Le neurotrofine e i loro specifici recettori nel cervello di Danio rerio

Neurotrophins include a family of growth factors acting on proliferation, differentiation and survival of developing neuronal poulations belonging to both central and peripheral nervous system. Particularly in the postnatal brain, they regulate plasticity and stability of synapses in the CNS, and the complex brain circuitry that regulates behavior. Under- or over-expression of neurotrophins cause neurodegenerative and psychiatric disorders. Neurotrophin signals are transduced by a class of receptors (Trks) with intracytoplasmic tyrosine-kinase domain. All neurotrophins and Trk receptors are well conserved during vertebrate phylogenesis. The aim of this study is to investigate neurotrophins and their Trk receptors in zebrafish or Danio rerio, a teleost fish used in laboratories throughout the world as animal model. It was originally employed for genetic developmental studies and, more recently, for human diseases purposes, included neurodegenerative pathologies. The brain of zebrafish, as in other fish, is extremely interesting for studies regarding adult neurogenesis and reparative processes, due to its high levels of neuronal precursor proliferation and differentiation, which are many times greater than in mammals. Immunochemical and immunocitochemical analysis have been used to investigate the presence and localization of nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), neurotrophin-4 (NT-4) and their specific receptors (TrkA, TrkB, TrkC). Western blottings analysis, performed on zebrafish brain homogenates, showed bands corresponding to expected molecular weights for zebrafish neurotrophins and Trk receptors. Single and double immunocitochemical labeling were carried out on microtomic paraffinic or vibratome agarose sections. Slides were observed at light or confocal microscope. Immunoreactivity to all tested neurotrophins and Trk receptors was widely diffused throughout the zebrafish brain. Particularly, it was localized in neurons and their dendrites/axons, in vessel walls and in different cell populations with glial charcteristics. Generally, cell identification was made on morphological criteria and by means of specific markers. The pattern of distribution of immunoreactivity, such as close proximity or colocalization of neurotrophin/ receptor, suggested an autocrine or paracrine mode of action and the presence of immunolabeling in numerous fibers is likely due to active retrograde and/or anterograde transport. Furthermore, their localization in cell populations with glial properties has been occurred in several neurogenic niches, leading to the hypothesis that these neurotrophic factors might play roles in regulating adult neurogenesis of zebrafish brain. Particularly for BDNF, often it was colocalized with S100β (a glial marker) and PCNA (proliferative marker) in cells of proliferative areas. These results were further confirmed by using two zebrafish transgenic lines: gfap-gfp and nestin-gfp zebrafish, kindly provided by Prof. U. Strahle of Tossicology and Genetic Institute, Forschungzentrum, Karlsruhe, Germany. In conclusion, the research gives ample morphological evidence regarding the involvement of neurotrophins in many biological functions, mostly cell proliferation, of zebrafish brain and represents a useful and promising starting point for future studies devoted to adult neurogenesis and brain repair

Oxidation and antioxidant status: effects on shelf-life of meat from Limousine cattle fed with supplements of alpha-tocopherol

The purpose of this study was to evaluate the antioxidant status of meat from cattle fed diets supplemented with vitamin E (α-tocopherol acetate) during the finishing period and to evaluate the effect of this treatment on meat shelf-life. Twenty purebred Limousine calves reared in the same farm, were randomly selected, divided into control group (n=10) and treated group (n=10) and fed a total mixed ration: treated group received a supplementation of vitamin E (900 mg/kg of CMF) for a period of 150 days before slaughter. Meat quality was evaluated by the following analyses: pH, water holding capacity (drip loss), colour (L*, a*, b*, C*, H*), chemical forms of myoglobin, substances reactive to thiobarbituric acid (MDA) and enzymatic antioxidant activity (superoxide dismutase, catalase and glutation peroxidase). Dietary vitamin E supplementation had a positive effect on water holding capacity; in the control group a considerable increase in drip loss from the 2nd to 6th day of conservation was observed (2.83% vs 7.54%), while in the treated group during the same time period this increase appeared to be much more gradual and occurred to a lesser degree (2.31% vs 4.15%). Moreover, administration of vitamin E led to greater stability of colorimetric coordinates and reduced discoloration of the longissimus dorsi muscle during conservation as indicated by the redness a* (control: 23.85 and 23.87 vs 19.34 at 2h, 2 and 6 days, respectively; treated: 24.88, 23.91 and 24.01 at 2h, 2 days and 6 days, respectively) and in the Chroma* (control: 26.89 and 26.77 vs 21.90 at 2h, 2 days and 6 days, respectively; treated: 27.67, 26.57 and 26.77 at 2h, 2 and 6 days, respectively). Superoxide dismutase activity was significantly greater in the meat from cattle treated with vitamin E compared to that observed in the meat from controls (0.204 vs 0.167). The study showed that vitamin E supplementation in the finishing diets of calves caused only slight modifications in the antioxidant status of the meat; however, it positively influenced several qualitative characteristics which appeared to be more stable over time, thus extending the shelf-life of the meat

