The experimental reconstruction of an Early Neolithic underground oven of Portonovo (Italy)

This contribution presents the experimental reconstruction of an underground oven replicated according to the archaeological evidence unearthed from the Early Neolithic site of Portonovo-Fosso Fontanaccia (Ancona-Italy). A domed structure, measuring 190x180 cm diameter at the base and 50 cm in height, was dug in 15 hours, in a sediment compatible with the geological formation that features the archaeological site. The experimental protocol presented in this article aims to reconstruct techniques, timing and tools needed to dig the peculiar underground structures of Portonovo used by Neolithic groups and understand key topics regarding the entire technical process such as energy investment for the community, seasonality and lifespan

A Multidisciplinary Approach to the Study of Early Neolithic Pyrotechnological Structures. The Case Study of Portonovo (Marche, Italy)

The introduction of agricultural practices fostered the development of specific technologies for the new subsistence practices and the production of new artefacts. Pyrotechnological structures such as ovens are part of the Neolithic equipment and accompanied the spread of agriculture from the Near East across Europe and the Mediterranean Sea. Ovens located within settlements – mainly domed, aboveground structures – have been traditionally linked to cooking and baking. The function is usually deduced from techno-morphological traits, although experimental approaches or ethnoarchaeological observations have often been used. This article aims to demonstrate the effectiveness of the multidisciplinary approach to understand the function of fire structures. An integrated methodology that combines archaeological analysis, archaeometry, and experimental archaeology has been applied to study the underground ovens of the Early Neolithic site of Portonovo (Marche, Italy) dated to the sixth millennium BCE. Samples of hardened sediment of archaeological ovens’ inner surface and selected pottery fragments were analysed through X-ray powder diffraction to estimate the temperature reached. A life-size replica of an underground oven was then created to perform firing experiments, including pottery firing. Samples of the oven’s walls and experimental vessels were analysed with the same method, and the values were compared. Our results indicate that the Portonovo ovens are potentially multifunctional structures, built for about 700 years, always with the same technique exploiting the natural soil’s insulating properties

Vascular Damage In Resistant Hypertension: Tnf-alpha Inhibition Effects On Endothelial Cells

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Inflammatory cytokines have been associated with the pathophysiology of hypertension and target organ damage (TOD). Resistant hypertensive patients (RHTN) are characterized by poor blood pressure control and higher prevalence of TOD. This study evaluated the relationship between plasma levels of TNF-alpha and arterial stiffness (pulse wave velocity-PWV) in 32 RHTN and 19 normotensive subjects. Moreover, we investigated the effect of TNF-alpha inhibition on human endothelial cells (HUVECs) incubated with serum from RHTN and normotensive subjects. HUVECs containing serum obtained from normotensive (n = 8) and hypertensive (n = 8) individuals were treated with TNF-alpha inhibitor (infliximab). Cell suspensions were used for measurement of DNA fragmentation and reactive oxygen species (ROS) content. RHTN patients showed higher levels of TNF-alpha compared to normotensive subjects, as well as higher PWV. Positive correlation was found between TNF-alpha levels and PWV measures in the whole group. HUVECs incubated with serum from RHTN showed increased cell apoptosis and higher ROS content compared to normotensive subjects. Infliximab attenuated the apoptosis of HUVECs incubated with serum from RHTN, but no effect in ROS production was observed. Our findings suggest that TNF-alpha might mediate, at least in part, vascular damage in resistant hypertension.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Laser Scanning Confocal Microscopy: Application in Manufacturing and Research of Corneal Stem Cells

Laser scanning confocal microscopes (LSCMs) are powerful devices used to acquire high definition optical images by choosing the required depth selectively. The presence of specific laser beams and features such as fluorescence recovery after photobleaching (FRAP), fluores\u2010 cence lifetime imaging microscopy (FLIM), and fluorescence resonance energy transfer (FRET) allow to: i. increase the quality of the image; ii. observe and analyze subcellular organelles; iii. track the localization of any given labeled molecule within the cell; iv. identify specific areas within a tissue/organ (Figure 1). In parallel, the development and manufacturing of fluorescent probes (=fluorophores) characterized by low toxicity profiles are allowing to perform the above mentioned studies using living cell cultures or tissues that are not fixed. Furthermore, fluorescent proteins such as the Green Fluorescent Protein (GFP) and its derivatives allow to detect how the biosynthetic machinery of the cell works or a transgene (driven by a plasmid or a genetically engineered virus) is expressed (Figure 2) or a chimeric protein interacts with other cellular components. The aim of this chapter is therefore to describe how LSCM functions and features have helped vision sciences and regenerative medicine applications in the field of ophthalmology. The next sections will analyze how LSCM-based analyses have helped to: 1. evaluate how the ocular surface is formed; 2. define the role of p63 as stem cell marker; 3. set up quality control assays required for clinical applications of limbal stem cells in patients with limbal stem cell deficiency (LSCD); 4. validate the use of impression citology as a diagnostic tool for LSCD; 5. study gene therapy-based potential ways to treat rare genetic disorders of the ocular surface

