51 research outputs found
Expression of VaChT and 5-HT in Ulcerative colitis dendritic cells
No full textUlcerative colitis is a chronic inflammatory condition of the gastrointestinal tract that can affect people of worldwide. In contrast with Crohn's disease, that can relate the entire thickness of the bowel wall, the inflammation of ulcerative colitis is limited to the colonic mucosa. Immune cells including activated T cells, plasma cells, mast cells, macrophages, and dendritic cells (DCs) trigger the inflammation. Furthermore, dendritic cells are antigen presenting cells involved in maintaining intestinal immune homeostasis. It has been described an increment of number in DCs colonic mucosa of patients with ulcerative colitis. The immune cells such as antigen-presenting cells can act as autocrine or paracrine modulators. Recent studies showed that dendritic cells synthetized and released classical neurotransmitters as glutamate, dopamine, acetylcholine, and serotonin. Paraformaldehyde-fixed intestinal tissues, obtained from the stricture sites of ten patients with ulcerative colitis were analyzed by immunostaining for Langerin/CD207, serotonin and vesicular acetylcholine transporter. As controls, unaffected (normal) portions of five patients were also investigated. Aim of this study was to characterize for the first time the human gut dendritic cells of ulcerative colitis patients, with Langerin/CD207 that is a c-type lectin expressed by different types of DCs and to colocalize in the same cells the expression of serotonin and vesicular acetylcholine transporter, showing the link between dendritic cells, gut enterochromaffin cells or autonomic nerves in immune activation and generation of intestinal inflammation
Incretin-based therapy and acute cholecystitis: a review of case reports and EudraVigilance spontaneous adverse drug reaction reporting database
No full textWHAT IS KNOWN AND OBJECTIVE:
Glucagon-like peptide-1 (GLP-1) receptor agonists delay gastric and bowel emptying. A similar inhibitory effect of GLP-1 on gallbladder motility has been suggested, possibly leading to an increased risk of cholecystitis related to incretin-based medications, which include GLP-1 antagonists. Our objective was to review evidence in EudraVigilance, the European spontaneous reporting database and the scientific literature on this issue.
COMMENT:
Increasing evidence suggests an association of incretins with gallbladder adverse events. Pharmacovigilance data from EudraVigilance includes 200 serious ADR reports concerning cholecystitis related to the use of incretin-based therapies. Several mechanisms may explain this increased risk of cholecystitis, including rapid weight loss, inhibition of gallbladder contraction and emptying, reduced bile acids production, modulation of inflammation.
WHAT IS NEW AND CONCLUSIONS:
The available data suggest the possibility of gallbladder disease in diabetic subjects treated with incretins and highlight the importance of evaluating risk factors for cholelithiasis and gallbladder diseases in patients with diabetes before starting this therapy
Histochemical and morphological aspect of fresh frozen bone after maxillary graft: a preliminary study
No full textBone grafts are being made using materials extracted from autogenous bone because of its excellent biocompatibility. Bone grafts are widely used, in dentistry, for the reconstruction of severely atrophic jaws ,caused by trauma, oncologic diseases, oral infections or congenitally missing teeth. Although autografts remain the “gold standard”, however, the amount of bone, donor site morbility and unpredictable graft resorption are limitations of using this bone source. There is an increased use of fresh frozen bone allografts (FFBs) in orthopedics and in dentistry.(1) The FFB is aseptically harvested from different skeletal sites of live or cadaveric donors, immediately frozen and stored at −80 °c(2). The rigorous protocol for bone processing, which eliminates living cells and consequently the risk of transmission of diseases, and the reduced immunological reaction. The fresh frozen bone provided with osteoconduction and osteoinduction properties The aim of the present work was to study the morphological characteristics of fresh frozen after 6 month of bone graft to evaluate new bone formation. For the study, we selected 10 patients which were undergone to bone graft after bone atro- phy; the study was carried out by histology, immunofluorescence and scanning electron microscopy techniques. Results have shown, after 6 month of graft, the presence of cellular elements demonstrating new vital bone apposition and supporting that fresh frozen grafts present regenerative capacities. These morphological data support the results prevalently clinical demonstrated in Literature. These are preliminary results which need further measurements in follow-up periods, histologic and ultrasctructural features and also experimental protocol on animal models
Can scanning near‐field optical microscopy be compared with confocal laser scanning microscopy? A preliminary study on α‐sarcoglycan and β1D‐integrin in human skeletal muscle
