Francesco Loy

Decellularized ovine arteries as small-diameter vascular grafts

Atherosclerosis and its complications still represent the leading cause of death in the developed countries. While autologous blood vessels may be regarded as the best solution for peripheral and coronary bypass, they are unavailable in most patients. Even though tissue engineering techniques are often applied to the development of small-diameter vascular grafts, limiting factors of this approach are represented by the lack of essential extracellular matrix proteins and/or poor biomechanical properties of the scaffolds used. Along these lines, the aim of this study was to develop a decellularization protocol for ovine carotids to be used as suitable small-diameter vascular grafts. Samples were treated either with sodium dodecyl sulphate (SDS) or with Trypsin and Triton X-100; a final nuclease digestion was performed for both protocols. Morphological analyses demonstrate complete removal of nuclei and cellular components in treated vessels, also confirmed by significant reduction in wall thickness and DNA content. Essential extracellular matrix proteins such as collagen, elastin, and fibronectin are well preserved after decellularization. From a mechanical point of view, Trypsin and Triton X-100 treated arteries show elastic modules and compliance comparable to native carotids, whereas the use of SDS makes samples stiffer, with a significant decrease in the compliance mean value and an increase in longitudinal and circumferential Youngs modules. It is demonstrated that the treatment where Trypsin and Triton X-100 are combined guarantees complete decellularization of carotids, with no significant alteration of biomechanical and structural properties, thus preserving a suitable environment for adhesion, proliferation, and migration of cells.

The evolution of anatomical illustration and wax modelling in Italy from the 16th to early 19th centuries

Although the contribution to anatomical illustration by Vesalius and his followers has received much attention, less credit has been given to Veslingius and particularly Fabricius. By 1600, Fabricius had amassed more than 300 paintings that together made the Tabulae Pictae, a great atlas of anatomy that was highly admired by his contemporaries. Many of his new observations were incorporated into subsequent books, including those by Casserius, Spighelius, Harvey and Veslingius. Also of importance were the Tabulae by Eustachius (1552), which, although only published in 1714, greatly influenced anatomical wax modelling. In 1742, Pope Benedict XIV established a Museum of Anatomy in Bologna, entrusting to Ercole Lelli the creation of several anatomical preparations in wax. Felice Fontana realised that the production of a large number of models by the casting method would make cadaveric specimens superfluous for anatomical teaching and in 1771 he asked the Grand Duke to fund a wax‐modelling workshop in Florence as part of the Natural History Museum, later known as La Specola. Fontana engaged Giuseppe Ferrini as his first modeller and then the 19‐year‐old Clemente Susini who, by his death in 1814, had superintended the production of, or personally made, more than 2000 models. In 1780, the Austrian Emperor Joseph II visited La Specola and ordered a great number of models for his Josephinum museum; these were made by Fontana with the help of Clemente Susini and supervised by the anatomist Paolo Mascagni. It is, however, in Cagliari that some of Susini’s greatest waxes are to be found. These were made when he was free of Fontana’s influence and were based on dissections made by Francesco Antonio Boi (University of Cagliari). Their distinctive anatomical features include the emphasis given to nerves and the absence of lymphatics in the brain, a mistake made on earlier waxes. The refined technical perfection of the anatomical details demonstrates the closeness of the cooperation between Susini and Boi, whereas the expressiveness of the faces and the harmony of colours make the models of Cagliari masterpieces of figurative art.

Diabetes causes morphological changes in human submandibular gland: a morphometric study.

BACKGROUND Dataon structural alterations in human diabetic salivary glands are scanty and conflicting. The goal of this study is based on the evaluation of the morphological changes in submandibular glands of subjects with well-controlled diabetes and without evident salivary malfunctions. METHODS Submandibular gland pieces from diabetic and non-diabetic patients were fixed, dehydrated, and processed to obtain sections for light and electron microscopy. Randomly selected micrographs were statistically analyzed to reveal variations in serous acini. RESULTS Morphometrical evaluation allowed us to reveal significant changes such as enlargement of acinar and granule size, reduction of mitochondrial size, increased density of microbuds and protrusions along luminal membranes. CONCLUSIONS The results indicate that diabetes affects submandibular gland structure even when glandular function appears unaltered and suggest that morphological changes reflect functional changes chiefly regarding the secretory activity.

