Immunohistochemistry detected and localized cannabinoid receptor type 2 in bovine fetal pancreas at late gestation

At present, data on the endocannabinoid system expression and distribution in the pancreatic gland appear scarce and controversial as descriptions are limited to humans and laboratory animals. Since the bovine pancreas is very similar to the human in endocrine portion development and control, studies on the fetal gland could prove to be very interesting, as an abnormal maternal condition during late pregnancy may be a predisposing trigger for adult metabolic disorders. The present investigation studied cannabinoid receptor type 2 presence and distribution in the bovine fetal pancreas towards the end of gestation. Histological analyses revealed numerous endocrinal cell clusters or islets which were distributed among exocrine adenomeri in connectival tissue. Immunohistochemistry showed that endocrine-islets contained some CB2-positive cells with a very peculiar localization that is a few primarily localized at the edges of islets and some of them also scattered in the center of the cluster. Characteristically, also the epithelium of the excretory ducts and the smooth muscle layers of the smaller arteries, in the interlobular glandular septa, tested positive for the CB2 endocannabinoid receptor. Conse quently, the endocannabinoid system, via the cannabinoid receptor type 2, was hypothesized to play a major role in controlling pancreas function from normal fetal development to correct metabolic functioning in adulthood

Further observations on the sensitive innervation of some bird’s proctodeum

The AA. studied the autonomic and sensitive somatic innervation of some female bird's proctodeum, through the properly modified Ruffini's gold chloride method. The vegetative component was constituted by ganglion cells of different size, isolated or grouped to form ganglia, found along the course of nerve trunks or in the concurrent point of different nerve bundles. The sensitive somatic innervation was represented by free and encapsulated endings differently distributed in the thickness of the wall. The former were composed of thin networks, while the latter, located more frequently in the muscular tunica and in the subadventitial connective, were composed of encapsulated receptors classified as Pacini, Pacini-like and Herbst corpuscles. The morphology of these receptors was described and hypotheses were brought up about their probable functional role. The AA. also found, even if very rarely, helicoidal collagen fibres around nerve fascicles

Multiple endocrine neoplasia type 2 syndromes (MEN 2): results from the ItaMEN network analysis on the prevalence of different genotypes and phenotypes.

OBJECTIVE: Multiple endocrine neoplasia type 2 (MEN 2) is a genetic disease characterized by medullary thyroid carcinoma (MTC) associated (MEN 2A and 2B) or not familial MTC (FMTC) with other endocrine neoplasia due to germline RET gene mutations. The prevalence of these rare genetic diseases and their corresponding RET mutations are unknown due to the small size of the study population. METHODS: We collected data on germline RET mutations of 250 families with hereditary MTC followed in 20 different Italian centres. RESULTS AND CONCLUSIONS: The most frequent RET amino acid substitution was Val804Met (19.6%) followed by Cys634Arg (13.6%). A total of 40 different germline RET mutations were present. Six families (2.4%) were negative for germline RET mutations. The comparison of the prevalence of RET germline mutations in the present study with those published by other European studies showed a higher prevalence of Val804Met and Ser891Ala mutations and a lower prevalence of Leu790Phe and Tyr791Phe (P<0.0001). A statistically significant higher prevalence of mutations affecting non-cysteine codons was also found (P<0.0001). Furthermore, the phenotype data collection showed an unexpected higher prevalence of FMTC (57.6%) with respect to other MEN 2 syndromes (34% MEN 2A and 6.8% of MEN 2B). In conclusion, we observed a statistically significant different pattern of RET mutations in Italian MEN 2 families with respect to other European studies and a higher prevalence of FMTC phenotype. The different ethnic origins of the patients and the particular attention given to analysing apparently sporadic MTC for RET germline mutations may explain these findings