No full textThe dystrophin-glycoprotein complex and the vinculin-talin-integrin system constitute, together a protein machinery, called costameres. The dystrophin-glycoprotein complex contains, among other proteins, also dystrophin and the sarcoglycans subcomplex, proteins playing a key role in the pathogenesis of many muscular dystrophies and linking the cytoplasmic myofibrillar contractile elements to the signal transducing molecules of the extracellular matrix, also providing structural support to the sarcolemma. The vinculin-talin-integrin system connects some components of the extracellular matrix with intermediate filaments of desmin, forming transverse bridges between Z and M lines. In our previous reports we always studied these systems by confocal laser scanning microscopy (CLSM). In this paper we report on the first applications of optical near-field fluorescence microscopy to the spatial localization of alpha-sarcoglycan and beta 1D-integrin in human skeletal muscle fibres in order to better compare and test the images obtained with conventional CLSM and with scanning near-field optical microscopy (SNOM). In addition, the analysis of the surface morphology, and the comparison with the fluorescence map is put forward and analyzed for the first time on human muscle fibres. In aperture-SNOM the sample is excited through the nanometre-scale aperture produced at the apex of an optical fibre after tapering and subsequent metal coating. The acquisition of the topography map, simultaneously to the optical signal, by SNOM, permits to exactly overlap the fluorescence images obtained from the two consecutive scans needed for the double localization. Besides, the differences between the topography and the optical spatial patterns permit to assess the absence of artefacts in the fluorescence maps. Although the SNOM represented a good method of analysis, this technique remains a complementary method to the CLSM and it can be accepted in order to confirm the hypothesis advanced by CLSM
Sarcoglycan sub-complex and Satellite cells in masseter muscle of crossbite patients
No full textCrossbite is a lateral misalignment of the dental arches. Subjects with unilateral posterior crossbite exhibit different kinematics of the mandible during mastication when chewing on the affected side, resulting in an increased frequency of reverse chewing cycles; moreover, the masseter of the affected side results less active than the counterpart (Piancino et al., 2006). An our immunofluorescence study on masseter muscle of crossbite patients has shown that in the affected side there is a lower expression of muscle specific integrins than the counterpart (Cutroneo et al., 2012). So, we hypothesized that integrins could be correlated with contraction activity in mastication, maybe promoting the production of new muscle fibers. Another our tractography study on crossbite patients has shown an increment of muscle fibers number in non affected side. On this basis in the present work we investigated the number of satellite cells in masseter muscle of crossbite patients, both in affected and non affected side. Since the existence of a bidirectional signaling between integrins and sarcoglycans we have also investigated the expression of the sarcoglycan subcomplex. Results show that the number of satellite cells in non affected side is higher than affected side. Moreover, in according with our previous results about integrins, even the sarcoglycan sub-complex show to be less expressed in the affected side than the counterpart. These data support that in non affected masseter muscle a new fibers production activity takes place which, in turns, determines a muscular hyperplasia. Based on our results, the myogenesis in masseter muscle could be regulated by sarcoglycan and integrin protein systems
Supravital exposure to propidium iodide identifies apoptosis on adherent cells
No full text: Several studies indicate that plasma membrane changes during apoptosis are a general phenomenon. Among the flow cytometric methods to measure apoptosis, the Annexin V assay that detects the membrane exposure of phosphatidylserine (PS) is one of the most commonly used. However, the various treatments used for the detachment of adherent cells generally interfere with the binding of Annexin V to membrane PS, making apoptosis measurement a technical problem. Materials and Methods Apoptosis of different cell lines was investigated by fluorescence microscopy and multiple flow assays designed to assess loss of membrane integrity, translocation of PS, DNA fragmentation, and light scatter changes. Results and Conclusions We show that supravital propidium iodide (PI) assay stains adherent apoptotic cells, allowing flow cytometric quantification. Moreover, supravital exposure to PI without prior permeabilization identifies apoptotic cells as well as Annexin V and permits the simultaneous surface staining by FITC- and PE-conjugated monoclonal antibodies. As in the case of necrotic or permeabilized cells, fluorescence microscopy has revealed that PI staining of apoptotic cells is localized in the nucleus. This suggests that the binding of PI to the DNA/RNA structures is stable enough to withstand the trypsinization and/or washing procedures necessary to detach adherent cells
Preliminary study on sarcoglycan sub-complex in rat cerebral and cerebellar cortex