Oxidation and antioxidant status: effects on shelf-life of meat from Limousine cattle fed with supplements of α-tocopherol

The purpose of this study was to evaluate the antioxidant status of meat from cattle fed diets supplemented with vitamin E (α-tocopherol acetate) during the finishing period and to evaluate the effect of this treatment on meat shelf-life. Twenty purebred Limousine calves reared in the same farm, were randomly selected, divided into control group (n=10) and treated group (n=10) and fed a total mixed ration: treated group received a supplementation of vitamin E (900 mg/kg of CMF) for a period of 150 days before slaughter. Meat quality was evaluated by the following analyses: pH, water holding capacity (drip loss), colour (L*, a*, b*, C*, H*), chemical forms of myoglobin, substances reactive to thiobarbituric acid (MDA) and enzymatic antioxidant activity (superoxide dismutase, catalase and glutation peroxidase). Dietary vitamin E supplementation had a positive effect on water holding capacity; in the control group a considerable increase in drip loss from the 2nd to 6th day of conservation was observed (2.83% vs 7.54%), while in the treated group during the same time period this increase appeared to be much more gradual and occurred to a lesser degree (2.31% vs 4.15%). Moreover, administration of vitamin E led to greater stability of colorimetric coordinates and reduced discoloration of the longissimus dorsi muscle during conservation as indicated by the redness a* (control: 23.85 and 23.87 vs 19.34 at 2h, 2 and 6 days, respectively; treated: 24.88, 23.91 and 24.01 at 2h, 2 days and 6 days, respectively) and in the Chroma* (control: 26.89 and 26.77 vs 21.90 at 2h, 2 days and 6 days, respectively; treated: 27.67, 26.57 and 26.77 at 2h, 2 and 6 days, respectively). Superoxide dismutase activity was significantly greater in the meat from cattle treated with vitamin E compared to that observed in the meat from controls (0.204 vs 0.167). The study showed that vitamin E supplementation in the finishing diets of calves caused only slight modifications in the antioxidant status of the meat; however, it positively influenced several qualitative characteristics which appeared to be more stable over time, thus extending the shelf-life of the meat

Neurotrophins in Zebrafish Taste Buds

SIMPLE SUMMARY: Zebrafish is a powerful vertebrate model organism, whose similarities with mammals are fundamental to validate its use for experimental purposes. In this study, the authors demonstrate the presence of neurotrophic factors, namely neurotrophins, in numerous taste bud cells of this fish. The reported results suggest an essential role of these factors in taste bud function. Interestingly, the results described in this study are in accordance with those reported in some mammalian species. Therefore, despite the different anatomical characteristics of the anterior digestive tract in mammals and fish, the taste buds maintain similarities in both shape and functional mechanisms in the two classes. ABSTRACT: The neurotrophin family is composed of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), Neurotrophin 3 (NT3) and NT4. These neurotrophins regulate several crucial functions through the activation of two types of transmembrane receptors, namely p75, which binds all neurotrophins with a similar affinity, and tyrosine kinase (Trk) receptors. Neurotrophins, besides their well-known pivotal role in the development and maintenance of the nervous system, also display the ability to regulate the development of taste buds in mammals. Therefore, the aim of this study is to investigate if NGF, BDNF, NT3 and NT4 are also present in the taste buds of zebrafish (Danio rerio), a powerful vertebrate model organism. Morphological analyses carried out on adult zebrafish showed the presence of neurotrophins in taste bud cells of the oropharyngeal cavity, also suggesting that BDNF positive cells are the prevalent cell population in the posterior part of the oropharyngeal region. In conclusion, by suggesting that all tested neurotrophins are present in zebrafish sensory cells, our results lead to the assumption that taste bud cells in this fish species contain the same homologous neurotrophins reported in mammals, further confirming the high impact of the zebrafish model in translational research

The Potential Impact of a Dog Training Program on the Animal Adoptions in an Italian Shelter

SIMPLE SUMMARY: Human–dog relationships are mainly focused on the physical and emotional wellbeing, and has much evolved in the last decades, becoming even more intense over time. Such a novel conception should also be applied to the life that takes place in kennels, which have been mistakenly regarded as a landfill or a burden on society for too many years. Here, we took into consideration an Italian shelter, and analyzed 555 adopted dogs who underwent a well-detailed behavioral training program, to assess the potential impact of the education upon the adoption of attitude. We documented a higher increase in the number of adoptions both for adult and senior animals when compared to the age-matched untrained dogs. Collectively, our data highlight the importance of a proper training, mainly accomplished with reward methods, to get a more suitable and balanced owner-dog attachment. ABSTRACT: One of the main concerns of the human–dog relationship is today associated with the quality life inside the kennels, which are very often regarded as animal dump where dogs are exiled, representing a burden on society. In the present study we sought to investigate the importance of performing an appropriate behavioral program on the adoption chances within an Italian shelter, near Naples (Ottaviano). In this respect, we enrolled 555 adopted dogs of different ages, who followed a tailored-4-month lasting training program between 2018 and 2020. Once entered there, they were carefully examined by the veterinary behaviorist, and directed towards a suited training program, to improve living conditions. We documented a higher number of both adult and senior dogs who left the kennel and were adopted, compared to the age-matched untrained animals (n = 479), housed in the same kennel from 2015 to 2017. Taken together, the present data highlight an important role for training in improving the natural attitudes of the companion dogs, thus pointing towards a better human–animal bond