Morphological evidence that pentagastrin regulates secretion in the human parotid gland

Salivary secretion is principally regulated by autonomic nerves. However, recent evidence from in vivo animal experiments suggests that gastrointestinal peptide hormones can also influence saliva production. The aim of the present study was to define the secretagogue activity of the gastrin‐analogue pentagastrin in human salivary glands. For this purpose, parotid tissues were exposed to pentagastrin in vitro. Morphological techniques were used to evaluate modifications to serous acinar cells associated with secretion. Using a variant of the osmium maceration method, high resolution scanning electron microscopy allowed assessment of the morphology of the cytoplasmic aspect of the plasmalemma to demonstrate secretory activity. To quantify responses to pentagastrin, we recorded morphometric data on microvilli, microbuds, and protrusions. Dose‐dependent morphological changes were observed, whereas protein concentration increased in the incubate. The use of selective receptor antagonists showed pentagastrin to act principally via cholecystokinin‐A receptors. The morphological responses observed following exposure to pentagastrin differed from those elicited following exposure to the pan‐muscarinic agonist carbachol. This study provides the first demonstration of a direct secretory action of gastrointestinal peptides on salivary glands in humans.

Morpho-functional identification of abdominal olfactory receptors in the midge Culicoides imicola

The aim of the present study was to examine the presence and the possible role of abdominal olfactory sensilla in Culicoides imicola mediating the search for potential hosts and oviposition sites, by means of a morphological, electrophysiological and behavioural approach. The results reported here show that in the midge C. imicola the whole abdomen, comprising the ovipositor, are endowed with three morphotypes of multiporous sensilla that display olfactory sensitivity towards kairomones related to the host-animal skin such as l-(+)-lactic acid and 1-octen-3-ol, to the host-animal urine such as 3-ethylphenol and 4-propylphenol, and to the potent attractant sesame seed oil. Electrophysiological and behavioural data for the first time suggest in the midge the involvement of abdominal olfactory structures in the choice of the oviposition sites and allow in discussing their possible role in the host-animal localisation. Field experiments showed that light traps baited with the aforementioned compounds elicited a stronger degree of attractiveness on midges with respect to the unbaited traps (control), although to a different extent. Our results, while implying a number of considerations concerning the role of molecules tested as kairomones, also suggest their use in the control of the midge C. imicola population.

Diabetes reduces statherin in human parotid: immunogold study and comparison with submandibular gland.

BACKGROUND AND OBJECTIVE Alteration of salivary gland secretion is one of the consequences of diabetes. In a recent study on the submandibular gland of diabetic subjects, we found changed expression of statherin, a salivary protein of fundamental importance in preserving tooth integrity, whose reduction was related with the high incidence of oral diseases in patients with diabetes. The goal of this report is to extend the study to human parotid gland and to compare the effects of diabetes on statherin expression with those previously described in submandibular gland. MATERIALS AND METHODS Fragments of parotid glands obtained from diabetic and non-diabetic patients were fixed, dehydrated, embedded in Epon Resin and processed for the immunogold histochemistry. The staining density was expressed as number of gold particles per μm(2) and statistically evaluated. RESULTS AND CONCLUSIONS In all samples, statherin reactivity was specifically localized in secretory granules of acinar cells. The statistical analysis showed that labelling density was significantly lower in diabetic than in non-diabetic parotid glands and that diabetes affects protein expression at identical extent in parotid and submandibular glands. The results strengthen the hypothesis that a reduced statherin secretion may be responsible for the higher incidence of oral disorders in diabetic subjects.

Diabetes affects statherin expression in human labial glands.