Retinitis Pigmentosa: Genes and Disease Mechanisms

Retinitis pigmentosa (RP) is a group of inherited disorders affecting 1 in 3000-7000 people and characterized by abnormalities of the photoreceptors (rods and cones) or the retinal pigment epithelium of the retina which lead to progressive visual loss. RP can be inherited in an autosomal dominant, autosomal recessive or X-linked manner. While usually limited to the eye, RP may also occur as part of a syndrome as in the Usher syndrome and Bardet-Biedl syndrome. Over 40 genes have been associated with RP so far, with the majority of them expressed in either the photoreceptors or the retinal pigment epithelium. The tremendous heterogeneity of the disease makes the genetics of RP complicated, thus rendering genotype-phenotype correlations not fully applicable yet. In addition to the multiplicity of mutations, in fact, different mutations in the same gene may cause different diseases. We will here review which genes are involved in the genesis of RP and how mutations can lead to retinal degeneration. In the future, a more thorough analysis of genetic and clinical data together with a better understanding of the genotype-phenotype correlation might allow to reveal important information with respect to the likelihood of disease development and choices of therapy

Generation of a transgene-free human induced pluripotent stem cell line (UNIPDi001-A) from oral mucosa epithelial stem cells.

Abstract Human oral mucosa epithelial stem cells (hOMESCs) were obtained from a fresh oral biopsy collected from a healthy subject at the Fondazione Banca degli Occhi del Veneto (FBOV). An integration-free reprogramming protocol was applied exploiting episomal plasmids transfected into cells using a Nucleofector device. Around day 20 post transfection, several human induced pluripotent stem cell (hiPSC) colonies were manually picked and expanded. One of these (UNIPDi001-A-hiPSCs) expressed undifferentiated state marker alkaline phosphatase along with a panel of pluripotency state markers and was able to differentiate into the derivatives of all the three germ layers

C/EBPδ regulates cell cycle and self-renewal of human limbal stem cells

Human limbal stem cells produce transit amplifying progenitors that migrate centripetally to regenerate the corneal epithelium. Coexpression of CCAAT enhancer binding protein δ (C/EBPδ), Bmi1, and ΔNp63α identifies mitotically quiescent limbal stem cells, which generate holoclones in culture. Upon corneal injury, a fraction of these cells switches off C/EBPδ and Bmi1, proliferates, and differentiates into mature corneal cells. Forced expression of C/EBPδ inhibits the growth of limbal colonies and increases the cell cycle length of primary limbal cells through the activity of p27Kip1 and p57Kip2. These effects are reversible; do not alter the limbal cell proliferative capacity; and are not due to apoptosis, senescence, or differentiation. C/EBPδ, but not ΔNp63α, indefinitely promotes holoclone self-renewal and prevents clonal evolution, suggesting that self-renewal and proliferation are distinct, albeit related, processes in limbal stem cells. C/EBPδ is recruited to the chromatin of positively (p27Kip1 and p57Kip2) and negatively (p16INK4A and involucrin) regulated gene loci, suggesting a direct role of this transcription factor in determining limbal stem cell identity

Vascular damage in resistant hypertension: Tnf-alpha inhibition effects on endothelial cells

Inflammatory cytokines have been associated with the pathophysiology of hypertension and target organ damage (TOD). Resistant hypertensive patients (RHTN) are characterized by poor blood pressure control and higher prevalence of TOD. This study evaluated the relationship between plasma levels of TNF-alpha and arterial stiffness (pulse wave velocity-PWV) in 32 RHTN and 19 normotensive subjects. Moreover, we investigated the effect of TNF-alpha inhibition on human endothelial cells (HUVECs) incubated with serum from RHTN and normotensive subjects. HUVECs containing serum obtained from normotensive (n = 8) and hypertensive (n = 8) individuals were treated with TNF-alpha inhibitor (infliximab). Cell suspensions were used for measurement of DNA fragmentation and reactive oxygen species (ROS) content. RHTN patients showed higher levels of TNF-alpha compared to normotensive subjects, as well as higher PWV. Positive correlation was found between TNF-alpha levels and PWV measures in the whole group. HUVECs incubated with serum from RHTN showed increased cell apoptosis and higher ROS content compared to normotensive subjects. Infliximab attenuated the apoptosis of HUVECs incubated with serum from RHTN, but no effect in ROS production was observed. Our findings suggest that TNF-alpha might mediate, at least in part, vascular damage in resistant hypertension2015CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPES