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Chronic renal disease (CKD) is a social problem affecting millions of patients characterized by loss of renal function and related to metabolic diseases. The approach from the dietary point of view to this problem could be a perfect strategy to slow down the progression of the disease and at the same time the problems of malnutrition typical of CKD. Several studies show that the Mediterranean diet (MD) may play a key role in the control of the early stages of the disease. Even if the MD showed to be the best diet for the control of metabolic diseases and for the general well-being, in the case of the patient with CKD, should be adapted in what is called the DASH diet, characterized by the typical roles of the MD but with a controlled intake of minerals and proteins.

‘We are what we eat’, said the philosopher Feuerbach. In fact, the quality of the food we eat affects our mind as well: the brain, which represents 2% of our body weight, consumes about 20% of the calories we eat each day. Follow a few rules could therefore help to feed properly our brain, so it works to the best of its ability. It is important to know properties of foods especially regarding their aminoacidic composition because aminoacids are components of neurotransmitters, molecules needed for brain transmission and function. It is also possible to choose specific food to prevent or support different diseases that affects nervous system.

The aim of this study was to isolate and characterize rat Adipose Derived Mesenchymal Stem Cells (AD-MSCs) in order toevaluate their proliferative potential and their ability to differentiate in different cell types. AD-MSCs and Derived MesenchymalStem Cells (BM-MSCs) have the same characteristics in terms of plasticity. The advantage of adipose tissue is that it is an easier accessible source and it offers a large amount of MSCs by less invasive surgical tecniques. MSCs were obtainedfrom subcutaneous adipose tissue of Wistar rats. First of all microbiological controls were made to exclude the presenceof bacteria or fungi in the tissue. Adipose tissue was mechanically and enzimatically fragmented and stomal cell fraction was seeded in adherent culture flasks in DMEM 20% FBS. After 48h the medium was replaced. Cells were characterized byevaluating: 1)their ability to adhere to the plastic; 2) the clonogenic potential by Colony Forming Unit (CFU) assay; 3) theirability to differentiate in 3 mesodermal lineages (adipocytes, osteocytes and chondrocytes). AD-MSCs are able to differentiate in adipocytes, osteocytes and chondrocytes as confirmed by Oil Red’O staining, von Kossa staining and histologicalanalysis respectively. This first characterization is essential for the second part of our study in which we are planning to useAD-MSCs in vivo to restore renal function after an induced ischemic damage in experimental animals.

Enteral nutrition (EN), as parenteral nutrition (PN), can be used in cases of patients whose medical conditions prevent the intake of food by mouth; unlike PN, EN keeps the functionality of the digestive tract and it makes home management of patients easier. However, the experience and literature have documented a number of serious complications, fortunately rare, which depend on the methods used in EN realization. We report in this paper our experience in 44 cases of percutaneous endoscopic gastrostomy (PEG), concluding that it is a safe and complications-free procedure. We believe that a nutritional intervention is indicated when, improving nutritional status, patients can obtain a better quality of life and have an average life expectancy.

The Mediterranean tradition offers a cousine rich in colors, aromas and memories, which support the taste and the spirit of those who live in harmony with nature. Everyone is talking about the Mediterranean diet, but few are those who do it properly, thus generating a lot of confusion in the reader. And so for some it coincides with the pizza, others identified it with the noodles with meat sauce, in a mixture of pseudo historical traditions and folklore that do not help to solve the question that is at the basis of any diet: combine and balance the food so as to satisfy the qualitative and quantitative needs of an individual and in a sense, preserves his health through the use of substances that help the body to perform normal vital functions. The purpose of our work is to demonstrate that the combination of taste and health is a goal that can be absolutely carried out by everybody, despite those who believe that only a generous caloric intake can guarantee the goodness of a dish and the satisfaction of the consumers. That should not be an absolute novelty, since the sound traditions of the Mediterranean cuisine we have used for some time in a wide variety of tasty gastronomic choices, from inviting colors and strong scents and absolutely in line with health.

The term "probiotic" comes from the greek "pro bios" and means "pro life": Nowadays, an increasing number of pharmaceutical preparations and functional foods are enriched with probiotics and for the patients it is increasingly important to receive information needed to know how to orient in the choice. The benefits from probiotics are many and include the modulation of the intestinal microflora (stimulation of beneficial bacteria and inhibition of pathogens), the support of bowel function and the stimulation of the immune system. This broad spectrum of beneficial effects to maintain efficient the intestinal ecosystem. Therefore, probiotics are an useful tool to prevent the formation of disorders and/or pathologies. The aim of this review is to describe the intestinal ecosystem and how probiotics could be effective in the treatment and prevention of possible alterations.

