adaptation of vertebrates, including fish, to their environment is strictly related to the capacity of feeding and is carried out by often severe modifications of the anatomy of the buccal cavity, especially of the tongue. The aim of this study is, using light and scanning electron microscopy and considering that no data are so far available about the morphology of the tongue in this species, to analyse the anatomical characteristics of the tongue, especially its dorsal surface. The tongue shows a triangular shape and an apex, a body and a root. By SEM the presence of several small denticles and filiform papillae on the latero-ventral body was demonstrated while no taste buds or other sensitive structures are observed. LM shows a squamous stratified epithelium, becoming simple cuboidal around the denticles. Therefore this study could add further data to the knowledges of the fish oral cavity morphology supporting the hypothesis that the modifications and evolution of the tongue anatomy are, also in fish, related to the environment and especially to the feeding habits.

Morphology of the European sea bass (Dicentrarchus labrax) tongue.

The European sea bass, a member of the Moronidae family, is a food fish, considered one of the first models for the intensive breeding in salt water. It has nowadays an important and increasing presence in the international fishing markets. Sea basses are carnivorous, feeding on little fishes and invertebrates. Considering the important role of the tongue during the intraoral transport and the swallowing of food, scarce data are present in literature about its morphology. The aim of this study was to analyze the morphology of the tongue by means of scanning electron and light microscopy. Adult sea basses were obtained from the aquarium of the Sicilian Center of Experimental Ichthyiopathology of the University of Messina. The fishes were anaesthetized with MS 222 and the heads were then quickly removed and processed for the paraffin embedding and SEM processing. Three different tongue regions could be distinguished: an apex, a body, and a root. Scanning electron and light microscopy showed the presence of numerous canine-like teeth, surrounded by taste buds and numerous fungiform and conical papillae. The teeth were curved and their tips were posteriorly oriented. The results confirm, in teleosts too, the fundamental role of the tongue in the mechanics of food ingestion. Moreover, the presence of taste buds demonstrates the interaction of food processing and taste. These data could be a potential source to identify new and better methods of nutrition in the breeding of this fish.