Dopamine-Induced Conformational Changes in Alpha-Synuclein

Background: Oligomerization and aggregation of α-synuclein molecules play a major role in neuronal dysfunction and loss in Parkinson's disease [1]. However, α-synuclein oligomerization and aggregation have mostly been detected indirectly in cells using detergent extraction methods [2], [3], [4]. A number of in vitro studies showed that dopamine can modulate the aggregation of α-synuclein by inhibiting the formation of or by disaggregating amyloid fibrils [5], [6], [7]. Methodology/Principal Findings: Here, we show that α-synuclein adopts a variety of conformations in primary neuronal cultures using fluorescence lifetime imaging microscopy (FLIM). Importantly, we found that dopamine, but not dopamine agonists, induced conformational changes in α-synuclein which could be prevented by blocking dopamine transport into the cell. Dopamine also induced conformational changes in α-synuclein expressed in neuronal cell lines, and these changes were also associated with alterations in oligomeric/aggregated species. Conclusion/Significance: Our results show, for the first time, a direct effect of dopamine on the conformation of α-synuclein in neurons, which may help explain the increased vulnerability of dopaminergic neurons in Parkinson's disease

Post translational changes to α-synuclein control iron and dopamine trafficking : a concept for neuron vulnerability in Parkinson's disease

Parkinson's disease is a multifactorial neurodegenerative disorder, the aetiology of which remains elusive. The primary clinical feature of progressively impaired motor control is caused by a loss of midbrain substantia nigra dopamine neurons that have a high α-synuclein (α-syn) and iron content. α-Syn is a neuronal protein that is highly modified post-translationally and central to the Lewy body neuropathology of the disease. This review provides an overview of findings on the role post translational modifications to α-syn have in membrane binding and intracellular vesicle trafficking. Furthermore, we propose a concept in which acetylation and phosphorylation of α-syn modulate endocytic import of iron and vesicle transport of dopamine during normal physiology. Disregulated phosphorylation and oxidation of α-syn mediate iron and dopamine dependent oxidative stress through impaired cellular location and increase propensity for α-syn aggregation. The proposition highlights a connection between α-syn, iron and dopamine, three pathological components associated with disease progression in sporadic Parkinson's disease

Modes of Aβ toxicity in Alzheimer’s disease

Alzheimer’s disease (AD) is reaching epidemic proportions, yet a cure is not yet available. While the genetic causes of the rare familial inherited forms of AD are understood, the causes of the sporadic forms of the disease are not. Histopathologically, these two forms of AD are indistinguishable: they are characterized by amyloid-β (Aβ) peptide-containing amyloid plaques and tau-containing neurofibrillary tangles. In this review we compare AD to frontotemporal dementia (FTD), a subset of which is characterized by tau deposition in the absence of overt plaques. A host of transgenic animal AD models have been established through the expression of human proteins with pathogenic mutations previously identified in familial AD and FTD. Determining how these mutant proteins cause disease in vivo should contribute to an understanding of the causes of the more frequent sporadic forms. We discuss the insight transgenic animal models have provided into Aβ and tau toxicity, also with regards to mitochondrial function and the crucial role tau plays in mediating Aβ toxicity. We also discuss the role of miRNAs in mediating the toxic effects of the Aβ peptide

EFFECT OF DIFFERENT PHYSICAL FORMS OF THE DIET FETO GROWING PIGS ON THE EXPRESSION OF CB1 AND CB2 IN THE MANDIBULAR GLAND