Stem cells have a key role in regenerative medicine and tissue engineering. Although not immortal, they are able to expand manyfold in culture retaining at the same time their growth and multilineage potential. They also show a migratory capacity when transplanted systemically in animal model with injuries. Thanks to their properties and their plasticity stem cells are of great importance since they can be used as a tool for repair damaged tissues and organs. Mesenchymal stem cells, in particular, have the ability to differentiate into lineages of mesodermal tissues, such as skeletal muscle, bone, tendons, cartilage, and fat under appropriate culturing conditions. Recent evidence suggest that the adipose tissue is a promising source of mesenchymal stem cells attracting the interest of researchers and clinicians. It is rich of pluripotent stromal cells, available in large amounts and more readily accessible than bone marrow. Furthermore, comparative analysis of mesenchymal stem cells of bone marrow and adipose tissue show that cells are not different regarding morphology, immune phenotype, success rate of isolating and differentiation ability. Our experience at the Experimental Zooprophylactic Institute of Sicily A. Mirri allows us to define a protocol for stem cells isolation of murine origin. We used 6 Winstar breed male rats whose average weight was 350 g. All animals were sedated with an intramuscular injection of midazolam and anesthesia was maintained with isoflurane and oxygen gas mixture administered with a mask. The adipose tissue has been taken from the root of the animal’s thigh with a small incision. Different steps are needed for processing and digesting the tissue. First it is washed several times in a solution enriched of antibiotics and then is mechanically fragmented. The homogenate is therefore digested enzymatically under permanent shaking. We obtain an heterogeneous population of cells that were subsequently selected through the plastic adhesion. These cells are able to grow and proliferate and show all the characteristics typical of stem cells. In conclusion, we report a multistep and reproducible technique for providing a substantial number of mesenchymal stem cells and for maintaining them in culture.

Objective. Biomaterial research for soft tissue augmentation is an increasing topic in aesthetic medicine. Hyaluronic acid (HA) fillers are widely used for their low invasiveness and easy application to correct aesthetic defects or traumatic injuries. Some complications as acute or chronic inflammation can occur in patients following the injection. Biocompatibility assays are required for medical devices intended for human use, in order to prevent damages or injuries in the host. In this study, nine HA fillers were tested in order to evaluate their cytotoxicity and their effects on L929 cell line, according to the UNI EN ISO 10993 regulation. Methods. Extracts were prepared from nine HA fillers, and MTS viability assay was performed after 24 h, 48 h, and 72 h of exposure of cells to extracts. Cells cultured with HA filler extracts were monitored for up to 72 h, counted, and stained with haematoxylin/eosin in order to evaluate the cell proliferation rate and morphology. Results. None of the filler tested showed a cytotoxic effect. Two samples showed a higher vitality percentage and higher cell number while two samples showed a lower vitality percentage and lower cell number at 72 h. Conclusion. Data obtained suggest that although examined fillers are not cytotoxic, they show different effects on the in vitro cell proliferation rate. In vitro studies of medical devices could lead to important implications since these could aid to predict effects about their in vivo application. These easy and rapid assays could be useful to test new materials intended for human use avoiding animal tests.

Intraoperative parathyroid hormone (PTH) monitoring in the setting of the operating room represents a valuable example of the rationale use of the laboratory diagnostic in a patient-oriented approach. Rapid intraoperative PTH (ioPTH) assay is a valid tool for an accurate evaluation of the success of parathyroid surgery. The reliability of the user-friendly portable systems as well as the collaboration between operators and surgical staff allow the one-site monitoring of the ioPTH decrements on the course of the surgical management of hyperparathyroidism. The rapid answer provided by an effective decrement of PTH during parathyroidectomy contributes dramatically to the efficacy of parathyroid surgery and the reduction of the number of re-operations. Therefore the dose of ioPTH is a valid and reliable support for the success of the intervention of parathyroidectomy at controlled costs.

