Gene expression of stem cells at different stages of ontological human development

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 <0.05. 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

An innovative method for the detection of contaminant viral genome in cell cultures

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

ENTERAL NUTRITION SUPPORT TO TREAT MALNUTRITION IN INFLAMMATORY BOWEL DISEASE

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

Investigating alkyl nitrates as nitric oxide releasing precursors of multitarget acetylcholinesterase-monoamine oxidase B inhibitors

Herein we envisaged the possibility of exploiting alkyl nitrates as precursors of alcohol-bearing dual inhibitors targeting acetylcholinesterase (AChE) and monoamine oxidase B (MAO B), key enzymes in neurodegenerative syndromes such as Alzheimer's disease (AD), through biotransformation unmasking an alcoholic function upon nitric oxide (NO) release. The cooperation to neuroprotection of low fluxes of NO and target enzymes’ inhibition by the alcohol metabolites might return a multitargeting effect. The in vitro screening towards ChEs and MAOs of a collection of 21 primary alcohols disclosed a subset of dual inhibitors, among which three diverse chemotypes were selected to study the corresponding nitrates. Nitrate 14 proved to be a brain permeant, potent AChE-MAO B inhibitor by itself. Moreover, it protected human SH-SY5Y lines against rotenone and hydrogen peroxide with a poor inherent cytotoxicity and showed a slow conversion profile to its alcohol metabolite 9d that still behaved as bimodal and neuroprotective molecule

Is secondary hyperparathyroidism-related myelofibrosis a negative prognostic factor for kidney transplant outcome?

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

THE MEDITERRANEAN DIET: A HISTORY OF HEALTH

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

Organismi geneticamente modificati: definizioni e finalità.

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

The influence of some dietary components on intestinal microbiota

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

THE INTESTINAL ECOSYSTEM AND PROBIOTICS

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

Protective effect of procyanidin-rich grape seed extract against Gram-negative virulence factors

Biofilm formation and lipopolysaccharide (LPS) are implicated in the pathogenesis of gastrointestinal (GI) diseases caused by Gram-negative bacteria. Grape seeds, wine industry by-products, have antioxidant and antimicrobial activity. In the present study, the protective effect of procyanidin-rich grape seed extract (prGSE), from unfermented pomace of Vitis vinifera L. cv Bellone, on bacterial LPS-induced oxidative stress and epithelial barrier integrity damage has been studied in a model of Caco-2 cells. The prGSE was characterized at the molecular level using HPLC and NMR. The in vitro activity of prGSE against formation of biofilm of Salmonella enterica subsp. enterica serovar Typhimurium and Escherichia coli was investigated. In vivo, prGSE activity using infected Galleria mellonella larvae has been evaluated. The results show that the prGSE, if administered with LPS, can significantly reduce the LPS-induced permeability alteration. Moreover, the ability of the extract to prevent Reactive Oxygen Species (ROS) production induced by the LPS treatment of Caco-2 cells was demonstrated. prGSE inhibited the biofilm formation of E. coli and S. Typhimurium. In terms of in vivo activity, an increase in survival of infected G. mellonella larvae after treatment with prGSE was demonstrated. In conclusion, grape seed extracts could be used to reduce GI damage caused by bacterial endotoxin and biofilms of Gram-negative bacteria