Diet-induced morphological changes of extra-enteral organs were associated with the physical forms of the diet [1]. The endocannabinoid system consists of a complexity of endogenous molecole, namely endocannabinoids ligands and receptors. There are two types of cannabinoid receptors, named type 1 receptor (CB1) and type 2 receptor (CB2). The investigation about the localization of these two receptors in the duct epithelial cells of the major salivary glands moved on the one hand from the different expression and localization of leptin and its receptor [2] and on the other hand from the decreased effect of endocannabinoids on the salivary secretion, observed in men exposed to exogenous cannabis [3]. The aim of the present study was to test whether different physical forms of diet can induce to different extents the expression of CB1 and CB2 in the mandibular glands in response to different mechanical stimulation perceived after the intake of feed differently ground (fine vs. coarse) and compacted (mash vs. pellet vs. extruded) by growing pigs. The experiment involved a total of 32 growing pigs fed ad libitum for 4 weeks with four experimental dietary treatments. One diet was differently processed to obtain different physical forms: FP - Finely ground pelleted diet (dMEAN, 0.46 mm); CM - Coarsely ground meal diet (dMEAN, 0.88 mm); CP - Coarsely ground pelleted diet (dMEAN, 0.84 mm); CE - Coarsely ground extruded (dMEAN, 0.66 mm) diet. At the end of the experimental feeding, all animals were euthanized and both mandibular glands immediately removed, weighed and fixed in buffered formaldehyde (2,5% v/v) for 24 h at room temperature. Samples were cut and automatically embedded in paraffin. The immunohistochemical reactions were visualized on 5 µm serial sections, utilising the primary goat polyclonal anti-CB1 and rabbit polyclonal anti-CB2 antibodies, the avidin-biotin-complex and the DAB as the chromogen. Sections in which the primary antibodies were omitted, represented the control of unspecific staining. A strong positivity for CB1 and CB2 in the mandibular glands of the animals fed with CP, FP and CE diets was pointed out in comparison with the animals of the CM group. In particular, the CB1 and CB2 immuno-positivity involved duct epithelial cells with a peculiar localization in the cytoplasm of some epithelial cells near or on the apical cell membrane. In the animal fed with CM diet the immuno-positivity no longer involved duct epithelial cells but in some samples as far as the serous cells in mixed acina. The connective tissue tested negative for CB1 and CB2. The CB1 and CB2 were differently espressed in the mandibular glands of pigs fed with different physical forms of the diet. These novel findings leads us to speculate on CB1 and CB2 role, as receptors involved in the control of pig salivary secretion via endocannabinoids ligands and that these molecole likely represent an important link between the physical form of the diet and salivation. However, whether they rule on amount of saliva produced or on its composition needs to be elucidated so far

Estimates of environmental exposure to radiofrequency electromagnetic fields and risk of lymphoma subtypes

We investigated the association between environmental exposure to radiofrequency electromagnetic fields (RF-EMF) and risk of lymphoma subtypes in a case-control study comprised of 322 patients and 444 individuals serving as controls in Sardinia, Italy in 1998–2004. Questionnaire information included the selfreported distance of the three longest held residential addresses from fixed radio-television transmitters and mobile phone base stations. We georeferenced the residential addresses of all study subjects and obtained the spatial coordinates of mobile phone base stations. For each address within a 500-meter radius from a mobile phone base station, we estimated the RF-EMF intensity using predictions from spatial models, and we performed RF-EMF measurements at the door in the subset of the longest held addresses within a 250meter radius. We calculated risk of lymphoma and its major subtypes associated with the RF-EMF exposure metrics with unconditional logistic regression, adjusting by age, gender and years of education. In the analysis of self-reported data, risk associated with residence in proximity (within 50 meters) to fixed radiotelevision transmitters was likewise elevated for lymphoma overall [odds ratio = 2.7, 95% confidence interval = 1.5–4.6], and for the major lymphoma subtypes. With reference to mobile phone base stations, we did not observe an association with either the self-reported, or the geocoded distance from mobile phone base stations. RF-EMF measurements did not vary by case-control status. By comparing the self-reports to the geocoded data, we discovered that the cases tended to underestimate the distance from mobile phone base stations differentially from the controls (P = 0.073). The interpretation of our findings is compromised by the limited study size, particularly in the analysis of the individual lymphoma subtypes, and the unavailability of the spatial coordinates of radio-television transmitters. Nonetheless, our results do not support the hypothesis of a link between environmental exposure to RF-EMF from mobile phone base stations and risk of lymphoma subtype