Malnutrition is a common consequence of inflammatory bowel disease (IBD). Diet has an important role in the management of IBD, as it prevents and corrects malnutrition. It is well known that diet may be implicated in the aethiology of IBD and that it plays a central role in the pathogenesis of gastrointestinal tract disease. Often oral nutrition alone is not sufficient in the pathogenesis of gastrointestinal tract disease. Often oral nutrition alone is not sufficient in the management of IBD patients, especially in children or elederly, and must be combined with oral supplementation or replaced with tube enteral nutrition. In this review, we describe several different approaches to enteral nutrition total-parenteral, oral supplementation and enteral tiube feeding in terms of results, patience compliance, risks and benefits. we also focus on the home enteral nutrition strategy as the future goal for treating IBD while focusing on patient wellness.

Secondary hyperparathyroidism (HP) presenting with hypocalcemia and subsequent increased parathormone (PTH), is mainly identified in patients with chronic renal failure, which has been associated with variable degrees of bone marrow fibrosis. For suitable patients with end-stage renal disease (ESRD), kidney transplantation is recognized as the therapy of choice, being superior to dialysis in terms of quality of life and long-term mortality risk; in this regard interesting data show that increased time on dialysis prior to kidney transplantation is associated with decreased graft and patient survival. In our opinion an important and until now underestimated determinant of graft survival is the proper activity of bone marrow because of the emerging role of hematopoietic stem cells (HSC) in repair of ischemia/reperfusion (IR) damage. We postulate that in ESRD patients, who usually undergo long dialytic treatment, a myelofibrosis caused by an overt secondary HP could drastically decrease the HSC potential for IR damage repair after kidney transplant; this could irremediably lead to a delay in graft function with all related complicances. If the curative role of bone marrow-derived stem cells was confirmed by more data obtained in experimental animal models, it could be possible to try a cellular-based therapeutic approach in the management of ESRD patients which are in waiting list for a kidney transplant.

With the high occurrence of cardiovascular disease and increasing numbers of patients requiring vascular access, there is a significant need for small-diameter (

The aim of this study was to isolate and characterize rat adipose Derived Mesenchymal Stem Cells (AD-MSCs) in order to evaluate their proliferative potential and their ability to different cell types. AD-MSCs and Derived Mesenchymal Stem Cells (BM-MSCs) have the same characteristic in terms of plasticity. The advantage of adipose tissue is that it is an easier accessible source and it offers a large amount of MSCs by less invasive surgical tecniques. MSCs were obtained from subcutaneous adipose tissue of Wistar rats. first of all microbiological controls were made to exclude the presence of bacteria of fungi in then tissue. Adipose tissue was mechanically and enzimatically fragmented and stromal cell fraction was seeded in adherent culture flasks in DMEM 20% FBS. After 48 h the medium was replaced. Cells were characterized by evaluating:1)their ability tho adhere to the plastic; 2) the clonogenic potential by Colony Forming Unit (CFU) assay, 3) their ability to differentiate in 3 mesodermal lineages (adipocytes, osteocytes and chondrocytes). AD-MSCs are able to differentiate in adipocytes, osteocytes and chondrocytes as confirmed by oil red'O staining, von Kossa staining and histological analuysis respectively. This first characterization is essential for the second part of our study in which we are planning to use AD-MSCs in vivo to restore renal function after an induced ischemic damage in experimental animals.

Objectives To compare multipotent mesenchymal stem cells (MSCs) obtained from chorionic villi (CV), amniotic fluid (AF) and placenta, with regard to their phenotype and gene expression, in order to understand if MSCs derived from different extra-embryonic tissues, at different stages of human ontological development, present distinct stemness characteristics. Study design MSCs obtained from 30 samples of CV, 30 of AF and 10 placentas (obtained from elective caesarean sections) were compared. MSCs at second confluence cultures were characterized by immunophenotypic analysis with flow cytometry using FACS CANTO II. The expression of the genes Oct-4 (Octamer-binding transcription factor 4, also known as POU5F1), Sox-2 (SRY box-containing factor 2), Nanog, Rex-1 (Zfp-42) and Pax-6 (Paired Box Protein-6), was analyzed. Real-time quantitative PCR was performed by ABI Prism 7700, after RNA isolation and retro-transcription in cDNA. Statistical analysis was performed using non-parametric test Kruskal–Wallis (XLSTAT 2011) and confirmed by REST software, to estimate fold changes between samples. Each gene was defined differentially expressed if p -value was Results Cells from all samples were negative for haematopoietic antigens CD45, CD34, CD117 and CD33 and positive for the typical MSCs antigens CD13, CD73 and CD90. Nevertheless, MSCs from AF and placentas showed different fluorescence intensity, reflecting the heterogeneity of these tissues. The gene expression of OCT-4, SOX-2, NANOG was not significantly different among the three groups. In AF, REX-1 and PAX-6 showed a higher expression in comparison to CV. Conclusions MSCs of different extra-embryonic tissues showed no differences in immunophenotype when collected from second confluence cultures. The expression of OCT-4, NANOG and SOX-2 was not significantly different, demonstrating that all fetal sources are suitable for obtaining MSCs. These results open new possibilities for the clinical use of MSCs derived from easily accessible sources, in order to develop new protocols for clinical and experimental research.

