Gene/longevity association studies at four autosomal loci (REN, THO, PARP, SOD2)

The possibility that four loci (REN, THO, PARP, SOD2) are associated with longevity was explored by comparing the genotypic pools of subjects older than 100 years with those of younger subjects matched for sex and geographic area (northern and southern Italy). The markers (all located within the respective gene) were HUMREN4; HUMTHO1; HUMPARP (gt)845nt; SOD2(C/T)401nt. In order to reduce the number of genotypes, multiallelic polymorphisms were recoded as diallelic according to allele size and frequency patterns (small: S, and large: L, alleles). A significant loss of LL homozygous genotypes was found at the THO locus in male but not in female centenarians with respect to matched controls. On the other hand no significant difference was found between case/control genotypic frequencies at REN, PARP, SOD2 loci. The latter loci therefore do not affect inter-individual variability in life expectancy (at least in terms of qualitative variants associated with the tested markers). However, the data is consistent with an association between the THO locus and longevity

Inflamm-aging of the stem cell niche: Breast cancer as a paradigmatic example: Breakdown of the multi-shell cytokine network fuels cancer in aged people.

Inflamm-aging is a relatively new terminology used to describe the age-related increase in the systemic pro-inflammatory status of humans. Here, we represent inflamm-aging as a breakdown in the multi-shell cytokine network, in which stem cells and stromal fibroblasts (referred to as the stem cell niche) become pro-inflammatory cytokine over-expressing cells due to the accumulation of DNA damage. Inflamm-aging self-propagates owing to the capability of pro-inflammatory cytokines to ignite the DNA-damage response in other cells surrounding DNA-damaged cells. Macrophages, the major cellular player in inflamm-aging, amplify the phenomenon, by broadcasting pro-inflammatory signals at both local and systemic levels. On the basis of this, we propose that inflamm-aging is a major contributor to the increase in cancer incidence and progression in aged people. Breast cancer will be presented as a paradigmatic example for this relationship

Valutazione dell'espressione dell'RNA messaggero del fattore di trascrizione GATA-4 in cellule H9c2

Il GATA- 4 \ue8 un membro della famiglia dei fattori di trascrizione GATA; queste proteine leganti il DNA hanno ruoli importanti nella regolazione del differenziamento, proliferazione e morfogenesi degli organi durante l\u2019embriogenesi e la vita adulta. Sono stati identificati sei membri della famiglia GATA nei vertebrati e queste proteine nucleari regolano la trascrizione attraverso un legame alla sequenza consenso 5\u2019 \u2013WGATAR-3\u2019 presente nei promotori dei target genetici; tre membri di questa famiglia, GATA 4/5/6, sono espressi nell\u2019embriogenesi del cuore. Inoltre sia GATA- 4 che GATA- 6 sono espressi nel cuore adulto e presumibilmente regolano il fenotipo differenziato dei cardiomiociti. Il presente studio e' stato finalazzato alla ricerca dell\u2019RNA messaggero del GATA- 4 in cardiomioblasti H9c2, una linea di cellule muscolari cardiache di ventricolo di embrione di ratto. Una volta isolato l\u2019RNA, \ue8 stata eseguita la retrotrascrizione in cDNA usando il Transcriptor First Strand cDNA Sinthesis Kit, Roche Diagnostics. E\u2019stata poi condotta una reazione di polimerizzazione a catena (PCR) attraverso una Real Time PCR, Light Cycler, Roche Diagnostics, che ha permesso di amplificare un segmento di DNA che si trova tra due regioni di sequenza nota e usando come punto di partenza della DNA polimerasi due oligonucleotidi, detti primer, complementari a due sequenze che fiancheggiano il segmento da amplificare. La tecnologia utilizzata \ue8 quella delle sonde TaqMan che, degradate durante la fase di amplificazione, rilasciano un fluoroforo rilevato in \u201ctempo reale\u201d dallo strumento. Il numero di cicli, o crossing point (CP), in cui inizia l\u2019amplificazione dei campioni \ue8 l\u2019indice quantitativo a cui ci si riferisce. La fluorescenza emessa dai campioni nel corso della reazione di amplificazione e' stata rappresentata da due curve ottenute utilizzando lo stesso campione e che sono quindi indicative del grado di riproducibilit\ue0 della misura. Allo scopo di verificare se l\u2019amplificazione \ue8 avvenuta in modo corretto nei riguardi dell\u2019espressione del gene target del GATA \u2013 4, si \ue8 provveduto a verificare, mediante elettroforesi su gel di agarosio, la presenza del DNA amplificato. Il dosaggio dell\u2019RNA messaggero del GATA- 4 e dell\u2019espressione delle proteine, pu\uf2 quindi fornire utili informazioni sia nelle ricerche nelle quali sono studiati i processi di differenziamento delle cellule staminali in cardiomiociti, sia per verificare nei cardiomiociti adulti il ruolo di questo fattore di trascrizione nei processi di sopravvivenza cellulare

Metabolic gene polymorphisms and p53 mutayions in healthy centenarians and younger controls