Combining cannabis and melatonin treatment with a rehabilitation program improved symptoms in a dog with compulsive disorder: A case report

: Compulsive disorder in dogs (CD) is characterized by constant and time-consuming repetition of behaviors, emancipated from the environment, that definitely compromise their everyday life activities. Here, we documented the efficacy of a novel approach to counteract the negative symptoms of CD in a 5-year-old mongrel dog, previously found to be resistant to the conventional antidepressant. The patient underwent an integrated and interdisciplinary approach, based on the cannabis and melatonin co-administration, together with a tailored 5-month-lasting behavioral program. Observational findings showed a lower rate of compulsive episodes and better management of the dog as well, when compared to the previous paroxetine treatment. We followed him for an additional four months of therapy, and the owners reported easier management of the dog, as reduction of abnormal behaviors to a level acceptable to the owners. Overall, our data so far collected in the CD dog may allow us to test more deeply the feasibility and safety of such an off-label approach, at both preclinical and clinical levels

Biophysical and biological characterization of a new line of hyaluronan-based dermal fillers: A scientific rationale to specific clinical indications

Chemico-physical and biological characterization of hyaluronan-based dermal fillers is of key importance to differentiate between numerous available products and to optimize their use. These studies on fillers are nowadays perceived as a reliable approach to predict their performance in vivo. The object of this paper is a recent line of hyaluronic acid (HA)-based dermal fillers, Aliaxin®, available in different formulations that claim a complete facial restoration. The aim of the study is to provide biophysical and biological data that may support the clinical indications and allow to predict performance possibly with respect to similar available products. Aliaxin® formulations were tested for their content in soluble HA, water uptake capacity, rheological behavior, stability to enzymatic degradation, and for in vitro capacity to stimulate extracellular matrix components production. The formulations were found to contain a low amount of soluble HA and were equivalent to each other regarding insoluble hydrogel concentration. The different crosslinking degree declared by the producer was consistent with the trend in water uptake capacity, rigidity, viscosity. No significant differences in stability to enzymatic hydrolysis were found. In vitro experiments, using a full thickness skin model, showed an increase in collagen production in the dermoepidermal junction. Results support the claims of different clinical indications, the classification of products regarding hydro-, lift-action and the specifically suggested needle gauge for the delivery. The biological outcomes also support products effectiveness in skin structure restoration. These data predicted a better performance regarding hydro-action, tissue integration, clinical management during delivery, and a high durability of the aesthetic effect when compared to data on marketed similar products

Immunolocalization of Nesfatin-1 in the Gastrointestinal Tract of the Common Bottlenose Dolphin Tursiops truncatus

First identified as an anorexigenic peptide, in the last decades, several studies have suggested that Nesfatin-1 (Nesf-1) is a pleiotropic hormone implicated in numerous regulatory processes in peripheral organs and tissues. In vertebrates, Nesf-1 is indeed expressed in the central nervous system and peripheral organs. In this study, we characterized the pattern of Nesf-1 distribution within the digestive tract of the common bottlenose dolphin (Tursiops truncatus), composed of three gastric chambers and and intestine without a clear subdivision in the small and large intestine, also lacking a caecum. Our results indicated that Nesf-1 is widely distributed in cells of the mucosal epithelium of the gastric chambers. Most of the immunoreactivity was observed in the second chamber, compared to the first and third chambers. Immunopositivity was also found in nerve fibers and neurons, scattered or/and clustered in ganglion structures along all the esaminaned gastrointestinal tracts. These observations add new data on the highly conserved role of Nesf-1 in the mammalian digestive system

Sequence-specific transcription factor NF-Y displays histone-like DNA binding and H2B-like ubiquitination

SummaryThe sequence-specific transcription factor NF-Y binds the CCAAT box, one of the sequence elements most frequently found in eukaryotic promoters. NF-Y is composed of the NF-YA and NF-YB/NF-YC subunits, the latter two hosting histone-fold domains (HFDs). The crystal structure of NF-Y bound to a 25 bp CCAAT oligonucleotide shows that the HFD dimer binds to the DNA sugar-phosphate backbone, mimicking the nucleosome H2A/H2B-DNA assembly. NF-YA both binds to NF-YB/NF-YC and inserts an α helix deeply into the DNA minor groove, providing sequence-specific contacts to the CCAAT box. Structural considerations and mutational data indicate that NF-YB ubiquitination at Lys138 precedes and is equivalent to H2B Lys120 monoubiquitination, important in transcriptional activation. Thus, NF-Y is a sequence-specific transcription factor with nucleosome-like properties of nonspecific DNA binding and helps establish permissive chromatin modifications at CCAAT promoters. Our findings suggest that other HFD-containing proteins may function in similar ways