Article 2 of the European Directive 2011/18/EC defines a genetically modified organism (GMO) a san organism in which the genetic material has been altered in a way that does not occuirr naturally by inserting DNA fragments from one organism's gene into the chromosomes of another, changing its genetic expression. In the field of animal and plants biotechnology there are different application areas of genetic engineering such as improving the quantity and quality of argicultural production, pharmaceutical production in more sustainable ways or applications in environmental field.

Marginal donors represent a poorly utilized source of organs for transplantation despite their availability. The key is to reduce the ischemic damage in the effort to improve organ quality. This study investigated the histologic effects after in situ perfusion of preservation with a two-layer method compared with the classic University of Wisconsin preservation in term of tissue integrity and number of viable exocrine cells in the rat pancreas both after exsanguination and at 8 weeks of cryopreservation. Pancreata harvested from 60 rats were collected using 3 methods: two-layer method following University of Wisconsin perfusion; exsanguination; and classic University of Wisconsin perfusion/storage. In addition to histologic analysis of collected pancreata, we analyzed the number of CK19(+) cells and their viability using chi-square tests with values P < .05 considered to be significant. Rat pancreas histology showed as University of Wisconsin in situ perfusion and preservation by the two-layer method to be more effective to maintain the morphologic integrity of both exocrine and endocrine tissues. There were a larger number of CK19(+) cells with good viability. Moreover, the effects of oxygenation were visible in pancreas biopsies preserved after exsanguination. In situ University of Wisconsin perfusion and preservation for 240 minutes with the two-layer method yielded greater numbers and viability of CK19(+) cells even after 8 weeks of cryopreservation.

Colonic leiomyoma is a mesenchymal tumor that arises from the muscularis mucosae or muscularis propria and is composed of well-differentiated smooth muscle cells with no atypia. It is often incidentally found since its growth affects the submucosal layer and the lesion is covered with normal epithelium. Endoscopic ultrasonography is recommended to define the grade of infiltration of the tumor and eventually lymph node involvement. Histological examination is critical to establish the nature of the tumor and its behaviour. In the case of a voluminous tumor surgical treatment is needed. we report case of a patient that underwent colonoscopy showing the presence of a neoformation at 70 cm from ileocecal valve occupying half lumen of transverse colon. A surgical resection was performed and histological analysis confirmed the presence of a leiomyoma.

Mesenchymal Stem Cells (MSCs) are multipotent cells that reside within various tissues, including adipose tissue and bone marrow. Research concerning the potential therapeutic role of MSCs has advanced significantly over the last decade. Many studies have demonstrated that adipose tissue is an excellent source of MSCs (AD-MSCs), easily accessible in experimental animals. AD-MSCs can be expanded in vitro over the short term and they can differentiate toward a variety of cell types of mesodermal lineage. Therefore, they are thought an attractive tool for cell therapy. Selection of appropriate animal models for preclinical evaluations is crucial for optimization and validation of therapeutic protocols in regenerative medicine. Therefore, in vitro studies are necessary to future in vivo applications of MSCs. Rat is an animal model usefull for a large number of in vitro studies. Aim of this work is the in vitro production of endothelial cells differentiated from rat ADMSCs. For this purpose, subcutaneous adipose tissue samples were collected from 5 Wistar rats, in accordance to the European Directives (86/609/CEE and 63/2010/EU) concerning the protection of experimental animals. These samples were processed for in vitro AD-MSCs isolation. After 2 days, nonadherent cells were removed and medium (α-MEM) was replaced. The adherent AD-MSC population was expanded for 5 passages. To assess their characteristics, the expression of stemness markers, such as OCT4, SOX2 and NANOG, was evaluated by Real Time Sybr Green PCR. The analysis of gene expression was performed from 0 to 5 passage. For this purpose, mRNA was extracted from each passage using a commercial kit. Successively, mRNA was retrotranscribed in cDNA and stemness genes were detected using specific primers pairs, designed by the use of Primer Express software. Gene expression analysis was normalised by the GADPH gene housekeeping expression. To induce endothelial differentiation, stem cells at passage 0 and 1 were cultivated in D-MEM medium for 18-24 h. Successively, D-MEM medium was replaced with an enriched medium with VEGF-C. Cells were cultivated for 15 days. To verify the begin of differentiation, gene expression analysis for endothelial markers CD31, eNOS and vWF was performed by Real Time Sybr Green PCR. Specific primers pairs were designed by the use of Primers Express software. Gene expression results show an increase of expression levels for stemness markers, up to the in vitro passage 3. The begin of the endotelial differentiation is proved by the initial increase of expression levels for CD31, eNOS and vWF, associated to a decrease in the expression levels for stemness markers. These are preliminary results of a project aimed to the in vitro production of endotelial differentiated cells from AD-MSC. Further studies are required to confirm the acquisition of the endothelial phenotype by AD-MSC cells, induced to differentiate.