No full textThe sarcoglycan sub-complex is a protein system which plays a key role in sarcolemma stabilization during muscle activity. Although numerous studies have been conducted on this system, there are few data about its localization in non-muscular tissues. On this basis we carried out an indirect immunofluorescence study on normal rat cerebral and cerebellar cortex. In particular, we carried out single localization reactions to analyze if these proteins are present in brain and double localization reactions between sarcoglycans and either SMI-32 or GFAP to verify if they are expressed both in neurons and glial cells. We found that all tested sarcoglycans are present both in cerebral and cerebellar cortex and that they are expressed both in neurons and glial cells. The typical staining pattern of all sarcoglycans is represented by “spot-like” fluorescence, with spots of 0.5-2 μm average diameter laid out mainly around the soma of the cells. The main difference about sarcoglycans expression between cerebral and cerebellar cortex is that in the cerebellar cortex the sarcoglycans positivity is detectable only in an area which is likely to correspond to Purkinje cells layer. The presence of sarcoglycans in cerebral and cerebellar cortex and their disposition mainly around the soma of the cells suggest a role of these proteins in cellular signalling and in regulating postsynaptic receptor assembly mainly in axo-somatic synapses
Sarcoglycan sub-complex in the adipose organ: a molecular and immunofluorescence study
No full textThe sarcoglycan sub-complex (SGC) is made up of six glycoproteins which connect the cytoskeleton to the extracellular matrix in skeletal muscle. Recent data show that this complex is also expressed in epithelial tissue such as gingival, breast and prostatic ones [1]. The adipose organ is organized in multiple fat depots consisting of white and brown adipose cells. White adipocytes can store energy in triglycerides; brown adipocytes can dissipate energy for thermogenesis. It has been demonstrated that white adipocytes transdifferentiate to brown adipocytes after cold exposure [3]. In this study we examined the expression of sarcoglycans (SGs) in the adipose organ from two groups of mice: the first group was exposed at 25 C°, as control, and the second one was exposed at low temperature (4 C°) for 24 hours and 4 days. Fat depots from the visceral and interscapular region, but also from the mammary gland, have been examined by histological, immunofluorescence and molecular techniques. Results have shown that SGC is expressed in the adipose organ, both in brown and white adipocytes of mice exposed at 25 C°. The main results is that the expression level of all sarcoglycans increase in cold exposure experiment. For the first time the expression of all SGs in the adipose organ has been demonstrated, both at mRNA and protein levels. Since we found an increase in SGs expression after transdifferentiation from white to brown adipocytes cold exposure induced, we hypothesize that sarcoglycans could be associated with β3 adrenergic receptor; sarcoglycans associations with other receptors, as GABAr, has been already demonstrated in central nervous system. Although that, the function of these glycoproteins in the adipose organ remain still unclear and further investigation are required
Sarcoglycan sub-complex in WAG/Rij rats, a model of absence epilepsy: an immunofluorescence study
No full textIt is known that even in Central Nervous System the Dystrophin Glycoprotein Complex (DGC) exists but differs in composition from the DGC core present in muscle for the presence of several isoforms of dystrophin and for the existence of a sarcoglycan sub-complex which is made up only for ε- and ζ-sarcoglycans; for these reasons it was called “DGC-like”. Although that, in our previous studies we have found that all sarcoglycans are present in human cerebral cortex and in different regions of rat’s brain, suggesting that the composition of the brain DGC is not different from the muscle DGC but differing just for the function. In fact, our previous data showing the colocalization between sarcoglycans and GABAA Rε receptors in rat’s cerebral and cerebellar cortex, thalamus and hippocampus suggested us that in brain sarcoglycans could be associated with synaptic neurotransmission. To better understand which kind of relationship between sarcoglycans and GABAA receptors exists, we aim to investigate α-,β-,γ-,δ-,ε- and ζ-sarcoglycans and the GABAA Rε in the brain of the WAG/Rij rats, a model of absence epilepsy, using immunofluorescence techniques; we have observed cerebral cortex, thalamus and hippocampus, the structures mainly involved in absence epilepsy, comparing this to normal rat’s brain. Results show that the GABAA Rε receptors staining pattern is different from the normal rat’s brain presenting an abnormal fluorescence distribution consisting in discontinuous clusters around the cellular body. Sarcoglycans, instead, have shown the typical “spot-like” staining pattern around the cellular body, sometimes colocalizing with GABAA Rε receptor; moreover, they seem to have an higher staining pattern than normal rat’s brain. In our opinion these results, showing an alteration of GABAA Rε receptor pattern distribution and a normal or increased sarcoglycans staining pattern, allow us to hypothesize that the sarcoglycans in absence epilepsy are likely to compensate for the alteration in GABAA Rε receptor distribution. That support the opinion about the involvement of sarcoglycans in cellular signalling and receptor assembly regulating indirectly synaptic neurotransmission both in normal and in pathological condition
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