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Objective: Previous studies suggested that polymorphisms in the coding region of the preproghrelin were involved in the etiology of obesity and might modulate glucose-induced insulin secretion. We evaluated the association of a new variation, -604C>T, in the promoter region of the ghrelin gene, of Leu72Met (247C>A) and of Gln90Leu (265A>T), all haplotype-tagging single nucleotide polymorphisms (SNPs), with measures of insulin sensitivity in 1420 adult individuals. Research methods: The three SNPs were genotyped using ABI PRISM 7900 HT Sequence Detection System. We used multiple linear regression analysis for quantitative traits and THESIAS software for haplotype analysis. Results: We observed a protective effect exerted by Met72 variant of Leu72Met SNP on insulin resistance parameters; a significant decreasing trend from Leu/Leu to Leu/Met and to Met/Met homozygous subjects in triglycerides, fasting insulin levels and HOMA-IR index (P=0.02, 0.01 and 0.003, respectively), and, consistently, an increase in ghrelin levels (P=0.003) was found. A significant decrease from CC to TC and to TT genotypes in insulin levels and HOMA-IR index was also detected (P=0.00l for both), but only in subjects homozygous for Leu72, where the protective effect of Met72 was not present. The haplotype analysis results supported the data obtained by the evaluation of each single SNP, showing the highest value of insulin levels and HOMA-IR index in the -604c247c haplotype intermediate value in -604T247C and lowest value in -604C247A. Conclusion: Our observations suggest a protective role of the Met72 variant and of -604 T allele in modulating insulin resistance. These SNPs or an unknown functional variant in linkage disequilibrium could increase ghrelin levels and probably insulin sensitivity.

Ices, silicates and carbonaceous materials have been detected in several astrophysical environments such as interstellar molecular clouds, comets, and planetary surfaces. These solids are continuously exposed to ion irradiation and UV photolysis. Our knowledge on the properties of solids and molecules and on the modification induced by fast ions (keV-MeV) and UV photons is mainly based on laboratory experiments and on the comparison of experimental results with observations. Here we will give a few examples of the role of laboratory experiments to our understanding of the physical and chemical properties of ices in space.

Role of galectin-3 as a receptor for advanced glycosylation end products. The advanced glycosylation end product (AGE)-binding proteins identified so far include the components of the AGE-receptor complex p60, p90 and galectin-3, receptor for advanced glycosylation end products (RAGE), and the macrophage scavenger receptor types I and II. Galectin-3 interacts with β-galactoside residues of several cell surface and matrix glycoproteins through the carbohydrate recognition domain and is also capable of peptide–peptide associations mediated by its N-terminus domain. These structural properties enable galectin-3 to exert multiple functions, including the modulation of cell adhesion, the control of cell cycle, and the mRNA splicing activity. Moreover, in macrophages, astrocytes, and endothelial cells, galectin-3 has been shown to exhibit a high-affinity binding for AGEs; the lack of a transmembrane anchor sequence or signal peptide suggests that it associates with other AGE-receptor components rather than playing an independent role as AGE-receptor. In tissues that are targets of diabetic vascular complications, such as the mesangium and the endothelium, galectin-3 is not expressed or only weakly expressed under basal conditions, at variance with p90 and p60 but becomes detectable with aging and is induced or up-regulated by the diabetic milieu, which only slightly affects the expression of p90 or p60. This (over)expression of galectin-3 may in turn modulate AGE-receptor-mediated events by modifying the function of the AGE-receptor complex, which could play a role in the pathogenesis of target tissue injury. Up-regulated galectin-3 expression may also exert direct effects on tissue remodeling, independently of AGE ligands, by virtue of its adhesive and growth regulating properties.

