An FTIR Spectroscopy Investigation on Different Methods of Lipid Extraction from HepG2 Cells

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Hematopoietic Stem/Progenitor Cells Express Functional Mitochondrial Energy-Dependent Cystic Fibrosis Transmembrane Conductance Regulator

Bone marrow-derived hematopoietic stem/progenitor cells (HSPCs) encompass a wide array of cell subsets with different capacities of engraftment and injured tissue-regenerating potential. The characterization/isolation of the stem cell subpopulations represents a major challenge to improve the efficacy of transplantation protocols used in regenerative medicine. Cystic fibrosis (CF) is one of the diseases whose hope of cure relies on the successful application of cell-based gene therapy. This study was aimed at characterizing murine HSPCs on the basis of their bioenergetic competence and CF transmembrane conductance regulator (CFTR) expression. Positively immunoselected Sca-1(+) HSPCs encompassed 2 populations distinguished by their different size, Sca-1 expression and mitochondrial content. The smaller were the cells, the higher was Sca-1 expression and the lower was the intracellular density of functional mitochondria. Reverse transcription-polymerase chain reaction and western blotting revealed that HSPCs expressed CFTR mRNA and protein, which was also functional, as assessed by spectrofluorimetric and patch-clamp techniques. Inhibition of mitochondrial oxidative phosphorylation by oligomycin resulted in a 70% decrease of both the intracelluar adenosine triphosphate content and CFTR-mediated channel activity. Finally, HSPCs with lower Sca-1 expression and higher mitochondrial content displayed higher CFTR levels. Our findings identify 2 subpopulations in HSPCs and unveil a so-far unappreciated relationship between bioenergetic metabolism and CFTR in HSPC biology

An FTIR Spectroscopy Investigation on Different Methods of Lipid Extraction from HepG2 Cells

Fourier transform infrared (FTIR) spectroscopy is a non-invasive technique that is largely used for studying lipidomics. Lipids are a primary class of biological molecules that play numerous vital roles in various processes. In the present work, we adopted FTIR spectroscopy for monitoring the lipid extraction efficiency of different methods used for extracting lipids from hepatocarcinoma cells. The spectra acquired from samples obtained with the selected methods showed the contributions of different functional groups. A qualitative comparison indicated that all the spectra exhibited similar lipid species profiles. The peak intensity attributed to the CH2 asymmetric stretching mode has been used for a quantitative comparison of the efficiency of the different extraction methods

Simplexide Induces CD1d-Dependent Cytokine and Chemokine Production from Human Monocytes

Monocytes are major effector cells of innate immunity and recognize several endogenous and exogenous molecules due to the expression of wide spectrum of receptors. Among them, the MHC class I-like molecule CD1d interacts with glycolipids and presents them to iNKT cells, mediating their activation. Simplexide belongs to a novel class of glycolipids isolated from marine sponges and is structurally distinct from other immunologically active glycolipids. In this study we have examined the effects of simplexide on cytokine and chemokine release from human monocytes. Simplexide induces a concentration- and time-dependent release of IL-6, CXCL8, TNF-α and IL-10 and increases the expression of IL6, CXCL8 and IL10 mRNA. Cytokine and chemokine release induced by simplexide from monocytes is dependent on CD1d since: i) a CD1d antagonist, 1,2-bis (diphenylphosphino) ethane [DPPE]- polyethylene glycolmonomethylether [PEG], specifically blocks simplexide-induced activation of monocytes; ii) CD1d knockdown inhibits monocyte activation by simplexide and iii) simplexide induces cytokine production from CD1d-transfected but not parental C1R cell line Finally, we have shown that simplexide also induces iNKT cell expansion in vitro. Our results demonstrate that simplexide, apart from activating iNKT cells, induces the production of cytokines and chemokines from human monocytes by direct interaction with CD1d

Expression of Leptin Receptor and Effects of Leptin on Papillary Thyroid Carcinoma Cells

Background. Obesity has been hypothesized to contribute to the aggressiveness of thyroid cancer through the production of abnormal levels of serum adipokines. Leptin receptor (OB-R) expression has also been documented in papillary thyroid cancer (PTC). Aim. In this translational study, we analyzed in vitro the effects of leptin on the growth and migration of thyroid cancer cells (TPC-1 and K1), the molecular mechanisms underlying leptin’s action, and the influence of prolonged leptin exposure on cell response to a protein kinase inhibitor lenvatinib. The expression levels of OB-R mRNA and protein were also investigated in vivo in a series of aggressive PTCs divided into two groups based on the presence of the BRAF mutation. Results. In TPC-1 and K1 cells, prolonged treatment with leptin (500 ng/ml for 96 h) resulted in a mild increase in the proliferation (about 20% over control only in K1 cells, p<0.05) and in the migration of both cancer cell lines. Immunoblot analysis revealed a slight increase in the phosphorylation of AKT, but no effect on β-catenin and phospho-ERK expressions. The inhibitory effects of lenvatinib on the viability of both cell lines were not influenced by the leptin treatment. OB-R transcript (in fresh tissues) and proteins (in formalin-fixed and paraffin-embedded specimens) were expressed in all PTC tissues examined, with no significant differences between BRAF-mutated and BRAF-wild-type tumors. Conclusions. These results demonstrate leptin’s role in mildly increasing the aggressive phenotype of PTC cells but without influencing the action of lenvatinib. Further studies will clarify whether it is possible to target OB-R, expressed in all aggressive PTCs, as an adjuvant treatment approach for these malignancies