The use of cell cultures involves different fields of biology, from diagnosis to research. Moreover, technologies based on animal cells represent a useful tool to the development of biological products for the prophylaxis and therapy in humans and animals. Therefore, it is necessary to perform quality controls, including virological tests. Several tests performed in research laboratories are able to discriminate one or more viral species, but it is not possible to demonstrate the presence of contaminant viral genome with one non-specific method. The aim of this work consisted on the realization of a biomolecular method able to detect and to identify by sequencing extraneous viral genome in cell cultures of animal and human origin in the absence of any specific information about the virus.

Purpose Pancreata from non-heart beating donors could represent an unlimited source of islets if their cell viability can be efficiently preserved during the time necessary to process the organs by the use of a better solution of preservation compared to the classic University of Wisconsin solution. The aim of this study was to determine whether it is possible to obtain functioning “alive islets” from non-heart-beating donors by comparing, on a porcine model, the classic “UW ice-store” method with a two-layer cold storage method (TLM) using oxygenated Perfluorocarbons (PFC) and UW. Methods Whole pancreata were harvested from 20 NHBDs female pigs with similar characteristics and preserved for 4 h in UW solution (n=10) or TLM (UW/PFC) solution (n=10). The isolated islets were then evaluated for number, viability, purity, and insulin secretion, also estimated after 8 weeks of cryopreservation. Results The total number of islets obtained from isolation, and their function assayed by the insulin stimulation index, before and after cryopreservation, showed a higher value in the TLM group. No significative differences in terms of purity and viability before and after cryopreservation were found when comparing the two groups. Conclusions TLM solution for NHBDs porcine pancreata with cold ischemia time lower than 4 h offers significant advantages over UW solution storage, thereby increasing the isolation yield and isolation success rate of the pancreatic porcine islets.

We obtained a nano-fibrillar scaffold starting from a polymeric solution which, through electrospinning, gave a biodegradable material with optimal mechanical features and the capacity to allow cell adhesion. In this paper we report the in-vivo application on a murine animal model of two electrospun biodegradable materials, specifically designed to create tubular structures. In one case PHEA-PLA was co-spun with silk fibroin (Fibro-PHEAPLA) by a parallel electrospinning process to obtain a scaffold with two different polymeric fibers. In the other case, PHEA-PLA was mixed with polycaprolactone (PCLPHEA-PLA) to obtain a hybrid fibers scaffold. The in-vitro assay showed fibroblast colonization in both materials. The scaffolds were implanted in the dorsal fascial pouch of rats to evaluate their in-vivo Biocompatibility and tissue integration. Histopathological findings showed that after implantation a neutrophilic reaction associated to colliquative necrosis was predominant, particularly for PCL-PHEA-PLA. Fibro-PCL-PHEA caused a non organized stromal reaction. Cell adhesion was confirmed at SEM scan. Both materials were totally absorbed after 40 days with an inflammatory reaction. This preliminary study showed that biocompatibility of the scaffolds needs further investigation. The capability of the materials to be functionalized could allow us to modulate the inflammatory host response.

The gut microbiota is the set of symbiotic microorganisms that resides in our intestine. The interaction between the gut microbiota and the mucosal immune system can be altered as a result of shifts in the composition or metabolic activity of the intestinal microflora. Recent studies establish that diet is one of the main involved factors in determining the microbial composition of the gut suggesting its role as external factor able to promote the onset of specific diseases by disrupting the immune homeostasis. Starting from the evidence that the 57% of the gut microbiota’s entire variation are due to dietary alterations this review aims at providing an overview of the positive impact of some dietary components on gut microbiota composition.