Results concerning the association of adiponectin gene polymorphisms (single-nucleotide polymorphisms, SNPs) with obesity, type 2 diabetes (T2DM), metabolic disorders and insulin resistance have not lead to definite conclusions. The aim of our study was to investigate a possible association between the -11391G>A and -11377C>G SNPs of adiponectin gene and measure of insulin sensitivity evaluated by the hyperinsulinemic-euglycemic clamp in a group of 'uncomplicated' obese subjects (with no associated comorbidities) (n=99, mean age 35 years) with a history of obesity lasting at least 10 years. The study of uncomplicated obese subjects, free of possible confounding factors that could interfere with insulin sensitivity, such as pharmacological treatment, provides a good model to assess insulin sensitivity per se. We observed that subjects homozygous for the G allele at locus -11391 had lower M (mg/kg min)/fat-free mass (FFM) index and adiponectin levels compared to subjects with GA+AA genotypes (P=0.002 and P=0.03, respectively) and subjects carrying the -11377G variant had lower M (mg/kg min)/FFM index and adiponectin levels compared to noncarriers (P=0.003 and P=0.03, respectively). Our results imply that the two promoter SNPs, -11391G>A and -11377C>G, of the adiponectin gene are associated with a reduced insulin sensitivity evaluated by hyperinsulinemic-euglycemic clamp in obese subjects.

The diabetic milieu modulates the advanced glycation end product-receptor complex in the mesangium by inducing or upregulating galectin-3 expression. G Pugliese , F Pricci , G Leto , L Amadio , C Iacobini , G Romeo , L Lenti , P Sale , R Gradini , F T Liu and U Di Mario Department of Clinical Sciences, La Sapienza University, Rome, Italy. demfound@tin.it Abstract Nonenzymatic glycation has been implicated in the pathogenesis of the dysregulated tissue remodeling that characterizes diabetic glomerulopathy, via the formation of advanced glycation end products (AGEs) and their binding to cell surface receptors. Several AGE-binding proteins have been identified so far, including p60, p90, and the adhesive and growth-regulating lectin galectin-3 (Gal-3), the components of the so-called AGE-receptor complex. This study aimed to evaluate the mesangial expression of the AGE-receptor complex and its modulation by the diabetic milieu, both in vivo, in non-diabetic versus streptozotocin-induced diabetic rats, and in vitro, in mesangial cells exposed to either normal glucose (NG) levels (5.5 mmol/l), as compared with high glucose (HG) levels (30 mmol/l) and iso-osmolar mannitol (M), or to native bovine serum albumin (BSA), as compared with glycated BSA with AGE formation (BSA-AGE) and glycated BSA in which AGE formation was prevented by aminoguanidine (BSA-AM). In vivo, Gal-3 protein and mRNA were not detectable in glomeruli from nondiabetic rats until 12 months after initiating the study. On the contrary, in diabetic rats, Gal-3 expression was observed at 2 months of disease duration, and it increased thereafter. Both p60 and p90 immunoreactivities were observed at the glomerular level with slightly increased expression of p90, but not p60, in diabetic versus nondiabetic animals. In vitro, Gal-3 was not detectable in mesangial cells cultured in NG (although it became evident after a certain number of passages in culture), whereas Gal-3 was detectable in cells grown on BSA. Prolonged exposure (2-4 weeks) of mesangial cells to HG but not to M, as well as growing cells on BSA-AGE and, to a lesser extent, BSA-AM, induced or significantly increased the expression of Gal-3, both protein (up to 2.65-fold) and mRNA (up to 3.10-fold) and its secretion in the medium (by approximately 50%). Both p60 and p90 were demonstrated in mesangial cells under NG conditions, and the expression of p90, but not p60, was upregulated by approximately 20% by HG or BSA-AGE. These results indicate that 1) under basal conditions, Gal-3, unlike p90 and p60, is not detectable in the mesangium but becomes expressed with aging and 2) the diabetic milieu induces or upregulates Gal-3 production, whereas it increases only slightly the expression of p90, but not p60. Gal-3 expression or overexpression may modulate the AGE-receptor-mediated events by modifying the function of the AGE-receptor complex. Additionally, it may exert direct effects on tissue remodeling by virtue of its adhesive and growth-regulating properties.