BACKGROUND AND OBJECTIVE Salivary statherin, which plays a special role in the defense of tooth integrity, is secreted by both major and minor salivary glands. A significantly reduced expression of this was recently found in human major salivary glands removed from diabetic subjects and was correlated with the high incidence of dental diseases occurring in patients with diabetes. In this study, we measured the density of gold particles indicating statherin immunoreactivity in labial glands to reveal a significant difference between diabetic and non-diabetic patients. MATERIALS AND METHODS Surgical samples of labial glands obtained from both diabetic and non-diabetic patients were fixed with a glutaraldehyde and paraformaldehyde mixture, embedded in Epon, and treated for immunogold histochemistry using a polyclonal antibody specific for statherin. RESULTS Statherin immunoreactivity was detected onto small vesicles diffused throughout the cytoplasm of serous cells. Statistical analysis revealed that the number of stained particles was significantly lower in the samples from diabetic subjects than from non-diabetic subjects. CONCLUSIONS The results indicate that diabetes affects statherin secretion in labial glands and support the hypothesis that the increased susceptibility to oral diseases associated with diabetes could be related with a reduced statherin secretion.

3-D structure of mitochondrial cristae in rat adrenal cortex varies after acute stimulation with ACTH and CRH.

We attempted to determine whether acute treatment with adrenocorticotropin hormone (ACTH) and corticotropin releasing hormone (CRH) affects mitochondrial morphology, as evaluated by the HRSEM and osmium maceration methods. We quantified CRH and ACTH effects on HRSEM images in rat glomerulosa and fasciculata. After ACTH or CRH treatment, mitochondrial cristae increased the number of globular expansions, whereas mitochondrial volume decreased in glomerulosa. As the morphological variations reported may be linked to increased hormonal production, further studies using parallel measurements of circulating and tissue hormones are now in progress, and may aid in clarifying their functional significance.

Cytomorphological study on human submandibular gland following treatment with secretagogue drugs

Using specimens of human submandibular glands, we have investigated in vitro the morphological modifications induced by clozapine, a dibenzodiazepine derivative that is used in psychotic patients and that provokes hypersalivation, a side-effect of therapy. The effects of the drug, used alone or in combination with carbachol, have been compared with those observed after treatment with drugs acting on specific receptors. To quantify the response to stimulation, we have calculated (with statistical methods) the number of microvilli and microbuds (corresponding to pits seen in images obtained by transmission electron microscopy) per square micrometre of the cytoplasmic surface of the intercellular canaliculi luminal membrane in images obtained by high-resolution scanning electron microscopy. Clozapine, when directly acting on human submandibular specimens, induces a small secretory response in serous cells; this is partially decreased by muscarinic and adrenergic antagonists and by combined incubation with carbachol, thus confirming its behaviour as a partial agonist to muscarinic receptors. We also suggests that the drug acts on the nerve terminals contained within the glandular specimens.

Fat storage in Drosophila suzukii is influenced by different dietary sugars in relation to their palatability

The peripheral sensitivity and palatability of different carbohydrates was evaluated and their nutritional value assessed in adult females of D. suzukii by means of an electrophysiological, behavioural and metabolic approach. The electrophysiological responses were recorded from the labellar “l” type sensilla stimulated with metabolizable mono- and disaccharides (glucose and maltose) and a non-metabolizable sugar (sucralose); the response rating and the palatability to the same sugars, evaluated by recording the proboscis extension reflex (PER), was maltose>glucose>sucralose. The nutritional value of carbohydrates was assessed by means of survival trials and fatty acids profile. Flies fed on a diet containing maltose had a longer lifespan than flies on monosaccharides, while flies fed on a diet containing sucralose had a shorter one. In addition, the ability to store fat seems to be influenced by the different sugars in the diet and is in relationship with their palatability. In fact, data showed a higher synthesis of palmitic and palmitoleic acids, most likely derived from de-novo lipogenesis with glucose as precursor, in flies fed with maltose and glucose than with non-metabolizable sucralose. In conclusion, these results suggest that the ability to select different sugars on the basis of their palatability may favour the storage of energy reserves such as fat by de-novo lipogenesis, determining a longer survival capability during prolonged periods of fasting.