Oral Administration of Oleuropein and Its Semisynthetic Peracetylated Derivative Prevents Hepatic Steatosis, Hyperinsulinemia, and Weight Gain in Mice Fed with High Fat Cafeteria Diet

The high consumption of olive tree products in the Mediterranean diet has been associated with a lower incidence of metabolic disorders and cardiovascular diseases. In particular, the protective effects of olive oil have been attributed to the presence of polyphenols such as oleuropein (Ole) and its derivatives. We have synthesized a peracetylated derivative of Ole (Ac-Ole) which has shown in vitro antioxidant and growth-inhibitory activity higher than the natural molecule. In this study, male C57BL/6JOlaHsd mice were fed with a standard (std), cafeteria (caf) diet, and caf diet supplemented with Ole (0.037 mmol/kg/day) and Ac-Ole (0.025 mmol/kg/day) for 15 weeks. We observed a significant reduction in the caf diet-induced body weight gain and increase of abdominal adipose tissue. Also, Ole and Ac-Ole prevented the development of hepatic steatosis. Finally, Ole and Ac-Ole determined a lower increase of HDL and LDL-cholesterol levels and corrected caf diet-induced elevation of plasma glucose concentrations by improving insulin sensitivity. The observed beneficial properties of Ole and Ac-Ole make these compounds and in particular Ac-Ole promising candidates for a potential pharmaceutic use in metabolic disorders

Chronic nonbacterial osteomyelitis may be associated with renal disease and bisphosphonates are a good option for the majority of patients

13noAIM: The aim of this Italian study was to describe the clinical features, treatment options and outcomes of a cohort of patients with chronic nonbacterial osteomyelitis (CNO). METHODS: This was a retrospective cohort study. Laboratory data, diagnostic imaging, histological features and clinical course are reported. RESULTS: We enrolled 47 patients diagnosed with CNO. Bone pain was the leading symptom, and multifocal disease was present in 87% of the patients. The majority of the bone lesions were located in the appendicular skeleton (58%). Extraosseous manifestations were present in 34% of the patients, and renal involvement was detected in four patients. Inflammatory indices were increased in 80%, and bone x-rays were negative in 15% of the patients. Nonsteroidal anti-inflammatory drugs (NSAIDs) were the first therapy for all patients, achieving clinical remission in 27%. A good response to NSAIDs was significantly associated with a better prognosis. Bisphosphonates were used in 26 patients, with remission in 73%. Only six patients (13%), all with spine involvement, developed sequelae. CONCLUSION: We found a possible association between CNO and renal disease. Bisphosphonates were more likely to lead to clinical remission when NSAIDs and corticosteroids had failed. Vertebral localisation was the only risk factor for potential sequelae.nonenonePastore, Serena; Ferrara, Giovanna; Monasta, Lorenzo; Meini, Antonella; Cattalini, Marco; Martino, Silvana; Alessio, Maria; La Torre, Francesco; Teruzzi, Barbara; Gerloni, Valeria; Breda, Luciana; Taddio, Andrea; Lepore, LoredanaPastore, Serena; Ferrara, Giovanna; Monasta, Lorenzo; Meini, Antonella; Cattalini, Marco; Martino, Silvana; Alessio, Maria; La Torre, Francesco; Teruzzi, Barbara; Gerloni, Valeria; Breda, Luciana; Taddio, Andrea; Lepore, Loredan

Le autorità locali in Africa : governance e pratiche di accesso alle risorse

Negli ultimi vent’anni, un dibattito molto ampio si è sviluppato sulla relazione tra democrazia, conflitti e sviluppo emersa dalle politiche di good governance che hanno spinto molti paesi dell’Africa subsahariana a intraprendere riforme di decentramento. Nell’attribuire ai governi locali un’importanza crescente nella promozione di politiche di sviluppo, queste riforme avrebbero dovuto rafforzare la democrazia, migliorare la trasparenza e l’accountability dei governanti e rendere più semplice la gestione non violenta dei conflitti. A questo processo corrisponde una più generale svolta localistica dei donatori internazionali, che sempre più adottano la nozione di comunità locale come quadro di riferimento amministrativo sia della rappresentanza politica delle popolazioni rurali, sia delle politiche partecipative di gestione della terra e dell’erogazione di servizi pubblici. A questa definizione prescrittiva del ruolo del governo locale e delle relazioni tra centro e località corrisponde una realtà africana molto più incerta e variegata. Questo volume raccoglie i contributi di una nuova generazione di studiosi emersa da una base formativa comune con l’obiettivo di andare oltre l’approccio normativo, esplorando attraverso casi empirici il ruolo dei governi locali nella gestione dei conflitti legati all’accesso alle risorse tanto nelle aree rurali quanto in quelle urbane