To obtain insights into the genetic mechanisms of ageing, we studied the frequency of the simultaneous presence of polymorphisms in phase I and phase II genes and of several p53 germline mutations in a group of 66 nonagenarians and centenarians in good health, selected from a larger sample of a multicentre Italian study in Northern Italy, and in a sample of 150 young healthy volunteers of the same ethnic group. We found a statistically significant difference in the frequency of the GSTT1 deletion and the p53 genotypes: the absence of any p53 polymorphisms and of GSTT1 deletion, and the simultaneous presence of the three p53 polymorphisms and of GSTT1 deletion, were much more frequent in young subjects than in centenarians (41.5% versus 26.9% and 8.8% versus 3.8%, respectively). One hypothesis to explain this difference is that subjects with both GSTT1 deletion and p53 polymorphisms may accumulate carcinogens and may have reduced DNA repair ability, and thus are more at risk for cancer. Another possible explanation is that both metabolic genes and p53 act on pathways related to cell ageing and death, and therefore certain composite genetic patterns could represent a generic mechanism of protection against ageing, not just against the development of chronic diseases. It is likely that longevity is related to a complex genetic trait as well as to certain environmental exposures

Epigenetic signature of early cardiac regulatory genes in human adipose-derived stem cells

Adipose-derived stem cells (ADSCs) are stromal mesenchymal stem cells isolated from lipoaspirates, and they display a broad potential to differentiate toward different lineages. The role of epigenetics in regulating the expression of their lineage-specific genes is under evaluation, however till date virtually nothing is known about the relative significance of cardiac-specific transcription factor genes in human ADSCs. The aim of this study was to investigate DNA promoter methylation and relevant histone modifications involving MEF-2C, GATA-4, and Nkx2.5 in native human ADSCs. CpG sites at the transcription start in their promoters were found unmethylated using methylation-specific PCR. Chromatin immunoprecipitation assay showed low levels of total acetylated H3 histone (acH3) and high levels of trimethylated lysine 27 in H3 histone (H3K27me3) which were associated with both GATA-4 and Nkx2.5 promoters, indicating their transcriptional repressive chromatin arrangement. On the other hand, the opposite was apparent for MEF-2C promoter. Accordingly, MEF-2C\u2014but not GATA-4 and Nkx2.5\u2014 transcripts were evidenced in native human ADSCs. These results suggest that the chromatin arrangement of these early cardiac regulatory genes could be explored as a level of intervention to address the differentiation of human ADSCs toward the cardiac lineage

Exploitation of C and X band SAR images for soil moisture change detection estimation in agricultural areas (Po valley-Italy)

This paper presents the analysis of C and X band images in the scope of soil moisture detection in agricultural fields. Archived data have been analyzed in order to understand the SAR signal behavior of vegetated fields in comparison to bare soils. The results indicate that the sensitivity to bare fields of C and X band signatures is very close, while it changes in presence of vegetation. In particular the effect is directly proportional to amount of vegetation that in this preliminary analysis has been evaluated through the NDVI variable. After this analysis, a statistical approach has been applied to SAR images to infer the information on the soil moisture values. Several experiments have been carried out by considering only C band data, only X band data and a combination of C and X band data. For bare soils, C and X band data determine very similar results and in good agreement to ground measurements. For vegetated fields, C band data tend to underestimate soil moisture due to the vegetation attenuation, while X band data, mainly influenced by vegetation, determine very poor results. Encouraging results are obtained by the combination of C and X band data, thus indicating that X band data can be used in combination to C band data in order to compensate the effect of vegetation

Comparison between stem cells harvested from wet and dry lipoaspirates

Adipose-derived stem cells (ASC) are usually isolated from lipoaspirates, but it is not known if the anesthetic solution injected into adipose tissue affects cell yield and functions. Two different samples were drawn from the abdominal region of female subjects. In the first, a physiological solution containing lidocaine/adrenaline was injected (wet liposuction, WL), while in the contralateral area, the sample was collected without injecting any solution (dry liposuction, DL). The aspirates were processed to investigate the yield of the stromal-vascular fraction (SVF) cells and ASC frequency, growth rate, apoptosis, and differentiation potential. The solid dried mass of fresh WL isolates was lower than that of DL isolates (p<0.01) due to the presence, in the former, of a liquid solution. As a consequence, the amount of WL-SVF cells was 18.7% lower than those obtained from DL (p < 0.01); this difference was also observed under culture conditions. In addition, the number of colony-forming unit-fibroblasts (CFU-Fs) obtained from 1 7 10(3) SVF cells was 25.5% lower in WL-aspirates than DL-aspirates (p < 0.05) owing, at least in part, to the observed presence of ASC in the liquid solution of the WL isolates. After WL and DL, no differences were observed in ASC growth rate, apoptosis, or differentiation potential toward adipogenic, osteogenic, and endothelial cell lineages. In conclusion, WL yields about 40% fewer ASC than DL due to the combined effect of tissue dilution and the reduced frequency of ASC in the SVF. The main biological features of ASC are suitable for cell-based therapies