In recent years, climate change has greatly affected rainfall and air temperature levels leading to a reduction in water resources in Southern Europe. This fact has emphasized the need to focus on the use of non-conventional water resources for agricultural irrigation. The reuse of treated wastewater (TWW) can represent a sustainable solution, reducing the consumption of freshwater (FW) and the need for mineral fertilisers. The main aim of this study was to assess, in a three-year period, the effects of TWW irrigation compared to FW on the biomass production of bermudagrass [ Cynodon dactylon (L.) Pers.] plants and soil characteristics and to estimate the nutritional input provided by TWW irrigation. TWW was obtained by a constructed wetland system (CWs) which was used to treat urban wastewater. The system had a total surface area of 100 m 2 . An experimental field of bermudagrass was set up close to the system in a Sicilian location (Italy), using a split-plot design for a two-factor experiment with three replications. Results highlighted a high organic pollutant removal [five days biochemical oxygen demand (BOD 5 ): 61%, chemical oxygen demand (COD): 65%] and a good efficiency in nutrients [total nitrogen (TN): 50%, total phosphorus (TP): 42%] of the CWs. Plants irrigated with TWW showed higher dry aboveground dry-weight (1259.3 kg ha -1 ) than those irrigated with FW (942.2 kg ha -1 ), on average. TWW irrigation approximately allowed a saving of 50.0 kg TN ha -1 year -1 , 24.0 kg TP ha -1 year -1 and 29.0 kg K ha -1 year -1 on average with respect to commonly used N-P-K fertilisation programme for bermudagrass in the Mediterranean region. Soil salinity increased significantly (p ≤ 0.01) over the years and was detected to be higher in TWW-irrigated plots (+6.34%) in comparison with FW-irrigated plots. Our findings demonstrate that medium-term TWW irrigation increases the biomass production of bermudagrass turf and contributes to save significant amounts of nutrients, providing a series of agronomic and environmental benefits.

Purpose Postprandial hyperglycemia and glycemic oscillations have been associated with increased oxidative stress. We sought to investigate the effect of two walking exercise protocols performed during lunchtime on glycemic control and oxidative stress in type 2 diabetic (T2D) patients. Methods Nine T2D patients participated in three randomized crossover trials; a control trial (Con), with participants having a standard lunch followed by their normal daily activities and two exercise trials (ContEx and Splitex). In ContEx, subjects performed 40 min of brisk walking 40 min after lunch, whereas in SplitEx the walking exercise was divided in two 20-min isoenergetic bouts, before and 40 min after meal. 24-h glycemic control was monitored by continuous glucose monitoring. 24-h urinary levels of 8-iso PGF2ɑ were measured as a marker of oxidative stress. Results SplitEx resulted in less time spent in moderate hyperglycemia after lunch vs ContEx (42.4 ± 38.7 % vs 68.2 ± 32.7 %, P  = 0.04). ContEx reduced hyperglycemic time after breakfast consumed the morning after the exercise session (58.3 ± 29.6 Con vs 40.2 ± 33.4 % ContEx, P  = 0.02). Compared with Con, 24-h urinary isoprostanes were decreased both in ContEx (−68 %, P  = 0.02) and SplitEx (−63 %, P  = 0.04). Conclusions Splitting an exercise session into two bouts, pre- and post-lunch, affects mainly the glycemic response to lunch, while a single-continuous isoenergetic session exerts its effect later in the 24-h period. Both exercise modalities effectively attenuate systemic oxidative stress with similar overall benefits.

The aim of our study was to evaluate the capability of a crude extract of phenols from extra virgin olive oil of Moraiolo cultivar to induce apoptosis and/or differentiation in sensitive and resistant HL60 cell lines to anticancer drugs (Typical Multidrug Resistance). Our data highlight that the crude extract is able to induce apoptosis on both sensitive and resistant cells, whereas the exposure to a number of anticancer drugs does not induce apoptosis in resistant cells. In differentiation experiments we investigated the capability of crude extract of phenols to induce the expression of CD11 granulocytic or CD14 monocytic cell surface antigen in sensitive and resistant HL60 cell lines. At IC50 dose level (17 ug/ml and 32 ug/ml respectively for sensitive and resistant cell lines), the crude extract induced differentiation associated with the expression of CD14 monocytic cell lines but not that of CD11 granulocityc cell surface antigen. Further investigations are in progress to better clarify the mechanism by which olive oil phenols induce diffentiation on this cell line.